Biostimulants from the phytomicrobiome

Microbe-to-plant signals as low input crop management technologies

Donald L SmithPlant Science

McGill University

Need for Enhanced Food Productionbull Growing global population

bull Headed for 9-10 billionbull Changes in diet

bull More meat

01234567

1 200 400 600 800 1000 1200 1400 1600 1800 2000Time (Years)

Billi

ons o

f Peo

ple

bull Meltingbull Mountain glaciersbull Greenland ice sheetbull Arctic sea ice

bull 5 to 8 oC warmer in Canadabull Birds return sooner in spring amp flowering

soonerbull More extreme weather conditions more often

bull Droughtbull Heatbull Salinitybull Flooding 1990 2001

Triftgletscher Switzerland

Threat of Climate Change

Need for Enhanced Food Productionbull Growing global population

bull Headed for 9-10 billionbull Changes in diet

bull More meat

01234567

1 200 400 600 800 1000 1200 1400 1600 1800 2000Time (Years)

Billi

ons o

f Peo

ple

bull Meltingbull Mountain glaciersbull Greenland ice sheetbull Arctic sea ice

bull 5 to 8 oC warmer in Canadabull Birds return sooner in spring amp flowering

soonerbull More extreme weather conditions more often

bull Droughtbull Heatbull Salinitybull Flooding 1990 2001

Triftgletscher Switzerland

Threat of Climate Change

bull Meltingbull Mountain glaciersbull Greenland ice sheetbull Arctic sea ice

bull 5 to 8 oC warmer in Canadabull Birds return sooner in spring amp flowering

soonerbull More extreme weather conditions more often

bull Droughtbull Heatbull Salinitybull Flooding 1990 2001

Triftgletscher Switzerland

Threat of Climate Change

Biomass for Advanced Biofuels and the Bioeconomy

bull A key step in the supply chain is reliable high quality biomass

bull Can bebull Crop or forestry residuesbull Purpose grown

bull Fast growing grasses bull Plantation forestrybull Generally grown on more marginal

lands so more often stressedbull If food crops more biomass residue and more

foodbull Food and fuel not food versus fuel

bull

The Phytomicrobiomebull Plants like mammals have a microbiome

bull Phytomicrobiomebull Plant + phytomicrobiome is holobiont

bull Present through all of terrestrial plant evolutionbull Community of microbes associated with all plant parts

bull Roots in humid soil have most well developedbull Help in a wide range of ways

bull Nutrient mobilizationbull Hormone productionbull Disease controlbull Signals

bull Hormones of the holobiont

Adapting to Climate Changebull Develop inputs that adapt agriculture to worsening climate change conditions

bull Improve plant stress tolerancebull Climate change resilient crop production

bull Reduced N2O and CO2 emissions

The Approachbull Biological inputs

bull Addition of small amounts of inexpensive materials that enhance crop growth

bull Reduced costsbull Enhance stress resistance

bull Both abiotic and bioticbull Improve yield

B B

C

So Farbull N2 fixation inoculants for legumesbull Rhizobia (Rhizobium Bradyrhizobium Sinorhizobium etc)

bull Widely used for over a centurybull Mycorrhizae for P uptakebull Mobilization of P and Zn

N

PBB

Example Line of Researchbull It started with soybeanbull Soybean evolved in the tropics and subtropics

bull We agriculturalists have carried it further north and south ever since

bull Its metabolism is adapted to and set up for relatively high temperatures

bull The optimum temperature for soybean nodulation was known to be 25 to 30oC

bull Production in Quebec since late 1980s

Something Newbull We observed that soybean plants under Quebec field conditions frequently look pale green for several weeks after the nitrogen in the original seed was expended

bull After this they ldquoregreenedrdquobull We wondered if nodulation was inhibited by low soil temperatures early in the season

The Problembull Low root zone temperatures slowed onset of N2 fixation

bull 1-2 days oC-1 to 17 oCbull The cause was disruption in signaling during the first stage of nodulation

bull Symbiont recognition

Privileged and Confidential

A Solutionbull Adding an appropriate isoflavonoid signal (genistein) to Bradyrhizobium japonicum cells before application as inoculant switches on the Nod genes in the bacterial cells

bull The B japonicum cells then began to produce and excrete the return signals

bull Lipo-chitooligosaccharides (LCOs)

Genistein

N2 Fixation and Growth - Genistein

a

Genisteinconcentration

Nod

ule

wei

ght p

er p

lant

(mg)

June 17 sampling

a

- N2 fixation started 4 to 5 days sooner- Total N fixed without genistein (15N dilution estimate) was 53 kg ha-1 while with genistein it was 95 kg ha-1 80 increase

0

5

10

15

20

25

30

AC Bravor Maple Glen

0 microM20 microM

a

b

0

05

1

15

2

25

3

AC Bravor Maple Glen

0 microM20 microM

a

b ba

Gra

in y

ield

(t h

a-1 )

Genisteinconcentration

-Yield increases have been as high as 40 with most being in the 10 to 20 range

Commercializationbull Established a spin-off company

bull Bios Agriculture Incbull To keep a technician employed post recession

bull Spin-offs were quite new at McGill University

bull 5 share non-dilutablebull The usual things

bull Venture capital and other investmentsbull Regulatory requirements must be satisfied

Serendipity amp Follow Up bull Earlier emergence observed at field sites

where genistein appliedbull What did this meanbull Experiment

bull seeds in waterbull genistein alonebull Bradyrhizobium japonicum alonebull genistein and B japonicum togetherbull only the last treatment accelerated germination - active material was the LCO the bacteria-to-plant signal

New LCO ActivitybullWe also found that the return signals (lipo-chitooligosaccharides) also directly stimulate growth of non-legume plants and this can increase yields

Control LCO

Corn Seedlings

01020304050607080

Corn Cotton Beet Soybean

ControlLCO

Commercializationbull Bios Agricluture eventually taken up by Agribiotics (also a Canadian Company)

bull Agribiotics was purchased by EMDbull EMD became part of Novozymes

The Phytomicrobiomebull Plants like mammals have a microbiome

bull Phytomicrobiomebull Plant + phytomicrobiome is holobiont

bull Present through all of terrestrial plant evolutionbull Community of microbes associated with all plant parts

bull Roots in humid soil have most well developedbull Help in a wide range of ways

bull Nutrient mobilizationbull Hormone productionbull Disease controlbull Signals

bull Hormones of the holobiont

Adapting to Climate Changebull Develop inputs that adapt agriculture to worsening climate change conditions

bull Improve plant stress tolerancebull Climate change resilient crop production

bull Reduced N2O and CO2 emissions

The Approachbull Biological inputs

bull Addition of small amounts of inexpensive materials that enhance crop growth

bull Reduced costsbull Enhance stress resistance

bull Both abiotic and bioticbull Improve yield

B B

C

So Farbull N2 fixation inoculants for legumesbull Rhizobia (Rhizobium Bradyrhizobium Sinorhizobium etc)

bull Widely used for over a centurybull Mycorrhizae for P uptakebull Mobilization of P and Zn

N

PBB

Example Line of Researchbull It started with soybeanbull Soybean evolved in the tropics and subtropics

bull We agriculturalists have carried it further north and south ever since

bull Its metabolism is adapted to and set up for relatively high temperatures

bull The optimum temperature for soybean nodulation was known to be 25 to 30oC

bull Production in Quebec since late 1980s

Something Newbull We observed that soybean plants under Quebec field conditions frequently look pale green for several weeks after the nitrogen in the original seed was expended

bull After this they ldquoregreenedrdquobull We wondered if nodulation was inhibited by low soil temperatures early in the season

The Problembull Low root zone temperatures slowed onset of N2 fixation

bull 1-2 days oC-1 to 17 oCbull The cause was disruption in signaling during the first stage of nodulation

bull Symbiont recognition

Privileged and Confidential

A Solutionbull Adding an appropriate isoflavonoid signal (genistein) to Bradyrhizobium japonicum cells before application as inoculant switches on the Nod genes in the bacterial cells

bull The B japonicum cells then began to produce and excrete the return signals

bull Lipo-chitooligosaccharides (LCOs)

Genistein

N2 Fixation and Growth - Genistein

a

Genisteinconcentration

Nod

ule

wei

ght p

er p

lant

(mg)

June 17 sampling

a

- N2 fixation started 4 to 5 days sooner- Total N fixed without genistein (15N dilution estimate) was 53 kg ha-1 while with genistein it was 95 kg ha-1 80 increase

0

5

10

15

20

25

30

AC Bravor Maple Glen

0 microM20 microM

a

b

0

05

1

15

2

25

3

AC Bravor Maple Glen

0 microM20 microM

a

b ba

Gra

in y

ield

(t h

a-1 )

Genisteinconcentration

-Yield increases have been as high as 40 with most being in the 10 to 20 range

Commercializationbull Established a spin-off company

bull Bios Agriculture Incbull To keep a technician employed post recession

bull Spin-offs were quite new at McGill University

bull 5 share non-dilutablebull The usual things

bull Venture capital and other investmentsbull Regulatory requirements must be satisfied

Serendipity amp Follow Up bull Earlier emergence observed at field sites

where genistein appliedbull What did this meanbull Experiment

bull seeds in waterbull genistein alonebull Bradyrhizobium japonicum alonebull genistein and B japonicum togetherbull only the last treatment accelerated germination - active material was the LCO the bacteria-to-plant signal

New LCO ActivitybullWe also found that the return signals (lipo-chitooligosaccharides) also directly stimulate growth of non-legume plants and this can increase yields

Control LCO

Corn Seedlings

01020304050607080

Corn Cotton Beet Soybean

ControlLCO

Commercializationbull Bios Agricluture eventually taken up by Agribiotics (also a Canadian Company)

bull Agribiotics was purchased by EMDbull EMD became part of Novozymes

Adapting to Climate Changebull Develop inputs that adapt agriculture to worsening climate change conditions

bull Improve plant stress tolerancebull Climate change resilient crop production

bull Reduced N2O and CO2 emissions

The Approachbull Biological inputs

bull Addition of small amounts of inexpensive materials that enhance crop growth

bull Reduced costsbull Enhance stress resistance

bull Both abiotic and bioticbull Improve yield

B B

C

So Farbull N2 fixation inoculants for legumesbull Rhizobia (Rhizobium Bradyrhizobium Sinorhizobium etc)

bull Widely used for over a centurybull Mycorrhizae for P uptakebull Mobilization of P and Zn

N

PBB

Example Line of Researchbull It started with soybeanbull Soybean evolved in the tropics and subtropics

bull We agriculturalists have carried it further north and south ever since

bull Its metabolism is adapted to and set up for relatively high temperatures

bull The optimum temperature for soybean nodulation was known to be 25 to 30oC

bull Production in Quebec since late 1980s

Something Newbull We observed that soybean plants under Quebec field conditions frequently look pale green for several weeks after the nitrogen in the original seed was expended

bull After this they ldquoregreenedrdquobull We wondered if nodulation was inhibited by low soil temperatures early in the season

The Problembull Low root zone temperatures slowed onset of N2 fixation

bull 1-2 days oC-1 to 17 oCbull The cause was disruption in signaling during the first stage of nodulation

bull Symbiont recognition

Privileged and Confidential

A Solutionbull Adding an appropriate isoflavonoid signal (genistein) to Bradyrhizobium japonicum cells before application as inoculant switches on the Nod genes in the bacterial cells

bull The B japonicum cells then began to produce and excrete the return signals

bull Lipo-chitooligosaccharides (LCOs)

Genistein

N2 Fixation and Growth - Genistein

a

Genisteinconcentration

Nod

ule

wei

ght p

er p

lant

(mg)

June 17 sampling

a

- N2 fixation started 4 to 5 days sooner- Total N fixed without genistein (15N dilution estimate) was 53 kg ha-1 while with genistein it was 95 kg ha-1 80 increase

0

5

10

15

20

25

30

AC Bravor Maple Glen

0 microM20 microM

a

b

0

05

1

15

2

25

3

AC Bravor Maple Glen

0 microM20 microM

a

b ba

Gra

in y

ield

(t h

a-1 )

Genisteinconcentration

-Yield increases have been as high as 40 with most being in the 10 to 20 range

Commercializationbull Established a spin-off company

bull Bios Agriculture Incbull To keep a technician employed post recession

bull Spin-offs were quite new at McGill University

bull 5 share non-dilutablebull The usual things

bull Venture capital and other investmentsbull Regulatory requirements must be satisfied

Serendipity amp Follow Up bull Earlier emergence observed at field sites

where genistein appliedbull What did this meanbull Experiment

bull seeds in waterbull genistein alonebull Bradyrhizobium japonicum alonebull genistein and B japonicum togetherbull only the last treatment accelerated germination - active material was the LCO the bacteria-to-plant signal

New LCO ActivitybullWe also found that the return signals (lipo-chitooligosaccharides) also directly stimulate growth of non-legume plants and this can increase yields

Control LCO

Corn Seedlings

01020304050607080

Corn Cotton Beet Soybean

ControlLCO

Commercializationbull Bios Agricluture eventually taken up by Agribiotics (also a Canadian Company)

bull Agribiotics was purchased by EMDbull EMD became part of Novozymes

The Approachbull Biological inputs

bull Addition of small amounts of inexpensive materials that enhance crop growth

bull Reduced costsbull Enhance stress resistance

bull Both abiotic and bioticbull Improve yield

B B

C

So Farbull N2 fixation inoculants for legumesbull Rhizobia (Rhizobium Bradyrhizobium Sinorhizobium etc)

bull Widely used for over a centurybull Mycorrhizae for P uptakebull Mobilization of P and Zn

N

PBB

Example Line of Researchbull It started with soybeanbull Soybean evolved in the tropics and subtropics

bull We agriculturalists have carried it further north and south ever since

bull Its metabolism is adapted to and set up for relatively high temperatures

bull The optimum temperature for soybean nodulation was known to be 25 to 30oC

bull Production in Quebec since late 1980s

Something Newbull We observed that soybean plants under Quebec field conditions frequently look pale green for several weeks after the nitrogen in the original seed was expended

bull After this they ldquoregreenedrdquobull We wondered if nodulation was inhibited by low soil temperatures early in the season

The Problembull Low root zone temperatures slowed onset of N2 fixation

bull 1-2 days oC-1 to 17 oCbull The cause was disruption in signaling during the first stage of nodulation

bull Symbiont recognition

Privileged and Confidential

A Solutionbull Adding an appropriate isoflavonoid signal (genistein) to Bradyrhizobium japonicum cells before application as inoculant switches on the Nod genes in the bacterial cells

bull The B japonicum cells then began to produce and excrete the return signals

bull Lipo-chitooligosaccharides (LCOs)

Genistein

N2 Fixation and Growth - Genistein

a

Genisteinconcentration

Nod

ule

wei

ght p

er p

lant

(mg)

June 17 sampling

a

- N2 fixation started 4 to 5 days sooner- Total N fixed without genistein (15N dilution estimate) was 53 kg ha-1 while with genistein it was 95 kg ha-1 80 increase

0

5

10

15

20

25

30

AC Bravor Maple Glen

0 microM20 microM

a

b

0

05

1

15

2

25

3

AC Bravor Maple Glen

0 microM20 microM

a

b ba

Gra

in y

ield

(t h

a-1 )

Genisteinconcentration

-Yield increases have been as high as 40 with most being in the 10 to 20 range

Commercializationbull Established a spin-off company

bull Bios Agriculture Incbull To keep a technician employed post recession

bull Spin-offs were quite new at McGill University

bull 5 share non-dilutablebull The usual things

bull Venture capital and other investmentsbull Regulatory requirements must be satisfied

Serendipity amp Follow Up bull Earlier emergence observed at field sites

where genistein appliedbull What did this meanbull Experiment

bull seeds in waterbull genistein alonebull Bradyrhizobium japonicum alonebull genistein and B japonicum togetherbull only the last treatment accelerated germination - active material was the LCO the bacteria-to-plant signal

New LCO ActivitybullWe also found that the return signals (lipo-chitooligosaccharides) also directly stimulate growth of non-legume plants and this can increase yields

Control LCO

Corn Seedlings

01020304050607080

Corn Cotton Beet Soybean

ControlLCO

Commercializationbull Bios Agricluture eventually taken up by Agribiotics (also a Canadian Company)

bull Agribiotics was purchased by EMDbull EMD became part of Novozymes

So Farbull N2 fixation inoculants for legumesbull Rhizobia (Rhizobium Bradyrhizobium Sinorhizobium etc)

bull Widely used for over a centurybull Mycorrhizae for P uptakebull Mobilization of P and Zn

N

PBB

Example Line of Researchbull It started with soybeanbull Soybean evolved in the tropics and subtropics

bull We agriculturalists have carried it further north and south ever since

bull Its metabolism is adapted to and set up for relatively high temperatures

bull The optimum temperature for soybean nodulation was known to be 25 to 30oC

bull Production in Quebec since late 1980s

Something Newbull We observed that soybean plants under Quebec field conditions frequently look pale green for several weeks after the nitrogen in the original seed was expended

bull After this they ldquoregreenedrdquobull We wondered if nodulation was inhibited by low soil temperatures early in the season

The Problembull Low root zone temperatures slowed onset of N2 fixation

bull 1-2 days oC-1 to 17 oCbull The cause was disruption in signaling during the first stage of nodulation

bull Symbiont recognition

Privileged and Confidential

A Solutionbull Adding an appropriate isoflavonoid signal (genistein) to Bradyrhizobium japonicum cells before application as inoculant switches on the Nod genes in the bacterial cells

bull The B japonicum cells then began to produce and excrete the return signals

bull Lipo-chitooligosaccharides (LCOs)

Genistein

N2 Fixation and Growth - Genistein

a

Genisteinconcentration

Nod

ule

wei

ght p

er p

lant

(mg)

June 17 sampling

a

- N2 fixation started 4 to 5 days sooner- Total N fixed without genistein (15N dilution estimate) was 53 kg ha-1 while with genistein it was 95 kg ha-1 80 increase

0

5

10

15

20

25

30

AC Bravor Maple Glen

0 microM20 microM

a

b

0

05

1

15

2

25

3

AC Bravor Maple Glen

0 microM20 microM

a

b ba

Gra

in y

ield

(t h

a-1 )

Genisteinconcentration

-Yield increases have been as high as 40 with most being in the 10 to 20 range

Commercializationbull Established a spin-off company

bull Bios Agriculture Incbull To keep a technician employed post recession

bull Spin-offs were quite new at McGill University

bull 5 share non-dilutablebull The usual things

bull Venture capital and other investmentsbull Regulatory requirements must be satisfied

Serendipity amp Follow Up bull Earlier emergence observed at field sites

where genistein appliedbull What did this meanbull Experiment

bull seeds in waterbull genistein alonebull Bradyrhizobium japonicum alonebull genistein and B japonicum togetherbull only the last treatment accelerated germination - active material was the LCO the bacteria-to-plant signal

New LCO ActivitybullWe also found that the return signals (lipo-chitooligosaccharides) also directly stimulate growth of non-legume plants and this can increase yields

Control LCO

Corn Seedlings

01020304050607080

Corn Cotton Beet Soybean

ControlLCO

Commercializationbull Bios Agricluture eventually taken up by Agribiotics (also a Canadian Company)

bull Agribiotics was purchased by EMDbull EMD became part of Novozymes

Example Line of Researchbull It started with soybeanbull Soybean evolved in the tropics and subtropics

bull We agriculturalists have carried it further north and south ever since

bull Its metabolism is adapted to and set up for relatively high temperatures

bull The optimum temperature for soybean nodulation was known to be 25 to 30oC

bull Production in Quebec since late 1980s

Something Newbull We observed that soybean plants under Quebec field conditions frequently look pale green for several weeks after the nitrogen in the original seed was expended

bull After this they ldquoregreenedrdquobull We wondered if nodulation was inhibited by low soil temperatures early in the season

The Problembull Low root zone temperatures slowed onset of N2 fixation

bull 1-2 days oC-1 to 17 oCbull The cause was disruption in signaling during the first stage of nodulation

bull Symbiont recognition

Privileged and Confidential

A Solutionbull Adding an appropriate isoflavonoid signal (genistein) to Bradyrhizobium japonicum cells before application as inoculant switches on the Nod genes in the bacterial cells

bull The B japonicum cells then began to produce and excrete the return signals

bull Lipo-chitooligosaccharides (LCOs)

Genistein

N2 Fixation and Growth - Genistein

a

Genisteinconcentration

Nod

ule

wei

ght p

er p

lant

(mg)

June 17 sampling

a

- N2 fixation started 4 to 5 days sooner- Total N fixed without genistein (15N dilution estimate) was 53 kg ha-1 while with genistein it was 95 kg ha-1 80 increase

0

5

10

15

20

25

30

AC Bravor Maple Glen

0 microM20 microM

a

b

0

05

1

15

2

25

3

AC Bravor Maple Glen

0 microM20 microM

a

b ba

Gra

in y

ield

(t h

a-1 )

Genisteinconcentration

-Yield increases have been as high as 40 with most being in the 10 to 20 range

Commercializationbull Established a spin-off company

bull Bios Agriculture Incbull To keep a technician employed post recession

bull Spin-offs were quite new at McGill University

bull 5 share non-dilutablebull The usual things

bull Venture capital and other investmentsbull Regulatory requirements must be satisfied

Serendipity amp Follow Up bull Earlier emergence observed at field sites

where genistein appliedbull What did this meanbull Experiment

bull seeds in waterbull genistein alonebull Bradyrhizobium japonicum alonebull genistein and B japonicum togetherbull only the last treatment accelerated germination - active material was the LCO the bacteria-to-plant signal

New LCO ActivitybullWe also found that the return signals (lipo-chitooligosaccharides) also directly stimulate growth of non-legume plants and this can increase yields

Control LCO

Corn Seedlings

01020304050607080

Corn Cotton Beet Soybean

ControlLCO

Commercializationbull Bios Agricluture eventually taken up by Agribiotics (also a Canadian Company)

bull Agribiotics was purchased by EMDbull EMD became part of Novozymes

Something Newbull We observed that soybean plants under Quebec field conditions frequently look pale green for several weeks after the nitrogen in the original seed was expended

bull After this they ldquoregreenedrdquobull We wondered if nodulation was inhibited by low soil temperatures early in the season

The Problembull Low root zone temperatures slowed onset of N2 fixation

bull 1-2 days oC-1 to 17 oCbull The cause was disruption in signaling during the first stage of nodulation

bull Symbiont recognition

Privileged and Confidential

A Solutionbull Adding an appropriate isoflavonoid signal (genistein) to Bradyrhizobium japonicum cells before application as inoculant switches on the Nod genes in the bacterial cells

bull The B japonicum cells then began to produce and excrete the return signals

bull Lipo-chitooligosaccharides (LCOs)

Genistein

N2 Fixation and Growth - Genistein

a

Genisteinconcentration

Nod

ule

wei

ght p

er p

lant

(mg)

June 17 sampling

a

- N2 fixation started 4 to 5 days sooner- Total N fixed without genistein (15N dilution estimate) was 53 kg ha-1 while with genistein it was 95 kg ha-1 80 increase

0

5

10

15

20

25

30

AC Bravor Maple Glen

0 microM20 microM

a

b

0

05

1

15

2

25

3

AC Bravor Maple Glen

0 microM20 microM

a

b ba

Gra

in y

ield

(t h

a-1 )

Genisteinconcentration

-Yield increases have been as high as 40 with most being in the 10 to 20 range

Commercializationbull Established a spin-off company

bull Bios Agriculture Incbull To keep a technician employed post recession

bull Spin-offs were quite new at McGill University

bull 5 share non-dilutablebull The usual things

bull Venture capital and other investmentsbull Regulatory requirements must be satisfied

Serendipity amp Follow Up bull Earlier emergence observed at field sites

where genistein appliedbull What did this meanbull Experiment

bull seeds in waterbull genistein alonebull Bradyrhizobium japonicum alonebull genistein and B japonicum togetherbull only the last treatment accelerated germination - active material was the LCO the bacteria-to-plant signal

New LCO ActivitybullWe also found that the return signals (lipo-chitooligosaccharides) also directly stimulate growth of non-legume plants and this can increase yields

Control LCO

Corn Seedlings

01020304050607080

Corn Cotton Beet Soybean

ControlLCO

Commercializationbull Bios Agricluture eventually taken up by Agribiotics (also a Canadian Company)

bull Agribiotics was purchased by EMDbull EMD became part of Novozymes

The Problembull Low root zone temperatures slowed onset of N2 fixation

bull 1-2 days oC-1 to 17 oCbull The cause was disruption in signaling during the first stage of nodulation

bull Symbiont recognition

Privileged and Confidential

A Solutionbull Adding an appropriate isoflavonoid signal (genistein) to Bradyrhizobium japonicum cells before application as inoculant switches on the Nod genes in the bacterial cells

bull The B japonicum cells then began to produce and excrete the return signals

bull Lipo-chitooligosaccharides (LCOs)

Genistein

N2 Fixation and Growth - Genistein

a

Genisteinconcentration

Nod

ule

wei

ght p

er p

lant

(mg)

June 17 sampling

a

- N2 fixation started 4 to 5 days sooner- Total N fixed without genistein (15N dilution estimate) was 53 kg ha-1 while with genistein it was 95 kg ha-1 80 increase

0

5

10

15

20

25

30

AC Bravor Maple Glen

0 microM20 microM

a

b

0

05

1

15

2

25

3

AC Bravor Maple Glen

0 microM20 microM

a

b ba

Gra

in y

ield

(t h

a-1 )

Genisteinconcentration

-Yield increases have been as high as 40 with most being in the 10 to 20 range

Commercializationbull Established a spin-off company

bull Bios Agriculture Incbull To keep a technician employed post recession

bull Spin-offs were quite new at McGill University

bull 5 share non-dilutablebull The usual things

bull Venture capital and other investmentsbull Regulatory requirements must be satisfied

Serendipity amp Follow Up bull Earlier emergence observed at field sites

where genistein appliedbull What did this meanbull Experiment

bull seeds in waterbull genistein alonebull Bradyrhizobium japonicum alonebull genistein and B japonicum togetherbull only the last treatment accelerated germination - active material was the LCO the bacteria-to-plant signal

New LCO ActivitybullWe also found that the return signals (lipo-chitooligosaccharides) also directly stimulate growth of non-legume plants and this can increase yields

Control LCO

Corn Seedlings

01020304050607080

Corn Cotton Beet Soybean

ControlLCO

Commercializationbull Bios Agricluture eventually taken up by Agribiotics (also a Canadian Company)

bull Agribiotics was purchased by EMDbull EMD became part of Novozymes

A Solutionbull Adding an appropriate isoflavonoid signal (genistein) to Bradyrhizobium japonicum cells before application as inoculant switches on the Nod genes in the bacterial cells

bull The B japonicum cells then began to produce and excrete the return signals

bull Lipo-chitooligosaccharides (LCOs)

Genistein

N2 Fixation and Growth - Genistein

a

Genisteinconcentration

Nod

ule

wei

ght p

er p

lant

(mg)

June 17 sampling

a

- N2 fixation started 4 to 5 days sooner- Total N fixed without genistein (15N dilution estimate) was 53 kg ha-1 while with genistein it was 95 kg ha-1 80 increase

0

5

10

15

20

25

30

AC Bravor Maple Glen

0 microM20 microM

a

b

0

05

1

15

2

25

3

AC Bravor Maple Glen

0 microM20 microM

a

b ba

Gra

in y

ield

(t h

a-1 )

Genisteinconcentration

-Yield increases have been as high as 40 with most being in the 10 to 20 range

Commercializationbull Established a spin-off company

bull Bios Agriculture Incbull To keep a technician employed post recession

bull Spin-offs were quite new at McGill University

bull 5 share non-dilutablebull The usual things

bull Venture capital and other investmentsbull Regulatory requirements must be satisfied

Serendipity amp Follow Up bull Earlier emergence observed at field sites

where genistein appliedbull What did this meanbull Experiment

bull seeds in waterbull genistein alonebull Bradyrhizobium japonicum alonebull genistein and B japonicum togetherbull only the last treatment accelerated germination - active material was the LCO the bacteria-to-plant signal

New LCO ActivitybullWe also found that the return signals (lipo-chitooligosaccharides) also directly stimulate growth of non-legume plants and this can increase yields

Control LCO

Corn Seedlings

01020304050607080

Corn Cotton Beet Soybean

ControlLCO

Commercializationbull Bios Agricluture eventually taken up by Agribiotics (also a Canadian Company)

bull Agribiotics was purchased by EMDbull EMD became part of Novozymes

N2 Fixation and Growth - Genistein

a

Genisteinconcentration

Nod

ule

wei

ght p

er p

lant

(mg)

June 17 sampling

a

- N2 fixation started 4 to 5 days sooner- Total N fixed without genistein (15N dilution estimate) was 53 kg ha-1 while with genistein it was 95 kg ha-1 80 increase

0

5

10

15

20

25

30

AC Bravor Maple Glen

0 microM20 microM

a

b

0

05

1

15

2

25

3

AC Bravor Maple Glen

0 microM20 microM

a

b ba

Gra

in y

ield

(t h

a-1 )

Genisteinconcentration

-Yield increases have been as high as 40 with most being in the 10 to 20 range

Commercializationbull Established a spin-off company

bull Bios Agriculture Incbull To keep a technician employed post recession

bull Spin-offs were quite new at McGill University

bull 5 share non-dilutablebull The usual things

bull Venture capital and other investmentsbull Regulatory requirements must be satisfied

Serendipity amp Follow Up bull Earlier emergence observed at field sites

where genistein appliedbull What did this meanbull Experiment

bull seeds in waterbull genistein alonebull Bradyrhizobium japonicum alonebull genistein and B japonicum togetherbull only the last treatment accelerated germination - active material was the LCO the bacteria-to-plant signal

New LCO ActivitybullWe also found that the return signals (lipo-chitooligosaccharides) also directly stimulate growth of non-legume plants and this can increase yields

Control LCO

Corn Seedlings

01020304050607080

Corn Cotton Beet Soybean

ControlLCO

Commercializationbull Bios Agricluture eventually taken up by Agribiotics (also a Canadian Company)

bull Agribiotics was purchased by EMDbull EMD became part of Novozymes

Commercializationbull Established a spin-off company

bull Bios Agriculture Incbull To keep a technician employed post recession

bull Spin-offs were quite new at McGill University

bull 5 share non-dilutablebull The usual things

bull Venture capital and other investmentsbull Regulatory requirements must be satisfied

Serendipity amp Follow Up bull Earlier emergence observed at field sites

where genistein appliedbull What did this meanbull Experiment

bull seeds in waterbull genistein alonebull Bradyrhizobium japonicum alonebull genistein and B japonicum togetherbull only the last treatment accelerated germination - active material was the LCO the bacteria-to-plant signal

New LCO ActivitybullWe also found that the return signals (lipo-chitooligosaccharides) also directly stimulate growth of non-legume plants and this can increase yields

Control LCO

Corn Seedlings

01020304050607080

Corn Cotton Beet Soybean

ControlLCO

Commercializationbull Bios Agricluture eventually taken up by Agribiotics (also a Canadian Company)

bull Agribiotics was purchased by EMDbull EMD became part of Novozymes

Serendipity amp Follow Up bull Earlier emergence observed at field sites

where genistein appliedbull What did this meanbull Experiment

bull seeds in waterbull genistein alonebull Bradyrhizobium japonicum alonebull genistein and B japonicum togetherbull only the last treatment accelerated germination - active material was the LCO the bacteria-to-plant signal

New LCO ActivitybullWe also found that the return signals (lipo-chitooligosaccharides) also directly stimulate growth of non-legume plants and this can increase yields

Control LCO

Corn Seedlings

01020304050607080

Corn Cotton Beet Soybean

ControlLCO

Commercializationbull Bios Agricluture eventually taken up by Agribiotics (also a Canadian Company)

bull Agribiotics was purchased by EMDbull EMD became part of Novozymes

New LCO ActivitybullWe also found that the return signals (lipo-chitooligosaccharides) also directly stimulate growth of non-legume plants and this can increase yields

Control LCO

Corn Seedlings

01020304050607080

Corn Cotton Beet Soybean

ControlLCO

Commercializationbull Bios Agricluture eventually taken up by Agribiotics (also a Canadian Company)

bull Agribiotics was purchased by EMDbull EMD became part of Novozymes

Commercializationbull Bios Agricluture eventually taken up by Agribiotics (also a Canadian Company)

bull Agribiotics was purchased by EMDbull EMD became part of Novozymes

Jasmonates ndash An Alternativebull Flavonoids have two problems

bull Extremely expensive (up to $1000 per mg)bull Damaging to rhizobial cells (20 microM slows

growth)bull A paper suggested that jasmonic acid might also activate nod genes

bull Much less expensivebull Not damaging to rhizobial cells bull Generally involved in stress related signaling

bull Biotic and abiotic stressesbull In this case signal to the microbe that the plant is stressed and

the microbe responds with a signal to manage stressbull Both jasmonates and flavonoids stress realted in plant

Findingsbull Jasmonates induced the expression of nod genes in B japonicum

bull Also induce return LCO signal production

bull However the strain specificity is very different from isoflavonoids

bull Accelerates nodulation at low root zone temperature

bull Commercialized

Concentrations (microM)0 20 40 60 80 100 120B

-gal

acto

sida

se a

ctiv

ity a

ssay

(Mill

er U

nits

)

0

1000

2000

3000

4000

5000

6000

Figure 1 Effect of various concentrations of genistein JA MeJA SAand ASA on the induction of nod genes in Bradyrhizobium japonicumstrain USDA3 as measured by B-galactosidase activity (Miller Units)

ASA SA JA MeJAGenistein

Jasmonates

Genistein

Effect of Methyl Jasmonate on Yield

AA

B

C

D

Two sites two strains at each this is typical data ndash strain 532Cat a sandy-loam site

Increase over control 4 12 19 17

3313233343536373839

441

Soybean Yield (t per ha)

ControlInocGen 5uMMeJA 20uMGen+MeJA

Environment - Temperature and Responsebull In the lab responses could be variable and frustrating

bull In the field responses were strongest in cold springs on heavy soils

bull Over the last few years have shown that low temperatures make the lab results clearer amp consistent

0

20

40

60

80

100

120

0 h 24 h 36 h 48 h 60 hTime in hours after stratification

g

erm

inati

on

Control

LCO A

LCO B

TH A

TH B

0

10

20

30

40

50

60

70

80

90

100

0 h 96 h 108 h 120 h 132 h 144 h

g

erm

ina

tio

n

Time in hours after stratification

Control LCO A LCO B TH A TH B

Arabidopsis at 22 oC

Arabidopsis at 10 oC

Set 3 normal arab

Control

LCO A

LCO B

TH A

TH B

Time in hours after preincubation at 4 deg C in dark

germination

Arabidopsis set 3 Normal temp

0

0

0

0

0

567825361513

619359959893

497094402277

609031198686

723514911015

839780953196

817360182709

793480194497

827134646962

898298460798

891825960469

930634469697

906368595825

869663382594

956048581049

New line graph

Control

LCO A

LCO B

TH A

TH B

Time in hours after stratification

germination

0

0

0

0

0

0

0

0

0

0

567825361513

619359959893

497094402277

609031198686

723514911015

839780953196

817360182709

793480194497

827134646962

898298460798

891825960469

930634469697

906368595825

869663382594

956048581049

Sheet1

Sheet2

FW set 3 Arab normal

FW (mg)

Treatments

Weight in mg

Fresh weight Arabidopsis Normal temp set 3

1104666666667

121075

767

145425

137025

Dry weight Arab set 3 Normal

DW (mg)

Treatments

Weight in mg

Dry weight of Arabidopsis set 3 Normal temp

61353333333

429

372375

57405

5767

Sheet3

Set 3 Low temp LCO TH

Control

LCO A

LCO B

TH A

TH B

Time in hours after stratification

germination

0

0

0

0

0

09615384615

185238095238

180776259336

200991842788

215801790802

100432437932

354880952381

343139202288

402271041898

397858160358

607626170126

736428571429

671569601144

646366332963

593594831095

778958078958

781428571429

773220244716

696421950315

741035816036

885134310134

875476190476

808318766884

81318131257

78457977208

Low temp

Control

LCO A

LCO B

TH A

TH B

Time in hours after stratification

germination

Low temp

0

0

0

0

0

09615384615

185238095238

180776259336

200991842788

215801790802

100432437932

354880952381

343139202288

402271041898

397858160358

607626170126

736428571429

671569601144

646366332963

593594831095

778958078958

781428571429

773220244716

696421950315

741035816036

Sheet1

Sheet2

Sheet3

Salt amp Drought Stress

bullArabidopsisbullSeeds on petriplates with signals

bullControl and 150 mM NaCl

bullSignals improved growth under salt stress

0 NaCl 150 mM NaCl

0 10-6 10-8 10-9 10-11 M LCO Thuricin 17

Hormones of the Holobiontbull Effective at very low concentrationsbull Regulate plant activity

bull Large changes in gene espression protein production hormone profile

bull Regulate other microbesbull Powerful plant productivity effectsbull Inexpensive to applybull Low environmental impact

bull Small amountbull Already produced in environment

bull Very poorly understoodbull Enormous potential

bull New networkbull gt 10 strains in lab now

Phytomicrobiome Potentialbull Wide range of species in the phytomicrobiomebull Most powerful control factor is the plant

bull Developmentbull Stressbull Nutrition

bull Are key to plant growth and productivitybull Management

bull Inoculationbull Hub species bull Signals

bull Microbe-to-plantbull Plant-to-microbebull Microbe-to-microbe

Development of Biologicals - Whobull Scope is largebull Collaboration

bull Basic research labs bull Academicbull Government

bull Industry laboratories

The Approachbull Sample plant-associated microbes

bull The phytomicrobiomebull Efficientrapid screening to identify microbes of interest

bull Biological assessmentsbull Agronomic assessments

Control Thuricin

Industrial Collaboratorsbull Synagribull EVLbull BASFbull RavenQuestbull Ulyssebull Agroworld Natural Productsbull Inocucorbull Axter

BioFuelNet - NCE

Past and Present Students Dr Feng Zhang Dr Fazli Mabood Dr Narjes Dashti Keldeagh Lindsay Dr Pan Bo Dr Sowmya Subramanian Dr Yuming Bai Dr Xiaomin Zhou Elizabeth Gray Nan Wang

PDFs amp Research Associates Dr Xiaomin Zhou Dr Balakrishna Prithiviraj Dr Alfred Souleimanov Dr Kung Dong Lee Dr Woo Jung Mike Lewis Stewart Leibovitch

Acknowledgements

ALSO Trevor Charles at U Waterloo

The End

Thank You

• Biostimulants from the phytomicrobiome Microbe-to-plant signals as low input crop management technologies
• Need for Enhanced Food Production
• Threat of Climate Change
• Biomass for Advanced Biofuels and the Bioeconomy
• The Phytomicrobiome
• Adapting to Climate Change
• The Approach
• So Far
• Example Line of Research
• Something New
• The Problem
• A Solution
• N2 Fixation and Growth - Genistein
• Commercialization
• Serendipity amp Follow Up
• New LCO Activity
• Commercialization
• Jasmonates ndash An Alternative
• Findings
• Effect of Methyl Jasmonate on Yield
• Environment - Temperature and Response
• Salt amp Drought Stress
• Hormones of the Holobiont
• Phytomicrobiome Potential
• Development of Biologicals - Who
• The Approach
• Industrial Collaborators
• BioFuelNet - NCE
• Diapositive numeacutero 29
• The End

Control		96h		000
Control		96h		385
Control		96h		000
Control		96h		000
Control		108h		667
Control		108h		1538
Control		108h		741
Control		108h		1071
Control		120h		5000
Control		120h		7308
Control		120h		5926
Control		120h		6071
Control		132h		6333
Control		132h		8846
Control		132h		7407
Control		132h		8571
Control		144h		8000
Control		144h		9231
Control		144h		8889
Control		144h		9286
LCO A		96h		1600
LCO A		96h		1667
LCO A		96h		2000
LCO A		96h		2143
LCO A		108h		3600
LCO A		108h		2667
LCO A		108h		4000
LCO A		108h		3929
LCO A		120h		7600
LCO A		120h		6333
LCO A		120h		7667
LCO A		120h		7857
LCO A		132h		8400
LCO A		132h		7000
LCO A		132h		8000
LCO A		132h		7857
LCO A		144h		8400
LCO A		144h		8000
LCO A		144h		9333
LCO A		144h		9286
LCO B		96h		1429
LCO B		96h		1935
LCO B		96h		1724
LCO B		96h		2143
LCO B		108h		2857
LCO B		108h		4194
LCO B		108h		3103
LCO B		108h		3571
LCO B		120h		6786
LCO B		120h		7097
LCO B		120h		6552
LCO B		120h		6429
LCO B		132h		7143
LCO B		132h		9032
LCO B		132h		6897
LCO B		132h		7857
LCO B		144h		7857
LCO B		144h		9032
LCO B		144h		7586
LCO B		144h		7857
TH A		96h		2258
TH A		96h		1379
TH A		96h		2069
TH A		96h		2333
TH A		108h		4516
TH A		108h		3448
TH A		108h		3793
TH A		108h		4333
TH A		120h		6774
TH A		120h		6552
TH A		120h		5862
TH A		120h		6667
TH A		132h		7742
TH A		132h		6552
TH A		132h		6897
TH A		132h		6667
TH A		144h		9355
TH A		144h		7241
TH A		144h		7931
TH A		144h		8000
TH B		96h		2000
TH B		96h		1923
TH B		96h		1852
TH B		96h		2857
TH B		108h		3667
TH B		108h		4615
TH B		108h		3704
TH B		108h		3929
TH B		120h		6333
TH B		120h		6154
TH B		120h		5185
TH B		120h		6071
TH B		132h		7000
TH B		132h		8462
TH B		132h		7037
TH B		132h		7143
TH B		144h		8000
TH B		144h		8846
TH B		144h		7037
TH B		144h		7500
LCOA+THA		96h		2500
LCOA+THA		96h		2308
LCOA+THA		96h		1852
LCOA+THA		96h		2800
LCOA+THA		108h		3929
LCOA+THA		108h		3462
LCOA+THA		108h		4444
LCOA+THA		108h		5200
LCOA+THA		120h		7143
LCOA+THA		120h		5385
LCOA+THA		120h		7037
LCOA+THA		120h		7200
LCOA+THA		132h		7500
LCOA+THA		132h		7308
LCOA+THA		132h		8519
LCOA+THA		132h		8400
LCOA+THA		144h		8929
LCOA+THA		144h		8077
LCOA+THA		144h		8519
LCOA+THA		144h		8400
LCOB+THB		96h		1852
LCOB+THB		96h		1724
LCOB+THB		96h		3103
LCOB+THB		96h		1875
LCOB+THB		108h		3704
LCOB+THB		108h		3793
LCOB+THB		108h		6207
LCOB+THB		108h		4063
LCOB+THB		120h		6296
LCOB+THB		120h		6207
LCOB+THB		120h		8276
LCOB+THB		120h		5625
LCOB+THB		132h		7037
LCOB+THB		132h		6552
LCOB+THB		132h		8966
LCOB+THB		132h		7188
LCOB+THB		144h		8519
LCOB+THB		144h		7586
LCOB+THB		144h		9310
LCOB+THB		144h		8750

Arabidopsis low temperature set 3
Treatment		No seeds		96 h		germ		Std		SE		108 h		germ		Std		SE		120 h		germ		Std		SE		132 h		germ		Std		SE		144 h		germ		Std		SE
Control 1		30		0		000				2		667				15		5000				19		6333				24		8000
2		26		1		385				4		1538				19		7308				23		8846				24		9231
3		27		0		000				2		741				16		5926				20		7407				24		8889
4		28		0		000				3		1071				17		6071				24		8571				26		9286
	111		025		096		192		096		275		1004		397		199		1675		6076		948		474		215		7790		1154		577		245		8851		594		297
LCO A 1		25		4		1600				9		3600				19		7600				21		8400				21		8400
2		30		5		1667				8		2667				19		6333				21		7000				24		8000
3		30		6		2000				12		4000				23		7667				24		8000				28		9333
4		28		6		2143				11		3929				22		7857				22		7857				26		9286
	113		525		1852		261		130		10		3549		613		307		2075		7364		696		110		22		7814		589		295		2475		8755		661		331
LCO B 1		28		4		1429				8		2857				19		6786				20		7143				22		7857
2		31		6		1935				13		4194				22		7097				28		9032				28		9032
3		29		5		1724				9		3103				19		6552				20		6897				22		7586
4		28		6		2143				10		3571				18		6429				22		7857				22		7857
	116		525		1808		305		153		10		3431		588		294		195		6716		294		147		225		7732		958		479		235		8083		645		323
TH A 1		31		7		2258				14		4516				21		6774				24		7742				29		9355
2		29		4		1379				10		3448				19		6552				19		6552				21		7241
3		29		6		2069				11		3793				17		5862				20		6897				23		7931
4		30		7		2333				13		4333				20		6667				20		6667				24		8000
	119		6		2010		435		217		12		4023		491		245		1925		6464		411		206		2075		6964		538		269		2425		8132		884		442
TH B 1		30		6		2000				11		3667				19		6333				21		7000				24		8000
2		26		5		1923				12		4615				16		6154				22		8462				23		8846
3		27		5		1852				10		3704				14		5185				19		7037				19		7037
4		28		8		2857				11		3929				17		6071				20		7143				21		7500
	111		6		2158		470		235		11		3979		440		220		165		5936		512		256		205		7410		703		352		2175		7846		774		387
LCOA+THA1		28		7		2500				11		3929				20		7143				21		7500				25		8929
2		26		6		2308				9		3462				14		5385				19		7308				21		8077
3		27		5		1852				12		4444				19		7037				23		8519				23		8519
4		25		7		2800				13		5200				18		7200				21		8400				21		8400
	106		625		2365		398		199		1125		4259		745		372		1775		6691		874		437		21		7932		616		308		225		8481		352		176
LCOB+THB1		27		5		1852				10		3704				17		6296				19		7037				23		8519
2		29		5		1724				11		3793				18		6207				19		6552				22		7586
3		29		9		3103				18		6207				24		8276				26		8966				27		9310
4		32		6		1875				13		4063				18		5625				23		7188				28		8750
	11700		625		2139		647		323		1300		4442		1187		593		1925		6601		1156		183		2175		7435		1056		528		2500		8541		718		359
germination
Treatment		0 h		96 h		108 h		120 h		132 h		144 h
Control		0		096		1004		6076		7790		8851
LCO A		0		1852		3549		7364		7814		8755
LCO B		0		1808		3431		6716		7732		8083
TH A		0		2010		4023		6464		6964		8132
TH B		0		2158		3979		5936		7410		7846
LCOA+THA		0		2365		4259		6691		7932		8481
LCOB+THB		0		2139		4442		6601		7435		8541
SE
	0 h		96 h		108 h		120 h		132 h		144 h
Control		0		096		199		474		577		297
LCO A		0		130		307		110		295		331
LCO B		0		153		294		147		479		323
TH A		0		217		245		206		269		442
TH B		0		235		220		256		352		387
LCOA+THA		0		199		372		437		308		176
LCOB+THB		0		323		593		183		528		359

0 h		0 h		0 h		0 h		0 h
96 h		96 h		96 h		96 h		96 h
108 h		108 h		108 h		108 h		108 h
120 h		120 h		120 h		120 h		120 h
132 h		132 h		132 h		132 h		132 h

0 h		0 h		0 h		0 h		0 h		0		0		0		0		0		0		0		0		0		0
96 h		96 h		96 h		96 h		96 h		09615384615		09615384615		13049704387		13049704387		15258333938		15258333938		2174332839		2174332839		23499650247		2174332839
108 h		108 h		108 h		108 h		108 h		19859811656		19859811656		30667415327		30667415327		29408073778		29408073778		24539813637		24539813637		2200128559		24539813637
120 h		120 h		120 h		120 h		120 h		47411187081		47411187081		11002902798		11002902798		14704036889		14704036889		20561084936		20561084936		25615660866		20561084936
132 h		132 h		132 h		132 h		132 h		57694206278		57694206278		29473844048		29473844048		4788349344		4788349344		26896579581		26896579581		35169817799		26896579581
144 h		144 h		144 h		144 h		144 h		29704619111		29704619111		33067820092		33067820092		32273789905		32273789905		4421868687		4421868687		3871003991		4421868687

Weight of seedlings
Treatment		FW (mg)		STD		SE			DW (mg)		STD		SE					Treatment		FW (mg)		STD		SE			DW (mg)		STD		SE
Control 1		39850					2924							Control 1
2		11940					854							2		11940					854
3		10680					468							3		10680					468
4		10520					518							4		10520					518
	11047		778		449			614		210		121						11047		778		449			614		210		121
LCO A 1		15860					899							LCO A 1
2		11320					449							2		11320					449
3		10640					437							3		10640					437
4		10610					400							4		10610					400
	12108		2523		1262			546		236		118						12108		2523		1262			429		026		015
LCO B 1		8040					416							LCO B 1		8040					416
2		7560					333							2		7560					333
3		8020					378							3		8020					378
4		7060					363							4		7060					363
	7670		463		232			372		035		017						7670		463		232			372		035		017
TH A 1		14760					632							TH A 1		14760					632
2		12180					457							2		12180					457
3		14450					469							3		14450					469
4		16780					739							4		16780					739
	14543		1884		942			574		136		068						14543		1884		942			574		136		068
TH B 1		10980					391							TH B 1		10980					391
2		15910					701							2		15910					701
3		17040					815							3		17040					815
4		10880					400							4		10880					400
	13703		3235		1617			577		215		107						13703		3235		1617			577		215		107
Treatment		FW (mg)		SE			Treatment		DW (mg)		SE
Control		11047		449			Control		614		121
LCO A		12108		1262			LCO A		429		016
LCO B		7670		232			LCO B		372		017
TH A		14543		942			TH A		574		068
TH B		13703		1617			TH B		577		107

Control		12125587729
LCO A		0155
LCO B		01725714224
TH A		0678241169
TH B		10731213818

Control		44904837651
LCO A		126152932982
LCO B		2315887159
TH A		94179239574
TH B		161736604288

Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B

Arabidopsis normal temperature set 3 continuous
Treatment		No seeds		24 h		germ		Std		SE		36 h		germ		Std		SE		48 h		germ		Std		SE		60 h		germ		Std		SE
Control 1		31		0					16		5161				23		7419				26		8387
2		29		0					19		6552				27		9310				28		9655
3		25		0					15		6000				21		8400				21		8400
4		26		0					13		5000				22		8462				24		9231
	111						1575		5678		729		364		2325		8398		773		387		2475		8918		630		315
LCO A 1		34		0					20		5882				26		7647				34		10000
2		32		0					19		5938				27		8438				30		9375
3		33		0					18		5455				28		8485				29		8788
4		32		0					24		7500				26		8125				29		9063
	131						2025		6194		897		449		2675		8174		386		193		305		9306		521		260
LCO B 1		31		0					14		4516				23		7419				26		8387
2		34		0					15		4412				31		9118				34		10000
3		32		0					20		6250				27		8438				28		8750
4		34		0					16		4706				23		6765				31		9118
	131						1625		4971		861		431		26		7935		1047		523		2975		9064		692		346
TH A 1		28		0					18		6429				23		8214				23		8214
2		29		0					19		6552				24		8276				27		9310
3		30		0					17		5667				23		7667				25		8333
4		28		0					16		5714				25		8929				25		8929
	115						175		6090		465		232		2375		8271		517		258		25		8697		515		257
TH B 1		33		0					23		6970				28		8485				32		9697
2		28		0					19		6786				25		8929				26		9286
3		27		0					21		7778				26		9630				27		10000
4		27		0					20		7407				24		8889				25		9259
	115						2075		7235		446		223		2575		8983		475		238		275		9560		355		177
germination
Treatment		0 h		24 h		36 h		48 h		60 h
Control		0		0		5678		8398		8918
LCO A		0		0		6194		8174		9306
LCO B		0		0		4971		7935		9064
TH A		0		0		6090		8271		8697
TH B		0		0		7235		8983		9560
SE
Control		0		0		364		387		315
LCO A		0		0		449		193		260
LCO B		0		0		431		523		346
TH A		0		0		232		258		257
TH B		0		0		223		238		177

0 h		0 h		0 h		0 h		0 h		0		0		0		0		0		0		0		0		0		0
24 h		24 h		24 h		24 h		24 h		0		0		0		0		0		0		0		0		0		0
36 h		36 h		36 h		36 h		36 h		3644345509		3644345509		44864095916		44864095916		3644345509		3644345509		23241337778		23241337778		2229677279		2229677279
48 h		48 h		48 h		48 h		48 h		38666958625		38666958625		19281389988		19281389988		38666958625		38666958625		2583374136		2583374136		23771554774		23771554774
60 h		60 h		60 h		60 h		60 h		31509650968		31509650968		26047046621		26047046621		31509650968		31509650968		25740422067		25740422067		1774911735		1774911735

24 h		24 h		24 h		24 h		24 h		0		0		0		0		0		0		0		0		0		0
36 h		36 h		36 h		36 h		36 h		3644345509		3644345509		44864095916		44864095916		43067583186		43067583186		23241337778		23241337778		2229677279		2229677279
48 h		48 h		48 h		48 h		48 h		38666958625		38666958625		19281389988		19281389988		52334169948		52334169948		2583374136		2583374136		23771554774		23771554774
60 h		60 h		60 h		60 h		60 h		31509650968		31509650968		26047046621		26047046621		34590070704		34590070704		25740422067		25740422067		1774911735		1774911735

Findingsbull Jasmonates induced the expression of nod genes in B japonicum

bull Also induce return LCO signal production

bull However the strain specificity is very different from isoflavonoids

bull Accelerates nodulation at low root zone temperature

bull Commercialized

Concentrations (microM)0 20 40 60 80 100 120B

-gal

acto

sida

se a

ctiv

ity a

ssay

(Mill

er U

nits

)

0

1000

2000

3000

4000

5000

6000

Figure 1 Effect of various concentrations of genistein JA MeJA SAand ASA on the induction of nod genes in Bradyrhizobium japonicumstrain USDA3 as measured by B-galactosidase activity (Miller Units)

ASA SA JA MeJAGenistein

Jasmonates

Genistein

Effect of Methyl Jasmonate on Yield

AA

B

C

D

Two sites two strains at each this is typical data ndash strain 532Cat a sandy-loam site

Increase over control 4 12 19 17

3313233343536373839

441

Soybean Yield (t per ha)

ControlInocGen 5uMMeJA 20uMGen+MeJA

Environment - Temperature and Responsebull In the lab responses could be variable and frustrating

bull In the field responses were strongest in cold springs on heavy soils

bull Over the last few years have shown that low temperatures make the lab results clearer amp consistent

0

20

40

60

80

100

120

0 h 24 h 36 h 48 h 60 hTime in hours after stratification

g

erm

inati

on

Control

LCO A

LCO B

TH A

TH B

0

10

20

30

40

50

60

70

80

90

100

0 h 96 h 108 h 120 h 132 h 144 h

g

erm

ina

tio

n

Time in hours after stratification

Control LCO A LCO B TH A TH B

Arabidopsis at 22 oC

Arabidopsis at 10 oC

Set 3 normal arab

Control

LCO A

LCO B

TH A

TH B

Time in hours after preincubation at 4 deg C in dark

germination

Arabidopsis set 3 Normal temp

0

0

0

0

0

567825361513

619359959893

497094402277

609031198686

723514911015

839780953196

817360182709

793480194497

827134646962

898298460798

891825960469

930634469697

906368595825

869663382594

956048581049

New line graph

Control

LCO A

LCO B

TH A

TH B

Time in hours after stratification

germination

0

0

0

0

0

0

0

0

0

0

567825361513

619359959893

497094402277

609031198686

723514911015

839780953196

817360182709

793480194497

827134646962

898298460798

891825960469

930634469697

906368595825

869663382594

956048581049

Sheet1

Sheet2

FW set 3 Arab normal

FW (mg)

Treatments

Weight in mg

Fresh weight Arabidopsis Normal temp set 3

1104666666667

121075

767

145425

137025

Dry weight Arab set 3 Normal

DW (mg)

Treatments

Weight in mg

Dry weight of Arabidopsis set 3 Normal temp

61353333333

429

372375

57405

5767

Sheet3

Set 3 Low temp LCO TH

Control

LCO A

LCO B

TH A

TH B

Time in hours after stratification

germination

0

0

0

0

0

09615384615

185238095238

180776259336

200991842788

215801790802

100432437932

354880952381

343139202288

402271041898

397858160358

607626170126

736428571429

671569601144

646366332963

593594831095

778958078958

781428571429

773220244716

696421950315

741035816036

885134310134

875476190476

808318766884

81318131257

78457977208

Low temp

Control

LCO A

LCO B

TH A

TH B

Time in hours after stratification

germination

Low temp

0

0

0

0

0

09615384615

185238095238

180776259336

200991842788

215801790802

100432437932

354880952381

343139202288

402271041898

397858160358

607626170126

736428571429

671569601144

646366332963

593594831095

778958078958

781428571429

773220244716

696421950315

741035816036

Sheet1

Sheet2

Sheet3

Salt amp Drought Stress

bullArabidopsisbullSeeds on petriplates with signals

bullControl and 150 mM NaCl

bullSignals improved growth under salt stress

0 NaCl 150 mM NaCl

0 10-6 10-8 10-9 10-11 M LCO Thuricin 17

Hormones of the Holobiontbull Effective at very low concentrationsbull Regulate plant activity

bull Large changes in gene espression protein production hormone profile

bull Regulate other microbesbull Powerful plant productivity effectsbull Inexpensive to applybull Low environmental impact

bull Small amountbull Already produced in environment

bull Very poorly understoodbull Enormous potential

bull New networkbull gt 10 strains in lab now

Phytomicrobiome Potentialbull Wide range of species in the phytomicrobiomebull Most powerful control factor is the plant

bull Developmentbull Stressbull Nutrition

bull Are key to plant growth and productivitybull Management

bull Inoculationbull Hub species bull Signals

bull Microbe-to-plantbull Plant-to-microbebull Microbe-to-microbe

Development of Biologicals - Whobull Scope is largebull Collaboration

bull Basic research labs bull Academicbull Government

bull Industry laboratories

The Approachbull Sample plant-associated microbes

bull The phytomicrobiomebull Efficientrapid screening to identify microbes of interest

bull Biological assessmentsbull Agronomic assessments

Control Thuricin

Industrial Collaboratorsbull Synagribull EVLbull BASFbull RavenQuestbull Ulyssebull Agroworld Natural Productsbull Inocucorbull Axter

BioFuelNet - NCE

Past and Present Students Dr Feng Zhang Dr Fazli Mabood Dr Narjes Dashti Keldeagh Lindsay Dr Pan Bo Dr Sowmya Subramanian Dr Yuming Bai Dr Xiaomin Zhou Elizabeth Gray Nan Wang

PDFs amp Research Associates Dr Xiaomin Zhou Dr Balakrishna Prithiviraj Dr Alfred Souleimanov Dr Kung Dong Lee Dr Woo Jung Mike Lewis Stewart Leibovitch

Acknowledgements

ALSO Trevor Charles at U Waterloo

The End

Thank You

• Biostimulants from the phytomicrobiome Microbe-to-plant signals as low input crop management technologies
• Need for Enhanced Food Production
• Threat of Climate Change
• Biomass for Advanced Biofuels and the Bioeconomy
• The Phytomicrobiome
• Adapting to Climate Change
• The Approach
• So Far
• Example Line of Research
• Something New
• The Problem
• A Solution
• N2 Fixation and Growth - Genistein
• Commercialization
• Serendipity amp Follow Up
• New LCO Activity
• Commercialization
• Jasmonates ndash An Alternative
• Findings
• Effect of Methyl Jasmonate on Yield
• Environment - Temperature and Response
• Salt amp Drought Stress
• Hormones of the Holobiont
• Phytomicrobiome Potential
• Development of Biologicals - Who
• The Approach
• Industrial Collaborators
• BioFuelNet - NCE
• Diapositive numeacutero 29
• The End

Control		96h		000
Control		96h		385
Control		96h		000
Control		96h		000
Control		108h		667
Control		108h		1538
Control		108h		741
Control		108h		1071
Control		120h		5000
Control		120h		7308
Control		120h		5926
Control		120h		6071
Control		132h		6333
Control		132h		8846
Control		132h		7407
Control		132h		8571
Control		144h		8000
Control		144h		9231
Control		144h		8889
Control		144h		9286
LCO A		96h		1600
LCO A		96h		1667
LCO A		96h		2000
LCO A		96h		2143
LCO A		108h		3600
LCO A		108h		2667
LCO A		108h		4000
LCO A		108h		3929
LCO A		120h		7600
LCO A		120h		6333
LCO A		120h		7667
LCO A		120h		7857
LCO A		132h		8400
LCO A		132h		7000
LCO A		132h		8000
LCO A		132h		7857
LCO A		144h		8400
LCO A		144h		8000
LCO A		144h		9333
LCO A		144h		9286
LCO B		96h		1429
LCO B		96h		1935
LCO B		96h		1724
LCO B		96h		2143
LCO B		108h		2857
LCO B		108h		4194
LCO B		108h		3103
LCO B		108h		3571
LCO B		120h		6786
LCO B		120h		7097
LCO B		120h		6552
LCO B		120h		6429
LCO B		132h		7143
LCO B		132h		9032
LCO B		132h		6897
LCO B		132h		7857
LCO B		144h		7857
LCO B		144h		9032
LCO B		144h		7586
LCO B		144h		7857
TH A		96h		2258
TH A		96h		1379
TH A		96h		2069
TH A		96h		2333
TH A		108h		4516
TH A		108h		3448
TH A		108h		3793
TH A		108h		4333
TH A		120h		6774
TH A		120h		6552
TH A		120h		5862
TH A		120h		6667
TH A		132h		7742
TH A		132h		6552
TH A		132h		6897
TH A		132h		6667
TH A		144h		9355
TH A		144h		7241
TH A		144h		7931
TH A		144h		8000
TH B		96h		2000
TH B		96h		1923
TH B		96h		1852
TH B		96h		2857
TH B		108h		3667
TH B		108h		4615
TH B		108h		3704
TH B		108h		3929
TH B		120h		6333
TH B		120h		6154
TH B		120h		5185
TH B		120h		6071
TH B		132h		7000
TH B		132h		8462
TH B		132h		7037
TH B		132h		7143
TH B		144h		8000
TH B		144h		8846
TH B		144h		7037
TH B		144h		7500
LCOA+THA		96h		2500
LCOA+THA		96h		2308
LCOA+THA		96h		1852
LCOA+THA		96h		2800
LCOA+THA		108h		3929
LCOA+THA		108h		3462
LCOA+THA		108h		4444
LCOA+THA		108h		5200
LCOA+THA		120h		7143
LCOA+THA		120h		5385
LCOA+THA		120h		7037
LCOA+THA		120h		7200
LCOA+THA		132h		7500
LCOA+THA		132h		7308
LCOA+THA		132h		8519
LCOA+THA		132h		8400
LCOA+THA		144h		8929
LCOA+THA		144h		8077
LCOA+THA		144h		8519
LCOA+THA		144h		8400
LCOB+THB		96h		1852
LCOB+THB		96h		1724
LCOB+THB		96h		3103
LCOB+THB		96h		1875
LCOB+THB		108h		3704
LCOB+THB		108h		3793
LCOB+THB		108h		6207
LCOB+THB		108h		4063
LCOB+THB		120h		6296
LCOB+THB		120h		6207
LCOB+THB		120h		8276
LCOB+THB		120h		5625
LCOB+THB		132h		7037
LCOB+THB		132h		6552
LCOB+THB		132h		8966
LCOB+THB		132h		7188
LCOB+THB		144h		8519
LCOB+THB		144h		7586
LCOB+THB		144h		9310
LCOB+THB		144h		8750

Arabidopsis low temperature set 3
Treatment		No seeds		96 h		germ		Std		SE		108 h		germ		Std		SE		120 h		germ		Std		SE		132 h		germ		Std		SE		144 h		germ		Std		SE
Control 1		30		0		000				2		667				15		5000				19		6333				24		8000
2		26		1		385				4		1538				19		7308				23		8846				24		9231
3		27		0		000				2		741				16		5926				20		7407				24		8889
4		28		0		000				3		1071				17		6071				24		8571				26		9286
	111		025		096		192		096		275		1004		397		199		1675		6076		948		474		215		7790		1154		577		245		8851		594		297
LCO A 1		25		4		1600				9		3600				19		7600				21		8400				21		8400
2		30		5		1667				8		2667				19		6333				21		7000				24		8000
3		30		6		2000				12		4000				23		7667				24		8000				28		9333
4		28		6		2143				11		3929				22		7857				22		7857				26		9286
	113		525		1852		261		130		10		3549		613		307		2075		7364		696		110		22		7814		589		295		2475		8755		661		331
LCO B 1		28		4		1429				8		2857				19		6786				20		7143				22		7857
2		31		6		1935				13		4194				22		7097				28		9032				28		9032
3		29		5		1724				9		3103				19		6552				20		6897				22		7586
4		28		6		2143				10		3571				18		6429				22		7857				22		7857
	116		525		1808		305		153		10		3431		588		294		195		6716		294		147		225		7732		958		479		235		8083		645		323
TH A 1		31		7		2258				14		4516				21		6774				24		7742				29		9355
2		29		4		1379				10		3448				19		6552				19		6552				21		7241
3		29		6		2069				11		3793				17		5862				20		6897				23		7931
4		30		7		2333				13		4333				20		6667				20		6667				24		8000
	119		6		2010		435		217		12		4023		491		245		1925		6464		411		206		2075		6964		538		269		2425		8132		884		442
TH B 1		30		6		2000				11		3667				19		6333				21		7000				24		8000
2		26		5		1923				12		4615				16		6154				22		8462				23		8846
3		27		5		1852				10		3704				14		5185				19		7037				19		7037
4		28		8		2857				11		3929				17		6071				20		7143				21		7500
	111		6		2158		470		235		11		3979		440		220		165		5936		512		256		205		7410		703		352		2175		7846		774		387
LCOA+THA1		28		7		2500				11		3929				20		7143				21		7500				25		8929
2		26		6		2308				9		3462				14		5385				19		7308				21		8077
3		27		5		1852				12		4444				19		7037				23		8519				23		8519
4		25		7		2800				13		5200				18		7200				21		8400				21		8400
	106		625		2365		398		199		1125		4259		745		372		1775		6691		874		437		21		7932		616		308		225		8481		352		176
LCOB+THB1		27		5		1852				10		3704				17		6296				19		7037				23		8519
2		29		5		1724				11		3793				18		6207				19		6552				22		7586
3		29		9		3103				18		6207				24		8276				26		8966				27		9310
4		32		6		1875				13		4063				18		5625				23		7188				28		8750
	11700		625		2139		647		323		1300		4442		1187		593		1925		6601		1156		183		2175		7435		1056		528		2500		8541		718		359
germination
Treatment		0 h		96 h		108 h		120 h		132 h		144 h
Control		0		096		1004		6076		7790		8851
LCO A		0		1852		3549		7364		7814		8755
LCO B		0		1808		3431		6716		7732		8083
TH A		0		2010		4023		6464		6964		8132
TH B		0		2158		3979		5936		7410		7846
LCOA+THA		0		2365		4259		6691		7932		8481
LCOB+THB		0		2139		4442		6601		7435		8541
SE
	0 h		96 h		108 h		120 h		132 h		144 h
Control		0		096		199		474		577		297
LCO A		0		130		307		110		295		331
LCO B		0		153		294		147		479		323
TH A		0		217		245		206		269		442
TH B		0		235		220		256		352		387
LCOA+THA		0		199		372		437		308		176
LCOB+THB		0		323		593		183		528		359

0 h		0 h		0 h		0 h		0 h
96 h		96 h		96 h		96 h		96 h
108 h		108 h		108 h		108 h		108 h
120 h		120 h		120 h		120 h		120 h
132 h		132 h		132 h		132 h		132 h

0 h		0 h		0 h		0 h		0 h		0		0		0		0		0		0		0		0		0		0
96 h		96 h		96 h		96 h		96 h		09615384615		09615384615		13049704387		13049704387		15258333938		15258333938		2174332839		2174332839		23499650247		2174332839
108 h		108 h		108 h		108 h		108 h		19859811656		19859811656		30667415327		30667415327		29408073778		29408073778		24539813637		24539813637		2200128559		24539813637
120 h		120 h		120 h		120 h		120 h		47411187081		47411187081		11002902798		11002902798		14704036889		14704036889		20561084936		20561084936		25615660866		20561084936
132 h		132 h		132 h		132 h		132 h		57694206278		57694206278		29473844048		29473844048		4788349344		4788349344		26896579581		26896579581		35169817799		26896579581
144 h		144 h		144 h		144 h		144 h		29704619111		29704619111		33067820092		33067820092		32273789905		32273789905		4421868687		4421868687		3871003991		4421868687

Weight of seedlings
Treatment		FW (mg)		STD		SE			DW (mg)		STD		SE					Treatment		FW (mg)		STD		SE			DW (mg)		STD		SE
Control 1		39850					2924							Control 1
2		11940					854							2		11940					854
3		10680					468							3		10680					468
4		10520					518							4		10520					518
	11047		778		449			614		210		121						11047		778		449			614		210		121
LCO A 1		15860					899							LCO A 1
2		11320					449							2		11320					449
3		10640					437							3		10640					437
4		10610					400							4		10610					400
	12108		2523		1262			546		236		118						12108		2523		1262			429		026		015
LCO B 1		8040					416							LCO B 1		8040					416
2		7560					333							2		7560					333
3		8020					378							3		8020					378
4		7060					363							4		7060					363
	7670		463		232			372		035		017						7670		463		232			372		035		017
TH A 1		14760					632							TH A 1		14760					632
2		12180					457							2		12180					457
3		14450					469							3		14450					469
4		16780					739							4		16780					739
	14543		1884		942			574		136		068						14543		1884		942			574		136		068
TH B 1		10980					391							TH B 1		10980					391
2		15910					701							2		15910					701
3		17040					815							3		17040					815
4		10880					400							4		10880					400
	13703		3235		1617			577		215		107						13703		3235		1617			577		215		107
Treatment		FW (mg)		SE			Treatment		DW (mg)		SE
Control		11047		449			Control		614		121
LCO A		12108		1262			LCO A		429		016
LCO B		7670		232			LCO B		372		017
TH A		14543		942			TH A		574		068
TH B		13703		1617			TH B		577		107

Control		12125587729
LCO A		0155
LCO B		01725714224
TH A		0678241169
TH B		10731213818

Control		44904837651
LCO A		126152932982
LCO B		2315887159
TH A		94179239574
TH B		161736604288

Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B

Arabidopsis normal temperature set 3 continuous
Treatment		No seeds		24 h		germ		Std		SE		36 h		germ		Std		SE		48 h		germ		Std		SE		60 h		germ		Std		SE
Control 1		31		0					16		5161				23		7419				26		8387
2		29		0					19		6552				27		9310				28		9655
3		25		0					15		6000				21		8400				21		8400
4		26		0					13		5000				22		8462				24		9231
	111						1575		5678		729		364		2325		8398		773		387		2475		8918		630		315
LCO A 1		34		0					20		5882				26		7647				34		10000
2		32		0					19		5938				27		8438				30		9375
3		33		0					18		5455				28		8485				29		8788
4		32		0					24		7500				26		8125				29		9063
	131						2025		6194		897		449		2675		8174		386		193		305		9306		521		260
LCO B 1		31		0					14		4516				23		7419				26		8387
2		34		0					15		4412				31		9118				34		10000
3		32		0					20		6250				27		8438				28		8750
4		34		0					16		4706				23		6765				31		9118
	131						1625		4971		861		431		26		7935		1047		523		2975		9064		692		346
TH A 1		28		0					18		6429				23		8214				23		8214
2		29		0					19		6552				24		8276				27		9310
3		30		0					17		5667				23		7667				25		8333
4		28		0					16		5714				25		8929				25		8929
	115						175		6090		465		232		2375		8271		517		258		25		8697		515		257
TH B 1		33		0					23		6970				28		8485				32		9697
2		28		0					19		6786				25		8929				26		9286
3		27		0					21		7778				26		9630				27		10000
4		27		0					20		7407				24		8889				25		9259
	115						2075		7235		446		223		2575		8983		475		238		275		9560		355		177
germination
Treatment		0 h		24 h		36 h		48 h		60 h
Control		0		0		5678		8398		8918
LCO A		0		0		6194		8174		9306
LCO B		0		0		4971		7935		9064
TH A		0		0		6090		8271		8697
TH B		0		0		7235		8983		9560
SE
Control		0		0		364		387		315
LCO A		0		0		449		193		260
LCO B		0		0		431		523		346
TH A		0		0		232		258		257
TH B		0		0		223		238		177

0 h		0 h		0 h		0 h		0 h		0		0		0		0		0		0		0		0		0		0
24 h		24 h		24 h		24 h		24 h		0		0		0		0		0		0		0		0		0		0
36 h		36 h		36 h		36 h		36 h		3644345509		3644345509		44864095916		44864095916		3644345509		3644345509		23241337778		23241337778		2229677279		2229677279
48 h		48 h		48 h		48 h		48 h		38666958625		38666958625		19281389988		19281389988		38666958625		38666958625		2583374136		2583374136		23771554774		23771554774
60 h		60 h		60 h		60 h		60 h		31509650968		31509650968		26047046621		26047046621		31509650968		31509650968		25740422067		25740422067		1774911735		1774911735

24 h		24 h		24 h		24 h		24 h		0		0		0		0		0		0		0		0		0		0
36 h		36 h		36 h		36 h		36 h		3644345509		3644345509		44864095916		44864095916		43067583186		43067583186		23241337778		23241337778		2229677279		2229677279
48 h		48 h		48 h		48 h		48 h		38666958625		38666958625		19281389988		19281389988		52334169948		52334169948		2583374136		2583374136		23771554774		23771554774
60 h		60 h		60 h		60 h		60 h		31509650968		31509650968		26047046621		26047046621		34590070704		34590070704		25740422067		25740422067		1774911735		1774911735

Effect of Methyl Jasmonate on Yield

AA

B

C

D

Two sites two strains at each this is typical data ndash strain 532Cat a sandy-loam site

Increase over control 4 12 19 17

3313233343536373839

441

Soybean Yield (t per ha)

ControlInocGen 5uMMeJA 20uMGen+MeJA

Environment - Temperature and Responsebull In the lab responses could be variable and frustrating

bull In the field responses were strongest in cold springs on heavy soils

bull Over the last few years have shown that low temperatures make the lab results clearer amp consistent

0

20

40

60

80

100

120

0 h 24 h 36 h 48 h 60 hTime in hours after stratification

g

erm

inati

on

Control

LCO A

LCO B

TH A

TH B

0

10

20

30

40

50

60

70

80

90

100

0 h 96 h 108 h 120 h 132 h 144 h

g

erm

ina

tio

n

Time in hours after stratification

Control LCO A LCO B TH A TH B

Arabidopsis at 22 oC

Arabidopsis at 10 oC

Set 3 normal arab

Control

LCO A

LCO B

TH A

TH B

Time in hours after preincubation at 4 deg C in dark

germination

Arabidopsis set 3 Normal temp

0

0

0

0

0

567825361513

619359959893

497094402277

609031198686

723514911015

839780953196

817360182709

793480194497

827134646962

898298460798

891825960469

930634469697

906368595825

869663382594

956048581049

New line graph

Control

LCO A

LCO B

TH A

TH B

Time in hours after stratification

germination

0

0

0

0

0

0

0

0

0

0

567825361513

619359959893

497094402277

609031198686

723514911015

839780953196

817360182709

793480194497

827134646962

898298460798

891825960469

930634469697

906368595825

869663382594

956048581049

Sheet1

Sheet2

FW set 3 Arab normal

FW (mg)

Treatments

Weight in mg

Fresh weight Arabidopsis Normal temp set 3

1104666666667

121075

767

145425

137025

Dry weight Arab set 3 Normal

DW (mg)

Treatments

Weight in mg

Dry weight of Arabidopsis set 3 Normal temp

61353333333

429

372375

57405

5767

Sheet3

Set 3 Low temp LCO TH

Control

LCO A

LCO B

TH A

TH B

Time in hours after stratification

germination

0

0

0

0

0

09615384615

185238095238

180776259336

200991842788

215801790802

100432437932

354880952381

343139202288

402271041898

397858160358

607626170126

736428571429

671569601144

646366332963

593594831095

778958078958

781428571429

773220244716

696421950315

741035816036

885134310134

875476190476

808318766884

81318131257

78457977208

Low temp

Control

LCO A

LCO B

TH A

TH B

Time in hours after stratification

germination

Low temp

0

0

0

0

0

09615384615

185238095238

180776259336

200991842788

215801790802

100432437932

354880952381

343139202288

402271041898

397858160358

607626170126

736428571429

671569601144

646366332963

593594831095

778958078958

781428571429

773220244716

696421950315

741035816036

Sheet1

Sheet2

Sheet3

Salt amp Drought Stress

bullArabidopsisbullSeeds on petriplates with signals

bullControl and 150 mM NaCl

bullSignals improved growth under salt stress

0 NaCl 150 mM NaCl

0 10-6 10-8 10-9 10-11 M LCO Thuricin 17

Hormones of the Holobiontbull Effective at very low concentrationsbull Regulate plant activity

bull Large changes in gene espression protein production hormone profile

bull Regulate other microbesbull Powerful plant productivity effectsbull Inexpensive to applybull Low environmental impact

bull Small amountbull Already produced in environment

bull Very poorly understoodbull Enormous potential

bull New networkbull gt 10 strains in lab now

Phytomicrobiome Potentialbull Wide range of species in the phytomicrobiomebull Most powerful control factor is the plant

bull Developmentbull Stressbull Nutrition

bull Are key to plant growth and productivitybull Management

bull Inoculationbull Hub species bull Signals

bull Microbe-to-plantbull Plant-to-microbebull Microbe-to-microbe

Development of Biologicals - Whobull Scope is largebull Collaboration

bull Basic research labs bull Academicbull Government

bull Industry laboratories

The Approachbull Sample plant-associated microbes

bull The phytomicrobiomebull Efficientrapid screening to identify microbes of interest

bull Biological assessmentsbull Agronomic assessments

Control Thuricin

Industrial Collaboratorsbull Synagribull EVLbull BASFbull RavenQuestbull Ulyssebull Agroworld Natural Productsbull Inocucorbull Axter

BioFuelNet - NCE

Past and Present Students Dr Feng Zhang Dr Fazli Mabood Dr Narjes Dashti Keldeagh Lindsay Dr Pan Bo Dr Sowmya Subramanian Dr Yuming Bai Dr Xiaomin Zhou Elizabeth Gray Nan Wang

PDFs amp Research Associates Dr Xiaomin Zhou Dr Balakrishna Prithiviraj Dr Alfred Souleimanov Dr Kung Dong Lee Dr Woo Jung Mike Lewis Stewart Leibovitch

Acknowledgements

ALSO Trevor Charles at U Waterloo

The End

Thank You

• Biostimulants from the phytomicrobiome Microbe-to-plant signals as low input crop management technologies
• Need for Enhanced Food Production
• Threat of Climate Change
• Biomass for Advanced Biofuels and the Bioeconomy
• The Phytomicrobiome
• Adapting to Climate Change
• The Approach
• So Far
• Example Line of Research
• Something New
• The Problem
• A Solution
• N2 Fixation and Growth - Genistein
• Commercialization
• Serendipity amp Follow Up
• New LCO Activity
• Commercialization
• Jasmonates ndash An Alternative
• Findings
• Effect of Methyl Jasmonate on Yield
• Environment - Temperature and Response
• Salt amp Drought Stress
• Hormones of the Holobiont
• Phytomicrobiome Potential
• Development of Biologicals - Who
• The Approach
• Industrial Collaborators
• BioFuelNet - NCE
• Diapositive numeacutero 29
• The End

Control		96h		000
Control		96h		385
Control		96h		000
Control		96h		000
Control		108h		667
Control		108h		1538
Control		108h		741
Control		108h		1071
Control		120h		5000
Control		120h		7308
Control		120h		5926
Control		120h		6071
Control		132h		6333
Control		132h		8846
Control		132h		7407
Control		132h		8571
Control		144h		8000
Control		144h		9231
Control		144h		8889
Control		144h		9286
LCO A		96h		1600
LCO A		96h		1667
LCO A		96h		2000
LCO A		96h		2143
LCO A		108h		3600
LCO A		108h		2667
LCO A		108h		4000
LCO A		108h		3929
LCO A		120h		7600
LCO A		120h		6333
LCO A		120h		7667
LCO A		120h		7857
LCO A		132h		8400
LCO A		132h		7000
LCO A		132h		8000
LCO A		132h		7857
LCO A		144h		8400
LCO A		144h		8000
LCO A		144h		9333
LCO A		144h		9286
LCO B		96h		1429
LCO B		96h		1935
LCO B		96h		1724
LCO B		96h		2143
LCO B		108h		2857
LCO B		108h		4194
LCO B		108h		3103
LCO B		108h		3571
LCO B		120h		6786
LCO B		120h		7097
LCO B		120h		6552
LCO B		120h		6429
LCO B		132h		7143
LCO B		132h		9032
LCO B		132h		6897
LCO B		132h		7857
LCO B		144h		7857
LCO B		144h		9032
LCO B		144h		7586
LCO B		144h		7857
TH A		96h		2258
TH A		96h		1379
TH A		96h		2069
TH A		96h		2333
TH A		108h		4516
TH A		108h		3448
TH A		108h		3793
TH A		108h		4333
TH A		120h		6774
TH A		120h		6552
TH A		120h		5862
TH A		120h		6667
TH A		132h		7742
TH A		132h		6552
TH A		132h		6897
TH A		132h		6667
TH A		144h		9355
TH A		144h		7241
TH A		144h		7931
TH A		144h		8000
TH B		96h		2000
TH B		96h		1923
TH B		96h		1852
TH B		96h		2857
TH B		108h		3667
TH B		108h		4615
TH B		108h		3704
TH B		108h		3929
TH B		120h		6333
TH B		120h		6154
TH B		120h		5185
TH B		120h		6071
TH B		132h		7000
TH B		132h		8462
TH B		132h		7037
TH B		132h		7143
TH B		144h		8000
TH B		144h		8846
TH B		144h		7037
TH B		144h		7500
LCOA+THA		96h		2500
LCOA+THA		96h		2308
LCOA+THA		96h		1852
LCOA+THA		96h		2800
LCOA+THA		108h		3929
LCOA+THA		108h		3462
LCOA+THA		108h		4444
LCOA+THA		108h		5200
LCOA+THA		120h		7143
LCOA+THA		120h		5385
LCOA+THA		120h		7037
LCOA+THA		120h		7200
LCOA+THA		132h		7500
LCOA+THA		132h		7308
LCOA+THA		132h		8519
LCOA+THA		132h		8400
LCOA+THA		144h		8929
LCOA+THA		144h		8077
LCOA+THA		144h		8519
LCOA+THA		144h		8400
LCOB+THB		96h		1852
LCOB+THB		96h		1724
LCOB+THB		96h		3103
LCOB+THB		96h		1875
LCOB+THB		108h		3704
LCOB+THB		108h		3793
LCOB+THB		108h		6207
LCOB+THB		108h		4063
LCOB+THB		120h		6296
LCOB+THB		120h		6207
LCOB+THB		120h		8276
LCOB+THB		120h		5625
LCOB+THB		132h		7037
LCOB+THB		132h		6552
LCOB+THB		132h		8966
LCOB+THB		132h		7188
LCOB+THB		144h		8519
LCOB+THB		144h		7586
LCOB+THB		144h		9310
LCOB+THB		144h		8750

Arabidopsis low temperature set 3
Treatment		No seeds		96 h		germ		Std		SE		108 h		germ		Std		SE		120 h		germ		Std		SE		132 h		germ		Std		SE		144 h		germ		Std		SE
Control 1		30		0		000				2		667				15		5000				19		6333				24		8000
2		26		1		385				4		1538				19		7308				23		8846				24		9231
3		27		0		000				2		741				16		5926				20		7407				24		8889
4		28		0		000				3		1071				17		6071				24		8571				26		9286
	111		025		096		192		096		275		1004		397		199		1675		6076		948		474		215		7790		1154		577		245		8851		594		297
LCO A 1		25		4		1600				9		3600				19		7600				21		8400				21		8400
2		30		5		1667				8		2667				19		6333				21		7000				24		8000
3		30		6		2000				12		4000				23		7667				24		8000				28		9333
4		28		6		2143				11		3929				22		7857				22		7857				26		9286
	113		525		1852		261		130		10		3549		613		307		2075		7364		696		110		22		7814		589		295		2475		8755		661		331
LCO B 1		28		4		1429				8		2857				19		6786				20		7143				22		7857
2		31		6		1935				13		4194				22		7097				28		9032				28		9032
3		29		5		1724				9		3103				19		6552				20		6897				22		7586
4		28		6		2143				10		3571				18		6429				22		7857				22		7857
	116		525		1808		305		153		10		3431		588		294		195		6716		294		147		225		7732		958		479		235		8083		645		323
TH A 1		31		7		2258				14		4516				21		6774				24		7742				29		9355
2		29		4		1379				10		3448				19		6552				19		6552				21		7241
3		29		6		2069				11		3793				17		5862				20		6897				23		7931
4		30		7		2333				13		4333				20		6667				20		6667				24		8000
	119		6		2010		435		217		12		4023		491		245		1925		6464		411		206		2075		6964		538		269		2425		8132		884		442
TH B 1		30		6		2000				11		3667				19		6333				21		7000				24		8000
2		26		5		1923				12		4615				16		6154				22		8462				23		8846
3		27		5		1852				10		3704				14		5185				19		7037				19		7037
4		28		8		2857				11		3929				17		6071				20		7143				21		7500
	111		6		2158		470		235		11		3979		440		220		165		5936		512		256		205		7410		703		352		2175		7846		774		387
LCOA+THA1		28		7		2500				11		3929				20		7143				21		7500				25		8929
2		26		6		2308				9		3462				14		5385				19		7308				21		8077
3		27		5		1852				12		4444				19		7037				23		8519				23		8519
4		25		7		2800				13		5200				18		7200				21		8400				21		8400
	106		625		2365		398		199		1125		4259		745		372		1775		6691		874		437		21		7932		616		308		225		8481		352		176
LCOB+THB1		27		5		1852				10		3704				17		6296				19		7037				23		8519
2		29		5		1724				11		3793				18		6207				19		6552				22		7586
3		29		9		3103				18		6207				24		8276				26		8966				27		9310
4		32		6		1875				13		4063				18		5625				23		7188				28		8750
	11700		625		2139		647		323		1300		4442		1187		593		1925		6601		1156		183		2175		7435		1056		528		2500		8541		718		359
germination
Treatment		0 h		96 h		108 h		120 h		132 h		144 h
Control		0		096		1004		6076		7790		8851
LCO A		0		1852		3549		7364		7814		8755
LCO B		0		1808		3431		6716		7732		8083
TH A		0		2010		4023		6464		6964		8132
TH B		0		2158		3979		5936		7410		7846
LCOA+THA		0		2365		4259		6691		7932		8481
LCOB+THB		0		2139		4442		6601		7435		8541
SE
	0 h		96 h		108 h		120 h		132 h		144 h
Control		0		096		199		474		577		297
LCO A		0		130		307		110		295		331
LCO B		0		153		294		147		479		323
TH A		0		217		245		206		269		442
TH B		0		235		220		256		352		387
LCOA+THA		0		199		372		437		308		176
LCOB+THB		0		323		593		183		528		359

0 h		0 h		0 h		0 h		0 h
96 h		96 h		96 h		96 h		96 h
108 h		108 h		108 h		108 h		108 h
120 h		120 h		120 h		120 h		120 h
132 h		132 h		132 h		132 h		132 h

0 h		0 h		0 h		0 h		0 h		0		0		0		0		0		0		0		0		0		0
96 h		96 h		96 h		96 h		96 h		09615384615		09615384615		13049704387		13049704387		15258333938		15258333938		2174332839		2174332839		23499650247		2174332839
108 h		108 h		108 h		108 h		108 h		19859811656		19859811656		30667415327		30667415327		29408073778		29408073778		24539813637		24539813637		2200128559		24539813637
120 h		120 h		120 h		120 h		120 h		47411187081		47411187081		11002902798		11002902798		14704036889		14704036889		20561084936		20561084936		25615660866		20561084936
132 h		132 h		132 h		132 h		132 h		57694206278		57694206278		29473844048		29473844048		4788349344		4788349344		26896579581		26896579581		35169817799		26896579581
144 h		144 h		144 h		144 h		144 h		29704619111		29704619111		33067820092		33067820092		32273789905		32273789905		4421868687		4421868687		3871003991		4421868687

Weight of seedlings
Treatment		FW (mg)		STD		SE			DW (mg)		STD		SE					Treatment		FW (mg)		STD		SE			DW (mg)		STD		SE
Control 1		39850					2924							Control 1
2		11940					854							2		11940					854
3		10680					468							3		10680					468
4		10520					518							4		10520					518
	11047		778		449			614		210		121						11047		778		449			614		210		121
LCO A 1		15860					899							LCO A 1
2		11320					449							2		11320					449
3		10640					437							3		10640					437
4		10610					400							4		10610					400
	12108		2523		1262			546		236		118						12108		2523		1262			429		026		015
LCO B 1		8040					416							LCO B 1		8040					416
2		7560					333							2		7560					333
3		8020					378							3		8020					378
4		7060					363							4		7060					363
	7670		463		232			372		035		017						7670		463		232			372		035		017
TH A 1		14760					632							TH A 1		14760					632
2		12180					457							2		12180					457
3		14450					469							3		14450					469
4		16780					739							4		16780					739
	14543		1884		942			574		136		068						14543		1884		942			574		136		068
TH B 1		10980					391							TH B 1		10980					391
2		15910					701							2		15910					701
3		17040					815							3		17040					815
4		10880					400							4		10880					400
	13703		3235		1617			577		215		107						13703		3235		1617			577		215		107
Treatment		FW (mg)		SE			Treatment		DW (mg)		SE
Control		11047		449			Control		614		121
LCO A		12108		1262			LCO A		429		016
LCO B		7670		232			LCO B		372		017
TH A		14543		942			TH A		574		068
TH B		13703		1617			TH B		577		107

Control		12125587729
LCO A		0155
LCO B		01725714224
TH A		0678241169
TH B		10731213818

Control		44904837651
LCO A		126152932982
LCO B		2315887159
TH A		94179239574
TH B		161736604288

Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B

Arabidopsis normal temperature set 3 continuous
Treatment		No seeds		24 h		germ		Std		SE		36 h		germ		Std		SE		48 h		germ		Std		SE		60 h		germ		Std		SE
Control 1		31		0					16		5161				23		7419				26		8387
2		29		0					19		6552				27		9310				28		9655
3		25		0					15		6000				21		8400				21		8400
4		26		0					13		5000				22		8462				24		9231
	111						1575		5678		729		364		2325		8398		773		387		2475		8918		630		315
LCO A 1		34		0					20		5882				26		7647				34		10000
2		32		0					19		5938				27		8438				30		9375
3		33		0					18		5455				28		8485				29		8788
4		32		0					24		7500				26		8125				29		9063
	131						2025		6194		897		449		2675		8174		386		193		305		9306		521		260
LCO B 1		31		0					14		4516				23		7419				26		8387
2		34		0					15		4412				31		9118				34		10000
3		32		0					20		6250				27		8438				28		8750
4		34		0					16		4706				23		6765				31		9118
	131						1625		4971		861		431		26		7935		1047		523		2975		9064		692		346
TH A 1		28		0					18		6429				23		8214				23		8214
2		29		0					19		6552				24		8276				27		9310
3		30		0					17		5667				23		7667				25		8333
4		28		0					16		5714				25		8929				25		8929
	115						175		6090		465		232		2375		8271		517		258		25		8697		515		257
TH B 1		33		0					23		6970				28		8485				32		9697
2		28		0					19		6786				25		8929				26		9286
3		27		0					21		7778				26		9630				27		10000
4		27		0					20		7407				24		8889				25		9259
	115						2075		7235		446		223		2575		8983		475		238		275		9560		355		177
germination
Treatment		0 h		24 h		36 h		48 h		60 h
Control		0		0		5678		8398		8918
LCO A		0		0		6194		8174		9306
LCO B		0		0		4971		7935		9064
TH A		0		0		6090		8271		8697
TH B		0		0		7235		8983		9560
SE
Control		0		0		364		387		315
LCO A		0		0		449		193		260
LCO B		0		0		431		523		346
TH A		0		0		232		258		257
TH B		0		0		223		238		177

0 h		0 h		0 h		0 h		0 h		0		0		0		0		0		0		0		0		0		0
24 h		24 h		24 h		24 h		24 h		0		0		0		0		0		0		0		0		0		0
36 h		36 h		36 h		36 h		36 h		3644345509		3644345509		44864095916		44864095916		3644345509		3644345509		23241337778		23241337778		2229677279		2229677279
48 h		48 h		48 h		48 h		48 h		38666958625		38666958625		19281389988		19281389988		38666958625		38666958625		2583374136		2583374136		23771554774		23771554774
60 h		60 h		60 h		60 h		60 h		31509650968		31509650968		26047046621		26047046621		31509650968		31509650968		25740422067		25740422067		1774911735		1774911735

24 h		24 h		24 h		24 h		24 h		0		0		0		0		0		0		0		0		0		0
36 h		36 h		36 h		36 h		36 h		3644345509		3644345509		44864095916		44864095916		43067583186		43067583186		23241337778		23241337778		2229677279		2229677279
48 h		48 h		48 h		48 h		48 h		38666958625		38666958625		19281389988		19281389988		52334169948		52334169948		2583374136		2583374136		23771554774		23771554774
60 h		60 h		60 h		60 h		60 h		31509650968		31509650968		26047046621		26047046621		34590070704		34590070704		25740422067		25740422067		1774911735		1774911735

Environment - Temperature and Responsebull In the lab responses could be variable and frustrating

bull In the field responses were strongest in cold springs on heavy soils

bull Over the last few years have shown that low temperatures make the lab results clearer amp consistent

0

20

40

60

80

100

120

0 h 24 h 36 h 48 h 60 hTime in hours after stratification

g

erm

inati

on

Control

LCO A

LCO B

TH A

TH B

0

10

20

30

40

50

60

70

80

90

100

0 h 96 h 108 h 120 h 132 h 144 h

g

erm

ina

tio

n

Time in hours after stratification

Control LCO A LCO B TH A TH B

Arabidopsis at 22 oC

Arabidopsis at 10 oC

Set 3 normal arab

Control

LCO A

LCO B

TH A

TH B

Time in hours after preincubation at 4 deg C in dark

germination

Arabidopsis set 3 Normal temp

0

0

0

0

0

567825361513

619359959893

497094402277

609031198686

723514911015

839780953196

817360182709

793480194497

827134646962

898298460798

891825960469

930634469697

906368595825

869663382594

956048581049

New line graph

Control

LCO A

LCO B

TH A

TH B

Time in hours after stratification

germination

0

0

0

0

0

0

0

0

0

0

567825361513

619359959893

497094402277

609031198686

723514911015

839780953196

817360182709

793480194497

827134646962

898298460798

891825960469

930634469697

906368595825

869663382594

956048581049

Sheet1

Sheet2

FW set 3 Arab normal

FW (mg)

Treatments

Weight in mg

Fresh weight Arabidopsis Normal temp set 3

1104666666667

121075

767

145425

137025

Dry weight Arab set 3 Normal

DW (mg)

Treatments

Weight in mg

Dry weight of Arabidopsis set 3 Normal temp

61353333333

429

372375

57405

5767

Sheet3

Set 3 Low temp LCO TH

Control

LCO A

LCO B

TH A

TH B

Time in hours after stratification

germination

0

0

0

0

0

09615384615

185238095238

180776259336

200991842788

215801790802

100432437932

354880952381

343139202288

402271041898

397858160358

607626170126

736428571429

671569601144

646366332963

593594831095

778958078958

781428571429

773220244716

696421950315

741035816036

885134310134

875476190476

808318766884

81318131257

78457977208

Low temp

Control

LCO A

LCO B

TH A

TH B

Time in hours after stratification

germination

Low temp

0

0

0

0

0

09615384615

185238095238

180776259336

200991842788

215801790802

100432437932

354880952381

343139202288

402271041898

397858160358

607626170126

736428571429

671569601144

646366332963

593594831095

778958078958

781428571429

773220244716

696421950315

741035816036

Sheet1

Sheet2

Sheet3

Salt amp Drought Stress

bullArabidopsisbullSeeds on petriplates with signals

bullControl and 150 mM NaCl

bullSignals improved growth under salt stress

0 NaCl 150 mM NaCl

0 10-6 10-8 10-9 10-11 M LCO Thuricin 17

Hormones of the Holobiontbull Effective at very low concentrationsbull Regulate plant activity

bull Large changes in gene espression protein production hormone profile

bull Regulate other microbesbull Powerful plant productivity effectsbull Inexpensive to applybull Low environmental impact

bull Small amountbull Already produced in environment

bull Very poorly understoodbull Enormous potential

bull New networkbull gt 10 strains in lab now

Phytomicrobiome Potentialbull Wide range of species in the phytomicrobiomebull Most powerful control factor is the plant

bull Developmentbull Stressbull Nutrition

bull Are key to plant growth and productivitybull Management

bull Inoculationbull Hub species bull Signals

bull Microbe-to-plantbull Plant-to-microbebull Microbe-to-microbe

Development of Biologicals - Whobull Scope is largebull Collaboration

bull Basic research labs bull Academicbull Government

bull Industry laboratories

The Approachbull Sample plant-associated microbes

bull The phytomicrobiomebull Efficientrapid screening to identify microbes of interest

bull Biological assessmentsbull Agronomic assessments

Control Thuricin

Industrial Collaboratorsbull Synagribull EVLbull BASFbull RavenQuestbull Ulyssebull Agroworld Natural Productsbull Inocucorbull Axter

BioFuelNet - NCE

Past and Present Students Dr Feng Zhang Dr Fazli Mabood Dr Narjes Dashti Keldeagh Lindsay Dr Pan Bo Dr Sowmya Subramanian Dr Yuming Bai Dr Xiaomin Zhou Elizabeth Gray Nan Wang

PDFs amp Research Associates Dr Xiaomin Zhou Dr Balakrishna Prithiviraj Dr Alfred Souleimanov Dr Kung Dong Lee Dr Woo Jung Mike Lewis Stewart Leibovitch

Acknowledgements

ALSO Trevor Charles at U Waterloo

The End

Thank You

• Biostimulants from the phytomicrobiome Microbe-to-plant signals as low input crop management technologies
• Need for Enhanced Food Production
• Threat of Climate Change
• Biomass for Advanced Biofuels and the Bioeconomy
• The Phytomicrobiome
• Adapting to Climate Change
• The Approach
• So Far
• Example Line of Research
• Something New
• The Problem
• A Solution
• N2 Fixation and Growth - Genistein
• Commercialization
• Serendipity amp Follow Up
• New LCO Activity
• Commercialization
• Jasmonates ndash An Alternative
• Findings
• Effect of Methyl Jasmonate on Yield
• Environment - Temperature and Response
• Salt amp Drought Stress
• Hormones of the Holobiont
• Phytomicrobiome Potential
• Development of Biologicals - Who
• The Approach
• Industrial Collaborators
• BioFuelNet - NCE
• Diapositive numeacutero 29
• The End

Control		96h		000
Control		96h		385
Control		96h		000
Control		96h		000
Control		108h		667
Control		108h		1538
Control		108h		741
Control		108h		1071
Control		120h		5000
Control		120h		7308
Control		120h		5926
Control		120h		6071
Control		132h		6333
Control		132h		8846
Control		132h		7407
Control		132h		8571
Control		144h		8000
Control		144h		9231
Control		144h		8889
Control		144h		9286
LCO A		96h		1600
LCO A		96h		1667
LCO A		96h		2000
LCO A		96h		2143
LCO A		108h		3600
LCO A		108h		2667
LCO A		108h		4000
LCO A		108h		3929
LCO A		120h		7600
LCO A		120h		6333
LCO A		120h		7667
LCO A		120h		7857
LCO A		132h		8400
LCO A		132h		7000
LCO A		132h		8000
LCO A		132h		7857
LCO A		144h		8400
LCO A		144h		8000
LCO A		144h		9333
LCO A		144h		9286
LCO B		96h		1429
LCO B		96h		1935
LCO B		96h		1724
LCO B		96h		2143
LCO B		108h		2857
LCO B		108h		4194
LCO B		108h		3103
LCO B		108h		3571
LCO B		120h		6786
LCO B		120h		7097
LCO B		120h		6552
LCO B		120h		6429
LCO B		132h		7143
LCO B		132h		9032
LCO B		132h		6897
LCO B		132h		7857
LCO B		144h		7857
LCO B		144h		9032
LCO B		144h		7586
LCO B		144h		7857
TH A		96h		2258
TH A		96h		1379
TH A		96h		2069
TH A		96h		2333
TH A		108h		4516
TH A		108h		3448
TH A		108h		3793
TH A		108h		4333
TH A		120h		6774
TH A		120h		6552
TH A		120h		5862
TH A		120h		6667
TH A		132h		7742
TH A		132h		6552
TH A		132h		6897
TH A		132h		6667
TH A		144h		9355
TH A		144h		7241
TH A		144h		7931
TH A		144h		8000
TH B		96h		2000
TH B		96h		1923
TH B		96h		1852
TH B		96h		2857
TH B		108h		3667
TH B		108h		4615
TH B		108h		3704
TH B		108h		3929
TH B		120h		6333
TH B		120h		6154
TH B		120h		5185
TH B		120h		6071
TH B		132h		7000
TH B		132h		8462
TH B		132h		7037
TH B		132h		7143
TH B		144h		8000
TH B		144h		8846
TH B		144h		7037
TH B		144h		7500
LCOA+THA		96h		2500
LCOA+THA		96h		2308
LCOA+THA		96h		1852
LCOA+THA		96h		2800
LCOA+THA		108h		3929
LCOA+THA		108h		3462
LCOA+THA		108h		4444
LCOA+THA		108h		5200
LCOA+THA		120h		7143
LCOA+THA		120h		5385
LCOA+THA		120h		7037
LCOA+THA		120h		7200
LCOA+THA		132h		7500
LCOA+THA		132h		7308
LCOA+THA		132h		8519
LCOA+THA		132h		8400
LCOA+THA		144h		8929
LCOA+THA		144h		8077
LCOA+THA		144h		8519
LCOA+THA		144h		8400
LCOB+THB		96h		1852
LCOB+THB		96h		1724
LCOB+THB		96h		3103
LCOB+THB		96h		1875
LCOB+THB		108h		3704
LCOB+THB		108h		3793
LCOB+THB		108h		6207
LCOB+THB		108h		4063
LCOB+THB		120h		6296
LCOB+THB		120h		6207
LCOB+THB		120h		8276
LCOB+THB		120h		5625
LCOB+THB		132h		7037
LCOB+THB		132h		6552
LCOB+THB		132h		8966
LCOB+THB		132h		7188
LCOB+THB		144h		8519
LCOB+THB		144h		7586
LCOB+THB		144h		9310
LCOB+THB		144h		8750

Arabidopsis low temperature set 3
Treatment		No seeds		96 h		germ		Std		SE		108 h		germ		Std		SE		120 h		germ		Std		SE		132 h		germ		Std		SE		144 h		germ		Std		SE
Control 1		30		0		000				2		667				15		5000				19		6333				24		8000
2		26		1		385				4		1538				19		7308				23		8846				24		9231
3		27		0		000				2		741				16		5926				20		7407				24		8889
4		28		0		000				3		1071				17		6071				24		8571				26		9286
	111		025		096		192		096		275		1004		397		199		1675		6076		948		474		215		7790		1154		577		245		8851		594		297
LCO A 1		25		4		1600				9		3600				19		7600				21		8400				21		8400
2		30		5		1667				8		2667				19		6333				21		7000				24		8000
3		30		6		2000				12		4000				23		7667				24		8000				28		9333
4		28		6		2143				11		3929				22		7857				22		7857				26		9286
	113		525		1852		261		130		10		3549		613		307		2075		7364		696		110		22		7814		589		295		2475		8755		661		331
LCO B 1		28		4		1429				8		2857				19		6786				20		7143				22		7857
2		31		6		1935				13		4194				22		7097				28		9032				28		9032
3		29		5		1724				9		3103				19		6552				20		6897				22		7586
4		28		6		2143				10		3571				18		6429				22		7857				22		7857
	116		525		1808		305		153		10		3431		588		294		195		6716		294		147		225		7732		958		479		235		8083		645		323
TH A 1		31		7		2258				14		4516				21		6774				24		7742				29		9355
2		29		4		1379				10		3448				19		6552				19		6552				21		7241
3		29		6		2069				11		3793				17		5862				20		6897				23		7931
4		30		7		2333				13		4333				20		6667				20		6667				24		8000
	119		6		2010		435		217		12		4023		491		245		1925		6464		411		206		2075		6964		538		269		2425		8132		884		442
TH B 1		30		6		2000				11		3667				19		6333				21		7000				24		8000
2		26		5		1923				12		4615				16		6154				22		8462				23		8846
3		27		5		1852				10		3704				14		5185				19		7037				19		7037
4		28		8		2857				11		3929				17		6071				20		7143				21		7500
	111		6		2158		470		235		11		3979		440		220		165		5936		512		256		205		7410		703		352		2175		7846		774		387
LCOA+THA1		28		7		2500				11		3929				20		7143				21		7500				25		8929
2		26		6		2308				9		3462				14		5385				19		7308				21		8077
3		27		5		1852				12		4444				19		7037				23		8519				23		8519
4		25		7		2800				13		5200				18		7200				21		8400				21		8400
	106		625		2365		398		199		1125		4259		745		372		1775		6691		874		437		21		7932		616		308		225		8481		352		176
LCOB+THB1		27		5		1852				10		3704				17		6296				19		7037				23		8519
2		29		5		1724				11		3793				18		6207				19		6552				22		7586
3		29		9		3103				18		6207				24		8276				26		8966				27		9310
4		32		6		1875				13		4063				18		5625				23		7188				28		8750
	11700		625		2139		647		323		1300		4442		1187		593		1925		6601		1156		183		2175		7435		1056		528		2500		8541		718		359
germination
Treatment		0 h		96 h		108 h		120 h		132 h		144 h
Control		0		096		1004		6076		7790		8851
LCO A		0		1852		3549		7364		7814		8755
LCO B		0		1808		3431		6716		7732		8083
TH A		0		2010		4023		6464		6964		8132
TH B		0		2158		3979		5936		7410		7846
LCOA+THA		0		2365		4259		6691		7932		8481
LCOB+THB		0		2139		4442		6601		7435		8541
SE
	0 h		96 h		108 h		120 h		132 h		144 h
Control		0		096		199		474		577		297
LCO A		0		130		307		110		295		331
LCO B		0		153		294		147		479		323
TH A		0		217		245		206		269		442
TH B		0		235		220		256		352		387
LCOA+THA		0		199		372		437		308		176
LCOB+THB		0		323		593		183		528		359

0 h		0 h		0 h		0 h		0 h
96 h		96 h		96 h		96 h		96 h
108 h		108 h		108 h		108 h		108 h
120 h		120 h		120 h		120 h		120 h
132 h		132 h		132 h		132 h		132 h

0 h		0 h		0 h		0 h		0 h		0		0		0		0		0		0		0		0		0		0
96 h		96 h		96 h		96 h		96 h		09615384615		09615384615		13049704387		13049704387		15258333938		15258333938		2174332839		2174332839		23499650247		2174332839
108 h		108 h		108 h		108 h		108 h		19859811656		19859811656		30667415327		30667415327		29408073778		29408073778		24539813637		24539813637		2200128559		24539813637
120 h		120 h		120 h		120 h		120 h		47411187081		47411187081		11002902798		11002902798		14704036889		14704036889		20561084936		20561084936		25615660866		20561084936
132 h		132 h		132 h		132 h		132 h		57694206278		57694206278		29473844048		29473844048		4788349344		4788349344		26896579581		26896579581		35169817799		26896579581
144 h		144 h		144 h		144 h		144 h		29704619111		29704619111		33067820092		33067820092		32273789905		32273789905		4421868687		4421868687		3871003991		4421868687

Weight of seedlings
Treatment		FW (mg)		STD		SE			DW (mg)		STD		SE					Treatment		FW (mg)		STD		SE			DW (mg)		STD		SE
Control 1		39850					2924							Control 1
2		11940					854							2		11940					854
3		10680					468							3		10680					468
4		10520					518							4		10520					518
	11047		778		449			614		210		121						11047		778		449			614		210		121
LCO A 1		15860					899							LCO A 1
2		11320					449							2		11320					449
3		10640					437							3		10640					437
4		10610					400							4		10610					400
	12108		2523		1262			546		236		118						12108		2523		1262			429		026		015
LCO B 1		8040					416							LCO B 1		8040					416
2		7560					333							2		7560					333
3		8020					378							3		8020					378
4		7060					363							4		7060					363
	7670		463		232			372		035		017						7670		463		232			372		035		017
TH A 1		14760					632							TH A 1		14760					632
2		12180					457							2		12180					457
3		14450					469							3		14450					469
4		16780					739							4		16780					739
	14543		1884		942			574		136		068						14543		1884		942			574		136		068
TH B 1		10980					391							TH B 1		10980					391
2		15910					701							2		15910					701
3		17040					815							3		17040					815
4		10880					400							4		10880					400
	13703		3235		1617			577		215		107						13703		3235		1617			577		215		107
Treatment		FW (mg)		SE			Treatment		DW (mg)		SE
Control		11047		449			Control		614		121
LCO A		12108		1262			LCO A		429		016
LCO B		7670		232			LCO B		372		017
TH A		14543		942			TH A		574		068
TH B		13703		1617			TH B		577		107

Control		12125587729
LCO A		0155
LCO B		01725714224
TH A		0678241169
TH B		10731213818

Control		44904837651
LCO A		126152932982
LCO B		2315887159
TH A		94179239574
TH B		161736604288

Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B

Arabidopsis normal temperature set 3 continuous
Treatment		No seeds		24 h		germ		Std		SE		36 h		germ		Std		SE		48 h		germ		Std		SE		60 h		germ		Std		SE
Control 1		31		0					16		5161				23		7419				26		8387
2		29		0					19		6552				27		9310				28		9655
3		25		0					15		6000				21		8400				21		8400
4		26		0					13		5000				22		8462				24		9231
	111						1575		5678		729		364		2325		8398		773		387		2475		8918		630		315
LCO A 1		34		0					20		5882				26		7647				34		10000
2		32		0					19		5938				27		8438				30		9375
3		33		0					18		5455				28		8485				29		8788
4		32		0					24		7500				26		8125				29		9063
	131						2025		6194		897		449		2675		8174		386		193		305		9306		521		260
LCO B 1		31		0					14		4516				23		7419				26		8387
2		34		0					15		4412				31		9118				34		10000
3		32		0					20		6250				27		8438				28		8750
4		34		0					16		4706				23		6765				31		9118
	131						1625		4971		861		431		26		7935		1047		523		2975		9064		692		346
TH A 1		28		0					18		6429				23		8214				23		8214
2		29		0					19		6552				24		8276				27		9310
3		30		0					17		5667				23		7667				25		8333
4		28		0					16		5714				25		8929				25		8929
	115						175		6090		465		232		2375		8271		517		258		25		8697		515		257
TH B 1		33		0					23		6970				28		8485				32		9697
2		28		0					19		6786				25		8929				26		9286
3		27		0					21		7778				26		9630				27		10000
4		27		0					20		7407				24		8889				25		9259
	115						2075		7235		446		223		2575		8983		475		238		275		9560		355		177
germination
Treatment		0 h		24 h		36 h		48 h		60 h
Control		0		0		5678		8398		8918
LCO A		0		0		6194		8174		9306
LCO B		0		0		4971		7935		9064
TH A		0		0		6090		8271		8697
TH B		0		0		7235		8983		9560
SE
Control		0		0		364		387		315
LCO A		0		0		449		193		260
LCO B		0		0		431		523		346
TH A		0		0		232		258		257
TH B		0		0		223		238		177

0 h		0 h		0 h		0 h		0 h		0		0		0		0		0		0		0		0		0		0
24 h		24 h		24 h		24 h		24 h		0		0		0		0		0		0		0		0		0		0
36 h		36 h		36 h		36 h		36 h		3644345509		3644345509		44864095916		44864095916		3644345509		3644345509		23241337778		23241337778		2229677279		2229677279
48 h		48 h		48 h		48 h		48 h		38666958625		38666958625		19281389988		19281389988		38666958625		38666958625		2583374136		2583374136		23771554774		23771554774
60 h		60 h		60 h		60 h		60 h		31509650968		31509650968		26047046621		26047046621		31509650968		31509650968		25740422067		25740422067		1774911735		1774911735

24 h		24 h		24 h		24 h		24 h		0		0		0		0		0		0		0		0		0		0
36 h		36 h		36 h		36 h		36 h		3644345509		3644345509		44864095916		44864095916		43067583186		43067583186		23241337778		23241337778		2229677279		2229677279
48 h		48 h		48 h		48 h		48 h		38666958625		38666958625		19281389988		19281389988		52334169948		52334169948		2583374136		2583374136		23771554774		23771554774
60 h		60 h		60 h		60 h		60 h		31509650968		31509650968		26047046621		26047046621		34590070704		34590070704		25740422067		25740422067		1774911735		1774911735

Set 3 normal arab

Control

LCO A

LCO B

TH A

TH B

Time in hours after preincubation at 4 deg C in dark

germination

Arabidopsis set 3 Normal temp

0

0

0

0

0

567825361513

619359959893

497094402277

609031198686

723514911015

839780953196

817360182709

793480194497

827134646962

898298460798

891825960469

930634469697

906368595825

869663382594

956048581049

New line graph

Control

LCO A

LCO B

TH A

TH B

Time in hours after stratification

germination

0

0

0

0

0

0

0

0

0

0

567825361513

619359959893

497094402277

609031198686

723514911015

839780953196

817360182709

793480194497

827134646962

898298460798

891825960469

930634469697

906368595825

869663382594

956048581049

Sheet1

Sheet2

FW set 3 Arab normal

FW (mg)

Treatments

Weight in mg

Fresh weight Arabidopsis Normal temp set 3

1104666666667

121075

767

145425

137025

Dry weight Arab set 3 Normal

DW (mg)

Treatments

Weight in mg

Dry weight of Arabidopsis set 3 Normal temp

61353333333

429

372375

57405

5767

Sheet3

Set 3 Low temp LCO TH

Control

LCO A

LCO B

TH A

TH B

Time in hours after stratification

germination

0

0

0

0

0

09615384615

185238095238

180776259336

200991842788

215801790802

100432437932

354880952381

343139202288

402271041898

397858160358

607626170126

736428571429

671569601144

646366332963

593594831095

778958078958

781428571429

773220244716

696421950315

741035816036

885134310134

875476190476

808318766884

81318131257

78457977208

Low temp

Control

LCO A

LCO B

TH A

TH B

Time in hours after stratification

germination

Low temp

0

0

0

0

0

09615384615

185238095238

180776259336

200991842788

215801790802

100432437932

354880952381

343139202288

402271041898

397858160358

607626170126

736428571429

671569601144

646366332963

593594831095

778958078958

781428571429

773220244716

696421950315

741035816036

Sheet1

Sheet2

Sheet3

Salt amp Drought Stress

bullArabidopsisbullSeeds on petriplates with signals

bullControl and 150 mM NaCl

bullSignals improved growth under salt stress

0 NaCl 150 mM NaCl

0 10-6 10-8 10-9 10-11 M LCO Thuricin 17

Hormones of the Holobiontbull Effective at very low concentrationsbull Regulate plant activity

bull Large changes in gene espression protein production hormone profile

bull Regulate other microbesbull Powerful plant productivity effectsbull Inexpensive to applybull Low environmental impact

bull Small amountbull Already produced in environment

bull Very poorly understoodbull Enormous potential

bull New networkbull gt 10 strains in lab now

Phytomicrobiome Potentialbull Wide range of species in the phytomicrobiomebull Most powerful control factor is the plant

bull Developmentbull Stressbull Nutrition

bull Are key to plant growth and productivitybull Management

bull Inoculationbull Hub species bull Signals

bull Microbe-to-plantbull Plant-to-microbebull Microbe-to-microbe

Development of Biologicals - Whobull Scope is largebull Collaboration

bull Basic research labs bull Academicbull Government

bull Industry laboratories

The Approachbull Sample plant-associated microbes

bull The phytomicrobiomebull Efficientrapid screening to identify microbes of interest

bull Biological assessmentsbull Agronomic assessments

Control Thuricin

Industrial Collaboratorsbull Synagribull EVLbull BASFbull RavenQuestbull Ulyssebull Agroworld Natural Productsbull Inocucorbull Axter

BioFuelNet - NCE

Past and Present Students Dr Feng Zhang Dr Fazli Mabood Dr Narjes Dashti Keldeagh Lindsay Dr Pan Bo Dr Sowmya Subramanian Dr Yuming Bai Dr Xiaomin Zhou Elizabeth Gray Nan Wang

PDFs amp Research Associates Dr Xiaomin Zhou Dr Balakrishna Prithiviraj Dr Alfred Souleimanov Dr Kung Dong Lee Dr Woo Jung Mike Lewis Stewart Leibovitch

Acknowledgements

ALSO Trevor Charles at U Waterloo

The End

Thank You

• Biostimulants from the phytomicrobiome Microbe-to-plant signals as low input crop management technologies
• Need for Enhanced Food Production
• Threat of Climate Change
• Biomass for Advanced Biofuels and the Bioeconomy
• The Phytomicrobiome
• Adapting to Climate Change
• The Approach
• So Far
• Example Line of Research
• Something New
• The Problem
• A Solution
• N2 Fixation and Growth - Genistein
• Commercialization
• Serendipity amp Follow Up
• New LCO Activity
• Commercialization
• Jasmonates ndash An Alternative
• Findings
• Effect of Methyl Jasmonate on Yield
• Environment - Temperature and Response
• Salt amp Drought Stress
• Hormones of the Holobiont
• Phytomicrobiome Potential
• Development of Biologicals - Who
• The Approach
• Industrial Collaborators
• BioFuelNet - NCE
• Diapositive numeacutero 29
• The End

Control		96h		000
Control		96h		385
Control		96h		000
Control		96h		000
Control		108h		667
Control		108h		1538
Control		108h		741
Control		108h		1071
Control		120h		5000
Control		120h		7308
Control		120h		5926
Control		120h		6071
Control		132h		6333
Control		132h		8846
Control		132h		7407
Control		132h		8571
Control		144h		8000
Control		144h		9231
Control		144h		8889
Control		144h		9286
LCO A		96h		1600
LCO A		96h		1667
LCO A		96h		2000
LCO A		96h		2143
LCO A		108h		3600
LCO A		108h		2667
LCO A		108h		4000
LCO A		108h		3929
LCO A		120h		7600
LCO A		120h		6333
LCO A		120h		7667
LCO A		120h		7857
LCO A		132h		8400
LCO A		132h		7000
LCO A		132h		8000
LCO A		132h		7857
LCO A		144h		8400
LCO A		144h		8000
LCO A		144h		9333
LCO A		144h		9286
LCO B		96h		1429
LCO B		96h		1935
LCO B		96h		1724
LCO B		96h		2143
LCO B		108h		2857
LCO B		108h		4194
LCO B		108h		3103
LCO B		108h		3571
LCO B		120h		6786
LCO B		120h		7097
LCO B		120h		6552
LCO B		120h		6429
LCO B		132h		7143
LCO B		132h		9032
LCO B		132h		6897
LCO B		132h		7857
LCO B		144h		7857
LCO B		144h		9032
LCO B		144h		7586
LCO B		144h		7857
TH A		96h		2258
TH A		96h		1379
TH A		96h		2069
TH A		96h		2333
TH A		108h		4516
TH A		108h		3448
TH A		108h		3793
TH A		108h		4333
TH A		120h		6774
TH A		120h		6552
TH A		120h		5862
TH A		120h		6667
TH A		132h		7742
TH A		132h		6552
TH A		132h		6897
TH A		132h		6667
TH A		144h		9355
TH A		144h		7241
TH A		144h		7931
TH A		144h		8000
TH B		96h		2000
TH B		96h		1923
TH B		96h		1852
TH B		96h		2857
TH B		108h		3667
TH B		108h		4615
TH B		108h		3704
TH B		108h		3929
TH B		120h		6333
TH B		120h		6154
TH B		120h		5185
TH B		120h		6071
TH B		132h		7000
TH B		132h		8462
TH B		132h		7037
TH B		132h		7143
TH B		144h		8000
TH B		144h		8846
TH B		144h		7037
TH B		144h		7500
LCOA+THA		96h		2500
LCOA+THA		96h		2308
LCOA+THA		96h		1852
LCOA+THA		96h		2800
LCOA+THA		108h		3929
LCOA+THA		108h		3462
LCOA+THA		108h		4444
LCOA+THA		108h		5200
LCOA+THA		120h		7143
LCOA+THA		120h		5385
LCOA+THA		120h		7037
LCOA+THA		120h		7200
LCOA+THA		132h		7500
LCOA+THA		132h		7308
LCOA+THA		132h		8519
LCOA+THA		132h		8400
LCOA+THA		144h		8929
LCOA+THA		144h		8077
LCOA+THA		144h		8519
LCOA+THA		144h		8400
LCOB+THB		96h		1852
LCOB+THB		96h		1724
LCOB+THB		96h		3103
LCOB+THB		96h		1875
LCOB+THB		108h		3704
LCOB+THB		108h		3793
LCOB+THB		108h		6207
LCOB+THB		108h		4063
LCOB+THB		120h		6296
LCOB+THB		120h		6207
LCOB+THB		120h		8276
LCOB+THB		120h		5625
LCOB+THB		132h		7037
LCOB+THB		132h		6552
LCOB+THB		132h		8966
LCOB+THB		132h		7188
LCOB+THB		144h		8519
LCOB+THB		144h		7586
LCOB+THB		144h		9310
LCOB+THB		144h		8750

Arabidopsis low temperature set 3
Treatment		No seeds		96 h		germ		Std		SE		108 h		germ		Std		SE		120 h		germ		Std		SE		132 h		germ		Std		SE		144 h		germ		Std		SE
Control 1		30		0		000				2		667				15		5000				19		6333				24		8000
2		26		1		385				4		1538				19		7308				23		8846				24		9231
3		27		0		000				2		741				16		5926				20		7407				24		8889
4		28		0		000				3		1071				17		6071				24		8571				26		9286
	111		025		096		192		096		275		1004		397		199		1675		6076		948		474		215		7790		1154		577		245		8851		594		297
LCO A 1		25		4		1600				9		3600				19		7600				21		8400				21		8400
2		30		5		1667				8		2667				19		6333				21		7000				24		8000
3		30		6		2000				12		4000				23		7667				24		8000				28		9333
4		28		6		2143				11		3929				22		7857				22		7857				26		9286
	113		525		1852		261		130		10		3549		613		307		2075		7364		696		110		22		7814		589		295		2475		8755		661		331
LCO B 1		28		4		1429				8		2857				19		6786				20		7143				22		7857
2		31		6		1935				13		4194				22		7097				28		9032				28		9032
3		29		5		1724				9		3103				19		6552				20		6897				22		7586
4		28		6		2143				10		3571				18		6429				22		7857				22		7857
	116		525		1808		305		153		10		3431		588		294		195		6716		294		147		225		7732		958		479		235		8083		645		323
TH A 1		31		7		2258				14		4516				21		6774				24		7742				29		9355
2		29		4		1379				10		3448				19		6552				19		6552				21		7241
3		29		6		2069				11		3793				17		5862				20		6897				23		7931
4		30		7		2333				13		4333				20		6667				20		6667				24		8000
	119		6		2010		435		217		12		4023		491		245		1925		6464		411		206		2075		6964		538		269		2425		8132		884		442
TH B 1		30		6		2000				11		3667				19		6333				21		7000				24		8000
2		26		5		1923				12		4615				16		6154				22		8462				23		8846
3		27		5		1852				10		3704				14		5185				19		7037				19		7037
4		28		8		2857				11		3929				17		6071				20		7143				21		7500
	111		6		2158		470		235		11		3979		440		220		165		5936		512		256		205		7410		703		352		2175		7846		774		387
LCOA+THA1		28		7		2500				11		3929				20		7143				21		7500				25		8929
2		26		6		2308				9		3462				14		5385				19		7308				21		8077
3		27		5		1852				12		4444				19		7037				23		8519				23		8519
4		25		7		2800				13		5200				18		7200				21		8400				21		8400
	106		625		2365		398		199		1125		4259		745		372		1775		6691		874		437		21		7932		616		308		225		8481		352		176
LCOB+THB1		27		5		1852				10		3704				17		6296				19		7037				23		8519
2		29		5		1724				11		3793				18		6207				19		6552				22		7586
3		29		9		3103				18		6207				24		8276				26		8966				27		9310
4		32		6		1875				13		4063				18		5625				23		7188				28		8750
	11700		625		2139		647		323		1300		4442		1187		593		1925		6601		1156		183		2175		7435		1056		528		2500		8541		718		359
germination
Treatment		0 h		96 h		108 h		120 h		132 h		144 h
Control		0		096		1004		6076		7790		8851
LCO A		0		1852		3549		7364		7814		8755
LCO B		0		1808		3431		6716		7732		8083
TH A		0		2010		4023		6464		6964		8132
TH B		0		2158		3979		5936		7410		7846
LCOA+THA		0		2365		4259		6691		7932		8481
LCOB+THB		0		2139		4442		6601		7435		8541
SE
	0 h		96 h		108 h		120 h		132 h		144 h
Control		0		096		199		474		577		297
LCO A		0		130		307		110		295		331
LCO B		0		153		294		147		479		323
TH A		0		217		245		206		269		442
TH B		0		235		220		256		352		387
LCOA+THA		0		199		372		437		308		176
LCOB+THB		0		323		593		183		528		359

0 h		0 h		0 h		0 h		0 h
96 h		96 h		96 h		96 h		96 h
108 h		108 h		108 h		108 h		108 h
120 h		120 h		120 h		120 h		120 h
132 h		132 h		132 h		132 h		132 h

0 h		0 h		0 h		0 h		0 h		0		0		0		0		0		0		0		0		0		0
96 h		96 h		96 h		96 h		96 h		09615384615		09615384615		13049704387		13049704387		15258333938		15258333938		2174332839		2174332839		23499650247		2174332839
108 h		108 h		108 h		108 h		108 h		19859811656		19859811656		30667415327		30667415327		29408073778		29408073778		24539813637		24539813637		2200128559		24539813637
120 h		120 h		120 h		120 h		120 h		47411187081		47411187081		11002902798		11002902798		14704036889		14704036889		20561084936		20561084936		25615660866		20561084936
132 h		132 h		132 h		132 h		132 h		57694206278		57694206278		29473844048		29473844048		4788349344		4788349344		26896579581		26896579581		35169817799		26896579581
144 h		144 h		144 h		144 h		144 h		29704619111		29704619111		33067820092		33067820092		32273789905		32273789905		4421868687		4421868687		3871003991		4421868687

Weight of seedlings
Treatment		FW (mg)		STD		SE			DW (mg)		STD		SE					Treatment		FW (mg)		STD		SE			DW (mg)		STD		SE
Control 1		39850					2924							Control 1
2		11940					854							2		11940					854
3		10680					468							3		10680					468
4		10520					518							4		10520					518
	11047		778		449			614		210		121						11047		778		449			614		210		121
LCO A 1		15860					899							LCO A 1
2		11320					449							2		11320					449
3		10640					437							3		10640					437
4		10610					400							4		10610					400
	12108		2523		1262			546		236		118						12108		2523		1262			429		026		015
LCO B 1		8040					416							LCO B 1		8040					416
2		7560					333							2		7560					333
3		8020					378							3		8020					378
4		7060					363							4		7060					363
	7670		463		232			372		035		017						7670		463		232			372		035		017
TH A 1		14760					632							TH A 1		14760					632
2		12180					457							2		12180					457
3		14450					469							3		14450					469
4		16780					739							4		16780					739
	14543		1884		942			574		136		068						14543		1884		942			574		136		068
TH B 1		10980					391							TH B 1		10980					391
2		15910					701							2		15910					701
3		17040					815							3		17040					815
4		10880					400							4		10880					400
	13703		3235		1617			577		215		107						13703		3235		1617			577		215		107
Treatment		FW (mg)		SE			Treatment		DW (mg)		SE
Control		11047		449			Control		614		121
LCO A		12108		1262			LCO A		429		016
LCO B		7670		232			LCO B		372		017
TH A		14543		942			TH A		574		068
TH B		13703		1617			TH B		577		107

Control		12125587729
LCO A		0155
LCO B		01725714224
TH A		0678241169
TH B		10731213818

Control		44904837651
LCO A		126152932982
LCO B		2315887159
TH A		94179239574
TH B		161736604288

Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B

Arabidopsis normal temperature set 3 continuous
Treatment		No seeds		24 h		germ		Std		SE		36 h		germ		Std		SE		48 h		germ		Std		SE		60 h		germ		Std		SE
Control 1		31		0					16		5161				23		7419				26		8387
2		29		0					19		6552				27		9310				28		9655
3		25		0					15		6000				21		8400				21		8400
4		26		0					13		5000				22		8462				24		9231
	111						1575		5678		729		364		2325		8398		773		387		2475		8918		630		315
LCO A 1		34		0					20		5882				26		7647				34		10000
2		32		0					19		5938				27		8438				30		9375
3		33		0					18		5455				28		8485				29		8788
4		32		0					24		7500				26		8125				29		9063
	131						2025		6194		897		449		2675		8174		386		193		305		9306		521		260
LCO B 1		31		0					14		4516				23		7419				26		8387
2		34		0					15		4412				31		9118				34		10000
3		32		0					20		6250				27		8438				28		8750
4		34		0					16		4706				23		6765				31		9118
	131						1625		4971		861		431		26		7935		1047		523		2975		9064		692		346
TH A 1		28		0					18		6429				23		8214				23		8214
2		29		0					19		6552				24		8276				27		9310
3		30		0					17		5667				23		7667				25		8333
4		28		0					16		5714				25		8929				25		8929
	115						175		6090		465		232		2375		8271		517		258		25		8697		515		257
TH B 1		33		0					23		6970				28		8485				32		9697
2		28		0					19		6786				25		8929				26		9286
3		27		0					21		7778				26		9630				27		10000
4		27		0					20		7407				24		8889				25		9259
	115						2075		7235		446		223		2575		8983		475		238		275		9560		355		177
germination
Treatment		0 h		24 h		36 h		48 h		60 h
Control		0		0		5678		8398		8918
LCO A		0		0		6194		8174		9306
LCO B		0		0		4971		7935		9064
TH A		0		0		6090		8271		8697
TH B		0		0		7235		8983		9560
SE
Control		0		0		364		387		315
LCO A		0		0		449		193		260
LCO B		0		0		431		523		346
TH A		0		0		232		258		257
TH B		0		0		223		238		177

0 h		0 h		0 h		0 h		0 h		0		0		0		0		0		0		0		0		0		0
24 h		24 h		24 h		24 h		24 h		0		0		0		0		0		0		0		0		0		0
36 h		36 h		36 h		36 h		36 h		3644345509		3644345509		44864095916		44864095916		3644345509		3644345509		23241337778		23241337778		2229677279		2229677279
48 h		48 h		48 h		48 h		48 h		38666958625		38666958625		19281389988		19281389988		38666958625		38666958625		2583374136		2583374136		23771554774		23771554774
60 h		60 h		60 h		60 h		60 h		31509650968		31509650968		26047046621		26047046621		31509650968		31509650968		25740422067		25740422067		1774911735		1774911735

24 h		24 h		24 h		24 h		24 h		0		0		0		0		0		0		0		0		0		0
36 h		36 h		36 h		36 h		36 h		3644345509		3644345509		44864095916		44864095916		43067583186		43067583186		23241337778		23241337778		2229677279		2229677279
48 h		48 h		48 h		48 h		48 h		38666958625		38666958625		19281389988		19281389988		52334169948		52334169948		2583374136		2583374136		23771554774		23771554774
60 h		60 h		60 h		60 h		60 h		31509650968		31509650968		26047046621		26047046621		34590070704		34590070704		25740422067		25740422067		1774911735		1774911735

New line graph

Control

LCO A

LCO B

TH A

TH B

Time in hours after stratification

germination

0

0

0

0

0

0

0

0

0

0

567825361513

619359959893

497094402277

609031198686

723514911015

839780953196

817360182709

793480194497

827134646962

898298460798

891825960469

930634469697

906368595825

869663382594

956048581049

Sheet1

Sheet2

FW set 3 Arab normal

FW (mg)

Treatments

Weight in mg

Fresh weight Arabidopsis Normal temp set 3

1104666666667

121075

767

145425

137025

Dry weight Arab set 3 Normal

DW (mg)

Treatments

Weight in mg

Dry weight of Arabidopsis set 3 Normal temp

61353333333

429

372375

57405

5767

Sheet3

Set 3 Low temp LCO TH

Control

LCO A

LCO B

TH A

TH B

Time in hours after stratification

germination

0

0

0

0

0

09615384615

185238095238

180776259336

200991842788

215801790802

100432437932

354880952381

343139202288

402271041898

397858160358

607626170126

736428571429

671569601144

646366332963

593594831095

778958078958

781428571429

773220244716

696421950315

741035816036

885134310134

875476190476

808318766884

81318131257

78457977208

Low temp

Control

LCO A

LCO B

TH A

TH B

Time in hours after stratification

germination

Low temp

0

0

0

0

0

09615384615

185238095238

180776259336

200991842788

215801790802

100432437932

354880952381

343139202288

402271041898

397858160358

607626170126

736428571429

671569601144

646366332963

593594831095

778958078958

781428571429

773220244716

696421950315

741035816036

Sheet1

Sheet2

Sheet3

Salt amp Drought Stress

bullArabidopsisbullSeeds on petriplates with signals

bullControl and 150 mM NaCl

bullSignals improved growth under salt stress

0 NaCl 150 mM NaCl

0 10-6 10-8 10-9 10-11 M LCO Thuricin 17

Hormones of the Holobiontbull Effective at very low concentrationsbull Regulate plant activity

bull Large changes in gene espression protein production hormone profile

bull Regulate other microbesbull Powerful plant productivity effectsbull Inexpensive to applybull Low environmental impact

bull Small amountbull Already produced in environment

bull Very poorly understoodbull Enormous potential

bull New networkbull gt 10 strains in lab now

Phytomicrobiome Potentialbull Wide range of species in the phytomicrobiomebull Most powerful control factor is the plant

bull Developmentbull Stressbull Nutrition

bull Are key to plant growth and productivitybull Management

bull Inoculationbull Hub species bull Signals

bull Microbe-to-plantbull Plant-to-microbebull Microbe-to-microbe

Development of Biologicals - Whobull Scope is largebull Collaboration

bull Basic research labs bull Academicbull Government

bull Industry laboratories

The Approachbull Sample plant-associated microbes

bull The phytomicrobiomebull Efficientrapid screening to identify microbes of interest

bull Biological assessmentsbull Agronomic assessments

Control Thuricin

Industrial Collaboratorsbull Synagribull EVLbull BASFbull RavenQuestbull Ulyssebull Agroworld Natural Productsbull Inocucorbull Axter

BioFuelNet - NCE

Past and Present Students Dr Feng Zhang Dr Fazli Mabood Dr Narjes Dashti Keldeagh Lindsay Dr Pan Bo Dr Sowmya Subramanian Dr Yuming Bai Dr Xiaomin Zhou Elizabeth Gray Nan Wang

PDFs amp Research Associates Dr Xiaomin Zhou Dr Balakrishna Prithiviraj Dr Alfred Souleimanov Dr Kung Dong Lee Dr Woo Jung Mike Lewis Stewart Leibovitch

Acknowledgements

ALSO Trevor Charles at U Waterloo

The End

Thank You

• Biostimulants from the phytomicrobiome Microbe-to-plant signals as low input crop management technologies
• Need for Enhanced Food Production
• Threat of Climate Change
• Biomass for Advanced Biofuels and the Bioeconomy
• The Phytomicrobiome
• Adapting to Climate Change
• The Approach
• So Far
• Example Line of Research
• Something New
• The Problem
• A Solution
• N2 Fixation and Growth - Genistein
• Commercialization
• Serendipity amp Follow Up
• New LCO Activity
• Commercialization
• Jasmonates ndash An Alternative
• Findings
• Effect of Methyl Jasmonate on Yield
• Environment - Temperature and Response
• Salt amp Drought Stress
• Hormones of the Holobiont
• Phytomicrobiome Potential
• Development of Biologicals - Who
• The Approach
• Industrial Collaborators
• BioFuelNet - NCE
• Diapositive numeacutero 29
• The End

Control		96h		000
Control		96h		385
Control		96h		000
Control		96h		000
Control		108h		667
Control		108h		1538
Control		108h		741
Control		108h		1071
Control		120h		5000
Control		120h		7308
Control		120h		5926
Control		120h		6071
Control		132h		6333
Control		132h		8846
Control		132h		7407
Control		132h		8571
Control		144h		8000
Control		144h		9231
Control		144h		8889
Control		144h		9286
LCO A		96h		1600
LCO A		96h		1667
LCO A		96h		2000
LCO A		96h		2143
LCO A		108h		3600
LCO A		108h		2667
LCO A		108h		4000
LCO A		108h		3929
LCO A		120h		7600
LCO A		120h		6333
LCO A		120h		7667
LCO A		120h		7857
LCO A		132h		8400
LCO A		132h		7000
LCO A		132h		8000
LCO A		132h		7857
LCO A		144h		8400
LCO A		144h		8000
LCO A		144h		9333
LCO A		144h		9286
LCO B		96h		1429
LCO B		96h		1935
LCO B		96h		1724
LCO B		96h		2143
LCO B		108h		2857
LCO B		108h		4194
LCO B		108h		3103
LCO B		108h		3571
LCO B		120h		6786
LCO B		120h		7097
LCO B		120h		6552
LCO B		120h		6429
LCO B		132h		7143
LCO B		132h		9032
LCO B		132h		6897
LCO B		132h		7857
LCO B		144h		7857
LCO B		144h		9032
LCO B		144h		7586
LCO B		144h		7857
TH A		96h		2258
TH A		96h		1379
TH A		96h		2069
TH A		96h		2333
TH A		108h		4516
TH A		108h		3448
TH A		108h		3793
TH A		108h		4333
TH A		120h		6774
TH A		120h		6552
TH A		120h		5862
TH A		120h		6667
TH A		132h		7742
TH A		132h		6552
TH A		132h		6897
TH A		132h		6667
TH A		144h		9355
TH A		144h		7241
TH A		144h		7931
TH A		144h		8000
TH B		96h		2000
TH B		96h		1923
TH B		96h		1852
TH B		96h		2857
TH B		108h		3667
TH B		108h		4615
TH B		108h		3704
TH B		108h		3929
TH B		120h		6333
TH B		120h		6154
TH B		120h		5185
TH B		120h		6071
TH B		132h		7000
TH B		132h		8462
TH B		132h		7037
TH B		132h		7143
TH B		144h		8000
TH B		144h		8846
TH B		144h		7037
TH B		144h		7500
LCOA+THA		96h		2500
LCOA+THA		96h		2308
LCOA+THA		96h		1852
LCOA+THA		96h		2800
LCOA+THA		108h		3929
LCOA+THA		108h		3462
LCOA+THA		108h		4444
LCOA+THA		108h		5200
LCOA+THA		120h		7143
LCOA+THA		120h		5385
LCOA+THA		120h		7037
LCOA+THA		120h		7200
LCOA+THA		132h		7500
LCOA+THA		132h		7308
LCOA+THA		132h		8519
LCOA+THA		132h		8400
LCOA+THA		144h		8929
LCOA+THA		144h		8077
LCOA+THA		144h		8519
LCOA+THA		144h		8400
LCOB+THB		96h		1852
LCOB+THB		96h		1724
LCOB+THB		96h		3103
LCOB+THB		96h		1875
LCOB+THB		108h		3704
LCOB+THB		108h		3793
LCOB+THB		108h		6207
LCOB+THB		108h		4063
LCOB+THB		120h		6296
LCOB+THB		120h		6207
LCOB+THB		120h		8276
LCOB+THB		120h		5625
LCOB+THB		132h		7037
LCOB+THB		132h		6552
LCOB+THB		132h		8966
LCOB+THB		132h		7188
LCOB+THB		144h		8519
LCOB+THB		144h		7586
LCOB+THB		144h		9310
LCOB+THB		144h		8750

Arabidopsis low temperature set 3
Treatment		No seeds		96 h		germ		Std		SE		108 h		germ		Std		SE		120 h		germ		Std		SE		132 h		germ		Std		SE		144 h		germ		Std		SE
Control 1		30		0		000				2		667				15		5000				19		6333				24		8000
2		26		1		385				4		1538				19		7308				23		8846				24		9231
3		27		0		000				2		741				16		5926				20		7407				24		8889
4		28		0		000				3		1071				17		6071				24		8571				26		9286
	111		025		096		192		096		275		1004		397		199		1675		6076		948		474		215		7790		1154		577		245		8851		594		297
LCO A 1		25		4		1600				9		3600				19		7600				21		8400				21		8400
2		30		5		1667				8		2667				19		6333				21		7000				24		8000
3		30		6		2000				12		4000				23		7667				24		8000				28		9333
4		28		6		2143				11		3929				22		7857				22		7857				26		9286
	113		525		1852		261		130		10		3549		613		307		2075		7364		696		110		22		7814		589		295		2475		8755		661		331
LCO B 1		28		4		1429				8		2857				19		6786				20		7143				22		7857
2		31		6		1935				13		4194				22		7097				28		9032				28		9032
3		29		5		1724				9		3103				19		6552				20		6897				22		7586
4		28		6		2143				10		3571				18		6429				22		7857				22		7857
	116		525		1808		305		153		10		3431		588		294		195		6716		294		147		225		7732		958		479		235		8083		645		323
TH A 1		31		7		2258				14		4516				21		6774				24		7742				29		9355
2		29		4		1379				10		3448				19		6552				19		6552				21		7241
3		29		6		2069				11		3793				17		5862				20		6897				23		7931
4		30		7		2333				13		4333				20		6667				20		6667				24		8000
	119		6		2010		435		217		12		4023		491		245		1925		6464		411		206		2075		6964		538		269		2425		8132		884		442
TH B 1		30		6		2000				11		3667				19		6333				21		7000				24		8000
2		26		5		1923				12		4615				16		6154				22		8462				23		8846
3		27		5		1852				10		3704				14		5185				19		7037				19		7037
4		28		8		2857				11		3929				17		6071				20		7143				21		7500
	111		6		2158		470		235		11		3979		440		220		165		5936		512		256		205		7410		703		352		2175		7846		774		387
LCOA+THA1		28		7		2500				11		3929				20		7143				21		7500				25		8929
2		26		6		2308				9		3462				14		5385				19		7308				21		8077
3		27		5		1852				12		4444				19		7037				23		8519				23		8519
4		25		7		2800				13		5200				18		7200				21		8400				21		8400
	106		625		2365		398		199		1125		4259		745		372		1775		6691		874		437		21		7932		616		308		225		8481		352		176
LCOB+THB1		27		5		1852				10		3704				17		6296				19		7037				23		8519
2		29		5		1724				11		3793				18		6207				19		6552				22		7586
3		29		9		3103				18		6207				24		8276				26		8966				27		9310
4		32		6		1875				13		4063				18		5625				23		7188				28		8750
	11700		625		2139		647		323		1300		4442		1187		593		1925		6601		1156		183		2175		7435		1056		528		2500		8541		718		359
germination
Treatment		0 h		96 h		108 h		120 h		132 h		144 h
Control		0		096		1004		6076		7790		8851
LCO A		0		1852		3549		7364		7814		8755
LCO B		0		1808		3431		6716		7732		8083
TH A		0		2010		4023		6464		6964		8132
TH B		0		2158		3979		5936		7410		7846
LCOA+THA		0		2365		4259		6691		7932		8481
LCOB+THB		0		2139		4442		6601		7435		8541
SE
	0 h		96 h		108 h		120 h		132 h		144 h
Control		0		096		199		474		577		297
LCO A		0		130		307		110		295		331
LCO B		0		153		294		147		479		323
TH A		0		217		245		206		269		442
TH B		0		235		220		256		352		387
LCOA+THA		0		199		372		437		308		176
LCOB+THB		0		323		593		183		528		359

0 h		0 h		0 h		0 h		0 h
96 h		96 h		96 h		96 h		96 h
108 h		108 h		108 h		108 h		108 h
120 h		120 h		120 h		120 h		120 h
132 h		132 h		132 h		132 h		132 h

0 h		0 h		0 h		0 h		0 h		0		0		0		0		0		0		0		0		0		0
96 h		96 h		96 h		96 h		96 h		09615384615		09615384615		13049704387		13049704387		15258333938		15258333938		2174332839		2174332839		23499650247		2174332839
108 h		108 h		108 h		108 h		108 h		19859811656		19859811656		30667415327		30667415327		29408073778		29408073778		24539813637		24539813637		2200128559		24539813637
120 h		120 h		120 h		120 h		120 h		47411187081		47411187081		11002902798		11002902798		14704036889		14704036889		20561084936		20561084936		25615660866		20561084936
132 h		132 h		132 h		132 h		132 h		57694206278		57694206278		29473844048		29473844048		4788349344		4788349344		26896579581		26896579581		35169817799		26896579581
144 h		144 h		144 h		144 h		144 h		29704619111		29704619111		33067820092		33067820092		32273789905		32273789905		4421868687		4421868687		3871003991		4421868687

Weight of seedlings
Treatment		FW (mg)		STD		SE			DW (mg)		STD		SE					Treatment		FW (mg)		STD		SE			DW (mg)		STD		SE
Control 1		39850					2924							Control 1
2		11940					854							2		11940					854
3		10680					468							3		10680					468
4		10520					518							4		10520					518
	11047		778		449			614		210		121						11047		778		449			614		210		121
LCO A 1		15860					899							LCO A 1
2		11320					449							2		11320					449
3		10640					437							3		10640					437
4		10610					400							4		10610					400
	12108		2523		1262			546		236		118						12108		2523		1262			429		026		015
LCO B 1		8040					416							LCO B 1		8040					416
2		7560					333							2		7560					333
3		8020					378							3		8020					378
4		7060					363							4		7060					363
	7670		463		232			372		035		017						7670		463		232			372		035		017
TH A 1		14760					632							TH A 1		14760					632
2		12180					457							2		12180					457
3		14450					469							3		14450					469
4		16780					739							4		16780					739
	14543		1884		942			574		136		068						14543		1884		942			574		136		068
TH B 1		10980					391							TH B 1		10980					391
2		15910					701							2		15910					701
3		17040					815							3		17040					815
4		10880					400							4		10880					400
	13703		3235		1617			577		215		107						13703		3235		1617			577		215		107
Treatment		FW (mg)		SE			Treatment		DW (mg)		SE
Control		11047		449			Control		614		121
LCO A		12108		1262			LCO A		429		016
LCO B		7670		232			LCO B		372		017
TH A		14543		942			TH A		574		068
TH B		13703		1617			TH B		577		107

Control		12125587729
LCO A		0155
LCO B		01725714224
TH A		0678241169
TH B		10731213818

Control		44904837651
LCO A		126152932982
LCO B		2315887159
TH A		94179239574
TH B		161736604288

Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B

Arabidopsis normal temperature set 3 continuous
Treatment		No seeds		24 h		germ		Std		SE		36 h		germ		Std		SE		48 h		germ		Std		SE		60 h		germ		Std		SE
Control 1		31		0					16		5161				23		7419				26		8387
2		29		0					19		6552				27		9310				28		9655
3		25		0					15		6000				21		8400				21		8400
4		26		0					13		5000				22		8462				24		9231
	111						1575		5678		729		364		2325		8398		773		387		2475		8918		630		315
LCO A 1		34		0					20		5882				26		7647				34		10000
2		32		0					19		5938				27		8438				30		9375
3		33		0					18		5455				28		8485				29		8788
4		32		0					24		7500				26		8125				29		9063
	131						2025		6194		897		449		2675		8174		386		193		305		9306		521		260
LCO B 1		31		0					14		4516				23		7419				26		8387
2		34		0					15		4412				31		9118				34		10000
3		32		0					20		6250				27		8438				28		8750
4		34		0					16		4706				23		6765				31		9118
	131						1625		4971		861		431		26		7935		1047		523		2975		9064		692		346
TH A 1		28		0					18		6429				23		8214				23		8214
2		29		0					19		6552				24		8276				27		9310
3		30		0					17		5667				23		7667				25		8333
4		28		0					16		5714				25		8929				25		8929
	115						175		6090		465		232		2375		8271		517		258		25		8697		515		257
TH B 1		33		0					23		6970				28		8485				32		9697
2		28		0					19		6786				25		8929				26		9286
3		27		0					21		7778				26		9630				27		10000
4		27		0					20		7407				24		8889				25		9259
	115						2075		7235		446		223		2575		8983		475		238		275		9560		355		177
germination
Treatment		0 h		24 h		36 h		48 h		60 h
Control		0		0		5678		8398		8918
LCO A		0		0		6194		8174		9306
LCO B		0		0		4971		7935		9064
TH A		0		0		6090		8271		8697
TH B		0		0		7235		8983		9560
SE
Control		0		0		364		387		315
LCO A		0		0		449		193		260
LCO B		0		0		431		523		346
TH A		0		0		232		258		257
TH B		0		0		223		238		177

0 h		0 h		0 h		0 h		0 h		0		0		0		0		0		0		0		0		0		0
24 h		24 h		24 h		24 h		24 h		0		0		0		0		0		0		0		0		0		0
36 h		36 h		36 h		36 h		36 h		3644345509		3644345509		44864095916		44864095916		3644345509		3644345509		23241337778		23241337778		2229677279		2229677279
48 h		48 h		48 h		48 h		48 h		38666958625		38666958625		19281389988		19281389988		38666958625		38666958625		2583374136		2583374136		23771554774		23771554774
60 h		60 h		60 h		60 h		60 h		31509650968		31509650968		26047046621		26047046621		31509650968		31509650968		25740422067		25740422067		1774911735		1774911735

Sheet1

Sheet2

FW set 3 Arab normal

FW (mg)

Treatments

Weight in mg

Fresh weight Arabidopsis Normal temp set 3

1104666666667

121075

767

145425

137025

Dry weight Arab set 3 Normal

DW (mg)

Treatments

Weight in mg

Dry weight of Arabidopsis set 3 Normal temp

61353333333

429

372375

57405

5767

Sheet3

Set 3 Low temp LCO TH

Control

LCO A

LCO B

TH A

TH B

Time in hours after stratification

germination

0

0

0

0

0

09615384615

185238095238

180776259336

200991842788

215801790802

100432437932

354880952381

343139202288

402271041898

397858160358

607626170126

736428571429

671569601144

646366332963

593594831095

778958078958

781428571429

773220244716

696421950315

741035816036

885134310134

875476190476

808318766884

81318131257

78457977208

Low temp

Control

LCO A

LCO B

TH A

TH B

Time in hours after stratification

germination

Low temp

0

0

0

0

0

09615384615

185238095238

180776259336

200991842788

215801790802

100432437932

354880952381

343139202288

402271041898

397858160358

607626170126

736428571429

671569601144

646366332963

593594831095

778958078958

781428571429

773220244716

696421950315

741035816036

Sheet1

Sheet2

Sheet3

Salt amp Drought Stress

bullArabidopsisbullSeeds on petriplates with signals

bullControl and 150 mM NaCl

bullSignals improved growth under salt stress

0 NaCl 150 mM NaCl

0 10-6 10-8 10-9 10-11 M LCO Thuricin 17

Hormones of the Holobiontbull Effective at very low concentrationsbull Regulate plant activity

bull Large changes in gene espression protein production hormone profile

bull Regulate other microbesbull Powerful plant productivity effectsbull Inexpensive to applybull Low environmental impact

bull Small amountbull Already produced in environment

bull Very poorly understoodbull Enormous potential

bull New networkbull gt 10 strains in lab now

Phytomicrobiome Potentialbull Wide range of species in the phytomicrobiomebull Most powerful control factor is the plant

bull Developmentbull Stressbull Nutrition

bull Are key to plant growth and productivitybull Management

bull Inoculationbull Hub species bull Signals

bull Microbe-to-plantbull Plant-to-microbebull Microbe-to-microbe

Development of Biologicals - Whobull Scope is largebull Collaboration

bull Basic research labs bull Academicbull Government

bull Industry laboratories

The Approachbull Sample plant-associated microbes

bull The phytomicrobiomebull Efficientrapid screening to identify microbes of interest

bull Biological assessmentsbull Agronomic assessments

Control Thuricin

Industrial Collaboratorsbull Synagribull EVLbull BASFbull RavenQuestbull Ulyssebull Agroworld Natural Productsbull Inocucorbull Axter

BioFuelNet - NCE

Past and Present Students Dr Feng Zhang Dr Fazli Mabood Dr Narjes Dashti Keldeagh Lindsay Dr Pan Bo Dr Sowmya Subramanian Dr Yuming Bai Dr Xiaomin Zhou Elizabeth Gray Nan Wang

PDFs amp Research Associates Dr Xiaomin Zhou Dr Balakrishna Prithiviraj Dr Alfred Souleimanov Dr Kung Dong Lee Dr Woo Jung Mike Lewis Stewart Leibovitch

Acknowledgements

ALSO Trevor Charles at U Waterloo

The End

Thank You

• Biostimulants from the phytomicrobiome Microbe-to-plant signals as low input crop management technologies
• Need for Enhanced Food Production
• Threat of Climate Change
• Biomass for Advanced Biofuels and the Bioeconomy
• The Phytomicrobiome
• Adapting to Climate Change
• The Approach
• So Far
• Example Line of Research
• Something New
• The Problem
• A Solution
• N2 Fixation and Growth - Genistein
• Commercialization
• Serendipity amp Follow Up
• New LCO Activity
• Commercialization
• Jasmonates ndash An Alternative
• Findings
• Effect of Methyl Jasmonate on Yield
• Environment - Temperature and Response
• Salt amp Drought Stress
• Hormones of the Holobiont
• Phytomicrobiome Potential
• Development of Biologicals - Who
• The Approach
• Industrial Collaborators
• BioFuelNet - NCE
• Diapositive numeacutero 29
• The End

Control		96h		000
Control		96h		385
Control		96h		000
Control		96h		000
Control		108h		667
Control		108h		1538
Control		108h		741
Control		108h		1071
Control		120h		5000
Control		120h		7308
Control		120h		5926
Control		120h		6071
Control		132h		6333
Control		132h		8846
Control		132h		7407
Control		132h		8571
Control		144h		8000
Control		144h		9231
Control		144h		8889
Control		144h		9286
LCO A		96h		1600
LCO A		96h		1667
LCO A		96h		2000
LCO A		96h		2143
LCO A		108h		3600
LCO A		108h		2667
LCO A		108h		4000
LCO A		108h		3929
LCO A		120h		7600
LCO A		120h		6333
LCO A		120h		7667
LCO A		120h		7857
LCO A		132h		8400
LCO A		132h		7000
LCO A		132h		8000
LCO A		132h		7857
LCO A		144h		8400
LCO A		144h		8000
LCO A		144h		9333
LCO A		144h		9286
LCO B		96h		1429
LCO B		96h		1935
LCO B		96h		1724
LCO B		96h		2143
LCO B		108h		2857
LCO B		108h		4194
LCO B		108h		3103
LCO B		108h		3571
LCO B		120h		6786
LCO B		120h		7097
LCO B		120h		6552
LCO B		120h		6429
LCO B		132h		7143
LCO B		132h		9032
LCO B		132h		6897
LCO B		132h		7857
LCO B		144h		7857
LCO B		144h		9032
LCO B		144h		7586
LCO B		144h		7857
TH A		96h		2258
TH A		96h		1379
TH A		96h		2069
TH A		96h		2333
TH A		108h		4516
TH A		108h		3448
TH A		108h		3793
TH A		108h		4333
TH A		120h		6774
TH A		120h		6552
TH A		120h		5862
TH A		120h		6667
TH A		132h		7742
TH A		132h		6552
TH A		132h		6897
TH A		132h		6667
TH A		144h		9355
TH A		144h		7241
TH A		144h		7931
TH A		144h		8000
TH B		96h		2000
TH B		96h		1923
TH B		96h		1852
TH B		96h		2857
TH B		108h		3667
TH B		108h		4615
TH B		108h		3704
TH B		108h		3929
TH B		120h		6333
TH B		120h		6154
TH B		120h		5185
TH B		120h		6071
TH B		132h		7000
TH B		132h		8462
TH B		132h		7037
TH B		132h		7143
TH B		144h		8000
TH B		144h		8846
TH B		144h		7037
TH B		144h		7500
LCOA+THA		96h		2500
LCOA+THA		96h		2308
LCOA+THA		96h		1852
LCOA+THA		96h		2800
LCOA+THA		108h		3929
LCOA+THA		108h		3462
LCOA+THA		108h		4444
LCOA+THA		108h		5200
LCOA+THA		120h		7143
LCOA+THA		120h		5385
LCOA+THA		120h		7037
LCOA+THA		120h		7200
LCOA+THA		132h		7500
LCOA+THA		132h		7308
LCOA+THA		132h		8519
LCOA+THA		132h		8400
LCOA+THA		144h		8929
LCOA+THA		144h		8077
LCOA+THA		144h		8519
LCOA+THA		144h		8400
LCOB+THB		96h		1852
LCOB+THB		96h		1724
LCOB+THB		96h		3103
LCOB+THB		96h		1875
LCOB+THB		108h		3704
LCOB+THB		108h		3793
LCOB+THB		108h		6207
LCOB+THB		108h		4063
LCOB+THB		120h		6296
LCOB+THB		120h		6207
LCOB+THB		120h		8276
LCOB+THB		120h		5625
LCOB+THB		132h		7037
LCOB+THB		132h		6552
LCOB+THB		132h		8966
LCOB+THB		132h		7188
LCOB+THB		144h		8519
LCOB+THB		144h		7586
LCOB+THB		144h		9310
LCOB+THB		144h		8750

Arabidopsis low temperature set 3
Treatment		No seeds		96 h		germ		Std		SE		108 h		germ		Std		SE		120 h		germ		Std		SE		132 h		germ		Std		SE		144 h		germ		Std		SE
Control 1		30		0		000				2		667				15		5000				19		6333				24		8000
2		26		1		385				4		1538				19		7308				23		8846				24		9231
3		27		0		000				2		741				16		5926				20		7407				24		8889
4		28		0		000				3		1071				17		6071				24		8571				26		9286
	111		025		096		192		096		275		1004		397		199		1675		6076		948		474		215		7790		1154		577		245		8851		594		297
LCO A 1		25		4		1600				9		3600				19		7600				21		8400				21		8400
2		30		5		1667				8		2667				19		6333				21		7000				24		8000
3		30		6		2000				12		4000				23		7667				24		8000				28		9333
4		28		6		2143				11		3929				22		7857				22		7857				26		9286
	113		525		1852		261		130		10		3549		613		307		2075		7364		696		110		22		7814		589		295		2475		8755		661		331
LCO B 1		28		4		1429				8		2857				19		6786				20		7143				22		7857
2		31		6		1935				13		4194				22		7097				28		9032				28		9032
3		29		5		1724				9		3103				19		6552				20		6897				22		7586
4		28		6		2143				10		3571				18		6429				22		7857				22		7857
	116		525		1808		305		153		10		3431		588		294		195		6716		294		147		225		7732		958		479		235		8083		645		323
TH A 1		31		7		2258				14		4516				21		6774				24		7742				29		9355
2		29		4		1379				10		3448				19		6552				19		6552				21		7241
3		29		6		2069				11		3793				17		5862				20		6897				23		7931
4		30		7		2333				13		4333				20		6667				20		6667				24		8000
	119		6		2010		435		217		12		4023		491		245		1925		6464		411		206		2075		6964		538		269		2425		8132		884		442
TH B 1		30		6		2000				11		3667				19		6333				21		7000				24		8000
2		26		5		1923				12		4615				16		6154				22		8462				23		8846
3		27		5		1852				10		3704				14		5185				19		7037				19		7037
4		28		8		2857				11		3929				17		6071				20		7143				21		7500
	111		6		2158		470		235		11		3979		440		220		165		5936		512		256		205		7410		703		352		2175		7846		774		387
LCOA+THA1		28		7		2500				11		3929				20		7143				21		7500				25		8929
2		26		6		2308				9		3462				14		5385				19		7308				21		8077
3		27		5		1852				12		4444				19		7037				23		8519				23		8519
4		25		7		2800				13		5200				18		7200				21		8400				21		8400
	106		625		2365		398		199		1125		4259		745		372		1775		6691		874		437		21		7932		616		308		225		8481		352		176
LCOB+THB1		27		5		1852				10		3704				17		6296				19		7037				23		8519
2		29		5		1724				11		3793				18		6207				19		6552				22		7586
3		29		9		3103				18		6207				24		8276				26		8966				27		9310
4		32		6		1875				13		4063				18		5625				23		7188				28		8750
	11700		625		2139		647		323		1300		4442		1187		593		1925		6601		1156		183		2175		7435		1056		528		2500		8541		718		359
germination
Treatment		0 h		96 h		108 h		120 h		132 h		144 h
Control		0		096		1004		6076		7790		8851
LCO A		0		1852		3549		7364		7814		8755
LCO B		0		1808		3431		6716		7732		8083
TH A		0		2010		4023		6464		6964		8132
TH B		0		2158		3979		5936		7410		7846
LCOA+THA		0		2365		4259		6691		7932		8481
LCOB+THB		0		2139		4442		6601		7435		8541
SE
	0 h		96 h		108 h		120 h		132 h		144 h
Control		0		096		199		474		577		297
LCO A		0		130		307		110		295		331
LCO B		0		153		294		147		479		323
TH A		0		217		245		206		269		442
TH B		0		235		220		256		352		387
LCOA+THA		0		199		372		437		308		176
LCOB+THB		0		323		593		183		528		359

0 h		0 h		0 h		0 h		0 h
96 h		96 h		96 h		96 h		96 h
108 h		108 h		108 h		108 h		108 h
120 h		120 h		120 h		120 h		120 h
132 h		132 h		132 h		132 h		132 h

0 h		0 h		0 h		0 h		0 h		0		0		0		0		0		0		0		0		0		0
96 h		96 h		96 h		96 h		96 h		09615384615		09615384615		13049704387		13049704387		15258333938		15258333938		2174332839		2174332839		23499650247		2174332839
108 h		108 h		108 h		108 h		108 h		19859811656		19859811656		30667415327		30667415327		29408073778		29408073778		24539813637		24539813637		2200128559		24539813637
120 h		120 h		120 h		120 h		120 h		47411187081		47411187081		11002902798		11002902798		14704036889		14704036889		20561084936		20561084936		25615660866		20561084936
132 h		132 h		132 h		132 h		132 h		57694206278		57694206278		29473844048		29473844048		4788349344		4788349344		26896579581		26896579581		35169817799		26896579581
144 h		144 h		144 h		144 h		144 h		29704619111		29704619111		33067820092		33067820092		32273789905		32273789905		4421868687		4421868687		3871003991		4421868687

Weight of seedlings
Treatment		FW (mg)		STD		SE			DW (mg)		STD		SE					Treatment		FW (mg)		STD		SE			DW (mg)		STD		SE
Control 1		39850					2924							Control 1
2		11940					854							2		11940					854
3		10680					468							3		10680					468
4		10520					518							4		10520					518
	11047		778		449			614		210		121						11047		778		449			614		210		121
LCO A 1		15860					899							LCO A 1
2		11320					449							2		11320					449
3		10640					437							3		10640					437
4		10610					400							4		10610					400
	12108		2523		1262			546		236		118						12108		2523		1262			429		026		015
LCO B 1		8040					416							LCO B 1		8040					416
2		7560					333							2		7560					333
3		8020					378							3		8020					378
4		7060					363							4		7060					363
	7670		463		232			372		035		017						7670		463		232			372		035		017
TH A 1		14760					632							TH A 1		14760					632
2		12180					457							2		12180					457
3		14450					469							3		14450					469
4		16780					739							4		16780					739
	14543		1884		942			574		136		068						14543		1884		942			574		136		068
TH B 1		10980					391							TH B 1		10980					391
2		15910					701							2		15910					701
3		17040					815							3		17040					815
4		10880					400							4		10880					400
	13703		3235		1617			577		215		107						13703		3235		1617			577		215		107
Treatment		FW (mg)		SE			Treatment		DW (mg)		SE
Control		11047		449			Control		614		121
LCO A		12108		1262			LCO A		429		016
LCO B		7670		232			LCO B		372		017
TH A		14543		942			TH A		574		068
TH B		13703		1617			TH B		577		107

Control		12125587729
LCO A		0155
LCO B		01725714224
TH A		0678241169
TH B		10731213818

Control		44904837651
LCO A		126152932982
LCO B		2315887159
TH A		94179239574
TH B		161736604288

Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B

Arabidopsis normal temperature set 3 continuous
Treatment		No seeds		24 h		germ		Std		SE		36 h		germ		Std		SE		48 h		germ		Std		SE		60 h		germ		Std		SE
Control 1		31		0					16		5161				23		7419				26		8387
2		29		0					19		6552				27		9310				28		9655
3		25		0					15		6000				21		8400				21		8400
4		26		0					13		5000				22		8462				24		9231
	111						1575		5678		729		364		2325		8398		773		387		2475		8918		630		315
LCO A 1		34		0					20		5882				26		7647				34		10000
2		32		0					19		5938				27		8438				30		9375
3		33		0					18		5455				28		8485				29		8788
4		32		0					24		7500				26		8125				29		9063
	131						2025		6194		897		449		2675		8174		386		193		305		9306		521		260
LCO B 1		31		0					14		4516				23		7419				26		8387
2		34		0					15		4412				31		9118				34		10000
3		32		0					20		6250				27		8438				28		8750
4		34		0					16		4706				23		6765				31		9118
	131						1625		4971		861		431		26		7935		1047		523		2975		9064		692		346
TH A 1		28		0					18		6429				23		8214				23		8214
2		29		0					19		6552				24		8276				27		9310
3		30		0					17		5667				23		7667				25		8333
4		28		0					16		5714				25		8929				25		8929
	115						175		6090		465		232		2375		8271		517		258		25		8697		515		257
TH B 1		33		0					23		6970				28		8485				32		9697
2		28		0					19		6786				25		8929				26		9286
3		27		0					21		7778				26		9630				27		10000
4		27		0					20		7407				24		8889				25		9259
	115						2075		7235		446		223		2575		8983		475		238		275		9560		355		177
germination
Treatment		0 h		24 h		36 h		48 h		60 h
Control		0		0		5678		8398		8918
LCO A		0		0		6194		8174		9306
LCO B		0		0		4971		7935		9064
TH A		0		0		6090		8271		8697
TH B		0		0		7235		8983		9560
SE
Control		0		0		364		387		315
LCO A		0		0		449		193		260
LCO B		0		0		431		523		346
TH A		0		0		232		258		257
TH B		0		0		223		238		177

Sheet2

FW set 3 Arab normal

FW (mg)

Treatments

Weight in mg

Fresh weight Arabidopsis Normal temp set 3

1104666666667

121075

767

145425

137025

Dry weight Arab set 3 Normal

DW (mg)

Treatments

Weight in mg

Dry weight of Arabidopsis set 3 Normal temp

61353333333

429

372375

57405

5767

Sheet3

Set 3 Low temp LCO TH

Control

LCO A

LCO B

TH A

TH B

Time in hours after stratification

germination

0

0

0

0

0

09615384615

185238095238

180776259336

200991842788

215801790802

100432437932

354880952381

343139202288

402271041898

397858160358

607626170126

736428571429

671569601144

646366332963

593594831095

778958078958

781428571429

773220244716

696421950315

741035816036

885134310134

875476190476

808318766884

81318131257

78457977208

Low temp

Control

LCO A

LCO B

TH A

TH B

Time in hours after stratification

germination

Low temp

0

0

0

0

0

09615384615

185238095238

180776259336

200991842788

215801790802

100432437932

354880952381

343139202288

402271041898

397858160358

607626170126

736428571429

671569601144

646366332963

593594831095

778958078958

781428571429

773220244716

696421950315

741035816036

Sheet1

Sheet2

Sheet3

Salt amp Drought Stress

bullArabidopsisbullSeeds on petriplates with signals

bullControl and 150 mM NaCl

bullSignals improved growth under salt stress

0 NaCl 150 mM NaCl

0 10-6 10-8 10-9 10-11 M LCO Thuricin 17

Hormones of the Holobiontbull Effective at very low concentrationsbull Regulate plant activity

bull Large changes in gene espression protein production hormone profile

bull Regulate other microbesbull Powerful plant productivity effectsbull Inexpensive to applybull Low environmental impact

bull Small amountbull Already produced in environment

bull Very poorly understoodbull Enormous potential

bull New networkbull gt 10 strains in lab now

Phytomicrobiome Potentialbull Wide range of species in the phytomicrobiomebull Most powerful control factor is the plant

bull Developmentbull Stressbull Nutrition

bull Are key to plant growth and productivitybull Management

bull Inoculationbull Hub species bull Signals

bull Microbe-to-plantbull Plant-to-microbebull Microbe-to-microbe

Development of Biologicals - Whobull Scope is largebull Collaboration

bull Basic research labs bull Academicbull Government

bull Industry laboratories

The Approachbull Sample plant-associated microbes

bull The phytomicrobiomebull Efficientrapid screening to identify microbes of interest

bull Biological assessmentsbull Agronomic assessments

Control Thuricin

Industrial Collaboratorsbull Synagribull EVLbull BASFbull RavenQuestbull Ulyssebull Agroworld Natural Productsbull Inocucorbull Axter

BioFuelNet - NCE

Past and Present Students Dr Feng Zhang Dr Fazli Mabood Dr Narjes Dashti Keldeagh Lindsay Dr Pan Bo Dr Sowmya Subramanian Dr Yuming Bai Dr Xiaomin Zhou Elizabeth Gray Nan Wang

PDFs amp Research Associates Dr Xiaomin Zhou Dr Balakrishna Prithiviraj Dr Alfred Souleimanov Dr Kung Dong Lee Dr Woo Jung Mike Lewis Stewart Leibovitch

Acknowledgements

ALSO Trevor Charles at U Waterloo

The End

Thank You

• Biostimulants from the phytomicrobiome Microbe-to-plant signals as low input crop management technologies
• Need for Enhanced Food Production
• Threat of Climate Change
• Biomass for Advanced Biofuels and the Bioeconomy
• The Phytomicrobiome
• Adapting to Climate Change
• The Approach
• So Far
• Example Line of Research
• Something New
• The Problem
• A Solution
• N2 Fixation and Growth - Genistein
• Commercialization
• Serendipity amp Follow Up
• New LCO Activity
• Commercialization
• Jasmonates ndash An Alternative
• Findings
• Effect of Methyl Jasmonate on Yield
• Environment - Temperature and Response
• Salt amp Drought Stress
• Hormones of the Holobiont
• Phytomicrobiome Potential
• Development of Biologicals - Who
• The Approach
• Industrial Collaborators
• BioFuelNet - NCE
• Diapositive numeacutero 29
• The End

Control		96h		000
Control		96h		385
Control		96h		000
Control		96h		000
Control		108h		667
Control		108h		1538
Control		108h		741
Control		108h		1071
Control		120h		5000
Control		120h		7308
Control		120h		5926
Control		120h		6071
Control		132h		6333
Control		132h		8846
Control		132h		7407
Control		132h		8571
Control		144h		8000
Control		144h		9231
Control		144h		8889
Control		144h		9286
LCO A		96h		1600
LCO A		96h		1667
LCO A		96h		2000
LCO A		96h		2143
LCO A		108h		3600
LCO A		108h		2667
LCO A		108h		4000
LCO A		108h		3929
LCO A		120h		7600
LCO A		120h		6333
LCO A		120h		7667
LCO A		120h		7857
LCO A		132h		8400
LCO A		132h		7000
LCO A		132h		8000
LCO A		132h		7857
LCO A		144h		8400
LCO A		144h		8000
LCO A		144h		9333
LCO A		144h		9286
LCO B		96h		1429
LCO B		96h		1935
LCO B		96h		1724
LCO B		96h		2143
LCO B		108h		2857
LCO B		108h		4194
LCO B		108h		3103
LCO B		108h		3571
LCO B		120h		6786
LCO B		120h		7097
LCO B		120h		6552
LCO B		120h		6429
LCO B		132h		7143
LCO B		132h		9032
LCO B		132h		6897
LCO B		132h		7857
LCO B		144h		7857
LCO B		144h		9032
LCO B		144h		7586
LCO B		144h		7857
TH A		96h		2258
TH A		96h		1379
TH A		96h		2069
TH A		96h		2333
TH A		108h		4516
TH A		108h		3448
TH A		108h		3793
TH A		108h		4333
TH A		120h		6774
TH A		120h		6552
TH A		120h		5862
TH A		120h		6667
TH A		132h		7742
TH A		132h		6552
TH A		132h		6897
TH A		132h		6667
TH A		144h		9355
TH A		144h		7241
TH A		144h		7931
TH A		144h		8000
TH B		96h		2000
TH B		96h		1923
TH B		96h		1852
TH B		96h		2857
TH B		108h		3667
TH B		108h		4615
TH B		108h		3704
TH B		108h		3929
TH B		120h		6333
TH B		120h		6154
TH B		120h		5185
TH B		120h		6071
TH B		132h		7000
TH B		132h		8462
TH B		132h		7037
TH B		132h		7143
TH B		144h		8000
TH B		144h		8846
TH B		144h		7037
TH B		144h		7500
LCOA+THA		96h		2500
LCOA+THA		96h		2308
LCOA+THA		96h		1852
LCOA+THA		96h		2800
LCOA+THA		108h		3929
LCOA+THA		108h		3462
LCOA+THA		108h		4444
LCOA+THA		108h		5200
LCOA+THA		120h		7143
LCOA+THA		120h		5385
LCOA+THA		120h		7037
LCOA+THA		120h		7200
LCOA+THA		132h		7500
LCOA+THA		132h		7308
LCOA+THA		132h		8519
LCOA+THA		132h		8400
LCOA+THA		144h		8929
LCOA+THA		144h		8077
LCOA+THA		144h		8519
LCOA+THA		144h		8400
LCOB+THB		96h		1852
LCOB+THB		96h		1724
LCOB+THB		96h		3103
LCOB+THB		96h		1875
LCOB+THB		108h		3704
LCOB+THB		108h		3793
LCOB+THB		108h		6207
LCOB+THB		108h		4063
LCOB+THB		120h		6296
LCOB+THB		120h		6207
LCOB+THB		120h		8276
LCOB+THB		120h		5625
LCOB+THB		132h		7037
LCOB+THB		132h		6552
LCOB+THB		132h		8966
LCOB+THB		132h		7188
LCOB+THB		144h		8519
LCOB+THB		144h		7586
LCOB+THB		144h		9310
LCOB+THB		144h		8750

Arabidopsis low temperature set 3
Treatment		No seeds		96 h		germ		Std		SE		108 h		germ		Std		SE		120 h		germ		Std		SE		132 h		germ		Std		SE		144 h		germ		Std		SE
Control 1		30		0		000				2		667				15		5000				19		6333				24		8000
2		26		1		385				4		1538				19		7308				23		8846				24		9231
3		27		0		000				2		741				16		5926				20		7407				24		8889
4		28		0		000				3		1071				17		6071				24		8571				26		9286
	111		025		096		192		096		275		1004		397		199		1675		6076		948		474		215		7790		1154		577		245		8851		594		297
LCO A 1		25		4		1600				9		3600				19		7600				21		8400				21		8400
2		30		5		1667				8		2667				19		6333				21		7000				24		8000
3		30		6		2000				12		4000				23		7667				24		8000				28		9333
4		28		6		2143				11		3929				22		7857				22		7857				26		9286
	113		525		1852		261		130		10		3549		613		307		2075		7364		696		110		22		7814		589		295		2475		8755		661		331
LCO B 1		28		4		1429				8		2857				19		6786				20		7143				22		7857
2		31		6		1935				13		4194				22		7097				28		9032				28		9032
3		29		5		1724				9		3103				19		6552				20		6897				22		7586
4		28		6		2143				10		3571				18		6429				22		7857				22		7857
	116		525		1808		305		153		10		3431		588		294		195		6716		294		147		225		7732		958		479		235		8083		645		323
TH A 1		31		7		2258				14		4516				21		6774				24		7742				29		9355
2		29		4		1379				10		3448				19		6552				19		6552				21		7241
3		29		6		2069				11		3793				17		5862				20		6897				23		7931
4		30		7		2333				13		4333				20		6667				20		6667				24		8000
	119		6		2010		435		217		12		4023		491		245		1925		6464		411		206		2075		6964		538		269		2425		8132		884		442
TH B 1		30		6		2000				11		3667				19		6333				21		7000				24		8000
2		26		5		1923				12		4615				16		6154				22		8462				23		8846
3		27		5		1852				10		3704				14		5185				19		7037				19		7037
4		28		8		2857				11		3929				17		6071				20		7143				21		7500
	111		6		2158		470		235		11		3979		440		220		165		5936		512		256		205		7410		703		352		2175		7846		774		387
LCOA+THA1		28		7		2500				11		3929				20		7143				21		7500				25		8929
2		26		6		2308				9		3462				14		5385				19		7308				21		8077
3		27		5		1852				12		4444				19		7037				23		8519				23		8519
4		25		7		2800				13		5200				18		7200				21		8400				21		8400
	106		625		2365		398		199		1125		4259		745		372		1775		6691		874		437		21		7932		616		308		225		8481		352		176
LCOB+THB1		27		5		1852				10		3704				17		6296				19		7037				23		8519
2		29		5		1724				11		3793				18		6207				19		6552				22		7586
3		29		9		3103				18		6207				24		8276				26		8966				27		9310
4		32		6		1875				13		4063				18		5625				23		7188				28		8750
	11700		625		2139		647		323		1300		4442		1187		593		1925		6601		1156		183		2175		7435		1056		528		2500		8541		718		359
germination
Treatment		0 h		96 h		108 h		120 h		132 h		144 h
Control		0		096		1004		6076		7790		8851
LCO A		0		1852		3549		7364		7814		8755
LCO B		0		1808		3431		6716		7732		8083
TH A		0		2010		4023		6464		6964		8132
TH B		0		2158		3979		5936		7410		7846
LCOA+THA		0		2365		4259		6691		7932		8481
LCOB+THB		0		2139		4442		6601		7435		8541
SE
	0 h		96 h		108 h		120 h		132 h		144 h
Control		0		096		199		474		577		297
LCO A		0		130		307		110		295		331
LCO B		0		153		294		147		479		323
TH A		0		217		245		206		269		442
TH B		0		235		220		256		352		387
LCOA+THA		0		199		372		437		308		176
LCOB+THB		0		323		593		183		528		359

0 h		0 h		0 h		0 h		0 h
96 h		96 h		96 h		96 h		96 h
108 h		108 h		108 h		108 h		108 h
120 h		120 h		120 h		120 h		120 h
132 h		132 h		132 h		132 h		132 h

0 h		0 h		0 h		0 h		0 h		0		0		0		0		0		0		0		0		0		0
96 h		96 h		96 h		96 h		96 h		09615384615		09615384615		13049704387		13049704387		15258333938		15258333938		2174332839		2174332839		23499650247		2174332839
108 h		108 h		108 h		108 h		108 h		19859811656		19859811656		30667415327		30667415327		29408073778		29408073778		24539813637		24539813637		2200128559		24539813637
120 h		120 h		120 h		120 h		120 h		47411187081		47411187081		11002902798		11002902798		14704036889		14704036889		20561084936		20561084936		25615660866		20561084936
132 h		132 h		132 h		132 h		132 h		57694206278		57694206278		29473844048		29473844048		4788349344		4788349344		26896579581		26896579581		35169817799		26896579581
144 h		144 h		144 h		144 h		144 h		29704619111		29704619111		33067820092		33067820092		32273789905		32273789905		4421868687		4421868687		3871003991		4421868687

Weight of seedlings
Treatment		FW (mg)		STD		SE			DW (mg)		STD		SE					Treatment		FW (mg)		STD		SE			DW (mg)		STD		SE
Control 1		39850					2924							Control 1
2		11940					854							2		11940					854
3		10680					468							3		10680					468
4		10520					518							4		10520					518
	11047		778		449			614		210		121						11047		778		449			614		210		121
LCO A 1		15860					899							LCO A 1
2		11320					449							2		11320					449
3		10640					437							3		10640					437
4		10610					400							4		10610					400
	12108		2523		1262			546		236		118						12108		2523		1262			429		026		015
LCO B 1		8040					416							LCO B 1		8040					416
2		7560					333							2		7560					333
3		8020					378							3		8020					378
4		7060					363							4		7060					363
	7670		463		232			372		035		017						7670		463		232			372		035		017
TH A 1		14760					632							TH A 1		14760					632
2		12180					457							2		12180					457
3		14450					469							3		14450					469
4		16780					739							4		16780					739
	14543		1884		942			574		136		068						14543		1884		942			574		136		068
TH B 1		10980					391							TH B 1		10980					391
2		15910					701							2		15910					701
3		17040					815							3		17040					815
4		10880					400							4		10880					400
	13703		3235		1617			577		215		107						13703		3235		1617			577		215		107
Treatment		FW (mg)		SE			Treatment		DW (mg)		SE
Control		11047		449			Control		614		121
LCO A		12108		1262			LCO A		429		016
LCO B		7670		232			LCO B		372		017
TH A		14543		942			TH A		574		068
TH B		13703		1617			TH B		577		107

Control		12125587729
LCO A		0155
LCO B		01725714224
TH A		0678241169
TH B		10731213818

Control		44904837651
LCO A		126152932982
LCO B		2315887159
TH A		94179239574
TH B		161736604288

Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
Control
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO A
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
LCO B
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH A
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B
TH B

FW set 3 Arab normal

FW (mg)

Treatments

Weight in mg

Fresh weight Arabidopsis Normal temp set 3

1104666666667

121075

767

145425

137025

Dry weight Arab set 3 Normal

DW (mg)

Treatments

Weight in mg

Dry weight of Arabidopsis set 3 Normal temp

61353333333

429

372375

57405

5767

Sheet3

Set 3 Low temp LCO TH

Control

LCO A

LCO B

TH A

TH B

Time in hours after stratification

germination

0

0

0

0

0

09615384615

185238095238

180776259336

200991842788

215801790802

100432437932

354880952381

343139202288

402271041898

397858160358

607626170126

736428571429

671569601144

646366332963

593594831095

778958078958

781428571429

773220244716

696421950315

741035816036

885134310134

875476190476

808318766884

81318131257

78457977208

Low temp

Control

LCO A

LCO B

TH A

TH B

Time in hours after stratification

germination

Low temp

0

0

0

0

0

09615384615

185238095238

180776259336

200991842788

215801790802

100432437932

354880952381

343139202288

402271041898

397858160358

607626170126

736428571429

671569601144

646366332963

593594831095

778958078958

781428571429

773220244716

696421950315

741035816036

Sheet1

Sheet2

Sheet3

Salt amp Drought Stress

bullArabidopsisbullSeeds on petriplates with signals

bullControl and 150 mM NaCl

bullSignals improved growth under salt stress

0 NaCl 150 mM NaCl

0 10-6 10-8 10-9 10-11 M LCO Thuricin 17

Hormones of the Holobiontbull Effective at very low concentrationsbull Regulate plant activity

bull Large changes in gene espression protein production hormone profile

bull Regulate other microbesbull Powerful plant productivity effectsbull Inexpensive to applybull Low environmental impact

bull Small amountbull Already produced in environment

bull Very poorly understoodbull Enormous potential

bull New networkbull gt 10 strains in lab now

Phytomicrobiome Potentialbull Wide range of species in the phytomicrobiomebull Most powerful control factor is the plant

bull Developmentbull Stressbull Nutrition

bull Are key to plant growth and productivitybull Management

bull Inoculationbull Hub species bull Signals

bull Microbe-to-plantbull Plant-to-microbebull Microbe-to-microbe

Development of Biologicals - Whobull Scope is largebull Collaboration

bull Basic research labs bull Academicbull Government

bull Industry laboratories

The Approachbull Sample plant-associated microbes

bull The phytomicrobiomebull Efficientrapid screening to identify microbes of interest

bull Biological assessmentsbull Agronomic assessments

Control Thuricin

Industrial Collaboratorsbull Synagribull EVLbull BASFbull RavenQuestbull Ulyssebull Agroworld Natural Productsbull Inocucorbull Axter

BioFuelNet - NCE

Past and Present Students Dr Feng Zhang Dr Fazli Mabood Dr Narjes Dashti Keldeagh Lindsay Dr Pan Bo Dr Sowmya Subramanian Dr Yuming Bai Dr Xiaomin Zhou Elizabeth Gray Nan Wang

PDFs amp Research Associates Dr Xiaomin Zhou Dr Balakrishna Prithiviraj Dr Alfred Souleimanov Dr Kung Dong Lee Dr Woo Jung Mike Lewis Stewart Leibovitch

Acknowledgements

ALSO Trevor Charles at U Waterloo

The End

Thank You

• Biostimulants from the phytomicrobiome Microbe-to-plant signals as low input crop management technologies
• Need for Enhanced Food Production
• Threat of Climate Change
• Biomass for Advanced Biofuels and the Bioeconomy
• The Phytomicrobiome
• Adapting to Climate Change
• The Approach
• So Far
• Example Line of Research
• Something New
• The Problem
• A Solution
• N2 Fixation and Growth - Genistein
• Commercialization
• Serendipity amp Follow Up
• New LCO Activity
• Commercialization
• Jasmonates ndash An Alternative
• Findings
• Effect of Methyl Jasmonate on Yield
• Environment - Temperature and Response
• Salt amp Drought Stress
• Hormones of the Holobiont
• Phytomicrobiome Potential
• Development of Biologicals - Who
• The Approach
• Industrial Collaborators
• BioFuelNet - NCE
• Diapositive numeacutero 29
• The End

Control		96h		000
Control		96h		385
Control		96h		000
Control		96h		000
Control		108h		667
Control		108h		1538
Control		108h		741
Control		108h		1071
Control		120h		5000
Control		120h		7308
Control		120h		5926
Control		120h		6071
Control		132h		6333
Control		132h		8846
Control		132h		7407
Control		132h		8571
Control		144h		8000
Control		144h		9231
Control		144h		8889
Control		144h		9286
LCO A		96h		1600
LCO A		96h		1667
LCO A		96h		2000
LCO A		96h		2143
LCO A		108h		3600
LCO A		108h		2667
LCO A		108h		4000
LCO A		108h		3929
LCO A		120h		7600
LCO A		120h		6333
LCO A		120h		7667
LCO A		120h		7857
LCO A		132h		8400
LCO A		132h		7000
LCO A		132h		8000
LCO A		132h		7857
LCO A		144h		8400
LCO A		144h		8000
LCO A		144h		9333
LCO A		144h		9286
LCO B		96h		1429
LCO B		96h		1935
LCO B		96h		1724
LCO B		96h		2143
LCO B		108h		2857
LCO B		108h		4194
LCO B		108h		3103
LCO B		108h		3571
LCO B		120h		6786
LCO B		120h		7097
LCO B		120h		6552
LCO B		120h		6429
LCO B		132h		7143
LCO B		132h		9032
LCO B		132h		6897
LCO B		132h		7857
LCO B		144h		7857
LCO B		144h		9032
LCO B		144h		7586
LCO B		144h		7857
TH A		96h		2258
TH A		96h		1379
TH A		96h		2069
TH A		96h		2333
TH A		108h		4516
TH A		108h		3448
TH A		108h		3793
TH A		108h		4333
TH A		120h		6774
TH A		120h		6552
TH A		120h		5862
TH A		120h		6667
TH A		132h		7742
TH A		132h		6552
TH A		132h		6897
TH A		132h		6667
TH A		144h		9355
TH A		144h		7241
TH A		144h		7931
TH A		144h		8000
TH B		96h		2000
TH B		96h		1923
TH B		96h		1852
TH B		96h		2857
TH B		108h		3667
TH B		108h		4615
TH B		108h		3704
TH B		108h		3929
TH B		120h		6333
TH B		120h		6154
TH B		120h		5185
TH B		120h		6071
TH B		132h		7000
TH B		132h		8462
TH B		132h		7037
TH B		132h		7143
TH B		144h		8000
TH B		144h		8846
TH B		144h		7037
TH B		144h		7500
LCOA+THA		96h		2500
LCOA+THA		96h		2308
LCOA+THA		96h		1852
LCOA+THA		96h		2800
LCOA+THA		108h		3929
LCOA+THA		108h		3462
LCOA+THA		108h		4444
LCOA+THA		108h		5200
LCOA+THA		120h		7143
LCOA+THA		120h		5385
LCOA+THA		120h		7037
LCOA+THA		120h		7200
LCOA+THA		132h		7500
LCOA+THA		132h		7308
LCOA+THA		132h		8519
LCOA+THA		132h		8400
LCOA+THA		144h		8929
LCOA+THA		144h		8077
LCOA+THA		144h		8519
LCOA+THA		144h		8400
LCOB+THB		96h		1852
LCOB+THB		96h		1724
LCOB+THB		96h		3103
LCOB+THB		96h		1875
LCOB+THB		108h		3704
LCOB+THB		108h		3793
LCOB+THB		108h		6207
LCOB+THB		108h		4063
LCOB+THB		120h		6296
LCOB+THB		120h		6207
LCOB+THB		120h		8276
LCOB+THB		120h		5625
LCOB+THB		132h		7037
LCOB+THB		132h		6552
LCOB+THB		132h		8966
LCOB+THB		132h		7188
LCOB+THB		144h		8519
LCOB+THB		144h		7586
LCOB+THB		144h		9310
LCOB+THB		144h		8750

Arabidopsis low temperature set 3
Treatment		No seeds		96 h		germ		Std		SE		108 h		germ		Std		SE		120 h		germ		Std		SE		132 h		germ		Std		SE		144 h		germ		Std		SE
Control 1		30		0		000				2		667				15		5000				19		6333				24		8000
2		26		1		385				4		1538				19		7308				23		8846				24		9231
3		27		0		000				2		741				16		5926				20		7407				24		8889
4		28		0		000				3		1071				17		6071				24		8571				26		9286
	111		025		096		192		096		275		1004		397		199		1675		6076		948		474		215		7790		1154		577		245		8851		594		297
LCO A 1		25		4		1600				9		3600				19		7600				21		8400				21		8400
2		30		5		1667				8		2667				19		6333				21		7000				24		8000
3		30		6		2000				12		4000				23		7667				24		8000				28		9333
4		28		6		2143				11		3929				22		7857				22		7857				26		9286
	113		525		1852		261		130		10		3549		613		307		2075		7364		696		110		22		7814		589		295		2475		8755		661		331
LCO B 1		28		4		1429				8		2857				19		6786				20		7143				22		7857
2		31		6		1935				13		4194				22		7097				28		9032				28		9032
3		29		5		1724				9		3103				19		6552				20		6897				22		7586
4		28		6		2143				10		3571				18		6429				22		7857				22		7857
	116		525		1808		305		153		10		3431		588		294		195		6716		294		147		225		7732		958		479		235		8083		645		323
TH A 1		31		7		2258				14		4516				21		6774				24		7742				29		9355
2		29		4		1379				10		3448				19		6552				19		6552				21		7241
3		29		6		2069				11		3793				17		5862				20		6897				23		7931
4		30		7		2333				13		4333				20		6667				20		6667				24		8000
	119		6		2010		435		217		12		4023		491		245		1925		6464		411		206		2075		6964		538		269		2425		8132		884		442
TH B 1		30		6		2000				11		3667				19		6333				21		7000				24		8000
2		26		5		1923				12		4615				16		6154				22		8462				23		8846
3		27		5		1852				10		3704				14		5185				19		7037				19		7037
4		28		8		2857				11		3929				17		6071				20		7143				21		7500
	111		6		2158		470		235		11		3979		440		220		165		5936		512		256		205		7410		703		352		2175		7846		774		387
LCOA+THA1		28		7		2500				11		3929				20		7143				21		7500				25		8929
2		26		6		2308				9		3462				14		5385				19		7308				21		8077
3		27		5		1852				12		4444				19		7037				23		8519				23		8519
4		25		7		2800				13		5200				18		7200				21		8400				21		8400
	106		625		2365		398		199		1125		4259		745		372		1775		6691		874		437		21		7932		616		308		225		8481		352		176
LCOB+THB1		27		5		1852				10		3704				17		6296				19		7037				23		8519
2		29		5		1724				11		3793				18		6207				19		6552				22		7586
3		29		9		3103				18		6207				24		8276				26		8966				27		9310
4		32		6		1875				13		4063				18		5625				23		7188				28		8750
	11700		625		2139		647		323		1300		4442		1187		593		1925		6601		1156		183		2175		7435		1056		528		2500		8541		718		359
germination
Treatment		0 h		96 h		108 h		120 h		132 h		144 h
Control		0		096		1004		6076		7790		8851
LCO A		0		1852		3549		7364		7814		8755
LCO B		0		1808		3431		6716		7732		8083
TH A		0		2010		4023		6464		6964		8132
TH B		0		2158		3979		5936		7410		7846
LCOA+THA		0		2365		4259		6691		7932		8481
LCOB+THB		0		2139		4442		6601		7435		8541
SE
	0 h		96 h		108 h		120 h		132 h		144 h
Control		0		096		199		474		577		297
LCO A		0		130		307		110		295		331
LCO B		0		153		294		147		479		323
TH A		0		217		245		206		269		442
TH B		0		235		220		256		352		387
LCOA+THA		0		199		372		437		308		176
LCOB+THB		0		323		593		183		528		359

0 h		0 h		0 h		0 h		0 h
96 h		96 h		96 h		96 h		96 h
108 h		108 h		108 h		108 h		108 h
120 h		120 h		120 h		120 h		120 h
132 h		132 h		132 h		132 h		132 h

0 h		0 h		0 h		0 h		0 h		0		0		0		0		0		0		0		0		0		0
96 h		96 h		96 h		96 h		96 h		09615384615		09615384615		13049704387		13049704387		15258333938		15258333938		2174332839		2174332839		23499650247		2174332839
108 h		108 h		108 h		108 h		108 h		19859811656		19859811656		30667415327		30667415327		29408073778		29408073778		24539813637		24539813637		2200128559		24539813637
120 h		120 h		120 h		120 h		120 h		47411187081		47411187081		11002902798		11002902798		14704036889		14704036889		20561084936		20561084936		25615660866		20561084936
132 h		132 h		132 h		132 h		132 h		57694206278		57694206278		29473844048		29473844048		4788349344		4788349344		26896579581		26896579581		35169817799		26896579581
144 h		144 h		144 h		144 h		144 h		29704619111		29704619111		33067820092		33067820092		32273789905		32273789905		4421868687		4421868687		3871003991		4421868687

Weight of seedlings
Treatment		FW (mg)		STD		SE			DW (mg)		STD		SE					Treatment		FW (mg)		STD		SE			DW (mg)		STD		SE
Control 1		39850					2924							Control 1
2		11940					854							2		11940					854
3		10680					468							3		10680					468
4		10520					518							4		10520					518
	11047		778		449			614		210		121						11047		778		449			614		210		121
LCO A 1		15860					899							LCO A 1
2		11320					449							2		11320					449
3		10640					437							3		10640					437
4		10610					400							4		10610					400
	12108		2523		1262			546		236		118						12108		2523		1262			429		026		015
LCO B 1		8040					416							LCO B 1		8040					416
2		7560					333							2		7560					333
3		8020					378							3		8020					378
4		7060					363							4		7060					363
	7670		463		232			372		035		017						7670		463		232			372		035		017
TH A 1		14760					632							TH A 1		14760					632
2		12180					457							2		12180					457
3		14450					469							3		14450					469
4		16780					739							4		16780					739
	14543		1884		942			574		136		068						14543		1884		942			574		136		068
TH B 1		10980					391							TH B 1		10980					391
2		15910					701							2		15910					701
3		17040					815							3		17040					815
4		10880					400							4		10880					400
	13703		3235		1617			577		215		107						13703		3235		1617			577		215		107
Treatment		FW (mg)		SE			Treatment		DW (mg)		SE
Control		11047		449			Control		614		121
LCO A		12108		1262			LCO A		429		016
LCO B		7670		232			LCO B		372		017
TH A		14543		942			TH A		574		068
TH B		13703		1617			TH B		577		107

Control		12125587729
LCO A		0155
LCO B		01725714224
TH A		0678241169
TH B		10731213818

Control		44904837651
LCO A		126152932982
LCO B		2315887159
TH A		94179239574
TH B		161736604288

Dry weight Arab set 3 Normal

DW (mg)

Treatments

Weight in mg

Dry weight of Arabidopsis set 3 Normal temp

61353333333

429

372375

57405

5767

Sheet3

Set 3 Low temp LCO TH

Control

LCO A

LCO B

TH A

TH B

Time in hours after stratification

germination

0

0

0

0

0

09615384615

185238095238

180776259336

200991842788

215801790802

100432437932

354880952381

343139202288

402271041898

397858160358

607626170126

736428571429

671569601144

646366332963

593594831095

778958078958

781428571429

773220244716

696421950315

741035816036

885134310134

875476190476

808318766884

81318131257

78457977208

Low temp

Control

LCO A

LCO B

TH A

TH B

Time in hours after stratification

germination

Low temp

0

0

0

0

0

09615384615

185238095238

180776259336

200991842788

215801790802

100432437932

354880952381

343139202288

402271041898

397858160358

607626170126

736428571429

671569601144

646366332963

593594831095

778958078958

781428571429

773220244716

696421950315

741035816036

Sheet1

Sheet2

Sheet3

Salt amp Drought Stress

bullArabidopsisbullSeeds on petriplates with signals

bullControl and 150 mM NaCl

bullSignals improved growth under salt stress

0 NaCl 150 mM NaCl

0 10-6 10-8 10-9 10-11 M LCO Thuricin 17

Hormones of the Holobiontbull Effective at very low concentrationsbull Regulate plant activity

bull Large changes in gene espression protein production hormone profile

bull Regulate other microbesbull Powerful plant productivity effectsbull Inexpensive to applybull Low environmental impact

bull Small amountbull Already produced in environment

bull Very poorly understoodbull Enormous potential

bull New networkbull gt 10 strains in lab now

Phytomicrobiome Potentialbull Wide range of species in the phytomicrobiomebull Most powerful control factor is the plant

bull Developmentbull Stressbull Nutrition

bull Are key to plant growth and productivitybull Management

bull Inoculationbull Hub species bull Signals

bull Microbe-to-plantbull Plant-to-microbebull Microbe-to-microbe

Development of Biologicals - Whobull Scope is largebull Collaboration

bull Basic research labs bull Academicbull Government

bull Industry laboratories

The Approachbull Sample plant-associated microbes

bull The phytomicrobiomebull Efficientrapid screening to identify microbes of interest

bull Biological assessmentsbull Agronomic assessments

Control Thuricin

Industrial Collaboratorsbull Synagribull EVLbull BASFbull RavenQuestbull Ulyssebull Agroworld Natural Productsbull Inocucorbull Axter

BioFuelNet - NCE

Past and Present Students Dr Feng Zhang Dr Fazli Mabood Dr Narjes Dashti Keldeagh Lindsay Dr Pan Bo Dr Sowmya Subramanian Dr Yuming Bai Dr Xiaomin Zhou Elizabeth Gray Nan Wang

PDFs amp Research Associates Dr Xiaomin Zhou Dr Balakrishna Prithiviraj Dr Alfred Souleimanov Dr Kung Dong Lee Dr Woo Jung Mike Lewis Stewart Leibovitch

Acknowledgements

ALSO Trevor Charles at U Waterloo

The End

Thank You

• Biostimulants from the phytomicrobiome Microbe-to-plant signals as low input crop management technologies
• Need for Enhanced Food Production
• Threat of Climate Change
• Biomass for Advanced Biofuels and the Bioeconomy
• The Phytomicrobiome
• Adapting to Climate Change
• The Approach
• So Far
• Example Line of Research
• Something New
• The Problem
• A Solution
• N2 Fixation and Growth - Genistein
• Commercialization
• Serendipity amp Follow Up
• New LCO Activity
• Commercialization
• Jasmonates ndash An Alternative
• Findings
• Effect of Methyl Jasmonate on Yield
• Environment - Temperature and Response
• Salt amp Drought Stress
• Hormones of the Holobiont
• Phytomicrobiome Potential
• Development of Biologicals - Who
• The Approach
• Industrial Collaborators
• BioFuelNet - NCE
• Diapositive numeacutero 29
• The End

Control		96h		000
Control		96h		385
Control		96h		000
Control		96h		000
Control		108h		667
Control		108h		1538
Control		108h		741
Control		108h		1071
Control		120h		5000
Control		120h		7308
Control		120h		5926
Control		120h		6071
Control		132h		6333
Control		132h		8846
Control		132h		7407
Control		132h		8571
Control		144h		8000
Control		144h		9231
Control		144h		8889
Control		144h		9286
LCO A		96h		1600
LCO A		96h		1667
LCO A		96h		2000
LCO A		96h		2143
LCO A		108h		3600
LCO A		108h		2667
LCO A		108h		4000
LCO A		108h		3929
LCO A		120h		7600
LCO A		120h		6333
LCO A		120h		7667
LCO A		120h		7857
LCO A		132h		8400
LCO A		132h		7000
LCO A		132h		8000
LCO A		132h		7857
LCO A		144h		8400
LCO A		144h		8000
LCO A		144h		9333
LCO A		144h		9286
LCO B		96h		1429
LCO B		96h		1935
LCO B		96h		1724
LCO B		96h		2143
LCO B		108h		2857
LCO B		108h		4194
LCO B		108h		3103
LCO B		108h		3571
LCO B		120h		6786
LCO B		120h		7097
LCO B		120h		6552
LCO B		120h		6429
LCO B		132h		7143
LCO B		132h		9032
LCO B		132h		6897
LCO B		132h		7857
LCO B		144h		7857
LCO B		144h		9032
LCO B		144h		7586
LCO B		144h		7857
TH A		96h		2258
TH A		96h		1379
TH A		96h		2069
TH A		96h		2333
TH A		108h		4516
TH A		108h		3448
TH A		108h		3793
TH A		108h		4333
TH A		120h		6774
TH A		120h		6552
TH A		120h		5862
TH A		120h		6667
TH A		132h		7742
TH A		132h		6552
TH A		132h		6897
TH A		132h		6667
TH A		144h		9355
TH A		144h		7241
TH A		144h		7931
TH A		144h		8000
TH B		96h		2000
TH B		96h		1923
TH B		96h		1852
TH B		96h		2857
TH B		108h		3667
TH B		108h		4615
TH B		108h		3704
TH B		108h		3929
TH B		120h		6333
TH B		120h		6154
TH B		120h		5185
TH B		120h		6071
TH B		132h		7000
TH B		132h		8462
TH B		132h		7037
TH B		132h		7143
TH B		144h		8000
TH B		144h		8846
TH B		144h		7037
TH B		144h		7500
LCOA+THA		96h		2500
LCOA+THA		96h		2308
LCOA+THA		96h		1852
LCOA+THA		96h		2800
LCOA+THA		108h		3929
LCOA+THA		108h		3462
LCOA+THA		108h		4444
LCOA+THA		108h		5200
LCOA+THA		120h		7143
LCOA+THA		120h		5385
LCOA+THA		120h		7037
LCOA+THA		120h		7200
LCOA+THA		132h		7500
LCOA+THA		132h		7308
LCOA+THA		132h		8519
LCOA+THA		132h		8400
LCOA+THA		144h		8929
LCOA+THA		144h		8077
LCOA+THA		144h		8519
LCOA+THA		144h		8400
LCOB+THB		96h		1852
LCOB+THB		96h		1724
LCOB+THB		96h		3103
LCOB+THB		96h		1875
LCOB+THB		108h		3704
LCOB+THB		108h		3793
LCOB+THB		108h		6207
LCOB+THB		108h		4063
LCOB+THB		120h		6296
LCOB+THB		120h		6207
LCOB+THB		120h		8276
LCOB+THB		120h		5625
LCOB+THB		132h		7037
LCOB+THB		132h		6552
LCOB+THB		132h		8966
LCOB+THB		132h		7188
LCOB+THB		144h		8519
LCOB+THB		144h		7586
LCOB+THB		144h		9310
LCOB+THB		144h		8750

Arabidopsis low temperature set 3
Treatment		No seeds		96 h		germ		Std		SE		108 h		germ		Std		SE		120 h		germ		Std		SE		132 h		germ		Std		SE		144 h		germ		Std		SE
Control 1		30		0		000				2		667				15		5000				19		6333				24		8000
2		26		1		385				4		1538				19		7308				23		8846				24		9231
3		27		0		000				2		741				16		5926				20		7407				24		8889
4		28		0		000				3		1071				17		6071				24		8571				26		9286
	111		025		096		192		096		275		1004		397		199		1675		6076		948		474		215		7790		1154		577		245		8851		594		297
LCO A 1		25		4		1600				9		3600				19		7600				21		8400				21		8400
2		30		5		1667				8		2667				19		6333				21		7000				24		8000
3		30		6		2000				12		4000				23		7667				24		8000				28		9333
4		28		6		2143				11		3929				22		7857				22		7857				26		9286
	113		525		1852		261		130		10		3549		613		307		2075		7364		696		110		22		7814		589		295		2475		8755		661		331
LCO B 1		28		4		1429				8		2857				19		6786				20		7143				22		7857
2		31		6		1935				13		4194				22		7097				28		9032				28		9032
3		29		5		1724				9		3103				19		6552				20		6897				22		7586
4		28		6		2143				10		3571				18		6429				22		7857				22		7857
	116		525		1808		305		153		10		3431		588		294		195		6716		294		147		225		7732		958		479		235		8083		645		323
TH A 1		31		7		2258				14		4516				21		6774				24		7742				29		9355
2		29		4		1379				10		3448				19		6552				19		6552				21		7241
3		29		6		2069				11		3793				17		5862				20		6897				23		7931
4		30		7		2333				13		4333				20		6667				20		6667				24		8000
	119		6		2010		435		217		12		4023		491		245		1925		6464		411		206		2075		6964		538		269		2425		8132		884		442
TH B 1		30		6		2000				11		3667				19		6333				21		7000				24		8000
2		26		5		1923				12		4615				16		6154				22		8462				23		8846
3		27		5		1852				10		3704				14		5185				19		7037				19		7037
4		28		8		2857				11		3929				17		6071				20		7143				21		7500
	111		6		2158		470		235		11		3979		440		220		165		5936		512		256		205		7410		703		352		2175		7846		774		387
LCOA+THA1		28		7		2500				11		3929				20		7143				21		7500				25		8929
2		26		6		2308				9		3462				14		5385				19		7308				21		8077
3		27		5		1852				12		4444				19		7037				23		8519				23		8519
4		25		7		2800				13		5200				18		7200				21		8400				21		8400
	106		625		2365		398		199		1125		4259		745		372		1775		6691		874		437		21		7932		616		308		225		8481		352		176
LCOB+THB1		27		5		1852				10		3704				17		6296				19		7037				23		8519
2		29		5		1724				11		3793				18		6207				19		6552				22		7586
3		29		9		3103				18		6207				24		8276				26		8966				27		9310
4		32		6		1875				13		4063				18		5625				23		7188				28		8750
	11700		625		2139		647		323		1300		4442		1187		593		1925		6601		1156		183		2175		7435		1056		528		2500		8541		718		359
germination
Treatment		0 h		96 h		108 h		120 h		132 h		144 h
Control		0		096		1004		6076		7790		8851
LCO A		0		1852		3549		7364		7814		8755
LCO B		0		1808		3431		6716		7732		8083
TH A		0		2010		4023		6464		6964		8132
TH B		0		2158		3979		5936		7410		7846
LCOA+THA		0		2365		4259		6691		7932		8481
LCOB+THB		0		2139		4442		6601		7435		8541
SE
	0 h		96 h		108 h		120 h		132 h		144 h
Control		0		096		199		474		577		297
LCO A		0		130		307		110		295		331
LCO B		0		153		294		147		479		323
TH A		0		217		245		206		269		442
TH B		0		235		220		256		352		387
LCOA+THA		0		199		372		437		308		176
LCOB+THB		0		323		593		183		528		359

0 h		0 h		0 h		0 h		0 h
96 h		96 h		96 h		96 h		96 h
108 h		108 h		108 h		108 h		108 h
120 h		120 h		120 h		120 h		120 h
132 h		132 h		132 h		132 h		132 h

0 h		0 h		0 h		0 h		0 h		0		0		0		0		0		0		0		0		0		0
96 h		96 h		96 h		96 h		96 h		09615384615		09615384615		13049704387		13049704387		15258333938		15258333938		2174332839		2174332839		23499650247		2174332839
108 h		108 h		108 h		108 h		108 h		19859811656		19859811656		30667415327		30667415327		29408073778		29408073778		24539813637		24539813637		2200128559		24539813637
120 h		120 h		120 h		120 h		120 h		47411187081		47411187081		11002902798		11002902798		14704036889		14704036889		20561084936		20561084936		25615660866		20561084936
132 h		132 h		132 h		132 h		132 h		57694206278		57694206278		29473844048		29473844048		4788349344		4788349344		26896579581		26896579581		35169817799		26896579581
144 h		144 h		144 h		144 h		144 h		29704619111		29704619111		33067820092		33067820092		32273789905		32273789905		4421868687		4421868687		3871003991		4421868687

Weight of seedlings
Treatment		FW (mg)		STD		SE			DW (mg)		STD		SE					Treatment		FW (mg)		STD		SE			DW (mg)		STD		SE
Control 1		39850					2924							Control 1
2		11940					854							2		11940					854
3		10680					468							3		10680					468
4		10520					518							4		10520					518
	11047		778		449			614		210		121						11047		778		449			614		210		121
LCO A 1		15860					899							LCO A 1
2		11320					449							2		11320					449
3		10640					437							3		10640					437
4		10610					400							4		10610					400
	12108		2523		1262			546		236		118						12108		2523		1262			429		026		015
LCO B 1		8040					416							LCO B 1		8040					416
2		7560					333							2		7560					333
3		8020					378							3		8020					378
4		7060					363							4		7060					363
	7670		463		232			372		035		017						7670		463		232			372		035		017
TH A 1		14760					632							TH A 1		14760					632
2		12180					457							2		12180					457
3		14450					469							3		14450					469
4		16780					739							4		16780					739
	14543		1884		942			574		136		068						14543		1884		942			574		136		068
TH B 1		10980					391							TH B 1		10980					391
2		15910					701							2		15910					701
3		17040					815							3		17040					815
4		10880					400							4		10880					400
	13703		3235		1617			577		215		107						13703		3235		1617			577		215		107
Treatment		FW (mg)		SE			Treatment		DW (mg)		SE
Control		11047		449			Control		614		121
LCO A		12108		1262			LCO A		429		016
LCO B		7670		232			LCO B		372		017
TH A		14543		942			TH A		574		068
TH B		13703		1617			TH B		577		107

Control		12125587729
LCO A		0155
LCO B		01725714224
TH A		0678241169
TH B		10731213818

Sheet3

Set 3 Low temp LCO TH

Control

LCO A

LCO B

TH A

TH B

Time in hours after stratification

germination

0

0

0

0

0

09615384615

185238095238

180776259336

200991842788

215801790802

100432437932

354880952381

343139202288

402271041898

397858160358

607626170126

736428571429

671569601144

646366332963

593594831095

778958078958

781428571429

773220244716

696421950315

741035816036

885134310134

875476190476

808318766884

81318131257

78457977208

Low temp

Control

LCO A

LCO B

TH A

TH B

Time in hours after stratification

germination

Low temp

0

0

0

0

0

09615384615

185238095238

180776259336

200991842788

215801790802

100432437932

354880952381

343139202288

402271041898

397858160358

607626170126

736428571429

671569601144

646366332963

593594831095

778958078958

781428571429

773220244716

696421950315

741035816036

Sheet1

Sheet2

Sheet3

Salt amp Drought Stress

bullArabidopsisbullSeeds on petriplates with signals

bullControl and 150 mM NaCl

bullSignals improved growth under salt stress

0 NaCl 150 mM NaCl

0 10-6 10-8 10-9 10-11 M LCO Thuricin 17

Hormones of the Holobiontbull Effective at very low concentrationsbull Regulate plant activity

bull Large changes in gene espression protein production hormone profile

bull Regulate other microbesbull Powerful plant productivity effectsbull Inexpensive to applybull Low environmental impact

bull Small amountbull Already produced in environment

bull Very poorly understoodbull Enormous potential

bull New networkbull gt 10 strains in lab now

Phytomicrobiome Potentialbull Wide range of species in the phytomicrobiomebull Most powerful control factor is the plant

bull Developmentbull Stressbull Nutrition

bull Are key to plant growth and productivitybull Management

bull Inoculationbull Hub species bull Signals

bull Microbe-to-plantbull Plant-to-microbebull Microbe-to-microbe

Development of Biologicals - Whobull Scope is largebull Collaboration

bull Basic research labs bull Academicbull Government

bull Industry laboratories

The Approachbull Sample plant-associated microbes

bull The phytomicrobiomebull Efficientrapid screening to identify microbes of interest

bull Biological assessmentsbull Agronomic assessments

Control Thuricin

Industrial Collaboratorsbull Synagribull EVLbull BASFbull RavenQuestbull Ulyssebull Agroworld Natural Productsbull Inocucorbull Axter

BioFuelNet - NCE

Past and Present Students Dr Feng Zhang Dr Fazli Mabood Dr Narjes Dashti Keldeagh Lindsay Dr Pan Bo Dr Sowmya Subramanian Dr Yuming Bai Dr Xiaomin Zhou Elizabeth Gray Nan Wang

PDFs amp Research Associates Dr Xiaomin Zhou Dr Balakrishna Prithiviraj Dr Alfred Souleimanov Dr Kung Dong Lee Dr Woo Jung Mike Lewis Stewart Leibovitch

Acknowledgements

ALSO Trevor Charles at U Waterloo

The End

Thank You

• Biostimulants from the phytomicrobiome Microbe-to-plant signals as low input crop management technologies
• Need for Enhanced Food Production
• Threat of Climate Change
• Biomass for Advanced Biofuels and the Bioeconomy
• The Phytomicrobiome
• Adapting to Climate Change
• The Approach
• So Far
• Example Line of Research
• Something New
• The Problem
• A Solution
• N2 Fixation and Growth - Genistein
• Commercialization
• Serendipity amp Follow Up
• New LCO Activity
• Commercialization
• Jasmonates ndash An Alternative
• Findings
• Effect of Methyl Jasmonate on Yield
• Environment - Temperature and Response
• Salt amp Drought Stress
• Hormones of the Holobiont
• Phytomicrobiome Potential
• Development of Biologicals - Who
• The Approach
• Industrial Collaborators
• BioFuelNet - NCE
• Diapositive numeacutero 29
• The End

Control		96h		000
Control		96h		385
Control		96h		000
Control		96h		000
Control		108h		667
Control		108h		1538
Control		108h		741
Control		108h		1071
Control		120h		5000
Control		120h		7308
Control		120h		5926
Control		120h		6071
Control		132h		6333
Control		132h		8846
Control		132h		7407
Control		132h		8571
Control		144h		8000
Control		144h		9231
Control		144h		8889
Control		144h		9286
LCO A		96h		1600
LCO A		96h		1667
LCO A		96h		2000
LCO A		96h		2143
LCO A		108h		3600
LCO A		108h		2667
LCO A		108h		4000
LCO A		108h		3929
LCO A		120h		7600
LCO A		120h		6333
LCO A		120h		7667
LCO A		120h		7857
LCO A		132h		8400
LCO A		132h		7000
LCO A		132h		8000
LCO A		132h		7857
LCO A		144h		8400
LCO A		144h		8000
LCO A		144h		9333
LCO A		144h		9286
LCO B		96h		1429
LCO B		96h		1935
LCO B		96h		1724
LCO B		96h		2143
LCO B		108h		2857
LCO B		108h		4194
LCO B		108h		3103
LCO B		108h		3571
LCO B		120h		6786
LCO B		120h		7097
LCO B		120h		6552
LCO B		120h		6429
LCO B		132h		7143
LCO B		132h		9032
LCO B		132h		6897
LCO B		132h		7857
LCO B		144h		7857
LCO B		144h		9032
LCO B		144h		7586
LCO B		144h		7857
TH A		96h		2258
TH A		96h		1379
TH A		96h		2069
TH A		96h		2333
TH A		108h		4516
TH A		108h		3448
TH A		108h		3793
TH A		108h		4333
TH A		120h		6774
TH A		120h		6552
TH A		120h		5862
TH A		120h		6667
TH A		132h		7742
TH A		132h		6552
TH A		132h		6897
TH A		132h		6667
TH A		144h		9355
TH A		144h		7241
TH A		144h		7931
TH A		144h		8000
TH B		96h		2000
TH B		96h		1923
TH B		96h		1852
TH B		96h		2857
TH B		108h		3667
TH B		108h		4615
TH B		108h		3704
TH B		108h		3929
TH B		120h		6333
TH B		120h		6154
TH B		120h		5185
TH B		120h		6071
TH B		132h		7000
TH B		132h		8462
TH B		132h		7037
TH B		132h		7143
TH B		144h		8000
TH B		144h		8846
TH B		144h		7037
TH B		144h		7500
LCOA+THA		96h		2500
LCOA+THA		96h		2308
LCOA+THA		96h		1852
LCOA+THA		96h		2800
LCOA+THA		108h		3929
LCOA+THA		108h		3462
LCOA+THA		108h		4444
LCOA+THA		108h		5200
LCOA+THA		120h		7143
LCOA+THA		120h		5385
LCOA+THA		120h		7037
LCOA+THA		120h		7200
LCOA+THA		132h		7500
LCOA+THA		132h		7308
LCOA+THA		132h		8519
LCOA+THA		132h		8400
LCOA+THA		144h		8929
LCOA+THA		144h		8077
LCOA+THA		144h		8519
LCOA+THA		144h		8400
LCOB+THB		96h		1852
LCOB+THB		96h		1724
LCOB+THB		96h		3103
LCOB+THB		96h		1875
LCOB+THB		108h		3704
LCOB+THB		108h		3793
LCOB+THB		108h		6207
LCOB+THB		108h		4063
LCOB+THB		120h		6296
LCOB+THB		120h		6207
LCOB+THB		120h		8276
LCOB+THB		120h		5625
LCOB+THB		132h		7037
LCOB+THB		132h		6552
LCOB+THB		132h		8966
LCOB+THB		132h		7188
LCOB+THB		144h		8519
LCOB+THB		144h		7586
LCOB+THB		144h		9310
LCOB+THB		144h		8750

Arabidopsis low temperature set 3
Treatment		No seeds		96 h		germ		Std		SE		108 h		germ		Std		SE		120 h		germ		Std		SE		132 h		germ		Std		SE		144 h		germ		Std		SE
Control 1		30		0		000				2		667				15		5000				19		6333				24		8000
2		26		1		385				4		1538				19		7308				23		8846				24		9231
3		27		0		000				2		741				16		5926				20		7407				24		8889
4		28		0		000				3		1071				17		6071				24		8571				26		9286
	111		025		096		192		096		275		1004		397		199		1675		6076		948		474		215		7790		1154		577		245		8851		594		297
LCO A 1		25		4		1600				9		3600				19		7600				21		8400				21		8400
2		30		5		1667				8		2667				19		6333				21		7000				24		8000
3		30		6		2000				12		4000				23		7667				24		8000				28		9333
4		28		6		2143				11		3929				22		7857				22		7857				26		9286
	113		525		1852		261		130		10		3549		613		307		2075		7364		696		110		22		7814		589		295		2475		8755		661		331
LCO B 1		28		4		1429				8		2857				19		6786				20		7143				22		7857
2		31		6		1935				13		4194				22		7097				28		9032				28		9032
3		29		5		1724				9		3103				19		6552				20		6897				22		7586
4		28		6		2143				10		3571				18		6429				22		7857				22		7857
	116		525		1808		305		153		10		3431		588		294		195		6716		294		147		225		7732		958		479		235		8083		645		323
TH A 1		31		7		2258				14		4516				21		6774				24		7742				29		9355
2		29		4		1379				10		3448				19		6552				19		6552				21		7241
3		29		6		2069				11		3793				17		5862				20		6897				23		7931
4		30		7		2333				13		4333				20		6667				20		6667				24		8000
	119		6		2010		435		217		12		4023		491		245		1925		6464		411		206		2075		6964		538		269		2425		8132		884		442
TH B 1		30		6		2000				11		3667				19		6333				21		7000				24		8000
2		26		5		1923				12		4615				16		6154				22		8462				23		8846
3		27		5		1852				10		3704				14		5185				19		7037				19		7037
4		28		8		2857				11		3929				17		6071				20		7143				21		7500
	111		6		2158		470		235		11		3979		440		220		165		5936		512		256		205		7410		703		352		2175		7846		774		387
LCOA+THA1		28		7		2500				11		3929				20		7143				21		7500				25		8929
2		26		6		2308				9		3462				14		5385				19		7308				21		8077
3		27		5		1852				12		4444				19		7037				23		8519				23		8519
4		25		7		2800				13		5200				18		7200				21		8400				21		8400
	106		625		2365		398		199		1125		4259		745		372		1775		6691		874		437		21		7932		616		308		225		8481		352		176
LCOB+THB1		27		5		1852				10		3704				17		6296				19		7037				23		8519
2		29		5		1724				11		3793				18		6207				19		6552				22		7586
3		29		9		3103				18		6207				24		8276				26		8966				27		9310
4		32		6		1875				13		4063				18		5625				23		7188				28		8750
	11700		625		2139		647		323		1300		4442		1187		593		1925		6601		1156		183		2175		7435		1056		528		2500		8541		718		359
germination
Treatment		0 h		96 h		108 h		120 h		132 h		144 h
Control		0		096		1004		6076		7790		8851
LCO A		0		1852		3549		7364		7814		8755
LCO B		0		1808		3431		6716		7732		8083
TH A		0		2010		4023		6464		6964		8132
TH B		0		2158		3979		5936		7410		7846
LCOA+THA		0		2365		4259		6691		7932		8481
LCOB+THB		0		2139		4442		6601		7435		8541
SE
	0 h		96 h		108 h		120 h		132 h		144 h
Control		0		096		199		474		577		297
LCO A		0		130		307		110		295		331
LCO B		0		153		294		147		479		323
TH A		0		217		245		206		269		442
TH B		0		235		220		256		352		387
LCOA+THA		0		199		372		437		308		176
LCOB+THB		0		323		593		183		528		359

0 h		0 h		0 h		0 h		0 h
96 h		96 h		96 h		96 h		96 h
108 h		108 h		108 h		108 h		108 h
120 h		120 h		120 h		120 h		120 h
132 h		132 h		132 h		132 h		132 h

0 h		0 h		0 h		0 h		0 h		0		0		0		0		0		0		0		0		0		0
96 h		96 h		96 h		96 h		96 h		09615384615		09615384615		13049704387		13049704387		15258333938		15258333938		2174332839		2174332839		23499650247		2174332839
108 h		108 h		108 h		108 h		108 h		19859811656		19859811656		30667415327		30667415327		29408073778		29408073778		24539813637		24539813637		2200128559		24539813637
120 h		120 h		120 h		120 h		120 h		47411187081		47411187081		11002902798		11002902798		14704036889		14704036889		20561084936		20561084936		25615660866		20561084936
132 h		132 h		132 h		132 h		132 h		57694206278		57694206278		29473844048		29473844048		4788349344		4788349344		26896579581		26896579581		35169817799		26896579581
144 h		144 h		144 h		144 h		144 h		29704619111		29704619111		33067820092		33067820092		32273789905		32273789905		4421868687		4421868687		3871003991		4421868687

Weight of seedlings
Treatment		FW (mg)		STD		SE			DW (mg)		STD		SE					Treatment		FW (mg)		STD		SE			DW (mg)		STD		SE
Control 1		39850					2924							Control 1
2		11940					854							2		11940					854
3		10680					468							3		10680					468
4		10520					518							4		10520					518
	11047		778		449			614		210		121						11047		778		449			614		210		121
LCO A 1		15860					899							LCO A 1
2		11320					449							2		11320					449
3		10640					437							3		10640					437
4		10610					400							4		10610					400
	12108		2523		1262			546		236		118						12108		2523		1262			429		026		015
LCO B 1		8040					416							LCO B 1		8040					416
2		7560					333							2		7560					333
3		8020					378							3		8020					378
4		7060					363							4		7060					363
	7670		463		232			372		035		017						7670		463		232			372		035		017
TH A 1		14760					632							TH A 1		14760					632
2		12180					457							2		12180					457
3		14450					469							3		14450					469
4		16780					739							4		16780					739
	14543		1884		942			574		136		068						14543		1884		942			574		136		068
TH B 1		10980					391							TH B 1		10980					391
2		15910					701							2		15910					701
3		17040					815							3		17040					815
4		10880					400							4		10880					400
	13703		3235		1617			577		215		107						13703		3235		1617			577		215		107
Treatment		FW (mg)		SE			Treatment		DW (mg)		SE
Control		11047		449			Control		614		121
LCO A		12108		1262			LCO A		429		016
LCO B		7670		232			LCO B		372		017
TH A		14543		942			TH A		574		068
TH B		13703		1617			TH B		577		107

Set 3 Low temp LCO TH

Control

LCO A

LCO B

TH A

TH B

Time in hours after stratification

germination

0

0

0

0

0

09615384615

185238095238

180776259336

200991842788

215801790802

100432437932

354880952381

343139202288

402271041898

397858160358

607626170126

736428571429

671569601144

646366332963

593594831095

778958078958

781428571429

773220244716

696421950315

741035816036

885134310134

875476190476

808318766884

81318131257

78457977208

Low temp

Control

LCO A

LCO B

TH A

TH B

Time in hours after stratification

germination

Low temp

0

0

0

0

0

09615384615

185238095238

180776259336

200991842788

215801790802

100432437932

354880952381

343139202288

402271041898

397858160358

607626170126

736428571429

671569601144

646366332963

593594831095

778958078958

781428571429

773220244716

696421950315

741035816036

Sheet1

Sheet2

Sheet3

Salt amp Drought Stress

bullArabidopsisbullSeeds on petriplates with signals

bullControl and 150 mM NaCl

bullSignals improved growth under salt stress

0 NaCl 150 mM NaCl

0 10-6 10-8 10-9 10-11 M LCO Thuricin 17

Hormones of the Holobiontbull Effective at very low concentrationsbull Regulate plant activity

bull Large changes in gene espression protein production hormone profile

bull Regulate other microbesbull Powerful plant productivity effectsbull Inexpensive to applybull Low environmental impact

bull Small amountbull Already produced in environment

bull Very poorly understoodbull Enormous potential

bull New networkbull gt 10 strains in lab now

Phytomicrobiome Potentialbull Wide range of species in the phytomicrobiomebull Most powerful control factor is the plant

bull Developmentbull Stressbull Nutrition

bull Are key to plant growth and productivitybull Management

bull Inoculationbull Hub species bull Signals

bull Microbe-to-plantbull Plant-to-microbebull Microbe-to-microbe

Development of Biologicals - Whobull Scope is largebull Collaboration

bull Basic research labs bull Academicbull Government

bull Industry laboratories

The Approachbull Sample plant-associated microbes

bull The phytomicrobiomebull Efficientrapid screening to identify microbes of interest

bull Biological assessmentsbull Agronomic assessments

Control Thuricin

Industrial Collaboratorsbull Synagribull EVLbull BASFbull RavenQuestbull Ulyssebull Agroworld Natural Productsbull Inocucorbull Axter

BioFuelNet - NCE

Past and Present Students Dr Feng Zhang Dr Fazli Mabood Dr Narjes Dashti Keldeagh Lindsay Dr Pan Bo Dr Sowmya Subramanian Dr Yuming Bai Dr Xiaomin Zhou Elizabeth Gray Nan Wang

PDFs amp Research Associates Dr Xiaomin Zhou Dr Balakrishna Prithiviraj Dr Alfred Souleimanov Dr Kung Dong Lee Dr Woo Jung Mike Lewis Stewart Leibovitch

Acknowledgements

ALSO Trevor Charles at U Waterloo

The End

Thank You

• Biostimulants from the phytomicrobiome Microbe-to-plant signals as low input crop management technologies
• Need for Enhanced Food Production
• Threat of Climate Change
• Biomass for Advanced Biofuels and the Bioeconomy
• The Phytomicrobiome
• Adapting to Climate Change
• The Approach
• So Far
• Example Line of Research
• Something New
• The Problem
• A Solution
• N2 Fixation and Growth - Genistein
• Commercialization
• Serendipity amp Follow Up
• New LCO Activity
• Commercialization
• Jasmonates ndash An Alternative
• Findings
• Effect of Methyl Jasmonate on Yield
• Environment - Temperature and Response
• Salt amp Drought Stress
• Hormones of the Holobiont
• Phytomicrobiome Potential
• Development of Biologicals - Who
• The Approach
• Industrial Collaborators
• BioFuelNet - NCE
• Diapositive numeacutero 29
• The End

Control		96h		000
Control		96h		385
Control		96h		000
Control		96h		000
Control		108h		667
Control		108h		1538
Control		108h		741
Control		108h		1071
Control		120h		5000
Control		120h		7308
Control		120h		5926
Control		120h		6071
Control		132h		6333
Control		132h		8846
Control		132h		7407
Control		132h		8571
Control		144h		8000
Control		144h		9231
Control		144h		8889
Control		144h		9286
LCO A		96h		1600
LCO A		96h		1667
LCO A		96h		2000
LCO A		96h		2143
LCO A		108h		3600
LCO A		108h		2667
LCO A		108h		4000
LCO A		108h		3929
LCO A		120h		7600
LCO A		120h		6333
LCO A		120h		7667
LCO A		120h		7857
LCO A		132h		8400
LCO A		132h		7000
LCO A		132h		8000
LCO A		132h		7857
LCO A		144h		8400
LCO A		144h		8000
LCO A		144h		9333
LCO A		144h		9286
LCO B		96h		1429
LCO B		96h		1935
LCO B		96h		1724
LCO B		96h		2143
LCO B		108h		2857
LCO B		108h		4194
LCO B		108h		3103
LCO B		108h		3571
LCO B		120h		6786
LCO B		120h		7097
LCO B		120h		6552
LCO B		120h		6429
LCO B		132h		7143
LCO B		132h		9032
LCO B		132h		6897
LCO B		132h		7857
LCO B		144h		7857
LCO B		144h		9032
LCO B		144h		7586
LCO B		144h		7857
TH A		96h		2258
TH A		96h		1379
TH A		96h		2069
TH A		96h		2333
TH A		108h		4516
TH A		108h		3448
TH A		108h		3793
TH A		108h		4333
TH A		120h		6774
TH A		120h		6552
TH A		120h		5862
TH A		120h		6667
TH A		132h		7742
TH A		132h		6552
TH A		132h		6897
TH A		132h		6667
TH A		144h		9355
TH A		144h		7241
TH A		144h		7931
TH A		144h		8000
TH B		96h		2000
TH B		96h		1923
TH B		96h		1852
TH B		96h		2857
TH B		108h		3667
TH B		108h		4615
TH B		108h		3704
TH B		108h		3929
TH B		120h		6333
TH B		120h		6154
TH B		120h		5185
TH B		120h		6071
TH B		132h		7000
TH B		132h		8462
TH B		132h		7037
TH B		132h		7143
TH B		144h		8000
TH B		144h		8846
TH B		144h		7037
TH B		144h		7500
LCOA+THA		96h		2500
LCOA+THA		96h		2308
LCOA+THA		96h		1852
LCOA+THA		96h		2800
LCOA+THA		108h		3929
LCOA+THA		108h		3462
LCOA+THA		108h		4444
LCOA+THA		108h		5200
LCOA+THA		120h		7143
LCOA+THA		120h		5385
LCOA+THA		120h		7037
LCOA+THA		120h		7200
LCOA+THA		132h		7500
LCOA+THA		132h		7308
LCOA+THA		132h		8519
LCOA+THA		132h		8400
LCOA+THA		144h		8929
LCOA+THA		144h		8077
LCOA+THA		144h		8519
LCOA+THA		144h		8400
LCOB+THB		96h		1852
LCOB+THB		96h		1724
LCOB+THB		96h		3103
LCOB+THB		96h		1875
LCOB+THB		108h		3704
LCOB+THB		108h		3793
LCOB+THB		108h		6207
LCOB+THB		108h		4063
LCOB+THB		120h		6296
LCOB+THB		120h		6207
LCOB+THB		120h		8276
LCOB+THB		120h		5625
LCOB+THB		132h		7037
LCOB+THB		132h		6552
LCOB+THB		132h		8966
LCOB+THB		132h		7188
LCOB+THB		144h		8519
LCOB+THB		144h		7586
LCOB+THB		144h		9310
LCOB+THB		144h		8750

Arabidopsis low temperature set 3
Treatment		No seeds		96 h		germ		Std		SE		108 h		germ		Std		SE		120 h		germ		Std		SE		132 h		germ		Std		SE		144 h		germ		Std		SE
Control 1		30		0		000				2		667				15		5000				19		6333				24		8000
2		26		1		385				4		1538				19		7308				23		8846				24		9231
3		27		0		000				2		741				16		5926				20		7407				24		8889
4		28		0		000				3		1071				17		6071				24		8571				26		9286
	111		025		096		192		096		275		1004		397		199		1675		6076		948		474		215		7790		1154		577		245		8851		594		297
LCO A 1		25		4		1600				9		3600				19		7600				21		8400				21		8400
2		30		5		1667				8		2667				19		6333				21		7000				24		8000
3		30		6		2000				12		4000				23		7667				24		8000				28		9333
4		28		6		2143				11		3929				22		7857				22		7857				26		9286
	113		525		1852		261		130		10		3549		613		307		2075		7364		696		110		22		7814		589		295		2475		8755		661		331
LCO B 1		28		4		1429				8		2857				19		6786				20		7143				22		7857
2		31		6		1935				13		4194				22		7097				28		9032				28		9032
3		29		5		1724				9		3103				19		6552				20		6897				22		7586
4		28		6		2143				10		3571				18		6429				22		7857				22		7857
	116		525		1808		305		153		10		3431		588		294		195		6716		294		147		225		7732		958		479		235		8083		645		323
TH A 1		31		7		2258				14		4516				21		6774				24		7742				29		9355
2		29		4		1379				10		3448				19		6552				19		6552				21		7241
3		29		6		2069				11		3793				17		5862				20		6897				23		7931
4		30		7		2333				13		4333				20		6667				20		6667				24		8000
	119		6		2010		435		217		12		4023		491		245		1925		6464		411		206		2075		6964		538		269		2425		8132		884		442
TH B 1		30		6		2000				11		3667				19		6333				21		7000				24		8000
2		26		5		1923				12		4615				16		6154				22		8462				23		8846
3		27		5		1852				10		3704				14		5185				19		7037				19		7037
4		28		8		2857				11		3929				17		6071				20		7143				21		7500
	111		6		2158		470		235		11		3979		440		220		165		5936		512		256		205		7410		703		352		2175		7846		774		387
LCOA+THA1		28		7		2500				11		3929				20		7143				21		7500				25		8929
2		26		6		2308				9		3462				14		5385				19		7308				21		8077
3		27		5		1852				12		4444				19		7037				23		8519				23		8519
4		25		7		2800				13		5200				18		7200				21		8400				21		8400
	106		625		2365		398		199		1125		4259		745		372		1775		6691		874		437		21		7932		616		308		225		8481		352		176
LCOB+THB1		27		5		1852				10		3704				17		6296				19		7037				23		8519
2		29		5		1724				11		3793				18		6207				19		6552				22		7586
3		29		9		3103				18		6207				24		8276				26		8966				27		9310
4		32		6		1875				13		4063				18		5625				23		7188				28		8750
	11700		625		2139		647		323		1300		4442		1187		593		1925		6601		1156		183		2175		7435		1056		528		2500		8541		718		359
germination
Treatment		0 h		96 h		108 h		120 h		132 h		144 h
Control		0		096		1004		6076		7790		8851
LCO A		0		1852		3549		7364		7814		8755
LCO B		0		1808		3431		6716		7732		8083
TH A		0		2010		4023		6464		6964		8132
TH B		0		2158		3979		5936		7410		7846
LCOA+THA		0		2365		4259		6691		7932		8481
LCOB+THB		0		2139		4442		6601		7435		8541
SE
	0 h		96 h		108 h		120 h		132 h		144 h
Control		0		096		199		474		577		297
LCO A		0		130		307		110		295		331
LCO B		0		153		294		147		479		323
TH A		0		217		245		206		269		442
TH B		0		235		220		256		352		387
LCOA+THA		0		199		372		437		308		176
LCOB+THB		0		323		593		183		528		359

0 h		0 h		0 h		0 h		0 h
96 h		96 h		96 h		96 h		96 h
108 h		108 h		108 h		108 h		108 h
120 h		120 h		120 h		120 h		120 h
132 h		132 h		132 h		132 h		132 h

0 h		0 h		0 h		0 h		0 h		0		0		0		0		0		0		0		0		0		0
96 h		96 h		96 h		96 h		96 h		09615384615		09615384615		13049704387		13049704387		15258333938		15258333938		2174332839		2174332839		23499650247		2174332839
108 h		108 h		108 h		108 h		108 h		19859811656		19859811656		30667415327		30667415327		29408073778		29408073778		24539813637		24539813637		2200128559		24539813637
120 h		120 h		120 h		120 h		120 h		47411187081		47411187081		11002902798		11002902798		14704036889		14704036889		20561084936		20561084936		25615660866		20561084936
132 h		132 h		132 h		132 h		132 h		57694206278		57694206278		29473844048		29473844048		4788349344		4788349344		26896579581		26896579581		35169817799		26896579581
144 h		144 h		144 h		144 h		144 h		29704619111		29704619111		33067820092		33067820092		32273789905		32273789905		4421868687		4421868687		3871003991		4421868687

Low temp

Control

LCO A

LCO B

TH A

TH B

Time in hours after stratification

germination

Low temp

0

0

0

0

0

09615384615

185238095238

180776259336

200991842788

215801790802

100432437932

354880952381

343139202288

402271041898

397858160358

607626170126

736428571429

671569601144

646366332963

593594831095

778958078958

781428571429

773220244716

696421950315

741035816036

Sheet1

Sheet2

Sheet3

Salt amp Drought Stress

bullArabidopsisbullSeeds on petriplates with signals

bullControl and 150 mM NaCl

bullSignals improved growth under salt stress

0 NaCl 150 mM NaCl

0 10-6 10-8 10-9 10-11 M LCO Thuricin 17

Hormones of the Holobiontbull Effective at very low concentrationsbull Regulate plant activity

bull Large changes in gene espression protein production hormone profile

bull Regulate other microbesbull Powerful plant productivity effectsbull Inexpensive to applybull Low environmental impact

bull Small amountbull Already produced in environment

bull Very poorly understoodbull Enormous potential

bull New networkbull gt 10 strains in lab now

Phytomicrobiome Potentialbull Wide range of species in the phytomicrobiomebull Most powerful control factor is the plant

bull Developmentbull Stressbull Nutrition

bull Are key to plant growth and productivitybull Management

bull Inoculationbull Hub species bull Signals

bull Microbe-to-plantbull Plant-to-microbebull Microbe-to-microbe

Development of Biologicals - Whobull Scope is largebull Collaboration

bull Basic research labs bull Academicbull Government

bull Industry laboratories

The Approachbull Sample plant-associated microbes

bull The phytomicrobiomebull Efficientrapid screening to identify microbes of interest

bull Biological assessmentsbull Agronomic assessments

Control Thuricin

Industrial Collaboratorsbull Synagribull EVLbull BASFbull RavenQuestbull Ulyssebull Agroworld Natural Productsbull Inocucorbull Axter

BioFuelNet - NCE

Past and Present Students Dr Feng Zhang Dr Fazli Mabood Dr Narjes Dashti Keldeagh Lindsay Dr Pan Bo Dr Sowmya Subramanian Dr Yuming Bai Dr Xiaomin Zhou Elizabeth Gray Nan Wang

PDFs amp Research Associates Dr Xiaomin Zhou Dr Balakrishna Prithiviraj Dr Alfred Souleimanov Dr Kung Dong Lee Dr Woo Jung Mike Lewis Stewart Leibovitch

Acknowledgements

ALSO Trevor Charles at U Waterloo

The End

Thank You

• Biostimulants from the phytomicrobiome Microbe-to-plant signals as low input crop management technologies
• Need for Enhanced Food Production
• Threat of Climate Change
• Biomass for Advanced Biofuels and the Bioeconomy
• The Phytomicrobiome
• Adapting to Climate Change
• The Approach
• So Far
• Example Line of Research
• Something New
• The Problem
• A Solution
• N2 Fixation and Growth - Genistein
• Commercialization
• Serendipity amp Follow Up
• New LCO Activity
• Commercialization
• Jasmonates ndash An Alternative
• Findings
• Effect of Methyl Jasmonate on Yield
• Environment - Temperature and Response
• Salt amp Drought Stress
• Hormones of the Holobiont
• Phytomicrobiome Potential
• Development of Biologicals - Who
• The Approach
• Industrial Collaborators
• BioFuelNet - NCE
• Diapositive numeacutero 29
• The End

Control		96h		000
Control		96h		385
Control		96h		000
Control		96h		000
Control		108h		667
Control		108h		1538
Control		108h		741
Control		108h		1071
Control		120h		5000
Control		120h		7308
Control		120h		5926
Control		120h		6071
Control		132h		6333
Control		132h		8846
Control		132h		7407
Control		132h		8571
Control		144h		8000
Control		144h		9231
Control		144h		8889
Control		144h		9286
LCO A		96h		1600
LCO A		96h		1667
LCO A		96h		2000
LCO A		96h		2143
LCO A		108h		3600
LCO A		108h		2667
LCO A		108h		4000
LCO A		108h		3929
LCO A		120h		7600
LCO A		120h		6333
LCO A		120h		7667
LCO A		120h		7857
LCO A		132h		8400
LCO A		132h		7000
LCO A		132h		8000
LCO A		132h		7857
LCO A		144h		8400
LCO A		144h		8000
LCO A		144h		9333
LCO A		144h		9286
LCO B		96h		1429
LCO B		96h		1935
LCO B		96h		1724
LCO B		96h		2143
LCO B		108h		2857
LCO B		108h		4194
LCO B		108h		3103
LCO B		108h		3571
LCO B		120h		6786
LCO B		120h		7097
LCO B		120h		6552
LCO B		120h		6429
LCO B		132h		7143
LCO B		132h		9032
LCO B		132h		6897
LCO B		132h		7857
LCO B		144h		7857
LCO B		144h		9032
LCO B		144h		7586
LCO B		144h		7857
TH A		96h		2258
TH A		96h		1379
TH A		96h		2069
TH A		96h		2333
TH A		108h		4516
TH A		108h		3448
TH A		108h		3793
TH A		108h		4333
TH A		120h		6774
TH A		120h		6552
TH A		120h		5862
TH A		120h		6667
TH A		132h		7742
TH A		132h		6552
TH A		132h		6897
TH A		132h		6667
TH A		144h		9355
TH A		144h		7241
TH A		144h		7931
TH A		144h		8000
TH B		96h		2000
TH B		96h		1923
TH B		96h		1852
TH B		96h		2857
TH B		108h		3667
TH B		108h		4615
TH B		108h		3704
TH B		108h		3929
TH B		120h		6333
TH B		120h		6154
TH B		120h		5185
TH B		120h		6071
TH B		132h		7000
TH B		132h		8462
TH B		132h		7037
TH B		132h		7143
TH B		144h		8000
TH B		144h		8846
TH B		144h		7037
TH B		144h		7500
LCOA+THA		96h		2500
LCOA+THA		96h		2308
LCOA+THA		96h		1852
LCOA+THA		96h		2800
LCOA+THA		108h		3929
LCOA+THA		108h		3462
LCOA+THA		108h		4444
LCOA+THA		108h		5200
LCOA+THA		120h		7143
LCOA+THA		120h		5385
LCOA+THA		120h		7037
LCOA+THA		120h		7200
LCOA+THA		132h		7500
LCOA+THA		132h		7308
LCOA+THA		132h		8519
LCOA+THA		132h		8400
LCOA+THA		144h		8929
LCOA+THA		144h		8077
LCOA+THA		144h		8519
LCOA+THA		144h		8400
LCOB+THB		96h		1852
LCOB+THB		96h		1724
LCOB+THB		96h		3103
LCOB+THB		96h		1875
LCOB+THB		108h		3704
LCOB+THB		108h		3793
LCOB+THB		108h		6207
LCOB+THB		108h		4063
LCOB+THB		120h		6296
LCOB+THB		120h		6207
LCOB+THB		120h		8276
LCOB+THB		120h		5625
LCOB+THB		132h		7037
LCOB+THB		132h		6552
LCOB+THB		132h		8966
LCOB+THB		132h		7188
LCOB+THB		144h		8519
LCOB+THB		144h		7586
LCOB+THB		144h		9310
LCOB+THB		144h		8750

Arabidopsis low temperature set 3
Treatment		No seeds		96 h		germ		Std		SE		108 h		germ		Std		SE		120 h		germ		Std		SE		132 h		germ		Std		SE		144 h		germ		Std		SE
Control 1		30		0		000				2		667				15		5000				19		6333				24		8000
2		26		1		385				4		1538				19		7308				23		8846				24		9231
3		27		0		000				2		741				16		5926				20		7407				24		8889
4		28		0		000				3		1071				17		6071				24		8571				26		9286
	111		025		096		192		096		275		1004		397		199		1675		6076		948		474		215		7790		1154		577		245		8851		594		297
LCO A 1		25		4		1600				9		3600				19		7600				21		8400				21		8400
2		30		5		1667				8		2667				19		6333				21		7000				24		8000
3		30		6		2000				12		4000				23		7667				24		8000				28		9333
4		28		6		2143				11		3929				22		7857				22		7857				26		9286
	113		525		1852		261		130		10		3549		613		307		2075		7364		696		110		22		7814		589		295		2475		8755		661		331
LCO B 1		28		4		1429				8		2857				19		6786				20		7143				22		7857
2		31		6		1935				13		4194				22		7097				28		9032				28		9032
3		29		5		1724				9		3103				19		6552				20		6897				22		7586
4		28		6		2143				10		3571				18		6429				22		7857				22		7857
	116		525		1808		305		153		10		3431		588		294		195		6716		294		147		225		7732		958		479		235		8083		645		323
TH A 1		31		7		2258				14		4516				21		6774				24		7742				29		9355
2		29		4		1379				10		3448				19		6552				19		6552				21		7241
3		29		6		2069				11		3793				17		5862				20		6897				23		7931
4		30		7		2333				13		4333				20		6667				20		6667				24		8000
	119		6		2010		435		217		12		4023		491		245		1925		6464		411		206		2075		6964		538		269		2425		8132		884		442
TH B 1		30		6		2000				11		3667				19		6333				21		7000				24		8000
2		26		5		1923				12		4615				16		6154				22		8462				23		8846
3		27		5		1852				10		3704				14		5185				19		7037				19		7037
4		28		8		2857				11		3929				17		6071				20		7143				21		7500
	111		6		2158		470		235		11		3979		440		220		165		5936		512		256		205		7410		703		352		2175		7846		774		387
LCOA+THA1		28		7		2500				11		3929				20		7143				21		7500				25		8929
2		26		6		2308				9		3462				14		5385				19		7308				21		8077
3		27		5		1852				12		4444				19		7037				23		8519				23		8519
4		25		7		2800				13		5200				18		7200				21		8400				21		8400
	106		625		2365		398		199		1125		4259		745		372		1775		6691		874		437		21		7932		616		308		225		8481		352		176
LCOB+THB1		27		5		1852				10		3704				17		6296				19		7037				23		8519
2		29		5		1724				11		3793				18		6207				19		6552				22		7586
3		29		9		3103				18		6207				24		8276				26		8966				27		9310
4		32		6		1875				13		4063				18		5625				23		7188				28		8750
	11700		625		2139		647		323		1300		4442		1187		593		1925		6601		1156		183		2175		7435		1056		528		2500		8541		718		359
germination
Treatment		0 h		96 h		108 h		120 h		132 h		144 h
Control		0		096		1004		6076		7790		8851
LCO A		0		1852		3549		7364		7814		8755
LCO B		0		1808		3431		6716		7732		8083
TH A		0		2010		4023		6464		6964		8132
TH B		0		2158		3979		5936		7410		7846
LCOA+THA		0		2365		4259		6691		7932		8481
LCOB+THB		0		2139		4442		6601		7435		8541
SE
	0 h		96 h		108 h		120 h		132 h		144 h
Control		0		096		199		474		577		297
LCO A		0		130		307		110		295		331
LCO B		0		153		294		147		479		323
TH A		0		217		245		206		269		442
TH B		0		235		220		256		352		387
LCOA+THA		0		199		372		437		308		176
LCOB+THB		0		323		593		183		528		359

0 h		0 h		0 h		0 h		0 h
96 h		96 h		96 h		96 h		96 h
108 h		108 h		108 h		108 h		108 h
120 h		120 h		120 h		120 h		120 h
132 h		132 h		132 h		132 h		132 h

Sheet1

Sheet2

Sheet3

Salt amp Drought Stress

bullArabidopsisbullSeeds on petriplates with signals

bullControl and 150 mM NaCl

bullSignals improved growth under salt stress

0 NaCl 150 mM NaCl

0 10-6 10-8 10-9 10-11 M LCO Thuricin 17

Hormones of the Holobiontbull Effective at very low concentrationsbull Regulate plant activity

bull Large changes in gene espression protein production hormone profile

bull Regulate other microbesbull Powerful plant productivity effectsbull Inexpensive to applybull Low environmental impact

bull Small amountbull Already produced in environment

bull Very poorly understoodbull Enormous potential

bull New networkbull gt 10 strains in lab now

Phytomicrobiome Potentialbull Wide range of species in the phytomicrobiomebull Most powerful control factor is the plant

bull Developmentbull Stressbull Nutrition

bull Are key to plant growth and productivitybull Management

bull Inoculationbull Hub species bull Signals

bull Microbe-to-plantbull Plant-to-microbebull Microbe-to-microbe

Development of Biologicals - Whobull Scope is largebull Collaboration

bull Basic research labs bull Academicbull Government

bull Industry laboratories

The Approachbull Sample plant-associated microbes

bull The phytomicrobiomebull Efficientrapid screening to identify microbes of interest

bull Biological assessmentsbull Agronomic assessments

Control Thuricin

Industrial Collaboratorsbull Synagribull EVLbull BASFbull RavenQuestbull Ulyssebull Agroworld Natural Productsbull Inocucorbull Axter

BioFuelNet - NCE

Past and Present Students Dr Feng Zhang Dr Fazli Mabood Dr Narjes Dashti Keldeagh Lindsay Dr Pan Bo Dr Sowmya Subramanian Dr Yuming Bai Dr Xiaomin Zhou Elizabeth Gray Nan Wang

PDFs amp Research Associates Dr Xiaomin Zhou Dr Balakrishna Prithiviraj Dr Alfred Souleimanov Dr Kung Dong Lee Dr Woo Jung Mike Lewis Stewart Leibovitch

Acknowledgements

ALSO Trevor Charles at U Waterloo

The End

Thank You

• Biostimulants from the phytomicrobiome Microbe-to-plant signals as low input crop management technologies
• Need for Enhanced Food Production
• Threat of Climate Change
• Biomass for Advanced Biofuels and the Bioeconomy
• The Phytomicrobiome
• Adapting to Climate Change
• The Approach
• So Far
• Example Line of Research
• Something New
• The Problem
• A Solution
• N2 Fixation and Growth - Genistein
• Commercialization
• Serendipity amp Follow Up
• New LCO Activity
• Commercialization
• Jasmonates ndash An Alternative
• Findings
• Effect of Methyl Jasmonate on Yield
• Environment - Temperature and Response
• Salt amp Drought Stress
• Hormones of the Holobiont
• Phytomicrobiome Potential
• Development of Biologicals - Who
• The Approach
• Industrial Collaborators
• BioFuelNet - NCE
• Diapositive numeacutero 29
• The End

Control		96h		000
Control		96h		385
Control		96h		000
Control		96h		000
Control		108h		667
Control		108h		1538
Control		108h		741
Control		108h		1071
Control		120h		5000
Control		120h		7308
Control		120h		5926
Control		120h		6071
Control		132h		6333
Control		132h		8846
Control		132h		7407
Control		132h		8571
Control		144h		8000
Control		144h		9231
Control		144h		8889
Control		144h		9286
LCO A		96h		1600
LCO A		96h		1667
LCO A		96h		2000
LCO A		96h		2143
LCO A		108h		3600
LCO A		108h		2667
LCO A		108h		4000
LCO A		108h		3929
LCO A		120h		7600
LCO A		120h		6333
LCO A		120h		7667
LCO A		120h		7857
LCO A		132h		8400
LCO A		132h		7000
LCO A		132h		8000
LCO A		132h		7857
LCO A		144h		8400
LCO A		144h		8000
LCO A		144h		9333
LCO A		144h		9286
LCO B		96h		1429
LCO B		96h		1935
LCO B		96h		1724
LCO B		96h		2143
LCO B		108h		2857
LCO B		108h		4194
LCO B		108h		3103
LCO B		108h		3571
LCO B		120h		6786
LCO B		120h		7097
LCO B		120h		6552
LCO B		120h		6429
LCO B		132h		7143
LCO B		132h		9032
LCO B		132h		6897
LCO B		132h		7857
LCO B		144h		7857
LCO B		144h		9032
LCO B		144h		7586
LCO B		144h		7857
TH A		96h		2258
TH A		96h		1379
TH A		96h		2069
TH A		96h		2333
TH A		108h		4516
TH A		108h		3448
TH A		108h		3793
TH A		108h		4333
TH A		120h		6774
TH A		120h		6552
TH A		120h		5862
TH A		120h		6667
TH A		132h		7742
TH A		132h		6552
TH A		132h		6897
TH A		132h		6667
TH A		144h		9355
TH A		144h		7241
TH A		144h		7931
TH A		144h		8000
TH B		96h		2000
TH B		96h		1923
TH B		96h		1852
TH B		96h		2857
TH B		108h		3667
TH B		108h		4615
TH B		108h		3704
TH B		108h		3929
TH B		120h		6333
TH B		120h		6154
TH B		120h		5185
TH B		120h		6071
TH B		132h		7000
TH B		132h		8462
TH B		132h		7037
TH B		132h		7143
TH B		144h		8000
TH B		144h		8846
TH B		144h		7037
TH B		144h		7500
LCOA+THA		96h		2500
LCOA+THA		96h		2308
LCOA+THA		96h		1852
LCOA+THA		96h		2800
LCOA+THA		108h		3929
LCOA+THA		108h		3462
LCOA+THA		108h		4444
LCOA+THA		108h		5200
LCOA+THA		120h		7143
LCOA+THA		120h		5385
LCOA+THA		120h		7037
LCOA+THA		120h		7200
LCOA+THA		132h		7500
LCOA+THA		132h		7308
LCOA+THA		132h		8519
LCOA+THA		132h		8400
LCOA+THA		144h		8929
LCOA+THA		144h		8077
LCOA+THA		144h		8519
LCOA+THA		144h		8400
LCOB+THB		96h		1852
LCOB+THB		96h		1724
LCOB+THB		96h		3103
LCOB+THB		96h		1875
LCOB+THB		108h		3704
LCOB+THB		108h		3793
LCOB+THB		108h		6207
LCOB+THB		108h		4063
LCOB+THB		120h		6296
LCOB+THB		120h		6207
LCOB+THB		120h		8276
LCOB+THB		120h		5625
LCOB+THB		132h		7037
LCOB+THB		132h		6552
LCOB+THB		132h		8966
LCOB+THB		132h		7188
LCOB+THB		144h		8519
LCOB+THB		144h		7586
LCOB+THB		144h		9310
LCOB+THB		144h		8750

Arabidopsis low temperature set 3
Treatment		No seeds		96 h		germ		Std		SE		108 h		germ		Std		SE		120 h		germ		Std		SE		132 h		germ		Std		SE		144 h		germ		Std		SE
Control 1		30		0		000				2		667				15		5000				19		6333				24		8000
2		26		1		385				4		1538				19		7308				23		8846				24		9231
3		27		0		000				2		741				16		5926				20		7407				24		8889
4		28		0		000				3		1071				17		6071				24		8571				26		9286
	111		025		096		192		096		275		1004		397		199		1675		6076		948		474		215		7790		1154		577		245		8851		594		297
LCO A 1		25		4		1600				9		3600				19		7600				21		8400				21		8400
2		30		5		1667				8		2667				19		6333				21		7000				24		8000
3		30		6		2000				12		4000				23		7667				24		8000				28		9333
4		28		6		2143				11		3929				22		7857				22		7857				26		9286
	113		525		1852		261		130		10		3549		613		307		2075		7364		696		110		22		7814		589		295		2475		8755		661		331
LCO B 1		28		4		1429				8		2857				19		6786				20		7143				22		7857
2		31		6		1935				13		4194				22		7097				28		9032				28		9032
3		29		5		1724				9		3103				19		6552				20		6897				22		7586
4		28		6		2143				10		3571				18		6429				22		7857				22		7857
	116		525		1808		305		153		10		3431		588		294		195		6716		294		147		225		7732		958		479		235		8083		645		323
TH A 1		31		7		2258				14		4516				21		6774				24		7742				29		9355
2		29		4		1379				10		3448				19		6552				19		6552				21		7241
3		29		6		2069				11		3793				17		5862				20		6897				23		7931
4		30		7		2333				13		4333				20		6667				20		6667				24		8000
	119		6		2010		435		217		12		4023		491		245		1925		6464		411		206		2075		6964		538		269		2425		8132		884		442
TH B 1		30		6		2000				11		3667				19		6333				21		7000				24		8000
2		26		5		1923				12		4615				16		6154				22		8462				23		8846
3		27		5		1852				10		3704				14		5185				19		7037				19		7037
4		28		8		2857				11		3929				17		6071				20		7143				21		7500
	111		6		2158		470		235		11		3979		440		220		165		5936		512		256		205		7410		703		352		2175		7846		774		387
LCOA+THA1		28		7		2500				11		3929				20		7143				21		7500				25		8929
2		26		6		2308				9		3462				14		5385				19		7308				21		8077
3		27		5		1852				12		4444				19		7037				23		8519				23		8519
4		25		7		2800				13		5200				18		7200				21		8400				21		8400
	106		625		2365		398		199		1125		4259		745		372		1775		6691		874		437		21		7932		616		308		225		8481		352		176
LCOB+THB1		27		5		1852				10		3704				17		6296				19		7037				23		8519
2		29		5		1724				11		3793				18		6207				19		6552				22		7586
3		29		9		3103				18		6207				24		8276				26		8966				27		9310
4		32		6		1875				13		4063				18		5625				23		7188				28		8750
	11700		625		2139		647		323		1300		4442		1187		593		1925		6601		1156		183		2175		7435		1056		528		2500		8541		718		359
germination
Treatment		0 h		96 h		108 h		120 h		132 h		144 h
Control		0		096		1004		6076		7790		8851
LCO A		0		1852		3549		7364		7814		8755
LCO B		0		1808		3431		6716		7732		8083
TH A		0		2010		4023		6464		6964		8132
TH B		0		2158		3979		5936		7410		7846
LCOA+THA		0		2365		4259		6691		7932		8481
LCOB+THB		0		2139		4442		6601		7435		8541
SE
	0 h		96 h		108 h		120 h		132 h		144 h
Control		0		096		199		474		577		297
LCO A		0		130		307		110		295		331
LCO B		0		153		294		147		479		323
TH A		0		217		245		206		269		442
TH B		0		235		220		256		352		387
LCOA+THA		0		199		372		437		308		176
LCOB+THB		0		323		593		183		528		359

Sheet2

Sheet3

Salt amp Drought Stress

bullArabidopsisbullSeeds on petriplates with signals

bullControl and 150 mM NaCl

bullSignals improved growth under salt stress

0 NaCl 150 mM NaCl

0 10-6 10-8 10-9 10-11 M LCO Thuricin 17

Hormones of the Holobiontbull Effective at very low concentrationsbull Regulate plant activity

bull Large changes in gene espression protein production hormone profile

bull Regulate other microbesbull Powerful plant productivity effectsbull Inexpensive to applybull Low environmental impact

bull Small amountbull Already produced in environment

bull Very poorly understoodbull Enormous potential

bull New networkbull gt 10 strains in lab now

Phytomicrobiome Potentialbull Wide range of species in the phytomicrobiomebull Most powerful control factor is the plant

bull Developmentbull Stressbull Nutrition

bull Are key to plant growth and productivitybull Management

bull Inoculationbull Hub species bull Signals

bull Microbe-to-plantbull Plant-to-microbebull Microbe-to-microbe

Development of Biologicals - Whobull Scope is largebull Collaboration

bull Basic research labs bull Academicbull Government

bull Industry laboratories

The Approachbull Sample plant-associated microbes

bull The phytomicrobiomebull Efficientrapid screening to identify microbes of interest

bull Biological assessmentsbull Agronomic assessments

Control Thuricin

Industrial Collaboratorsbull Synagribull EVLbull BASFbull RavenQuestbull Ulyssebull Agroworld Natural Productsbull Inocucorbull Axter

BioFuelNet - NCE

Past and Present Students Dr Feng Zhang Dr Fazli Mabood Dr Narjes Dashti Keldeagh Lindsay Dr Pan Bo Dr Sowmya Subramanian Dr Yuming Bai Dr Xiaomin Zhou Elizabeth Gray Nan Wang

PDFs amp Research Associates Dr Xiaomin Zhou Dr Balakrishna Prithiviraj Dr Alfred Souleimanov Dr Kung Dong Lee Dr Woo Jung Mike Lewis Stewart Leibovitch

Acknowledgements

ALSO Trevor Charles at U Waterloo

The End

Thank You

• Biostimulants from the phytomicrobiome Microbe-to-plant signals as low input crop management technologies
• Need for Enhanced Food Production
• Threat of Climate Change
• Biomass for Advanced Biofuels and the Bioeconomy
• The Phytomicrobiome
• Adapting to Climate Change
• The Approach
• So Far
• Example Line of Research
• Something New
• The Problem
• A Solution
• N2 Fixation and Growth - Genistein
• Commercialization
• Serendipity amp Follow Up
• New LCO Activity
• Commercialization
• Jasmonates ndash An Alternative
• Findings
• Effect of Methyl Jasmonate on Yield
• Environment - Temperature and Response
• Salt amp Drought Stress
• Hormones of the Holobiont
• Phytomicrobiome Potential
• Development of Biologicals - Who
• The Approach
• Industrial Collaborators
• BioFuelNet - NCE
• Diapositive numeacutero 29
• The End

Control		96h		000
Control		96h		385
Control		96h		000
Control		96h		000
Control		108h		667
Control		108h		1538
Control		108h		741
Control		108h		1071
Control		120h		5000
Control		120h		7308
Control		120h		5926
Control		120h		6071
Control		132h		6333
Control		132h		8846
Control		132h		7407
Control		132h		8571
Control		144h		8000
Control		144h		9231
Control		144h		8889
Control		144h		9286
LCO A		96h		1600
LCO A		96h		1667
LCO A		96h		2000
LCO A		96h		2143
LCO A		108h		3600
LCO A		108h		2667
LCO A		108h		4000
LCO A		108h		3929
LCO A		120h		7600
LCO A		120h		6333
LCO A		120h		7667
LCO A		120h		7857
LCO A		132h		8400
LCO A		132h		7000
LCO A		132h		8000
LCO A		132h		7857
LCO A		144h		8400
LCO A		144h		8000
LCO A		144h		9333
LCO A		144h		9286
LCO B		96h		1429
LCO B		96h		1935
LCO B		96h		1724
LCO B		96h		2143
LCO B		108h		2857
LCO B		108h		4194
LCO B		108h		3103
LCO B		108h		3571
LCO B		120h		6786
LCO B		120h		7097
LCO B		120h		6552
LCO B		120h		6429
LCO B		132h		7143
LCO B		132h		9032
LCO B		132h		6897
LCO B		132h		7857
LCO B		144h		7857
LCO B		144h		9032
LCO B		144h		7586
LCO B		144h		7857
TH A		96h		2258
TH A		96h		1379
TH A		96h		2069
TH A		96h		2333
TH A		108h		4516
TH A		108h		3448
TH A		108h		3793
TH A		108h		4333
TH A		120h		6774
TH A		120h		6552
TH A		120h		5862
TH A		120h		6667
TH A		132h		7742
TH A		132h		6552
TH A		132h		6897
TH A		132h		6667
TH A		144h		9355
TH A		144h		7241
TH A		144h		7931
TH A		144h		8000
TH B		96h		2000
TH B		96h		1923
TH B		96h		1852
TH B		96h		2857
TH B		108h		3667
TH B		108h		4615
TH B		108h		3704
TH B		108h		3929
TH B		120h		6333
TH B		120h		6154
TH B		120h		5185
TH B		120h		6071
TH B		132h		7000
TH B		132h		8462
TH B		132h		7037
TH B		132h		7143
TH B		144h		8000
TH B		144h		8846
TH B		144h		7037
TH B		144h		7500
LCOA+THA		96h		2500
LCOA+THA		96h		2308
LCOA+THA		96h		1852
LCOA+THA		96h		2800
LCOA+THA		108h		3929
LCOA+THA		108h		3462
LCOA+THA		108h		4444
LCOA+THA		108h		5200
LCOA+THA		120h		7143
LCOA+THA		120h		5385
LCOA+THA		120h		7037
LCOA+THA		120h		7200
LCOA+THA		132h		7500
LCOA+THA		132h		7308
LCOA+THA		132h		8519
LCOA+THA		132h		8400
LCOA+THA		144h		8929
LCOA+THA		144h		8077
LCOA+THA		144h		8519
LCOA+THA		144h		8400
LCOB+THB		96h		1852
LCOB+THB		96h		1724
LCOB+THB		96h		3103
LCOB+THB		96h		1875
LCOB+THB		108h		3704
LCOB+THB		108h		3793
LCOB+THB		108h		6207
LCOB+THB		108h		4063
LCOB+THB		120h		6296
LCOB+THB		120h		6207
LCOB+THB		120h		8276
LCOB+THB		120h		5625
LCOB+THB		132h		7037
LCOB+THB		132h		6552
LCOB+THB		132h		8966
LCOB+THB		132h		7188
LCOB+THB		144h		8519
LCOB+THB		144h		7586
LCOB+THB		144h		9310
LCOB+THB		144h		8750

Sheet3

Salt amp Drought Stress

bullArabidopsisbullSeeds on petriplates with signals

bullControl and 150 mM NaCl

bullSignals improved growth under salt stress

0 NaCl 150 mM NaCl

0 10-6 10-8 10-9 10-11 M LCO Thuricin 17

Hormones of the Holobiontbull Effective at very low concentrationsbull Regulate plant activity

bull Large changes in gene espression protein production hormone profile

bull Regulate other microbesbull Powerful plant productivity effectsbull Inexpensive to applybull Low environmental impact

bull Small amountbull Already produced in environment

bull Very poorly understoodbull Enormous potential

bull New networkbull gt 10 strains in lab now

Phytomicrobiome Potentialbull Wide range of species in the phytomicrobiomebull Most powerful control factor is the plant

bull Developmentbull Stressbull Nutrition

bull Are key to plant growth and productivitybull Management

bull Inoculationbull Hub species bull Signals

bull Microbe-to-plantbull Plant-to-microbebull Microbe-to-microbe

Development of Biologicals - Whobull Scope is largebull Collaboration

bull Basic research labs bull Academicbull Government

bull Industry laboratories

The Approachbull Sample plant-associated microbes

bull The phytomicrobiomebull Efficientrapid screening to identify microbes of interest

bull Biological assessmentsbull Agronomic assessments

Control Thuricin

Industrial Collaboratorsbull Synagribull EVLbull BASFbull RavenQuestbull Ulyssebull Agroworld Natural Productsbull Inocucorbull Axter

BioFuelNet - NCE

Past and Present Students Dr Feng Zhang Dr Fazli Mabood Dr Narjes Dashti Keldeagh Lindsay Dr Pan Bo Dr Sowmya Subramanian Dr Yuming Bai Dr Xiaomin Zhou Elizabeth Gray Nan Wang

PDFs amp Research Associates Dr Xiaomin Zhou Dr Balakrishna Prithiviraj Dr Alfred Souleimanov Dr Kung Dong Lee Dr Woo Jung Mike Lewis Stewart Leibovitch

Acknowledgements

ALSO Trevor Charles at U Waterloo

The End

Thank You

• Biostimulants from the phytomicrobiome Microbe-to-plant signals as low input crop management technologies
• Need for Enhanced Food Production
• Threat of Climate Change
• Biomass for Advanced Biofuels and the Bioeconomy
• The Phytomicrobiome
• Adapting to Climate Change
• The Approach
• So Far
• Example Line of Research
• Something New
• The Problem
• A Solution
• N2 Fixation and Growth - Genistein
• Commercialization
• Serendipity amp Follow Up
• New LCO Activity
• Commercialization
• Jasmonates ndash An Alternative
• Findings
• Effect of Methyl Jasmonate on Yield
• Environment - Temperature and Response
• Salt amp Drought Stress
• Hormones of the Holobiont
• Phytomicrobiome Potential
• Development of Biologicals - Who
• The Approach
• Industrial Collaborators
• BioFuelNet - NCE
• Diapositive numeacutero 29
• The End

Salt amp Drought Stress

bullArabidopsisbullSeeds on petriplates with signals

bullControl and 150 mM NaCl

bullSignals improved growth under salt stress

0 NaCl 150 mM NaCl

0 10-6 10-8 10-9 10-11 M LCO Thuricin 17

Hormones of the Holobiontbull Effective at very low concentrationsbull Regulate plant activity

bull Large changes in gene espression protein production hormone profile

bull Regulate other microbesbull Powerful plant productivity effectsbull Inexpensive to applybull Low environmental impact

bull Small amountbull Already produced in environment

bull Very poorly understoodbull Enormous potential

bull New networkbull gt 10 strains in lab now

Phytomicrobiome Potentialbull Wide range of species in the phytomicrobiomebull Most powerful control factor is the plant

bull Developmentbull Stressbull Nutrition

bull Are key to plant growth and productivitybull Management

bull Inoculationbull Hub species bull Signals

bull Microbe-to-plantbull Plant-to-microbebull Microbe-to-microbe

Development of Biologicals - Whobull Scope is largebull Collaboration

bull Basic research labs bull Academicbull Government

bull Industry laboratories

The Approachbull Sample plant-associated microbes

bull The phytomicrobiomebull Efficientrapid screening to identify microbes of interest

bull Biological assessmentsbull Agronomic assessments

Control Thuricin

Industrial Collaboratorsbull Synagribull EVLbull BASFbull RavenQuestbull Ulyssebull Agroworld Natural Productsbull Inocucorbull Axter

BioFuelNet - NCE

Past and Present Students Dr Feng Zhang Dr Fazli Mabood Dr Narjes Dashti Keldeagh Lindsay Dr Pan Bo Dr Sowmya Subramanian Dr Yuming Bai Dr Xiaomin Zhou Elizabeth Gray Nan Wang

PDFs amp Research Associates Dr Xiaomin Zhou Dr Balakrishna Prithiviraj Dr Alfred Souleimanov Dr Kung Dong Lee Dr Woo Jung Mike Lewis Stewart Leibovitch

Acknowledgements

ALSO Trevor Charles at U Waterloo

The End

Thank You

• Biostimulants from the phytomicrobiome Microbe-to-plant signals as low input crop management technologies
• Need for Enhanced Food Production
• Threat of Climate Change
• Biomass for Advanced Biofuels and the Bioeconomy
• The Phytomicrobiome
• Adapting to Climate Change
• The Approach
• So Far
• Example Line of Research
• Something New
• The Problem
• A Solution
• N2 Fixation and Growth - Genistein
• Commercialization
• Serendipity amp Follow Up
• New LCO Activity
• Commercialization
• Jasmonates ndash An Alternative
• Findings
• Effect of Methyl Jasmonate on Yield
• Environment - Temperature and Response
• Salt amp Drought Stress
• Hormones of the Holobiont
• Phytomicrobiome Potential
• Development of Biologicals - Who
• The Approach
• Industrial Collaborators
• BioFuelNet - NCE
• Diapositive numeacutero 29
• The End

Hormones of the Holobiontbull Effective at very low concentrationsbull Regulate plant activity

bull Large changes in gene espression protein production hormone profile

bull Regulate other microbesbull Powerful plant productivity effectsbull Inexpensive to applybull Low environmental impact

bull Small amountbull Already produced in environment

bull Very poorly understoodbull Enormous potential

bull New networkbull gt 10 strains in lab now

Phytomicrobiome Potentialbull Wide range of species in the phytomicrobiomebull Most powerful control factor is the plant

bull Developmentbull Stressbull Nutrition

bull Are key to plant growth and productivitybull Management

bull Inoculationbull Hub species bull Signals

bull Microbe-to-plantbull Plant-to-microbebull Microbe-to-microbe

Development of Biologicals - Whobull Scope is largebull Collaboration

bull Basic research labs bull Academicbull Government

bull Industry laboratories

The Approachbull Sample plant-associated microbes

bull The phytomicrobiomebull Efficientrapid screening to identify microbes of interest

bull Biological assessmentsbull Agronomic assessments

Control Thuricin

Industrial Collaboratorsbull Synagribull EVLbull BASFbull RavenQuestbull Ulyssebull Agroworld Natural Productsbull Inocucorbull Axter

BioFuelNet - NCE

Past and Present Students Dr Feng Zhang Dr Fazli Mabood Dr Narjes Dashti Keldeagh Lindsay Dr Pan Bo Dr Sowmya Subramanian Dr Yuming Bai Dr Xiaomin Zhou Elizabeth Gray Nan Wang

PDFs amp Research Associates Dr Xiaomin Zhou Dr Balakrishna Prithiviraj Dr Alfred Souleimanov Dr Kung Dong Lee Dr Woo Jung Mike Lewis Stewart Leibovitch

Acknowledgements

ALSO Trevor Charles at U Waterloo

The End

Thank You

• Biostimulants from the phytomicrobiome Microbe-to-plant signals as low input crop management technologies
• Need for Enhanced Food Production
• Threat of Climate Change
• Biomass for Advanced Biofuels and the Bioeconomy
• The Phytomicrobiome
• Adapting to Climate Change
• The Approach
• So Far
• Example Line of Research
• Something New
• The Problem
• A Solution
• N2 Fixation and Growth - Genistein
• Commercialization
• Serendipity amp Follow Up
• New LCO Activity
• Commercialization
• Jasmonates ndash An Alternative
• Findings
• Effect of Methyl Jasmonate on Yield
• Environment - Temperature and Response
• Salt amp Drought Stress
• Hormones of the Holobiont
• Phytomicrobiome Potential
• Development of Biologicals - Who
• The Approach
• Industrial Collaborators
• BioFuelNet - NCE
• Diapositive numeacutero 29
• The End

Phytomicrobiome Potentialbull Wide range of species in the phytomicrobiomebull Most powerful control factor is the plant

bull Developmentbull Stressbull Nutrition

bull Are key to plant growth and productivitybull Management

bull Inoculationbull Hub species bull Signals

bull Microbe-to-plantbull Plant-to-microbebull Microbe-to-microbe

Development of Biologicals - Whobull Scope is largebull Collaboration

bull Basic research labs bull Academicbull Government

bull Industry laboratories

The Approachbull Sample plant-associated microbes

bull The phytomicrobiomebull Efficientrapid screening to identify microbes of interest

bull Biological assessmentsbull Agronomic assessments

Control Thuricin

Industrial Collaboratorsbull Synagribull EVLbull BASFbull RavenQuestbull Ulyssebull Agroworld Natural Productsbull Inocucorbull Axter

BioFuelNet - NCE

Past and Present Students Dr Feng Zhang Dr Fazli Mabood Dr Narjes Dashti Keldeagh Lindsay Dr Pan Bo Dr Sowmya Subramanian Dr Yuming Bai Dr Xiaomin Zhou Elizabeth Gray Nan Wang

PDFs amp Research Associates Dr Xiaomin Zhou Dr Balakrishna Prithiviraj Dr Alfred Souleimanov Dr Kung Dong Lee Dr Woo Jung Mike Lewis Stewart Leibovitch

Acknowledgements

ALSO Trevor Charles at U Waterloo

The End

Thank You

• Biostimulants from the phytomicrobiome Microbe-to-plant signals as low input crop management technologies
• Need for Enhanced Food Production
• Threat of Climate Change
• Biomass for Advanced Biofuels and the Bioeconomy
• The Phytomicrobiome
• Adapting to Climate Change
• The Approach
• So Far
• Example Line of Research
• Something New
• The Problem
• A Solution
• N2 Fixation and Growth - Genistein
• Commercialization
• Serendipity amp Follow Up
• New LCO Activity
• Commercialization
• Jasmonates ndash An Alternative
• Findings
• Effect of Methyl Jasmonate on Yield
• Environment - Temperature and Response
• Salt amp Drought Stress
• Hormones of the Holobiont
• Phytomicrobiome Potential
• Development of Biologicals - Who
• The Approach
• Industrial Collaborators
• BioFuelNet - NCE
• Diapositive numeacutero 29
• The End

Development of Biologicals - Whobull Scope is largebull Collaboration

bull Basic research labs bull Academicbull Government

bull Industry laboratories

The Approachbull Sample plant-associated microbes

bull The phytomicrobiomebull Efficientrapid screening to identify microbes of interest

bull Biological assessmentsbull Agronomic assessments

Control Thuricin

Industrial Collaboratorsbull Synagribull EVLbull BASFbull RavenQuestbull Ulyssebull Agroworld Natural Productsbull Inocucorbull Axter

BioFuelNet - NCE

Past and Present Students Dr Feng Zhang Dr Fazli Mabood Dr Narjes Dashti Keldeagh Lindsay Dr Pan Bo Dr Sowmya Subramanian Dr Yuming Bai Dr Xiaomin Zhou Elizabeth Gray Nan Wang

PDFs amp Research Associates Dr Xiaomin Zhou Dr Balakrishna Prithiviraj Dr Alfred Souleimanov Dr Kung Dong Lee Dr Woo Jung Mike Lewis Stewart Leibovitch

Acknowledgements

ALSO Trevor Charles at U Waterloo

The End

Thank You

• Biostimulants from the phytomicrobiome Microbe-to-plant signals as low input crop management technologies
• Need for Enhanced Food Production
• Threat of Climate Change
• Biomass for Advanced Biofuels and the Bioeconomy
• The Phytomicrobiome
• Adapting to Climate Change
• The Approach
• So Far
• Example Line of Research
• Something New
• The Problem
• A Solution
• N2 Fixation and Growth - Genistein
• Commercialization
• Serendipity amp Follow Up
• New LCO Activity
• Commercialization
• Jasmonates ndash An Alternative
• Findings
• Effect of Methyl Jasmonate on Yield
• Environment - Temperature and Response
• Salt amp Drought Stress
• Hormones of the Holobiont
• Phytomicrobiome Potential
• Development of Biologicals - Who
• The Approach
• Industrial Collaborators
• BioFuelNet - NCE
• Diapositive numeacutero 29
• The End

The Approachbull Sample plant-associated microbes

bull The phytomicrobiomebull Efficientrapid screening to identify microbes of interest

bull Biological assessmentsbull Agronomic assessments

Control Thuricin

Industrial Collaboratorsbull Synagribull EVLbull BASFbull RavenQuestbull Ulyssebull Agroworld Natural Productsbull Inocucorbull Axter

BioFuelNet - NCE

Past and Present Students Dr Feng Zhang Dr Fazli Mabood Dr Narjes Dashti Keldeagh Lindsay Dr Pan Bo Dr Sowmya Subramanian Dr Yuming Bai Dr Xiaomin Zhou Elizabeth Gray Nan Wang

PDFs amp Research Associates Dr Xiaomin Zhou Dr Balakrishna Prithiviraj Dr Alfred Souleimanov Dr Kung Dong Lee Dr Woo Jung Mike Lewis Stewart Leibovitch

Acknowledgements

ALSO Trevor Charles at U Waterloo

The End

Thank You

• Biostimulants from the phytomicrobiome Microbe-to-plant signals as low input crop management technologies
• Need for Enhanced Food Production
• Threat of Climate Change
• Biomass for Advanced Biofuels and the Bioeconomy
• The Phytomicrobiome
• Adapting to Climate Change
• The Approach
• So Far
• Example Line of Research
• Something New
• The Problem
• A Solution
• N2 Fixation and Growth - Genistein
• Commercialization
• Serendipity amp Follow Up
• New LCO Activity
• Commercialization
• Jasmonates ndash An Alternative
• Findings
• Effect of Methyl Jasmonate on Yield
• Environment - Temperature and Response
• Salt amp Drought Stress
• Hormones of the Holobiont
• Phytomicrobiome Potential
• Development of Biologicals - Who
• The Approach
• Industrial Collaborators
• BioFuelNet - NCE
• Diapositive numeacutero 29
• The End

Industrial Collaboratorsbull Synagribull EVLbull BASFbull RavenQuestbull Ulyssebull Agroworld Natural Productsbull Inocucorbull Axter

BioFuelNet - NCE

Past and Present Students Dr Feng Zhang Dr Fazli Mabood Dr Narjes Dashti Keldeagh Lindsay Dr Pan Bo Dr Sowmya Subramanian Dr Yuming Bai Dr Xiaomin Zhou Elizabeth Gray Nan Wang

PDFs amp Research Associates Dr Xiaomin Zhou Dr Balakrishna Prithiviraj Dr Alfred Souleimanov Dr Kung Dong Lee Dr Woo Jung Mike Lewis Stewart Leibovitch

Acknowledgements

ALSO Trevor Charles at U Waterloo

The End

Thank You

• Biostimulants from the phytomicrobiome Microbe-to-plant signals as low input crop management technologies
• Need for Enhanced Food Production
• Threat of Climate Change
• Biomass for Advanced Biofuels and the Bioeconomy
• The Phytomicrobiome
• Adapting to Climate Change
• The Approach
• So Far
• Example Line of Research
• Something New
• The Problem
• A Solution
• N2 Fixation and Growth - Genistein
• Commercialization
• Serendipity amp Follow Up
• New LCO Activity
• Commercialization
• Jasmonates ndash An Alternative
• Findings
• Effect of Methyl Jasmonate on Yield
• Environment - Temperature and Response
• Salt amp Drought Stress
• Hormones of the Holobiont
• Phytomicrobiome Potential
• Development of Biologicals - Who
• The Approach
• Industrial Collaborators
• BioFuelNet - NCE
• Diapositive numeacutero 29
• The End

BioFuelNet - NCE

Past and Present Students Dr Feng Zhang Dr Fazli Mabood Dr Narjes Dashti Keldeagh Lindsay Dr Pan Bo Dr Sowmya Subramanian Dr Yuming Bai Dr Xiaomin Zhou Elizabeth Gray Nan Wang

PDFs amp Research Associates Dr Xiaomin Zhou Dr Balakrishna Prithiviraj Dr Alfred Souleimanov Dr Kung Dong Lee Dr Woo Jung Mike Lewis Stewart Leibovitch

Acknowledgements

ALSO Trevor Charles at U Waterloo

The End

Thank You

• Biostimulants from the phytomicrobiome Microbe-to-plant signals as low input crop management technologies
• Need for Enhanced Food Production
• Threat of Climate Change
• Biomass for Advanced Biofuels and the Bioeconomy
• The Phytomicrobiome
• Adapting to Climate Change
• The Approach
• So Far
• Example Line of Research
• Something New
• The Problem
• A Solution
• N2 Fixation and Growth - Genistein
• Commercialization
• Serendipity amp Follow Up
• New LCO Activity
• Commercialization
• Jasmonates ndash An Alternative
• Findings
• Effect of Methyl Jasmonate on Yield
• Environment - Temperature and Response
• Salt amp Drought Stress
• Hormones of the Holobiont
• Phytomicrobiome Potential
• Development of Biologicals - Who
• The Approach
• Industrial Collaborators
• BioFuelNet - NCE
• Diapositive numeacutero 29
• The End

Past and Present Students Dr Feng Zhang Dr Fazli Mabood Dr Narjes Dashti Keldeagh Lindsay Dr Pan Bo Dr Sowmya Subramanian Dr Yuming Bai Dr Xiaomin Zhou Elizabeth Gray Nan Wang

PDFs amp Research Associates Dr Xiaomin Zhou Dr Balakrishna Prithiviraj Dr Alfred Souleimanov Dr Kung Dong Lee Dr Woo Jung Mike Lewis Stewart Leibovitch

Acknowledgements

ALSO Trevor Charles at U Waterloo

The End

Thank You

• Biostimulants from the phytomicrobiome Microbe-to-plant signals as low input crop management technologies
• Need for Enhanced Food Production
• Threat of Climate Change
• Biomass for Advanced Biofuels and the Bioeconomy
• The Phytomicrobiome
• Adapting to Climate Change
• The Approach
• So Far
• Example Line of Research
• Something New
• The Problem
• A Solution
• N2 Fixation and Growth - Genistein
• Commercialization
• Serendipity amp Follow Up
• New LCO Activity
• Commercialization
• Jasmonates ndash An Alternative
• Findings
• Effect of Methyl Jasmonate on Yield
• Environment - Temperature and Response
• Salt amp Drought Stress
• Hormones of the Holobiont
• Phytomicrobiome Potential
• Development of Biologicals - Who
• The Approach
• Industrial Collaborators
• BioFuelNet - NCE
• Diapositive numeacutero 29
• The End

The End

Thank You

• Biostimulants from the phytomicrobiome Microbe-to-plant signals as low input crop management technologies
• Need for Enhanced Food Production
• Threat of Climate Change
• Biomass for Advanced Biofuels and the Bioeconomy
• The Phytomicrobiome
• Adapting to Climate Change
• The Approach
• So Far
• Example Line of Research
• Something New
• The Problem
• A Solution
• N2 Fixation and Growth - Genistein
• Commercialization
• Serendipity amp Follow Up
• New LCO Activity
• Commercialization
• Jasmonates ndash An Alternative
• Findings
• Effect of Methyl Jasmonate on Yield
• Environment - Temperature and Response
• Salt amp Drought Stress
• Hormones of the Holobiont
• Phytomicrobiome Potential
• Development of Biologicals - Who
• The Approach
• Industrial Collaborators
• BioFuelNet - NCE
• Diapositive numeacutero 29
• The End