I went to the beach and brought back sand: a story of anoxygenic phototrophic bacteria. Lorena...
-
Upload
megan-hutchinson -
Category
Documents
-
view
215 -
download
1
Transcript of I went to the beach and brought back sand: a story of anoxygenic phototrophic bacteria. Lorena...
![Page 1: I went to the beach and brought back sand: a story of anoxygenic phototrophic bacteria. Lorena Rangel Microbial Diversity Course 2013 MBL.](https://reader036.fdocuments.net/reader036/viewer/2022070401/56649f1d5503460f94c33c31/html5/thumbnails/1.jpg)
I went to the beach and brought back sand: a story of anoxygenic phototrophic bacteria.
Lorena Rangel
Microbial Diversity Course 2013
MBL
![Page 2: I went to the beach and brought back sand: a story of anoxygenic phototrophic bacteria. Lorena Rangel Microbial Diversity Course 2013 MBL.](https://reader036.fdocuments.net/reader036/viewer/2022070401/56649f1d5503460f94c33c31/html5/thumbnails/2.jpg)
Introduction
Anoxygenic phototrophic bacteria
• chlorophyll-based photosynthetic energy
Overmann 2006. The Prokaryotes
www.nau.edu
![Page 3: I went to the beach and brought back sand: a story of anoxygenic phototrophic bacteria. Lorena Rangel Microbial Diversity Course 2013 MBL.](https://reader036.fdocuments.net/reader036/viewer/2022070401/56649f1d5503460f94c33c31/html5/thumbnails/3.jpg)
• Indole-3-acetic acid (IAA)– Plant hormone
• Signaling compound – Controls root growth, tropism, and plant senescence
– Microorganism capable of interfering with IAA signals– Synthesis and Degradation of IAA by bacteria– At least five IAA biosynthetic pathways identified in bacteria– Example: Pseudomonas savastanoi produces IAA to induce tumor formation
– Examples of bacterial response to IAA exposure• Increased bacterial tolerance
– IAA exposure to nonpathogenic E.coli upregulated genes involved in adaptation to unfavorable environments (Bianco et al. 2006. Arch Microbiol 185, 373)
• Optimizing conditions for bacterial entry– IAA opens stomata, even in dark (Huang et al. 2000. Plant Sci 156, 65)
Introduction
![Page 4: I went to the beach and brought back sand: a story of anoxygenic phototrophic bacteria. Lorena Rangel Microbial Diversity Course 2013 MBL.](https://reader036.fdocuments.net/reader036/viewer/2022070401/56649f1d5503460f94c33c31/html5/thumbnails/4.jpg)
Sippewissett Salt Marsh
DiatomsCyanobacteria
Purple Sulfur BacteriaGreen Sulfur Bacteria
Purple NonSulfur Bacteria
Overmann 2006. The Prokaryotes
![Page 5: I went to the beach and brought back sand: a story of anoxygenic phototrophic bacteria. Lorena Rangel Microbial Diversity Course 2013 MBL.](https://reader036.fdocuments.net/reader036/viewer/2022070401/56649f1d5503460f94c33c31/html5/thumbnails/5.jpg)
PNSB
GSB
PSB
All in 10ml Marine Phototrophic Base!
Add 1mM Na succinateIlluminate at 850nm
Add 5mM Na thiosulfateAdd 1mM Na2SIlluminate at 850nm
Add 3mM Na2SIlluminate at 750nm
![Page 6: I went to the beach and brought back sand: a story of anoxygenic phototrophic bacteria. Lorena Rangel Microbial Diversity Course 2013 MBL.](https://reader036.fdocuments.net/reader036/viewer/2022070401/56649f1d5503460f94c33c31/html5/thumbnails/6.jpg)
SHAKE TUBES
![Page 7: I went to the beach and brought back sand: a story of anoxygenic phototrophic bacteria. Lorena Rangel Microbial Diversity Course 2013 MBL.](https://reader036.fdocuments.net/reader036/viewer/2022070401/56649f1d5503460f94c33c31/html5/thumbnails/7.jpg)
GSB
PSB PNSB
![Page 8: I went to the beach and brought back sand: a story of anoxygenic phototrophic bacteria. Lorena Rangel Microbial Diversity Course 2013 MBL.](https://reader036.fdocuments.net/reader036/viewer/2022070401/56649f1d5503460f94c33c31/html5/thumbnails/8.jpg)
![Page 9: I went to the beach and brought back sand: a story of anoxygenic phototrophic bacteria. Lorena Rangel Microbial Diversity Course 2013 MBL.](https://reader036.fdocuments.net/reader036/viewer/2022070401/56649f1d5503460f94c33c31/html5/thumbnails/9.jpg)
Purple Sulfur Bacteria
Green Sulfur Bacteria
• Class Gammaproteobacteria• Order Chromatiales• BChl a or b• Internal sulfur granules
• Phylum Chlorobi• BChl c, d, or e and BChl a• External sulfur granules
![Page 10: I went to the beach and brought back sand: a story of anoxygenic phototrophic bacteria. Lorena Rangel Microbial Diversity Course 2013 MBL.](https://reader036.fdocuments.net/reader036/viewer/2022070401/56649f1d5503460f94c33c31/html5/thumbnails/10.jpg)
Purple Non-Sulfur Bacteria
• Highest metabolic flexibility of all phototrophs
• BChl a or b• Class Alphaproteobacteria
or Betaproteobacteria
![Page 11: I went to the beach and brought back sand: a story of anoxygenic phototrophic bacteria. Lorena Rangel Microbial Diversity Course 2013 MBL.](https://reader036.fdocuments.net/reader036/viewer/2022070401/56649f1d5503460f94c33c31/html5/thumbnails/11.jpg)
GSB
PNSB
PSB
ChlorinAbsorption maxima (nm)
Whole Cells
BChl a 375, 590, 805, 830-911
BChl b400, 605, 835-850, 986-1035
BChl c 457-460, 745-755
BChl d 450, 715-745
BChl e 460-462, 710-725
BChl g 375, 419, 575, 788
590.
2258
9.75
![Page 12: I went to the beach and brought back sand: a story of anoxygenic phototrophic bacteria. Lorena Rangel Microbial Diversity Course 2013 MBL.](https://reader036.fdocuments.net/reader036/viewer/2022070401/56649f1d5503460f94c33c31/html5/thumbnails/12.jpg)
? Rhodovulum sp. JA545Prosthecochloris
aestuarii DSM 271
![Page 13: I went to the beach and brought back sand: a story of anoxygenic phototrophic bacteria. Lorena Rangel Microbial Diversity Course 2013 MBL.](https://reader036.fdocuments.net/reader036/viewer/2022070401/56649f1d5503460f94c33c31/html5/thumbnails/13.jpg)
Acetate
Propionate
Citrate
Hypothesis1:
Under limited light conditions PNS bacteria will not grow at rates comparable to “natural” or full light exposure.
Treatments • 24L• 16L:8D• 0L
Hypothesis2:
PNS bacteria will grow at a greater rate when provided a less complex carbon source
Treatments • 10mM Citrate• 10mM Acetate• 10mM Propionate
![Page 14: I went to the beach and brought back sand: a story of anoxygenic phototrophic bacteria. Lorena Rangel Microbial Diversity Course 2013 MBL.](https://reader036.fdocuments.net/reader036/viewer/2022070401/56649f1d5503460f94c33c31/html5/thumbnails/14.jpg)
Propionate + Tryp
Propionate
Acetate + Tryp
Acetate
Citrate + Tryp
Citrate
Carbon Sources 24h Light
0
2
4
6
8
10
12
0 2 4 6 8 10
Days Post Inoculation
Con
cent
ratio
n (m
M)
Carbon Sources 16h Light
0
2
4
6
8
10
12
0 2 4 6 8 10
Days Post Inoculation
Con
cent
ratio
n (m
M)
Carbon Sources 0h Light
0
2
4
6
8
10
12
0 2 4 6 8 10
Days Post Inoculation
Con
cent
ratio
n (m
M)
![Page 15: I went to the beach and brought back sand: a story of anoxygenic phototrophic bacteria. Lorena Rangel Microbial Diversity Course 2013 MBL.](https://reader036.fdocuments.net/reader036/viewer/2022070401/56649f1d5503460f94c33c31/html5/thumbnails/15.jpg)
Do PNSB make IAA?
Hypothesis3:
PNS bacteria have the ability to make indole-acetic acid.
Rhodovulum sp. JA545
Rhodovulum sp. PH10:• Indole-3-glycerol phosphate
synthase• Tryptophan synthase
Ouyang et al 2000.. The Plant Journal 24: 327-333.
Chorismate
Anthranilate
PRA
CdRP
IGP
Indole
Tryptophan
IAA
ASA GI: 402500488ASB GI: 402500495
PAT GI: 402500496
PAIGI:495666825
IGSGI: 402500497
TSAGI: 402498195
TSBGI: 495666823
Trp-independent
Trp-depen
dent
Chorismate
Anthranilate
PRA
CdRP
IGP
Indole
Tryptophan
IAA
ASA GI: 402500488ASB GI: 402500495
PAT GI: 402500496
PAIGI:495666825
IGSGI: 402500497
TSAGI: 402498195
TSBGI: 495666823
Trp-independent
Trp-depen
dent
![Page 16: I went to the beach and brought back sand: a story of anoxygenic phototrophic bacteria. Lorena Rangel Microbial Diversity Course 2013 MBL.](https://reader036.fdocuments.net/reader036/viewer/2022070401/56649f1d5503460f94c33c31/html5/thumbnails/16.jpg)
Standard
- tryp
+ tryp
+ tryp
- tryp
C C CA A AP P P
24h 16h 0h
200µM 100µM 80µM 60µM 40µM 20µM 10µM 0µM
Standard
- tryp
+ tryp
+ tryp
- tryp
C C CA A AP P P
24h 16h 0h
200µM 100µM 80µM 60µM 40µM 20µM 10µM 0µM
![Page 17: I went to the beach and brought back sand: a story of anoxygenic phototrophic bacteria. Lorena Rangel Microbial Diversity Course 2013 MBL.](https://reader036.fdocuments.net/reader036/viewer/2022070401/56649f1d5503460f94c33c31/html5/thumbnails/17.jpg)
0
5
10
15
20
25
30
35
0 1 2 3 4 5 6 7 8 9 10
Day(s) Post Inoculation
Con
cent
rati
on (
uM) Citrate/NH4Cl 24h
Citrate/NH4Cl 16h
Citrate/NH4Cl 0h
Citrate/Tryp 24h
Citrate/Tryp 16h
Citrate/Tryp 0h
0
5
10
15
20
25
30
35
0 1 2 3 4 5 6 7 8 9 10
Day(s) Post Inoculation
Con
cent
rati
on (
uM) Propionate/NH4Cl 24h
Propionate/NH4Cl 16h
Propionate/NH4Cl 0h
Propionate/Tryp 24h
Propionate/Tryp 16h
Propionate/Tryp 0h
0
5
10
15
20
25
30
35
0 1 2 3 4 5 6 7 8 9 10
Day(s) Post Inoculation
Con
cent
rati
on (
uM) Acetate/NH4Cl 24h
Acetate/NH4Cl 16h
Acetate/NH4Cl 0h
Acetate/Tryp 24h
Acetate/Tryp 16h
Acetate/Tryp 0h
Salkowski Assay
Measures specifically for IAA, indolepyruvic acid, and indoleacetamide
Tryptophan IAA
![Page 18: I went to the beach and brought back sand: a story of anoxygenic phototrophic bacteria. Lorena Rangel Microbial Diversity Course 2013 MBL.](https://reader036.fdocuments.net/reader036/viewer/2022070401/56649f1d5503460f94c33c31/html5/thumbnails/18.jpg)
Tryptophan IAA
5-Hydroxyindoleacetic acidIndole-3-acetaldehyde
![Page 19: I went to the beach and brought back sand: a story of anoxygenic phototrophic bacteria. Lorena Rangel Microbial Diversity Course 2013 MBL.](https://reader036.fdocuments.net/reader036/viewer/2022070401/56649f1d5503460f94c33c31/html5/thumbnails/19.jpg)
Main Findings
Conclusions
• Enriched for GSB (Prosthecochloris), PSB and PNSB (Rhodovulum)
• Propionate encourages faster growth rates of PNSB
• PNSB may produce low concentrations of indole-like compounds
Conclusions
• Enriched for GSB (Prosthecochloris), PSB and PNSB (Rhodovulum)
• Propionate encourages faster growth rates of PNSB
• PNSB may produce low concentrations of indole-like compounds
Main Findings
Conclusions
• Enriched for GSB (Prosthecochloris), PSB and PNSB (Rhodovulum)
• Propionate encourages faster growth rates of PNSB
• PNSB may produce low concentrations of indole-like compounds
Main Findings
Conclusions
• Optimization of carbon sources for future PNSB enrichments
• First evidence of anoxygenic phototrophic bacteria producing indole
• Further work on the actual compound produced