Ing. Josef Patzak, PhD. - chizatecHumulenol II Humuladienone Bicyclogermacrene Aromadrene...
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Chmelařský Institut s.r.o., oddělení Biotechnologie Kadaňská 2525, 438 46 Žatec, Česká republika TEL: +420 415732109 mobil: +420 724929535 E-mail: [email protected] Ing. Josef Patzak, PhD.
Transcript of Ing. Josef Patzak, PhD. - chizatecHumulenol II Humuladienone Bicyclogermacrene Aromadrene...
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Chmelařský Institut s.r.o., oddělení Biotechnologie Kadaňská 2525, 438 46 Žatec, Česká republika TEL: +420 415732109 mobil: +420 724929535
E-mail: [email protected]
Ing. Josef Patzak, PhD.
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Cannabaceae < Urticales < Dicotyledons < Magnoliophyta vytrvalá, pnoucí, pravotočivá dvoudomá dlouhodenní 2n = 20 (9A+1G), 2C = 5,6 pg X a Y, typ Drosophila ♀G(2) ♂P5A5
Genetická konference GSGM, 24-26. září 2014, Průhonice
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Genetická konference GSGM, 24-26. září 2014, Průhonice
CH2OPPCH2OPP
CH2OPP
CH2OH
OH
CH2OH
O
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CO2CH3
O
CH2OPP
CH2OPP
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H
H
H H
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H
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HO
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OH
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SCoA
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SCoA
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SCoA
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SCoA
O
OH
O OH
SCoA
O
OH
O OHCH2OH
OH
O OHCH2OP OH
O OHCH2OPP
CH2OPP
O
OH
OH
OHO
OH
OH
OH
O
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OHOH
OH
O
OH
OH
O
O-Gly
OH
OH
O
OH
OH
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O-Gly
OHOH
O
OH
OMe
OHOH
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OH
OMe
OHOH
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OH
OH
OH
O
OH
OMe
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OH
O
OH
OH
O
CH2OPP
CH2OPP
CH2OPP
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CoAS
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CoAS
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OH
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OH
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SCoA
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SCoA
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Biosynthesis of hop resins, essential oils, flavanoids and polyphenols
Acetyl-CoA Acetyl-CoA
Acetyl-CoAAcetoacetyl-CoA
HMG-CoA
Mevalonic acid
Mevalonate-5-phosphate Mevalonate-5-pyrophosphate
IsopentenylpyrophosphateIPP
IPPDimethylallylpyrophosphateDMAPP
GeranylpyrophosphateIPP
FarnesylpyrophosphateCamphene α-Pinene
β-Pinene
α-Pinene oxide
Geraniol
Myrcene Ocimene
Linalool
Nerol
Methyl geranate
α-TerpineolLimonene
Carvone 8(9)-p-Methandienep-CymeneTerpinolene α-Terpinene γ-Terpinene
β-Farnesene
Farnesol
Pyruvate Pyruvate
Acetolactate
2,3-Dihydroxyisovalerate
2-Oxoisovalerate
Isobutyryl-CoA
Isobutyryl-CoA
2-Oxoisocaproate
Isovaleryl-CoA
Isovaleryl-CoA 3-Methylcrotonyl-CoA
3-Methylglutaconyl-CoA
Malonyl-CoA
Phloroisovelerophenone
DMAPP
DMAPP
DMAPP
Desoxyhumulone
Lupulone
Humulone
DMAPP
DMAPP
DMAPP
Malonyl-CoA
Phloroisobutyrophenone
Desoxycohumulone
Colupulone
Cohumulone
Germacrene D
δ-Cadinene
γ-Cadinene
α-Cadinene
α-Copaene
α-Ylangene
β-Ylangene
Cadina-1,4-diene
γ-Muurolene
α-Muurolene
α-Cubebene
β-Cubebene
ε-Muurolene
Nerolidol
β-Bisabolene
trans-Humulanecis-Humulane
β-Caryophyllene
Caryophyllene epoxide
α-Humulene
epoxide I epoxide II epoxide III
Humulenol I HumulolHumulenol II
Humuladienone
Bicyclogermacrene
AromadreneAlloaromadrene
Globulol
α-Gurjunene
Viridiflorene
Viridiflorol
epoxideepoxide
Ledol Alloglobulol
γ-Elemene
Germacrene A Hedycaryol Germacrene B
α-Selinene α-Eudesmol Selina-4(14),7,(11)-diene
β-Selinene β-Eudesmol Selina-3,7(11)-diene
γ-Eudesmol
β-Selinene epoxide
β-Selinenol α-Selinenol
1
2
3
4
5
6
7
8
88
8
8
8
8
8
9
9
10
11
12
13
14
14
15
16
17
18
18
Involved enzymes:
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acetoacetyl CoA thiolaseHMG-CoA synthaseHMG-CoA reductaseMVA kinasemevalonate-5-phosphate kinasemevalonate-5-diphosphate decarboxylaseIPP isomeraseprenyl transferaseacetyl-CoA carboxylaseacetohydroxyacid synthase2-hydroxy-3-oxoacid reductoisomerasedihydroxiacid dehydratase3 enzymes and precursors3-methyl-2-oxobutanoate dehydrogenase3-methylglutaconyl-CoA hydratasemethylcrotonyl-CoA carboxylaseacyl-CoA dehydrogenasephloroisovaleryl synthasephenylalanine ammonia-lyasechalcone synthasechalcone isomeraseflavanone 3-hydroxylasedihydroflavonol/one 4-reductaseflavonol synthase
flavan-3,4-diol 4-reductasecyclaseshydroxylasescytochrome P-450 dependent oxidase
G 3-O-glycosyltransferases
C
CC
C
C
O C
O O
O O
O
OO
O
C
C
C C
H
O
O
O
O
O
C
O
OH
H
C
H
H
O
C
C C
OH
H
H
H
HOH
H
OH
α-Cadinol τ-Cadinol Cubenol
O
O
C
C
O
O
R
SCoA
O
R
SCoA
OOH
R
OH
O
O
R
CHO
R
O
SCoA
O
R
SCoA
O
R
O
O
R
3 x
3 x
OH
OH
OH
O
OH
OH
OH
OH
OH
OH
O
OH
OH
OH
OH
OH
O
OH
OH
OH
OH
OH
OH
O
OH
OH
OH
OH
OH
O
OH
OH
OH
OH
O
OH
OH
OH
OH
OHOH
O
OH
OH
OH
O
OH
OH
O
OH
O
OH
OH
O
OHOH
O
OH
OH
SCoA
O
OHO
O O
CoAS+
OHOH
O
OH
OH
OH
O
OH
OH
O
OH
O
OH
OH
O
OHOH
O
OH
OH
O
OH
OH
OH
O
OH
OH
O
OH
OH
OH
OH
O
OHOH
OH
O
OH
OH
O
O-Gly
OH
OH
O
OH
OH
O
O-Gly
OHOH
O
OH
OMe
OHOH
O
OH
OMe
OHOH
O
OH
OH
OH
O
OH
OMe
O
OH
O
OH
OH
O
α-Bergamotene
Malonyl-CoA
9
Shikimate/Arogenate Pathway
Phenylalanine/HydrocynnamatePathway19
Hydroxycinnamic acid
Hydroxybenzoic acid
4-Coumaroyl-CoA
Naringenin chalcone
3 x
20
21
22
24
23
24
25
G
G
8
CH2OPP
CH2OPP
CH2OPP
CH2OPP
DMAPP
DMAPP
DMAPP
DMAPP
+
+
+
+
8
8
8
Naringenin
Desmethylxanthohumol
Xanthohumol
3’-Prenylchalconaringenin
5’-Prenylxanthohumol3’-Geranylchalconaringenin
8-Prenylnaringenin
6-Prenylnaringenin
Isoxanthohumol
6-Geranylnaringenin
Dihydroflavonol
Kaempferol
Quercetin
Kaempferol glycoside
Quercetin glycoside
Leucocyanidin
CatechinGallacatechin
Proanthocyanidin Proanthodelphinidin
Hop resins
Hop essential oils
Hop flavanoids
Hop polyphenols
α-bitter acids
β-bitter acids
Monoterpenes
Sesquiterpenes
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Genetická konference GSGM, 24-26. září 2014, Průhonice
H. lupulus lupuloidesvar.
H. lupulus neomexicanusvar.H. lupulus pubescensvar.
H. lupulus cordifolius +var.H. japonicus
H. lupulus lupulus var.
Centrum původu
H. japonicus
H. yunnanensis
1000736
11601300
9501300
1629
1650
17991822
1850
1850
1874
1880
1930
1920
1868
1880
1950
1934
1980
1929
ostatní druhy a variety
Legenda:
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Genetická konference GSGM, 24-26. září 2014, Průhonice
Brewers Gold[50]
X(American)
X
Eroica [8]
Galena [10]
X(75/5/3M)
Magnum[19]
X
X
X
Souther nBrewer
[21]
XCantenburyGolding
Northern Brewer
[33]
X
X
Sládek [52]
Osvald’s Clone 72
[55]
X(American)
X
Osvald’s Clone 72
[55]
Premiant[53]
Zattler (German)
X O.P.
X
Wye Nor thdown [2] X X
Wye Challenger[5]
X
Wye Target[6]
EastwellGolding
X
Neomex(American) X O.P.
X O.P.
X O.P.
Br amlingCross
[38]
XX
X
X
X Br amlingX O.P.
X
EnglishGolding X
Yeoman[25]
BBI Manitoba X O.P.
Legend:
O.P.
Female
Male
Crossing
Open pollinated
Number
Origin
Linkage
Landrace
X
[ ]
( )
Golding[35]
O.P.
X(81/8/13M)
Mer kur[60]
X
Taurus[24]
X O.P.
Zenith[62]
X
XO.P.(German)
Fuggle[34]
United Kingdom
X
X
XX
X
XO.P. Ear lyGreen
Nugget [13]
Centennial [29]
United States
X
X
X(German)
Hor izon [66]
X
X O.P.
X
XSerebr ianka
[16](Russia) (English)
X O.P.
Cascade [45]
X(Utah wild)
4n
Willamete [3] Petham
GoldingX
X
Chinnok [12]
X Sunshine
Comet [27]
X(21381M)
4n
X4n
Liber ty [41]
Mt. Hood [23]
Crystal [28]
X
Haller tauerMittelfr herü
X
X O.P.
X
X
O.P.
(German)
Vanguard [67]
X
X
Ster ling [63]
Saazer (Czech)
X4n
4nX
Santiam [64]
Tettnanger (German)
X
Columbus [61]
? ?
X
Glacier [65]
X
Elsasser(French)
X
X
(English)X
Bullion (English)
Bullion
Late Cluster [48]
X O.P.
California Cluster O.P. X X
Smooth Cone [47]
Callicross [51]
New Zealand
O.P.(English)
X O.P.
Pr ide of Kent
Pr ide ofRingwood
[44]
XO.P.X O.P.
Ringwood special
[43]
Australia
O.P.
Hersbruck[39]
Tetnang[36]
Haller tau[30]
Spalt[40]
Saaz
Str iesselspalt
[18](French)
Bor [56]
X
X O.P. X
O.P.
X
X
X
X
Agnus[57]
X
Per le [7]
(63/5/27M)
H ller Anfang
ü X(81t9)
X(54/33/11)
X
Haller tauer Gold X(75/15/106M)
Haller tauer Tradition
[22]
X
Spalter Select
[26]
(71/16/7M)(76/18/80)
Germany
Backa
[42](Yugoslavian)
Nadwislavsky[37]
Lubelski[31]
Izabela[17]
X(Yugoslavian)
X
X
Marynka[20]
Estera[14]
Czech Republic
Poland
XO.P. White VineSaazer X
Shinshuwase
JapanKirin II
[1]
Golden Star[9]
3n
3n 3n
3n
3n
X(3/3)
Atlas[46]
AhilApolonSavinski Golding[32]
X
(Sx502)
O.P.
X
Vojvodina[54]
XO.P.
(2/173)X
Buket[4]
AtlasX(1/9)
Blisk[15]
4n3n
(TG)X
Bobek[11]
(TG)X
Aurora[49]
X(3/3)
Cicero[58]
4n 3n
(105/58)X
Celeia[59]
4n 3n
(39-06)X
Outeniqua[68]
Slovenia
SouthAfrica
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Genetická konference GSGM, 24-26. září 2014, Průhonice
Rostlinná výroba 45: 165-172, 1999
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Genetická konference GSGM, 24-26. září 2014, Průhonice
Rostlinná Výroba 48: 318-321, 2002 Rostlinná Výroba 48: 343-350, 2002
M1 O B S P A M24382 4527 4715 4353676 pb
517 pb460 pb396 pb350 pb
222 pb179 pb
700 pb
500 pb600 pb
400 pb
300 pb
200 pb
222 pb179 pb
350 pb300 pb
200 pb
126 pb
M1 O B S P A M24382 4527 4715 4353
676 pb
517 pb460 pb396 pb350 pb
1605 pb
1198 pb1000 pb
800 pb700 pb
500 pb600 pb
400 pb
2645 pb
900 pb
1500 pb
M1 O B S P A M24382 4527 4715 4353
676 pb
517 pb460 pb396 pb350 pb
1605 pb
1198 pb
2645 pb
1000 pb
800 pb700 pb
500 pb600 pb
400 pb
900 pb
1500 pb
M1 O B S P A M24382 4527 4715 4353
a)
b)
c)
d)
M2 M11500 pb1000 pb
200 pb
800 pb
500 pb600 pb
400 pb300 pb
100 pb126 pb
222 pb179 pb
676 pb
1198 pb1605 pb
517 bp460 bp396 bp350 bp
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Genetická konference GSGM, 24-26. září 2014, Průhonice
Euphytica 121: 9-18, 2001 Euphytica 131: 343-350, 2003
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Genetická konference GSGM, 24-26. září 2014, Průhonice
145 SSR loci Osv
. klo
n 31
Osv
. klo
n 72
Osv
. klo
n 11
4B
orSl
ádek
Agn
usPr
emia
ntH
arm
onie
Rub
ínV
ital
Kaz
bek
4237
4784
4788
4849
4914
4915
4932
4964
4837
HlGA3
HlGT4
HlGA29
Genome 50: 15-25, 2007 Kvasný průmysl 57: 296-304, 2011
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Genetická konference GSGM, 24-26. září 2014, Průhonice
Plant Science 162: 1007-1018, 2002 Biologia Plantarum 46: 375-381, 2003 Journal of Agriculture and Food Chemistry 53: 4793-4798, 2005 Biologia Plantarum 50: 48-54, 2006 Journal of Agriculture and Food Chemistry 55: 7767-7776, 2007 Journal of Agriculture and Food Chemistry 58: 902-912, 2010 BMC Plant Biology 12: 27, 2012
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Genetická konference GSGM, 24-26. září 2014, Průhonice
http://www.ncbi.nlm.nih.gov ESTs: 75 672 932 (42,158,059,791) TSA: 52 596 023 (42,426,765,504) http://www.planttrichome.org/trichomedb/ ESTs: 825 370 Unigenes: 106 370 Suntory Liquors Limited, Osaka, Japonsko ESTs: 387 929 (915,421,000) 41 228 pravděpodobně genů (29 950 redukováno kvůli heterozygotnosti)
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Genetická konference GSGM, 24-26. září 2014, Průhonice
Biologia Plantarum 55: 761-765, 2011 Brewing Science 65: 96-102, 2012
ŽPČ
Bor
Slád
ekPr
emia
ntA
gnus
Har
mon
ieR
ubín
Vita
lK
azbe
kB
ohem
ieSa
az la
te
LAR1
WRKY75
Harmonie - sadba
CMPS
3 658 EST-SSR loci 17 128 SNPs 6 144 DART
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Genetická konference GSGM, 24-26. září 2014, Průhonice
Kvasný Průmysl 59: 2-13, 2013 Kvasný Průmysl 59: 296-305, 2013
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Genetická konference GSGM, 24-26. září 2014, Průhonice
Biochemical Systematics and Ecology 38: 136-145, 2010 Genome 53: 545-557, 2010
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Genetická konference GSGM, 24-26. září 2014, Průhonice
2,57 Gb odhad (2.05 Gb sekvenováno) cpDNA 180 204 bp, mtDNA 336 811 bp 13 776 946 – 22 012 445 SNPs
Natsume et al. Journal of Plant Physiology, prepared.
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Genetická konference GSGM, 24-26. září 2014, Průhonice
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Genetická konference GSGM, 24-26. září 2014, Průhonice
Bc. Alena Henychová BC AVČR v.v.i., ÚMBR, České Budějovice VÚRV v.v.i., Praha-Ruzyně EP7254, NAZV Mze, 1997-2000 EP9357, NAZV MZe, 1999-2001 521/99/1591, GAČR, 1999-2001 QC1336, NAZV MZe, 2001-2004 521/03/0072, GAČR, 2003-2005 521/08/0740, GAČR, 2008-2011 QH81052, NAZV MZe, 2008-2012 13-03037S, GAČR, 2013-2017
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Genetická konference GSGM, 24-26. září 2014, Průhonice
Od neznámého k přesné informaci – 15 let pokroku v genomice chmele (Humulus lupulus L.)Chmel otáčivý (Humulus lupulus l.)chemická továrna – lupulinová zrnaPěstování chmeleŠlechtění chmelePCR metody - RAPDPCR metody - STSPCR metody - AFLP a ISSRPCR - SSRAnalýza sekvencí genůSekvence – trankriptomPCR - EST-SSRHodnocení genetické příbuznostiHodnocení genetické příbuznostiSekvence – genomSekvence – genomPoděkování ��� Děkuji � � za �pozornost
