Post on 14-Sep-2018
transcript
Development of molecular markers for application in A stralian Canola application in Australian Canola
breedingg
C l M l l M k PCanola Molecular Marker Program(harsh.raman@industry.nsw.gov.au)
Mapped Blackleg Resistance lociMapped Blackleg Resistance lociSkipton/Ag Spectrum (191 lines) Skipton/Ag-Spectrum (191 lines) BLN2762/Surpass400 (139 lines)
C l b *3/W t 10 (76 li ) Columbus*3/Westar 10 (76 lines)
Maxol*1/Westar 10 (101 lines)
Ag-Castle/Topas Ag-Castle*4/Westar10 (173 lines) g ( )
Mapped blackleg resistance loci in the DH population from BLN2762/Surpass-400population from BLN2762/Surpass-400
Screened with three isolatesScreened with three isolatesD1 (AvrLM2,5,6,9)D7 (AvrLM1 3 5 6 8)
r2=70%D7 (AvrLM1,3,5,6,8)D10 (AvrLM5,6,8,9,S)
Rlm9M1
M2
A10A7
Mapping of Ag-Castle/Topas
Population is developed by International
Mapping of Ag Castle/Topas
Population is developed by International Blackleg Consortium, Canada
F -1 F1-1 F1-2 Mapped with SSR (Canada Wagga) and Mapped with SSR (Canada, Wagga) and
DArT markers (Wagga) Qualitative Resistance Genes Qualitative Resistance Genes Quantitative Resistance Genes
3-4 QTLs– 3-4 QTLs
Strong Evidence of novel genesStrong Evidence of novel genes
QTL on chromosome X: r2= 28% – Field conditions
QTL on chromosome X: r2= 25%– Seedling stage
Validation of Loci for BLR
Bi-parental populations– Evaluate two alleles/locus
•Skipton/Ag-Spectrum•Maxol/WestarA C l /• Known ancestry •Ag-Castle/Westar
•Surpass/Topas•BLN3347/Carousel-10
Association mapping– Evaluate more alleles/locus
• Historic recombination
Digital Image analysis
BR Scores of 188 Lines Using Differential SetBR Scores of 188 Lines Using Differential SetD1 D2 D3 D4 D5 D6 D7 D8 D9 D10
4.730140304 7.646925 7.128249 6.971014 1.836446 2.285203 2.115384 3.319144 4.757586 6.789557
7.097330341 6.618246 7.920466 6.133153 4.431464 6.812594 5.135307 3.122412 3.744606 7.372861
4.356579722 7.232666 7.533186 6.021005 4.989769 4.887729 5.961428 3.576325 3.821709 6.558984
3.668504598 6.394026 6.921822 6.13768 3.952241 5.087057 4.581889 3.161766 4.912204 6.454949
4.522183939 6.229967 6.028761 6.361742 2.957218 1.091836 2.472812 1.323677 5.722262 4.500186
6.015414023 6.040849 6.505169 6.073434 2.411565 2.724239 4.632233 3.208622 3.200387 5.417817
5.591654768 7.413355 7.361577 6.63768 3.654039 2.698503 2.850807 2.183013 5.962158 6.839883
4.423611703 6.555453 7.396042 5.633161 1.550735 3.589092 5.679019 3.309156 4.056738 6.270415
6.886679493 7.023896 7.984523 6.358386 5.292574 6.127917 4.005167 4.99292 6.186199 7.152023
6.135385064 7.462896 7.961975 6.679347 4.994903 6.960856 4.823716 4.126742 5.141677 7.428624
4.129040146 5.409193 6.290693 4.01268 2.825122 3.857382 1.85182 2.110125 5.153192 6.457673
5.431069152 5.915106 4.398745 1.204228 1.863305 4.074617 4.304964 2.816818 3.277779 4.295161
5.614302808 6.066947 7.552231 5.31672 3.201049 6.393255 4.565232 3.339089 5.958782 5.880304
2.826531339 6.369425 6.558874 4.349931 3.370806 3.242903 1.600677 3.854578 4.823295 3.60767
4.073260062 6.035585 7.189969 4.833161 3.093878 4.880359 5.526491 3.301663 2.586679 6.747776
3.520257892 6.004412 5.034623 6.961948 2.380899 5.700361 1.953367 1.297054 3.235428 5.006055
2.362642866 5.175833 6.251495 3.34907 3.678294 3.312643 3.117591 3.749729 3.562106 3.181641
Evaluated 188 lines for Adult Plant Resistance
AV-Garnet (Rlm1 9) AV Garnet (Rlm1,9) Monola76 (RlmS) ATR-Cobbler (Rlm 4,9)( , ) CB-JardeeHT (Rlm2,3) Hyola50 (Unknown)
Results: S B j d B Some B. juncea and B. carinata lines were resistant to all stubbles
Phenotypic Relationships for BLRGroup average
Resemblance: D1 Euclidean distance
100
ance
50
Dis
ta
0
Hyo
la50
Hyo
la76
AT
C931
84-1
AT
C940
44-1
CB
J-00
1S
eeth
aA
TR-S
igna
lA
v-Ru
byT
orna
doT
T45
C05
Bra
voT
TO
asis
CL
ATR
-Bar
raA
v-Ja
deT
awrif
ficT
TAT
R-S
umm
ittSA
RDI
607
Ag-C
omet
Ag-
Spec
trum
Car
ouse
l-10
Tar
cool
a46
C04
Sto
rmT
TS
urpa
ss40
2CS
urpa
ss40
046
Y78
ZY0
10B
LN27
37*C
O02
0305
-P71
-11
P624
Wes
bell
RO
Y33-
99P
3Ag
-Mus
ter
BLN
3347
CB-T
elfe
rR
OY
9712
5B
LN28
52-0
3W02
RO
Y33-
99P
2Iw
ao-n
atan
eA
v-O
pal
Hur
rican
eTT
Nor
in_22
RO
Y33-
99P
4B
LN33
03*C
R030
2B
LN36
14R
OY3
1-99
P1
Thu
nder
TT
Osc
arR
OY
9705
6SA
RDI
604
Lant
ern
Rot
tnes
tTT
CC
B_T
rilogy
Rive
tte
AT
R409
BLN
3343
*CO
0401
CB-A
rgyl
eSu
rpas
s501
TT
44C
73D
unke
ld45
C75
Mys
ticA
TR-
Cob
bler
Rock
etC
L45
Y77
AT
R-B
eaco
n44
Y06
War
riorC
LF
linde
rsT
TC
Hyo
la60
ATR
-Mar
linP6
1703
-p74
-4A
G-O
utba
ckW
A05
0088
WA
0500
89Ex
pand
erR
ainb
ow69
1198
04-p
34Su
rpas
s603
CL
ZY0
15A
v-S
apph
ireC
harlt
onW
A05
0097
ZY0
0503
-p74
-6C
B-B
oom
erZ
Y016
RO
Y98
279
03-p
74-3
Fan
023
Yu17
8R
OY
9700
3Ch
on-n
amZ
hong
shu-
angN
0.4(
3087
2)C
olt
Lisor
a03
-p74
-11
ZY0
13A
-198
90Z
Y001
RO
Y97
019
ZY0
14A
TR-
Stu
bby
GAS
GO
TTT
ERI
(OO
)R9
06-P
71-1
Dra
kkar
GSC
-5P3
083
RO
Y33-
99P
5D
ing47
4T
atyo
onQ
u110
4W
esre
o06
-P71
-2M
arno
oZ
Y003
ZY0
08Z
Y009
RO
Y33-
99P
1R
OY
9701
7M
aluka
ZY0
12F
an16
8N
indoo
BLN
3343
*CO
0402
46C
76C
B_P
ilbar
aA
TR-B
anjo
Skip
ton
RO
Y97
033
Scoo
pY
icka
dee
Zho
ngsh
u-an
gN0.
4(30
920)
RO
Y97
018
RO
Y97
021
Wes
bark
erR
OY3
3-99
P6
Wes
roon
aW
A05
0096
Eur
eka
RS
O94
-67(
98R
OY3
3-99
P8
Gro
use
ZY0
04A
TR-H
yden
BLN
1990
Zho
ngyo
u-za
ngN
o.8
Zhon
gyou
_821
Ding
110
CB-
Tan
ami
Fan
189
Bar
ossa
6711
09H
yola
61M
ajor
Mut
u-98
-1Z
Y002
Wes
way
95*3
06-3
10R
OY
9709
6Ce
scalj
arni
Mon
tyS
hiral
eeA
G-E
mbl
emT
ranb
yR
OY
9833
0Ra
nge
05-P
36_R
RO
Y97
020
CB-T
rigol
dR
OY
9707
3Z
Y007
N00
1-28
-246
Karo
oR
OY3
3-99
P7
Samples
0
Principal Component Analysis Principal Component Analysis 50
05-P71-1146Y78
BLN1990 P624
ZY010
0671109
05-P36_R
06-P71-1
44C73
44Y06
45C75
45Y77
46C04
46C76
95*306 310
AG-Emblem
Ag-Muster
ATR409
ATR-Banjo
ATR-Barra
ATR-Beacon
ATR-Cobbler
ATR-Hyden
ATR-Marlin
ATR-Stubby
ATR-Summitt
Av-Opal
Barossa
BLN1990
BLN2737*CO0203
BLN2852-03W02
BLN3303*CR0302
BLN3343*CO0401
BLN3343*CO0402
BLN3614
Carousel-10
CB-Argyle
CB_Pilbara
CB-Tanami
CB_Trilogy
Cescaljarni
Ding110
Ding474
Drakkar
Dunkeld
Eureka
ExpanderFan168
Fan189
FlindersTTC
Grouse
GSC-5
Hyola50
Hyola60
Hyola61Hyola76
Karoo
Lantern
Monty
Mystic
N001-28-246
Norin_22
Oscar
P3083 Rainbow
Range
RivetteRocketCL
RottnestTTC
ROY31-99P1
ROY33-99P1
ROY33-99P4
ROY33-99P5
ROY33-99P6
ROY33-99P7
ROY33-99P8
ROY97017
ROY97018ROY97020
ROY97021
ROY97056
ROY97073
ROY97125
ROY98330
RSO94-67(98SARDI604
SARDI607
Shiralee
StormTT
Surpass501TT
Surpass603CL
Tarcoola
Tatyoon
TawrifficTT
ThunderTTTranby
WA050096
WarriorCL
Wesbarker
Wesbell
Wesroona
WeswayYickadee
Zhongshu-angN0.4(30920)
Zhongyou_821
Zhongyou-zangNo.8
ZY004
ZY007
0.5%
)PC
2
69119803-p74-11
03-p74-3
03-p74-4
03-p74-6
04-p34
06-P71-2
45C05
46C0495*306-310
A-19890 Ag-Comet
AG-Outback
Ag-Spectrum ATC93184-1
ATC94044-1
Av-JadeAv-Ruby
Av-Sapphire
BLN3347
BravoTTCB-Boomer
CB-TelferCB-Trigold
CBJ-001Charlton
Chon-namColt
Expander
Fan023GASGOTT HurricaneTTIwao-natane
Lisora
Major Maluka
Marnoo
Mutu-98-1Nindoo
OasisCL
P617Qu1104
ROY33-99P2
ROY33-99P3ROY97003
ROY97017ROY97019ROY97033
ROY97096
ROY98279
Scoop
Seetha
Skipton
Surpass400
Surpass603CLTERI(OO)R9
TornadoTT
WA050088WA050089
WA050097
WesreoYu178
Zhongshu-angN0.4(30872)
ZY001
ZY002
ZY003
ZY005ZY008
ZY009
ZY012
ZY013ZY014
ZY016
(10
-50
ATR-SignalSurpass400
Surpass402C
ZY015
80
90
-100 30
40
50
60
70
80
Eig
enva
lue
%
-100 -50 0 50 100 150PC1(67.6)
1 1.5 2 2.5 3 3.5 4 4.5 5 5.5Component
0
10
20
Non-metric Multi-Dimensional ScalingNon metric Multi Dimensional ScalingResemblance: D1 Euclidean distance
DistanceSurpass402C
2D Stress: 0.11
60Surpass402C
03-p74-6
06-P71-2
44C73
45C05
46C76ATC93184-1
ATR409 ATR-Banjo
ATR-Barra ATR-Beacon
BLN3343*CO0401BLN3343*CO0402
BravoTT
Carousel-10CB_PilbaraCB_Trilogy CharltonDunkeld Fan168
L t MarnooOasisCL
Oscar
Qu1104RivetteRottnestTTC
ROY33-99P1ROY97017
ROY97033ROY97056
SARDI604
Scoop
Skipton
Tarcoola
TawrifficTT
Wesreo
ZY005 ZY008ZY009
ZY012
ZY015
67110903-p74-11 03-p74-3
03-p74-4
05-P36_R06-P71-1
44C73
44Y06
45C75
45Y77
46C04
95*306-310 A-19890Ag-Comet
AG-Emblem
Ag-Muster
AG-OutbackAg-SpectrumATC94044-1 ATR-CobblerATR-HydenATR-Signal
ATR-StubbyATR-Summitt
Av-Jade
Av-Opal
Av-Ruby
Av-Sapphire
Barossa
BLN1990
BLN2852-03W02
BLN3303*CR0302
BLN3347
BLN3614
CB-Argyle
CB-Boomer
CB-Tanami
CB-TelferCB-Trigold
CBJ-001
Cescaljarni
Chon-namColt
Ding110
Ding474
DrakkarEureka
Expander
Fan023
Fan189
GASGOTT
Grouse
GSC-5HurricaneTT
Hyola61
Iwao-natane
Karoo
Lantern
LisoraMajor
Maluka
Marnoo
Monty
Mutu-98-1
Mystic
N001-28-246
NindooNorin_22
P3083
P617
RainbowRangeRocketCL
ROY31-99P1
ROY33-99P2
ROY33-99P3
ROY33-99P4
ROY33-99P5
ROY33-99P6ROY33-99P7
ROY33-99P8
ROY97003ROY97018ROY97019
ROY97020ROY97021
ROY97073
ROY97096
ROY97125
ROY98279
ROY98330
RSO94-67(98
SARDI607
Seetha
Shiralee
StormTT
Surpass501TT
Tatyoon
TERI(OO)R9
ThunderTTTornadoTT
Tranby
WA050088
WA050089
WA050096
WA050097
WarriorCLWesbarker
WesroonaWesway
Yickadee
Yu178
Zhongshu-angN0.4(30872)
Zhongshu-angN0.4(30920)
Zhongyou-zangNo.8
ZY001
ZY002
ZY003
ZY004
ZY007
ZY008ZY012
ZY013
ZY014ZY016
691198
04-p34
05-P71-11
46Y78
ATR-Marlin
BLN2737*CO0203FlindersTTC
Hyola50
Hyola60
Hyola76
P617P624
Surpass400
Surpass603CLWesbell Zhongyou_821
ZY010
Characterisation of APR linesCharacterisation of APR lines
Differential set was used to Differential set was used to characterise APR linesAll li i t t t ll All lines were resistant to all isolates
Loci remains to be characterised
Conclusions
Mapped and validated known Rlm d th l i i t d ithand other loci associated with
blackleg resistanceCharacterised germplasm for
blackleg resistance at seedling andblackleg resistance at seedling and adult plant stages– Identified new sources and loci for
blackleg resistance
AcknowledgmentsgCanola Molecular Marker Program
Collaborators •Australian Canola
I&I NSW•Rosy Raman
Marker Program•Simon Diffey (NSWDPI)•Steve Marcroft (MGP)•Kurt Lindbeck (I&I NSW)
Breeding Programs
•International Blackleg Consortium, C d•Rosy Raman
•Belinda Taylor•Ata Rehman
Kurt Lindbeck (I&I NSW)•Angela Van de Wouw (UM)•Paul Eckerman (UA)
Canada
Ata Rehman•Neil Coombes•David Luckett
•Ray Cowley (I&I NSW) •Dave Roberts (I&I NSW)
UQ•Jacqui Batley•David Edwards
•Shanoor Hossain (VicDPI)•Phil Salisbury (UM)B b (Vi DPI)•David Edwards
•Jiri Stiller•Bob (VicDPI)