evolution de novo of stable, heritable resistance to leaf rust ( puccinia triticina eriks.) in...
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Evolution de novo of stable, heritable resistance to leaf rust(Puccinia triticina Eriks.) in susceptible wheat following
cycles of infection with WSMV (Wheat Streak Mosaic Virus)
Steve Haber1, D.L. Seifers2, P. Seto-Goh1 & B. McCallum1 1AAFC-Cereal Research Centre, 195 Dafoe Rd., Winnipeg MB R3T 2M92Agricultural Research Center, KSU-Hays, Hays KS 67601-9228, U.S.A.
……Yeah, looks good, Yeah, looks good, farmers like it,farmers like it, low polyphenol oxidase,low polyphenol oxidase, makes makes greatgreat Korean noodles… Korean noodles…
……BUT, terrible rust resistance,BUT, terrible rust resistance, and it sure gets hammered and it sure gets hammered by WSMV…by WSMV…
Whadda we gonna do???Whadda we gonna do???
Wheat streak mosaic Wheat streak mosaic virusvirus
• naturally transmitted by wheat curl mite (Aceria tosichella)
• experimentally transmitted by rub-inoculation
6
The The Tiller EffectTiller Effect
Variable expressionexpression of traits (e.g. resistance to WSMVWSMV) in the same plant
…expansion ofrange of variation??
ExtremeExtreme variation variation in response to WSMVWSMV:Generation grown from seed experiencing ‘Tiller Effect’‘Tiller Effect’
Could we harness this Could we harness this heritable variation in heritable variation in expression to expression to evolveevolve resistance to resistance to Leaf RustLeaf Rust de novode novo??????
Lakin‘s progressLakin‘s progress KSU-ARCH
‘‘natural WSM’natural WSM’ 11stst cycle cycle
100 healthy plants100 WSMWSM(x1)(x1) plants
( < 1% of plants in field of plants in field affected byaffected by WSM )
Seed from WSMWSM(x1)(x1) plants Seed from healthyhealthy plants
FieldField2001-022001-02
Lakin‘s progressLakin‘s progressFieldField
2001-022001-0211stst cycle cycle
22ndnd cycle cycle IndoorsIndoors2002-032002-03
Grow out plants
rub-inoculate @ 2-leaf stage
vernalize 6 weeks
572 surviving WSMWSM(x2)(x2) plants
572 WSMWSM(x2)(x2) sublines
Seed from WSMWSM(x1)(x1) plants
Lakin‘s progressLakin‘s progress
11stst cycle cycle
22ndnd cycle cycle
33rdrd cycle cycle
572 WSMWSM(x2)(x2) sublines
572 headrows Healthy controls
((littlelittle) ‘natural disease pressure’) ‘natural disease pressure’* Harvest seed from* Harvest seed from
200 headrows 200 rows
169g bulk seed Bulk seed
Seed from WSMWSM(x1)(x1) plants FieldField2001-022001-02
IndoorsIndoors2002-032002-03
FieldField2003-042003-04
12
Lakin‘s progressLakin‘s progress
33rdrd cycle cycle
44thth cycle cycle
169g bulk seed Bulk seed
2m x 60m plot 2m x 60m plotheavy disease pressureheavy disease pressure
* * Leaf RustLeaf Rust* * Stripe RustStripe Rust
** Identify individuals resisting Identify individuals resisting
RR11 RR22 RR33 RR44 RR55
RR11------n/an/a------
------n/an/a------
R1R1
FieldField2003-042003-04
FieldField2004-052004-05
Lakin‘s progressLakin‘s progress
Leaf Rust }Leaf Rust }
44thth cycle cycle
55thth cycle cycle2005-062005-06
indoorsindoors
RR11 Stripe Rust}Stripe Rust} resistantresistant
cont‘dcont‘d
34 RR11 seedvernalize 6 weeks
34 ’R’R1’1’ plants
30 RR11 seed
transplant21 ‘R‘R1’1’ plants
55 ’R’R1’1’ plants
survivors
grown indoors to maturitygrown indoors to maturity
55 ’R’R1-1-derivedderived’’ sublines
ProgressiveProgressiveNecrosisNecrosis5 plants
#2 #3 #1 #4‘Fred’
FieldField2004-052004-05
FieldField
Lakin‘s progressLakin‘s progress
2006-072006-0766thth cycle cycle
Indoor seedling testsIndoor seedling tests Field TestField Test
cont‘dcont‘d
55 ’R’R1-1-derivedderived’’ sublines
Leaf RustLeaf Rust
Stripe RustStripe Rust
allSusc.
12
mixed
35
allRes.
8855
allSusc.
mixedallRes.
Necrosis
12 35 882 22 28283 31 2121
#3‘Fred’ Lakin Lakin (orig.)
Lakin Lakin v R1R1-descendants-descendants
LineKSU-ARCHKSU-ARCH
Stripe Rust
Field test
KSU-ARCHKSU-ARCHL.Rust (MKPS)
Field test
CRC-WpgCRC-WpgLR (TDBG)
Seedling testNecrosis
Lakin (1-7-5) S S 15/15mesothetic (;/3+)
NO
2-6-2 n/a n/a 15/15Resistant (0;)
Uniform
2-4-5 R R 10/10Resistant (0;)
NO
4-3-3 R S8 mesothetic (;/3+)
7 Susceptible (3+)NO
4-12-3 R R13 Resistant (0;)
6 mesothetic (;/3+)
1 Susceptible (3+)
NO
...OK…Evolving ...OK…Evolving resistance to resistance to Leaf RustLeaf Rust de novode novo looks promising looks promising forfor LakinLakin……
...BUT…Will it work for ...BUT…Will it work for other wheats, including other wheats, including spring wheats???spring wheats???
...Let’s try it with...Let’s try it with Thatcher !?Thatcher !?
Thatcher‘s progressThatcher‘s progress
4-wk4-wk ‘quasi’vernalizationvernalization
11stst cycle cycle
CRC-Winnipeg
10 Thatcher (spring wheat) seeds
10 Thatcher seedlingsrub-inoculate WSMV WSMV @ tillering
Tillers expressing WSMV-resistanceWSMV-resistance
qvA071A071ws1.2m
IndoorsIndoors20082008
20
Thatcher‘s progressThatcher‘s progress
11stst cycle cycle
22ndnd cycle cycle
qvA071A071ws1.2m
Altered response to WSMVWSMV
30 seeds
30 seedlingsrub-inoculate WSMVrub-inoculate WSMV @ tillering
Tillers expressing WSMV-resistanceWSMV-resistance
A267A267ws3.1tB
IndoorsIndoors20082008
IndoorsIndoors2008-092008-09
21
Thatcher‘s progressThatcher‘s progress
A267A267ws3.1tB
IndoorsIndoors2008200811stst cycle cycle
22ndnd cycle cycle IndoorsIndoors2008-092008-09
33rdrd cycle cycle FieldField20092009
Altered response to WSMVqvA071A071ws1.2m
Altered response to WSMV
Head-row (21 seeds) FHBFHB nursery nurseryerFH09_i011erFH09_i011
SelectSelectspecificspecificheadsheads
A505A505
22
Thatcher‘s progressThatcher‘s progress44thth cycle cycle IndoorsIndoors
2009-102009-10
cont‘dcont‘d
A505A50524 seeds
24 seedlingsrub-inoculate WSMV WSMV @ tillering
ThatcherThatcherBreeder‘s seedBreeder‘s seed
ThatcherThatcherA505A505
22/24 plants express WSMV-resistanceWSMV-resistance
14
10
ThatcherThatcherA505ws1.07A505ws1.07
TDBGTDBG Leaf Rust Leaf Rust
21/24expressexpressAdult-plantAdult-plantResistanceResistance
Thatcher‘s progressThatcher‘s progress
44thth cycle cycle
55thth cycle cycle
cont‘dcont‘d
de novo WSMV resistancede novo WSMV resistancede novo Leaf Rust de novo Leaf Rust (TDBG)(TDBG) resistance resistance
A505A505rw1.07m
A699A699
14 seedlings
TPMKTPMK Stem Rust Stem Rust@ 2-leaf stage
A699rs1.2
Thatcher2+2+
;1-;1-
TDBGTDBG Leaf Rust Leaf Rust@ Adult-plant stage
Thatcher33-33-
A699rs1.20;0;
11/14 11/14 (more) (more) ResistantResistant
11/14 Resistant11/14 Resistant
IndoorsIndoors2009-102009-10
IndoorsIndoors20102010
Thatcher‘s progressThatcher‘s progress
55thth cycle cycle
66thth cycle cycle
cont‘dcont‘d
de novode novo Leaf Rust Leaf Rust ((TDBGTDBG)) resistance resistance (Adult plant)(Adult plant)de novode novo Stem Rust Stem Rust ((TPMKTPMK)) resistance resistance (seedling)(seedling)
A699rs1.2
A829A829
10 seedlings
0TDBGTDBG Leaf Rust Leaf Rust@ 2-leaf stage
rub-inoculate WSMV WSMV @ tillering
@ Adult-plant stage
IndoorsIndoors20102010
IndoorsIndoors20102010
Thatcher3+3+
A829rs1.71-2=1-2=
TDBGTDBG Leaf Rust Leaf Rust0
A829rs1.7;1-2=;1-2=
...OK…Evolving ...OK…Evolving resistance to resistance to Leaf RustLeaf Rustde novode novo appears to work appears to workforfor LakinLakin……ANDAND…… ThatcherThatcher……
BUT…it would sure helpBUT…it would sure helpthe case if we had an ideathe case if we had an ideahow the how the de novode novo Leaf RustLeaf Rust resistance is resistance isinherited !?inherited !?
...Yeah…...Yeah…
Gotta make crossesGotta make crossesand look at progeny!and look at progeny!
28
Inheritance of Inheritance of de novode novo LRLR-resistance-resistance
LineKSU-ARCHKSU-ARCH
Stripe Rust
Field test
KSU-ARCHKSU-ARCHL.Rust (MKPS)
Field test
CRC-WpgCRC-WpgLr (TDBG)
Seedling testNecrosis
4-3-3 R S8 mesothetic (;/3+)
7 Susceptible (3+)NO
4-12-3 R R13 Resistant (0;)
6 mesothetic (;/3+)
1 Susceptible (3+)
NO
......choosing parents for a cross
4-3-3 4-12-3
9935 9938
0
9935rs1.6 9938rw1.2(;1-/3+ mesothetic) (0; Resistant no WSMWSM symptoms)
XX
A124 (F1) (24/24 0;)LrLr--TDBGTDBG Resistant
Seed
Seedlings
LrLr--TDBGTDBG Leaf RustLeaf Rust
@ 2-leaf stage
WSMVWSMV@ tillering
(3/8 no WSMWSM symptoms)WSMV WSMV Resistant
A124 (F1)
Inheritance of Inheritance of de novode novo LrLr-resistance-resistance...Reassortment in daughter generations...Reassortment in daughter generations
(24/24 0;)
Resistant
MKPSMKPS
78 Res.29 Susc.
~ 3 : 1
TDBGTDBG
100 Res.7 mesothetic
18 Susc. (3+)
~ 3 : 1
F2
A124ws3.2tA
MKPSMKPS
F2 plants
@ 2-leaf stage
9 Res.4 Susc.
TDBGTDBG
11 Res.2 mesothetic1 Susc. (3+)
Seed from F1 plant
WSMVWSMV @ tillering
A368ws1.1mLr-Lr-TDBGTDBG Resistant
WSMV WSMV Resistant
LrLr--TDBGTDBG
Inheritance of combined Inheritance of combined de novode novoLr Lr & WSMVWSMV ResistanceResistance
...Example of selection for ...Example of selection for true-breedingtrue-breeding line line
A368ws1.1m
F3 plants
TDBGTDBG@ 2-leaf stage
15/15 (0;)
WSMVWSMV @ tillering
1/3 Res.
25 seeds 25 seeds toKSU-ARCH
2009/10 Trial Row14
Seed from F2 plant
A486
Lakin A486A486
LakinA486ws1.1A486ws1.1
Inheritance of combined Inheritance of combined de novode novoLr Lr & WSMVWSMV ResistanceResistance
...Example of selection for true-breeding line...Example of selection for true-breeding line
WSMVWSMV Half-row rub-inoculated Fall 2009Fall 2009
KSU-ARCHKSU-ARCH2009/10 Trial Row14
F3 plants
Seed from F2 plant
April 12 April 12 (2010)(2010)
Inheritance of combined Inheritance of combined de novode novoLr Lr & WSMVWSMV ResistanceResistance
...Example of selection for true-breeding line...Example of selection for true-breeding line
WSMVWSMV Half-row rub-inoculated Fall 2009Fall 2009
KSU-ARCHKSU-ARCH2009/10 Trial Row14
F3 plants
Seed from F2 plant
April 26April 26
Inheritance of combined Inheritance of combined de novode novoLr Lr & WSMVWSMV ResistanceResistance
...Example of selection for true-breeding line...Example of selection for true-breeding line
WSMVWSMV Half-row rub-inoculated Fall 2009Fall 2009
KSU-ARCHKSU-ARCH2009/10 Trial Row14
F3 plants
Seed from F2 plant
May 11May 11
Inheritance of combined Inheritance of combined de novode novoLr Lr & WSMVWSMV ResistanceResistance
...Example of selection for true-breeding line...Example of selection for true-breeding line
WSMVWSMV Half-row rub-inoculated Fall 2009Fall 2009
KSU-ARCHKSU-ARCH2009/10 Trial Row14
F3 plants
Seed from F2 plant
May 27May 27 Lakin Row14 8/8 Resistant
Inheritance of combined Inheritance of combined de novode novoLr Lr & WSMVWSMV ResistanceResistance
...Example of selection for true-breeding line...Example of selection for true-breeding line
WSMVWSMV Half-row
rub-inoculated Fall 2009Fall 2009
8/8 Resistant
KSU-ARCHKSU-ARCH2009/10 Trial Row14
F3 plants
Natur
al
Natur
al
inocu
lum
inocu
lum
Stripe RustStripe RustRow14 Lakin
Seed from F2 plant
May 27May 27
Inheritance of combined Inheritance of combined de novode novoLr Lr & WSMVWSMV ResistanceResistance
...Example of selection for true-breeding line...Example of selection for true-breeding line
WSMVWSMV Half-row
rub-inoculated Fall 2009Fall 2009
8/8 Resistant
KSU-ARCHKSU-ARCH2009/10 Trial Row14
F3 plants
0 Natur
al
Natur
al
inocu
lum
inocu
lum
Stripe RustStripe RustRow14 Lakin
Seed from F2 plant
May 27May 27
Lakin
Row14
Leaf RustLeaf Rust
June 3June 3
SummarySummary
Lakin ‘Row 14Row 14’
Leaf RustLeaf RustField test
(MKPS) (TDBG)
Stripe RustStripe RustField test
Seedling
WSMVWSMVIndoor test
Field test
MS(2) R(0)S(3+)S(3+)
R(0)R(0;)
S R
SS
RR
Thatcher‘A829A829’
Leaf RustLeaf Rust
(TDBG)Seedling
AdultR(0;1=)
WSMVWSMVIndoor testR S
Stem RustStem RustSeedling (TPMK)MR(;1=) MS(2+)
S(3+)
S(3+)MR(;12=)
ConclusionsConclusions
Epigenetics Epigenetics discoversdiscovers & & identifiesidentifies … …GeneticsGenetics fixesfixes && exploitsexploits
Modifying expression Modifying expression of existing gene alleles affords a new toola new tool to complement gene introgression gene introgression
SusceptibleSusceptible progenitor progenitor
ResistantResistant descendants descendants
Cyclesof
selfing
Selection and advanceunder pressure fromsystemic stress(es)
40
AcknowledgementsAcknowledgementsBrian Gillis
Jeff AckermanPat Seto-Goh
Taye Zegeye
KANSAS WHEAT COMMISSIONKANSAS WHEAT COMMISSION
for fundingfor fundingsupportsupport
……andand