evolution de novo of stable, heritable resistance to leaf rust ( puccinia triticina eriks.) in...

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