In Search of New Effective Genes for Crown Rust In Search of New Effective Genes for Crown Rust Resistance in Resistance in Avena sterilisAvena sterilis

James ChongJames Chong and Gang Chen and Gang Chen

Cereal Research Centre, Agriculture & Agri-Food CanadaCereal Research Centre, Agriculture & Agri-Food Canada

Winnipeg, MB, Canada R3T 2M9Winnipeg, MB, Canada R3T 2M9

BREEDING FOR RESISTANCEBREEDING FOR RESISTANCE

By the end of 1950s,By the end of 1950s,

all deployable sources of resistance fromall deployable sources of resistance from

cultivated oat had been overcome incultivated oat had been overcome in

North America (Martens & Dyck 1989).North America (Martens & Dyck 1989).

A large pool of genes A large pool of genes

for resistance to crown for resistance to crown

rust was discovered in rust was discovered in

Avena sterilisAvena sterilis,, native to native to

the Middle East and the Middle East and

North Africa in1960s.North Africa in1960s.

0

10

20

30

40

50

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Year

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Pc38

Pc39

Pc48

Pc68

Rapid Adaptation of Rapid Adaptation of Puccinia coronataPuccinia coronata to toA. sterilisA. sterilis-derived Resistance Genes-derived Resistance Genes

Leggett(Pc68,94)

DumontRobertRiel +

(Pc38,39)

AC AssiniboiaRonald +

(Pc38,39,68)

Fidler(Pc39)

TripleCrown(Pc48)

HiFi(Pc91)

Spring OatSpring Oat Canadian PrairiesCanadian Prairies(Chong)(Chong)

Spring OatSpring Oat ND, SD, NE, IA, ND, SD, NE, IA, MN, WIMN, WI(Carson)(Carson)

Winter OatWinter Oat TX, LA, MS, FL, GA, TX, LA, MS, FL, GA, AL, SCAL, SC(Carson)(Carson)

0

20

40

60

80

100

Pc

38

Pc

39

Pc

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Pc

45

Pc

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Pc

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Pc

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Pc

51

Pc

52

Pc

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Pc

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Pc

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Pc

91

Pc

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Pc

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Pc

97

Gene

% V

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20

40

60

80

100

Pc

38

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Pc

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Pc

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Pc

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Pc

91

Pc

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Pc

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Pc

97

Gene

% V

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20

40

60

80

100

Pc

38

Pc

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Pc

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Pc

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Pc

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Pc

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Pc

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Pc

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Pc

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Pc

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Pc

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Pc

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Pc

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Pc

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Pc

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Pc

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Gene

% V

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20052005

Winter OatWinter Oat(TX, LA, MS, FL, (TX, LA, MS, FL, GA, AL, SC)GA, AL, SC)(Carson)(Carson)

Spring OatSpring OatND, SD, NE, IA, ND, SD, NE, IA, MN, WIMN, WI(Carson)(Carson)

0

10

20

30

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60

70

80

90

100

Pc

14

Pc

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Pc

36

Pc

53

Pc

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Pc

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Gene%

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20052005

Country of originCountry of origin No. of accessionsNo. of accessions

Middle EastMiddle East

Israel, Iran, Libya …Israel, Iran, Libya … 32003200

AfricaAfrica

Algeria, Ethiopia, Tunisia …Algeria, Ethiopia, Tunisia … 200200

EuropeEurope

Spain, Turkey, Morocco …Spain, Turkey, Morocco … 640640

Avena sterilisAvena sterilis Accessions Screened for Crown Rust Accessions Screened for Crown RustReactions in Greenhouse / Field since 1999Reactions in Greenhouse / Field since 1999

Avena sterilisAvena sterilis Accession CAV1980 (Algeria) Accession CAV1980 (Algeria)

with Adult-plant Resistancewith Adult-plant Resistance

Susceptible checkSusceptible check

Flag leafFlag leaf

Avena sterilisAvena sterilis Accession PI334961 (Israel) Accession PI334961 (Israel) immune to Crown Rustimmune to Crown Rust

Primary leavesPrimary leavesImmune to rustImmune to rust

Number of Number of Avena sterilisAvena sterilis accessions accessions

Selected for Genetic StudiesSelected for Genetic Studies

Country of originCountry of origin NumberNumber

Middle EastMiddle East

IsraelIsrael 1010

LebanonLebanon 11

AfricaAfrica

AlgeriaAlgeria 33

EuropeEurope

PortugalPortugal 11

ItalyItaly 11

Main ObjectiveMain Objective

Identify new effective crown rust Identify new effective crown rust

resistance sources, particularly those resistance sources, particularly those

expressed only at the adult-plant stageexpressed only at the adult-plant stage

Materials & MethodsMaterials & Methods

• crosses/backcrosses to a susceptible cultivar.crosses/backcrosses to a susceptible cultivar.

• FF11 and BCF and BCF11 and progenies raised in growth and progenies raised in growth

chambers.chambers.

• panicles bagged to prevent out-crossing.panicles bagged to prevent out-crossing.

• rust tests at seedling and adult-plant stages to rust tests at seedling and adult-plant stages to

identify adult-plant resistance.identify adult-plant resistance.

INOCULUMINOCULUM

• greenhouse - individual crown rust isolates/races greenhouse - individual crown rust isolates/races

to identify number of genes involved.to identify number of genes involved.

• greenhouse and field - composites of crown rust greenhouse and field - composites of crown rust

isolates collected from annual surveys of isolates collected from annual surveys of

previous year to identify effective gene(s).previous year to identify effective gene(s).

Flood at the Glenlea Oat Nursery - 2005Flood at the Glenlea Oat Nursery - 2005

BeforeBefore AfterAfter

Harmon x PI 334961 (Israel)Harmon x PI 334961 (Israel)

No. of No. of FF33 lines lines

InoculumInoculum ResRes Seg (IT)Seg (IT) SusSus RatioRatio XX22 PP

CR181CR181 1717 23 (0 IT)23 (0 IT) 22 7:8:17:8:1 0.44220.4422 0.7-0.50.7-0.5

Bulk03Bulk03 1111 2121 1010 1:2:11:2:1 0.04760.0476 0.9-0.80.9-0.8

Bulk04Bulk04 1111 2121 1010 1:2:11:2:1 0.04760.0476 0.9-0.80.9-0.8

- - two dom. genes resistant to CR181.two dom. genes resistant to CR181.

- only one was effective to bulk03 and bulk04.- only one was effective to bulk03 and bulk04.

- ineffective gene most likely - ineffective gene most likely Pc48.Pc48.

HarmonHarmon22 x PI333699 (Israel) x PI333699 (Israel)

No. of plants or familiesNo. of plants or families

BCFBCF11 Resistant (IT)Resistant (IT) SusSus RatioRatio XX22 PP

- CR181- CR181 34 (0)34 (0) 1111 3:13:1 0.00740.0074 0.95-0.900.95-0.90

BCFBCF22 Segregating (IT)Segregating (IT) SusSus

- CR181- CR181 34 (0) 6 (;-;1)34 (0) 6 (;-;1) 55 6:1:16:1:1 0.09630.0963 0.98-0.950.98-0.95

- Bulk03- Bulk03 24 (0)24 (0) 2121 1:11:1 0.20.2 0.70-0.500.70-0.50

- one rec. and one dom. gene resistant to CR181.- one rec. and one dom. gene resistant to CR181.

- dom. gene effective to Bulk03.- dom. gene effective to Bulk03.

- within segregating lines, 462 res : 132 sus plants (X- within segregating lines, 462 res : 132 sus plants (X22 = 2.444, = 2.444, P P = 0.2-0.1).= 0.2-0.1).

HarmonHarmon22 x CAV1979 (Algeria) x CAV1979 (Algeria)

No. of familiesNo. of families

IsolateIsolate Segregating (IT)Segregating (IT) SusSus RatioRatio X2X2 PP

CR181CR181 43 (0;-;1-)43 (0;-;1-) 5757 1:11:1 0.25960.2596 0.70-0.500.70-0.50

CR240CR240 43 (0-;)43 (0-;) 5757 1:11:1

CR205CR205 43 (0;-;11+)43 (0;-;11+) 5757 1:11:1

CR192CR192 43 (0;-;1-)43 (0;-;1-) 5757 1:11:1

CR225CR225 43 (;1-12)43 (;1-12) 5757 1:11:1

CR223CR223 43 (;1-11+)43 (;1-11+) 5757 1:11:1

- - either a single gene (either a single gene (tempPc98tempPc98) or two tightly linked genes conditioned resistance.) or two tightly linked genes conditioned resistance.

- no putative recombinant was found in large number of segregating progenies. - no putative recombinant was found in large number of segregating progenies.

- resistant lines had 70MR reaction in New Zealand 2004-05.- resistant lines had 70MR reaction in New Zealand 2004-05.

HarmonHarmon22 x CAV1180 (Algeria) x CAV1180 (Algeria)

No. of plants or familiesNo. of plants or families

BCFBCF11 Resistant (IT)Resistant (IT) SusSus RatioRatio XX22 PP

- CR181- CR181 52 (;) 10 (;11+)52 (;) 10 (;11+) 3333 3:13:1 4.80344.8034 <0.05<0.05

BCFBCF22 Segregating (IT)Segregating (IT) SusSus

- CR181- CR181 71 (0;-;1)71 (0;-;1) 2222 7:17:1 10.582210.5822 <0.01<0.01

- CR223- CR223 61 (0;-;1)61 (0;-;1) 3232 3:13:1 4.39074.3907 0.05-0.020.05-0.02

- Bulk04- Bulk04 61 (0;-;1)61 (0;-;1) 3232 3:13:1 4.39074.3907 0.05-0.020.05-0.02

- CR225- CR225 40 (;-;1)40 (;-;1) 5252 1:11:1 1.56521.5652 0.30-0.200.30-0.20

- CR13- CR13 23 (;-;1)23 (;-;1) 7171 1:31:3 0.01410.0141 0.900.90

- one rec. and two loosely linked (dom. and p. dom) genes resistant to CR181.one rec. and two loosely linked (dom. and p. dom) genes resistant to CR181.

- p. dom. gene ineffective to CR223 and bulk04.p. dom. gene ineffective to CR223 and bulk04.

- rec. gene (rec. gene (pc97temppc97temp) ) resistant to CR225 and bulk04, suppressed by an inhibitor. resistant to CR225 and bulk04, suppressed by an inhibitor.

- pc97temppc97temp: 10 MR (Canadian Prairies); 10 R (New Zealand).: 10 MR (Canadian Prairies); 10 R (New Zealand).

AC Morgan x CAV1980 (Algeria)AC Morgan x CAV1980 (Algeria)

No. of No. of FF22 plants & F plants & F33 lines lines

Res (IT)Res (IT) SegSeg SusSus RatioRatio X2X2 PP

CR225CR225

FF22 seedlings seedlings 00 …… 127127

FF22 adults adults 32 (;1-;2)32 (;1-;2) …… 9595 1:31:3 0.00260.0026 0.98-0.950.98-0.95

FF33 lines lines 3535 5656 3232 1:2:11:2:1 1.13011.1301 0.70-0.500.70-0.50

CR181CR181

FF33 seedlings seedlings 2626 7070 3131 1:2:11:2:1 1.72441.7244 0.50-0.300.50-0.30

2004 field2004 field - F - F33 34 (0-20 RMR)34 (0-20 RMR) 93 93 1:31:3 0.21250.2125 0.70-0.500.70-0.50

Lines with rec. adult-plant resistance gene (Lines with rec. adult-plant resistance gene (pc99temppc99temp) identified by CR225 were ) identified by CR225 were

resistant in 2004 field test. resistant in 2004 field test.

Dom. gene resistant to CR181 was ineffective in the field.Dom. gene resistant to CR181 was ineffective in the field.

Summary of Crown Rust Resistance Genes IdentifiedSummary of Crown Rust Resistance Genes Identified

Accession &Accession & FieldField No. of genes*No. of genes*   

originorigin reactionreaction identifiedidentified effectiveeffective CommentsComments

IsraelIsrael

CAV1339CAV1339 trMSStrMSS 2 (D, r)2 (D, r)

PI333463PI333463 00 2 (D, r)2 (D, r) 1 (r)1 (r)

PI333699PI333699 00 2 (D, r)2 (D, r) 1 (D)1 (D)

PI334961PI334961 0-trS0-trS 2 (D)2 (D) 1 (D)1 (D)

PI374915PI374915 2MRMS-10MR2MRMS-10MR 2 (D)2 (D) 1 (D)1 (D) Field (10 RMR)Field (10 RMR)

PI375695PI375695 trMRMS-5MRtrMRMS-5MR 1 (r)1 (r) 1 (r)1 (r)

PI378740PI378740 10RMR-trS10RMR-trS 2 +2 + 1(r)1(r)

PI404589PI404589 5RMR-trS5RMR-trS 1 (D)1 (D) 1(D)1(D) inhibitorinhibitor

LebanonLebanon

   CAV1180CAV1180    2(D) 1(r)2(D) 1(r) 1(D) 1(r)1(D) 1(r) inhibitor of 'r‘; Field (10MR)inhibitor of 'r‘; Field (10MR)

*D = dominant or near-dominant, r = recessive or near-recessive.*D = dominant or near-dominant, r = recessive or near-recessive.

Summary of Crown Rust Resistance Genes Identified (cont.)Summary of Crown Rust Resistance Genes Identified (cont.)

AccessionAccession FieldField No. of genesNo. of genes

& origin& origin reactionreaction IdentifiedIdentified EffectiveEffective CommentsComments

AlgeriaAlgeria

CAV1975CAV1975 2 (D, r)2 (D, r) 1 (D)1 (D)

CAV1979CAV1979    1? (D-r)1? (D-r) 1? (D-r)1? (D-r) Field ( MR)Field ( MR)

CAV1980CAV1980 5RMR5RMR 2 (D, APR)2 (D, APR) 1 (APR)1 (APR) APR field (0-5RMR)APR field (0-5RMR)

PortugalPortugal

IB2433IB2433 1 (D), 2 (linked r)1 (D), 2 (linked r) 1 (r)1 (r) inhibitor of 'r'inhibitor of 'r'

ItalyItaly

CAV2568CAV2568 60R60R 1 (D) 2(r)1 (D) 2(r) 1 (D) 1(r)1 (D) 1(r)

PI497690PI497690 trR-5RMRtrR-5RMR 3?(0, ;1, 2 ITs)3?(0, ;1, 2 ITs) 1+ ?1+ ? progenies mostly sterileprogenies mostly sterile

(MN788788, Amagalon)(MN788788, Amagalon) inhibitor present ?inhibitor present ?

*D = dominant or near-dominant, r = recessive or near-recessive*D = dominant or near-dominant, r = recessive or near-recessive

GENERAL SUMMARYGENERAL SUMMARY

• generally at least two genes present in each generally at least two genes present in each

accession.accession.

• composite of isolates useful for identifying the composite of isolates useful for identifying the

putative, effective gene(s) in greenhouse tests.putative, effective gene(s) in greenhouse tests.

• effective adult-plant resistance rare.effective adult-plant resistance rare.

• gene inhibitors common.gene inhibitors common.

CURRENT EFFECTIVE CROWN RUST CURRENT EFFECTIVE CROWN RUST

RESISTANCE SOURCESRESISTANCE SOURCES

• Pc53 – A. sterilisPc53 – A. sterilis

• Pc91Pc91 – amagalon material – amagalon material

• Pc94Pc94 – – A. strigosaA. strigosa

• Pc96Pc96 – – A. sativaA. sativa

• Adult-plant resistance from MN841801Adult-plant resistance from MN841801

FURTHER STUDIESFURTHER STUDIES

• confirm segregation of genes in field studies.confirm segregation of genes in field studies.

• isolate genes in single-gene lines & gene isolate genes in single-gene lines & gene

postulation.postulation.

• verify effectiveness in multi-locations. verify effectiveness in multi-locations.

• determine allelic/linkage relationships of genes determine allelic/linkage relationships of genes

with known with known PcPc genes genes

Financial support provided in part from:Financial support provided in part from:- AAFC Matching Investment Initiative- AAFC Matching Investment Initiative

- Agriculture Research Development Initiative- Agriculture Research Development Initiative

- The Quaker Oats Company- The Quaker Oats Company

- The North American Millers Association- The North American Millers Association

Technical assistance:Technical assistance: Rebecca Dueck and Wayne MayertRebecca Dueck and Wayne Mayert

Avena sterilisAvena sterilis accessions from: accessions from:- - Original collection at AAFC, WinnipegOriginal collection at AAFC, Winnipeg

- National Small Grains Collection, Aberdeen- National Small Grains Collection, Aberdeen

- Plant Gene Resources of Canada, Saskatoon- Plant Gene Resources of Canada, Saskatoon

ACKNOWLEDGEMENTSACKNOWLEDGEMENTS