Race-typing in North American Rust Laboratories & Their Roles in the Global Cereal Rust Monitoring System

Yue Jin Cereal Disease Laboratory (Cereal Rust Lab)

US Department of Agriculture-Agricultural Research Service St. Paul, Minnesota, USA

Tom Fetch, Jr. Cereal Research Centre (The Dominion Rust Research Lab)

Agriculture and Agri-Food Canada Winnipeg, Manitoba, Canada

Functions of the NA Rust Labs

Pathogen biology, genetics, disease epidemiology: •  Puccinia graminis •  Puccinia triticina •  Puccinia coronata •  Puccinia hordei •  Fusarium graminearum

Host resistance: •  Sources and genetics of rust resistance •  Developing new germplasm •  Rust resistance breeding

Training: •  In collaborations with universities

Functions of the NA Rust Labs

Systematic surveys of cereal rusts since 1920s •  New virulences & implications to crop production •  Race dynamics & causes •  Rust epidemiology: inoculum, movement, environments •  Genetic resources: population genetics, genomics

isolates for germplasm screening

National Rust Surveys

Stem Rust Races Used in Screening Race Utility Avirulence Virulence

MCCFC Panel/historical 6 8a 9b 9d 9e 11 24 30 31 36 38 5 7b 9a 9g 10 17 Tmp McN

QFCSC Panel/dominant 6 7b 9b 9e 11 24 30 31 36 38 Tmp 5 8a 9a 9d 9g 10 17 21 McN

QTHJC Panel/unique v 7b 9a 9e 24 30 31 36 Tmp 5 6 8a 9b 9d 9g 10 11 17 21 38 McN

RCRSC Panel 6 8a 9e 11 24 30 31 Tmp 5 7b 9a 9b 9d 9g 10 17 21 38 McN

RKQQC Panel/unique v 9e 10 11 17 24 30 31 38 Tmp 5 6 7b 8a 9a 9b 9d 9g 21 McN

TPMKC Panel/historical 6 9a 9b 24 30 31 38 5 7b 8a 9a 9d 9e 9g 10 11 17 21 36 Tmp McN

TTTTF Panel/broad v 24 31 5 6 7b 8a 9a 9b 9d 9e 9g 10 11 17 21 30 36 38 McN

QCCSM Unique v-Sr24 6 7b 8a 9b 9e 11 30 31 36 38 Tmp 5 9a 9d 9g 10 17 21 24 McN

SCCSC Unique v-Sr9e+13 6 7b 8a 9b 11 24 30 31 36 38 Tmp 5 9a 9d 9e 9g 10 17 21 McN

TTKSK Significant v 24 36 Tmp 5 6 7b 8a 9a 9b 9d 9e 9g 10 11 17 21 30 31 38 McN

TTKST Significant v 36 Tmp 5 6 7b 8a 9a 9b 9d 9e 9g 10 11 17 21 24 30 31 38 McN

TTTSK Significant v 24 Tmp 5 6 7b 8a 9a 9b 9d 9e 9g 10 11 17 21 30 31 36 38 McN

RRTTF Significant v 8a 9e 24 31 5 6 7b 9b 9d 9g 10 11 17 21 30 36 38 McN

TRTTF Significant v 8a 24 31 5 6 7b 9b 9d 9e 9g 10 11 17 21 30 36 38 McN

Rust Pathogen Deposits at CDL

30,800 accessions in liquid nitrogen storage, representing: 19 different genera 69 different species 53 different countries Collections dating back to 1956

~1,000 accessions in temporary storage

Facilities & Operating Seasons

Laboratory Race-typing season for foreign rust collections ARS CDL: December to February ARS MD: Year-round, receiving and reviving samples UMN-BLS3 November to March-mostly Ug99 screening Morden Lab October to April Ottawa Lab: October to April (with year-round potential)

Sampling

Inoculating differentials

Deriving pure isolates

Race-typing

Confirmation

Testing on additional SR genes

Procedure in Race-typing

Permanent culture collections  

Seedling Infection Types

Low IT High IT

1 ;1 ;1- ; 0; 4 3 2+ 2

Ug99 Race Group Set Line Sr gene 04KEN156 06KEN19v 09ZIM01-4 07KEN24-1 1 ISr5Ra 5 4 4 4 4 T.m.deri. 21 3+ 3+ 3 3+ Vernstein 9e 4 4 4 4 ISr7bRa 7b 3 3+ 4 4

2 ISr11Ra 11 4 4 4 4 ISr6Ra 6 4 4 4 4 ISr8Ra 8a 4 4 4 4 CnsSr9g 9g 4 4 4 4

3 W2691SrTt-1 36 0 0 0 4 W2691Sr9b 9b 4 4 4 3+ BtSr30Wst 30 4 3+ 4 4 Comb. VII 17(+13) 2++ 2++ 2+ 2++

4 ISr9aRa 9a 4 4 4 4 ISr9dRa 9d 4 4 4 4 W2691Sr10 10 4 4 4 4 CnsSrTmp Tmp 2+ 2+ 2+ 2+ 5  LcSr24Ag 24 2 3 2 2 Sr31/LMPG 31 4 4 1 4 VPM1 38 4 4 4 4 McNair701 McN 4 4 4 4

Race: TTKSK TTKST TTKSF TTTSK

Set Line Sr gene Virulence 6 SwSr22T.B. 22 - Agatha/9*LMPG 25 + Eagle (Aus) 26 - 73,214,3-1/9*LMPG 27 +

7 Federation*4/Kavkaz 31 + ER 5155 32 - Tetra Canthatch/Ae. sqr 33 (+) Mq(2)5XG2919 35 +

8 W3563 37 + RL6080 39 - RL6088 40 - TAF 2 44 (+)

9 DAS15 47 - Satu Satu + TAM 107 1A.1R + Fed*3/Gabo*51BL_1RS-1 R (+?) 10 Iumillo 9g,12,+ - Leeds 9e,13,+ - ST464 13 + Q21861 Rpg1,4,5 - 11 Siouxland 24+31 + X08069B-F8-10-1 24+31 + Sisson 31+36 + X08067B-F8-16-3 31+36 +

Additional Sr-genes Tested

Stem Rust Samples Race-typed at CDL Country Season Isolates/samples Race Identified Target (sampling) Kenya 2004-2013 ~300/500 TTKSK, TTKST, TTTSK Njoro, Mt Kenya Ethiopia 2005-2013 ~288/210 TTKSK, TRTTF, JRCQC Debre Zeit, N Shewa Yemen 2006 ~8/30 TTKSK, TRTTF (Yarim Wadi Arab) Sudan 2006 ~2/26 TTKSK (New Halfa) Tanzania 2006,2009 39/20 TTKSK, TTKST, TTTSK Northern region Pakistan 2011-13 2/150 RRTTF National survey

Turkey 2013 7/18 TKNTC, TK_TC (?Sr9b:P,T) Nursery samples Chile 2011 6 QFCSC Culture collection Uruguay 2013 20 QTTTF, RHTTC, SPLKC… Culture collection

Stem Rust Samples Race-typed at AAFC Country Season Isolates/samples Race Identified Target (sampling) Kenya 2007-2012 87/277 TTKSK, TTKST, TTTSK, Njoro, Survey

PTKST, PTKSK Ethiopia 2007-2010 94/144 TTKSK, RRTTF, SRHQF Debre Zeit, Survey

PTKSK, PTKST, SRHSF Yemen 2007-2010 65/212 TTKSK, PTKSK, SRHQF Survey

RRTTF, SRHSF Pakistan 2009 8/20 RRTTF National survey

Eritrea 2010-2012 46/150 TTKST, PTKST, JRCSC National survey

JRHSC, JRHSF

Stem Rust (D) Samples Genotyped Country Year Samples UG99-RG + Predicted races Kenya 2011-13 152 131 TTKSK, TTKST, TTTSK Ethiopia 2012-13 93 62 TTKSK, TTTSK Uganda 2012 52 39 TTKSK, TTKST, TTTSK Eritrea 2012 23 1 TTKSK Zimbabwe 2011-12 42 33 TTKSF, PTKST South Africa 2012 26 9 TTKSF Yemen 2011 25 14 TTKSK Iraq 2011 3 0 Pakistan 2012 24 0

-­‐-­‐from  Les  Szabo,  CDL  

Future Work at CDL •  Monitoring/typing Pgt races of international nurseries -- Njoro, Debre Zeit

•  Confirming critical Pgt samples/isolates -- national surveys, special studies, etc

•  Working with samples/isolates associated with alternate hosts -- in conjunction with barberry surveys

•  Continuing the analysis of global Pgt collections using molecular diagnostic assay

•  Identifying rusts on barberry -- Eastern Africa, South Asia, Central and South America

•  Developing/distributing genetic stocks -- stem and leaf rust differentials (ARS Genebank, Aberdeen) -- ID series, UR series (in progress) -- stocks of new resistance genes (everyone’s responsibility)

Future work at AAFC

•  Conduct new international stem rust virulence survey as last one was in early 70’s (Luig)

•  Interested collaborators to contact Dr. Fetch •  Countries with capacity do the rust race typing •  Countries without capacity send isolates to Dr. Fetch for

race typing •  Results to be presented in booklet form and put onto

RUSTSPORE webpage •  Representative rust isolates also will be typed on

molecular basis and compared to phenotype (Szabo, Park, Hambleton labs)

Set Line Sr gene 1 ISr5Ra 5 T.m.deri. 21 Vernstein 9e ISr7bRa 7b 2 ISr11Ra 11 ISr6Ra 6 ISr8Ra 8a CnsSr9g 9g 3 W2691SrTt-1 36 W2691Sr9b 9b BtSr30Wst 30 Comb. VII 17+13 4 ISr9aRa 9a ISr9dRa 9d W2691Sr10 10 CnsSrTmp Tmp

Ug99 Reaction on NA Differentials

Ug99 Reaction on Differentials

Set Line Sr gene 04KEN156 1 ISr5Ra 5 4 T.m.deri. 21 3+ Vernstein 9e 4 ISr7bRa 7b 3 2 ISr11Ra 11 4 ISr6Ra 6 4 ISr8Ra 8a 4 CnsSr9g 9g 4 3 W2691SrTt-1 36 0 W2691Sr9b 9b 4 BtSr30Wst 30 4 Comb. VII 17+13 2++ 4 ISr9aRa 9a 4 ISr9dRa 9d 4 W2691Sr10 10 4 CnsSrTmp Tmp 2+

Ug99 Reaction on Differentials

Set Line Sr gene 04KEN156 1 ISr5Ra 5 4 T.m.deri. 21 3+ T Vernstein 9e 4 ISr7bRa 7b 3 2 ISr11Ra 11 4 ISr6Ra 6 4 T ISr8Ra 8a 4 CnsSr9g 9g 4 3 W2691SrTt-1 36 0 W2691Sr9b 9b 4 K BtSr30Wst 30 4 Comb. VII 17+13 2++ 4 ISr9aRa 9a 4 ISr9dRa 9d 4 S W2691Sr10 10 4 CnsSrTmp Tmp 2+

B L L L L C L L L H D L L H L F L L H H G L H L L H L H L H J L H H L K L H H H L H L L L M H L L H N H L H L P H L H H Q H H L L R H H L H S H H H L T H H H H