Detection of Ug99 (TTKSK) race of stem rust fungus in Egypt

Borlaug Global Rust Initiative (BGRI) Technical Workshop, Sydney, Australia, September 17-20, 2015

Atef Shahin1, Moustafa Azab2 and Doaa El-Nagar1

Plant Pathology Research Institute1 and Wheat Research Program2, ARC, Egypt

International collaborators• Ravi Singh, CIMMYT• Mogens Hovmøller, GRRC• Kumarse Nazari, ICARDA• Dave Hodson, CIMMYT• Yue Jin, USDA-ARS• Les J. Szabo, USDA, ARS• Mehran Patpour, GRRC • Maria Newcomb Minnesota, US• Pablo Olivera, Minnesota, US

Cooperation between the wheat pathologists and breeders in Egypt and International research center and labs

Outline• Wheat in Egypt• Historical and current importance to stem rust

in Egypt• Race identification of stem rust in Egypt• Resistance of the cultivars to stem rust• Detection of Ug99 (TTKSK) race in Egypt • Conclusion • Future plan

Bread wheat improvement in Egypt

Hendi 62

Giza 139

Giza 150

Giza 156

Sakha 157

Sakha 69

Sakha 162

Sakha 164

Gemm

eiza 1

Sids 4

Sids 7

Gemm

eiza 5

Sakha 93

Gemm

eiza 9

Gemm

eiza 10

0

1

2

3

4

5

6

7

8

9

10Grain yield; 8.9

Grai

n yi

eld

(ton/

ha)

Cultivars

Adel Hagras 2014. J. Agric. Res. Kafrelsheikh Univ., 40 (3):517-539.

Released wheat varieties in Egypt(1920 - 2013)

1920-1950

1950-1970

1970-1990

1990-2013

Total

0

5

10

15

20

25

30

35

40

45

3 4

11

25

43

0 03 4

7

BWDW

Wheat Production Trend in Egypt (1960 – 2014)

Source: USDA (http://www.indexmundi.com/agriculture/?country=et&commodity=wheat&graph=production)

0

1000

2000

3000

4000

5000

6000

7000

8000

9000 Production; 8300

Prod

uctio

n (,0

00 M

T)

Production Year

Giza 167Gemmeiza5Sids1 Beni Sweif1Sakha93Shandaweel1Beni Sweif6

Sakha61Gemmeiza10Gemmeiza11Sids13Beni Sweif4Beni Sweif5Sohag3

Giza168Sakha94

Gemmeiza7

Sids12Giza 171

Misr1Misr2

Some representative Egyptian wheat cultivars

Wheat Rust in Egypt

• Wheat in Egypt suffers from wheat rust diseases– Yellow rust or Stripe rust– Stem rust or Black rust– Leaf rust or Brown rust

• Egypt wheat breeder and pathologist are interested in searching for new sources of resistance to wheat rust

• Control is through genetic resistance , and foliar fungicides

Variety Year of release

Year of epidemic

Rust

Hindi D & Tosson 1921 1947 Stem rust

Giza 139 1945 1954 Leaf rust

Giza 144 1958 1967 Yellow rust

Sakha 8 1976 1986 Yellow rust

Sakha 8 & Sakha79 1976 1987 Stem rust

Gemmiza 1& Giza 163 1994 & 1989 1995 Yellow rust

Sakha 69 1980 1997 Yellow rust

Epidemics of Wheat rusts in Egypt

In Egypt wheat stem rust disease was epidemic in 1987, for Sakha-79 and Sakha-8, which the loss in grain yield due to early stem rust infection recorded 33.5%, Bassiouni et al. (1987).

Historical background and current importance to stem rust in Egypt

In Egypt, the first reports on physiologic specialization of wheat stem rust incited by Puccinia graminis tritici was carried out in 1949 (Abdel-Hak, 1953), who identified 15 physiologic races at different locations in Egypt i.e. 9, 11, 14, 17, 19, 21, 24, 42, 53, 59, 69, 88, 123, 279 (E2) and (E1). Later on (Abdel-Hak and Kamel, 1966) so far, recorded races i.e. 117, 133, 186, 194 and 249 in Egypt.

Greenhouse• Seedling evaluation of wheat

accessions against stem rust.• Rust spore /isolate increase and

maintenance • Wheat stem rust race identification • Gene efficacy and virulence formula for

the resulted race.• Gene postulation within wheat

cultivars.• Identification of Sr’s genes using

molecular marker.

Field and greenhouse activities for stem rust • Losses due to stem rust • Evaluation of commercial wheat

varieties and monogenic lines. • Close cooperation with the wheat

research program. • Cooperation with ICARDA and

CIMMYT.• Evaluation of the effective wheat

rusts fungicides.

Field

Egyptian Wheat Trap Rust Nursery (EWTRN)Locations: 16Entries : 130

Standard and supplemental differential varieties

The race nomenclature systemThis system was adopted by (Roelfs and Martens, 1988 and McVey et al., 1997) the set included 20 differential hosts each with single stem rust gene i.e. in 5 subsets in the following order,(1) Sr5, Sr21, Sr9e and Sr7b, (2) Sr11, Sr6, Sr8a and Sr9g, (3) Sr36, Sr9b, Sr30 and Sr17, (4) Sr9a, Sr9d, Sr10, SrTmp and (5) Sr24, Sr31, Sr38, and SrMcN. Races were designated by a five letters code [Pgt-code] such as TTKSK, TTKTK,….Infection types (IT) were recorded for each line in the form avirulence/virulence of pathogen cultures which were determined by low (IT) [L]and high (IT) [H]. Determination of races were based on the reaction of the inoculated differential hosts.

Pgt-code for the 20 [Pgt] differential hosts for (Puccinia graminis f. sp. tritici)

Low IT [L] High IT [H]

Stem Rust reaction record as infection types [ITs] and scoring 14 days after inoculation using the 0 – 4 scale adopted by Stakman et al., 1962.

Infection Types

Pgt-Code

Virulence (ineffective Sr genes)No. of

resistance response

No. of susceptible response

Virulence(%)

BBBPB 9b,9a,10,Tmp 16 4 20BBBBC McN 19 1 5CCGBC 7b, 9g,9b,McN 16 4 20GHMLB 21,6,9a,36,17,9e 14 6 30NGRBB 5,9e,7b,6,36,9b 14 6 30LDBBB 5,8a 18 2 10LCBDC 5,9g,10, McN 16 4 20LCPDB 5,9g,36,30,17,10 14 6 30NGRBB 5,9e,6,36,9b,17 14 6 30TKSTB 5,21,9e,7b,6,8a,9g,36,9b,30,9a,9d,10,Tmp 6 14 70TTKTF 5,21,9e,7b,11,6,8a,9g,9b,30,17,9a,9d,10,Tmp,38,McN 3 17 85TTKTC 5,21,9e,7b,11,6,8a,9g,9b,30,17,9a,9d,10,Tmp,McN 4 16 80

Some identified Pgt races and its virulence formula in Egypt, 2014/2015 growing season.

Variety Pedigree/Cross NameSeason/Stem rust responses2012/13 2014/15

Misr 1 Oasis/SKauz//4*Bcn/3/2*Pastor 10R 30SMisr 2 Skauz/Bav92 Skauz/Bav92 10R 20SGiza 168  MRL/BUC//Seri.CM93046-8M-0Y-0M-2Y-0B 5MS 20SGiza 171 Sakha93/Gemmeiza9 S 6-1GZ-2GZ-2GZ-0S 5R 10MRSakha 93 Sakha 92 / TR810328 10S 10SSakha 94 OPATA / RAYON // KAUZ 5S 10SGemmeiza-7 CMH74A.630/5X//SERI 82/3/AGENT 5MS 10SGemmeiza-10  Maya 74"S"/on / 1160- 147 /3/ Bb / G11 /4/ chat "S"/5/ crow "S" 10MS-S 10SGemmeiza-11  BOW''S'' /KVZ''S''// 7C/SERI82/3/GIZA168 /SKHA61 R 5SSids-6  Maya”S”/Mon”S”//CMH74A.592/3/Sakha8*2SD/10002 60S 50SSids-12  BUC//7C/ALD/5/MAYA74/ON//1160-147/3/BB/GLL/4/

CHAT''S''/6/MAYA/VUL//CMH74A.630//4*SX.10S 20S

Sids-13  AMAZ 19=KAUZ"S"//TSI/SNB"S". R RBeni Sweif-4  DIPPERZ/BUSHEN3 R 10SBeni Sweif-5  DIPPERZ/BUSHEN3CDSS92B1281-M-OY-OM-OY-3B-OY-OSD. R 5SBeni Sweif-6  Jo”s”/AA”s”//Fg”s”. R TrSSandawel-1  SITE/ MO/4/NAC//3*PVN/3/MiRLO R 10S

Response of some Egyptian wheat cultivars to stem rust reaction under field conditions during 2010 and 2015

Cultivar  Gene

Season / Stem rust responses

2012 2013 2014

ISr5-Ra CI 14159 Sr5 5S 30S 5SISr6-Ra CI 14163 Sr6 30S 40S 30SNa 101/6*Marquis Sr7a 10S 40S 30SISr7b-Ra CI 14165 Sr7b 10S 20S 30SCI 14167/9*LMPG-6 DK04 Sr8a 5MS-S 10S 40SBarieta Benvenuto (CI 14196) Sr8b 5S 20S 60SISr9a-Ra CI 14169 Sr9a 20S 30S 10SPrelude*4/2/ Marquis*6/Kenya 117A Sr9b 5S 60S 30SISr9d-Ra CI 14177 Sr9d 30S 20S 30SVernstein PI 442914 Sr9e 10S 60S 40SChinese Spring*7/Marquis 2B Sr9g 10MS-S 30S TrSW2691Sr10 CI 17388 Sr10 20MS-S 20S 30SLee/6*LMPG-6 DK37 Sr11 TrS 40S 20SChinese Spring*5/Thatcher 3B Sr12 40S 60S 60SPrelude*4/2/ Marquis*6/Khapstein Sr13 TrS 30S 20SW2691*2/ Khapstein Sr14 20S 60S 20SPrelude*2/Norka Sr15 20MR-MS 50S 50SThatcher/CS (CI 14173) Sr16 10S 60S -Prelude/8*Marquis*2/2/Esp 518/9 Sr17 10S 40S 30SLittle Clube/Sr18Mq Marquis"A" Sr18 30S 50S 50S94A 236-1 Marquis"B" Sr19 10S 40S 40S94A 237-1 Marquis"C" Sr20 30S 50S 50SMcNair 701 SrMcN 20S 20S 60S

Stem rust responses of differential genotypes in Egypt at adult stage over three cropping seasons (2012 - 2014) at Sakha Agric. Res. Stn. In Egypt

Cultivar - Gene

Season/Stem rust responses

2012 2013 2014

T. Monococcum/8*LMPG-6 DK13 Sr21 TrS 10S 40SMq*6//Stewart*3/RL 5244 Sr22 10S 50S 50SExchange CI 12635 Sr23 10S 30S 10SLcSr24AG Sr24 5MR 40MR TrR-MRAgatha (CI 14048)/9*LMPG-6 DK16 Sr25 10MR 10S 30SEagle Sr26 McIntosh Sr26 20MR 30MR 5MR-MSWRT 238-5 (1984) Roelfs Sr27 5MR 40MR 20SKota RL71 Sr28 20S 50S 20SPrelude/8*Marquis/2/Etiole de Choisy Sr29 20S 50S 20SSelection from Webster F3:F4 #6 Sr30 5S 30S 50SSr31 (Benno)/6*LMPG-6 DK42 Sr31 5MR 10MR-MS 40MR-MSER5155 S-203 (1995) Roelfs Sr32 10R 40MR 10MSRL 5405 (1192) Kerber Sr33 5MS 10S 30SRL 6098 (1997) Dyck Sr34 10S 40S 20SRL 6099 (1995) Dyck Sr35 TrS 20S 40SW2691SrTt-1 CI 17385 Sr36 5S 30S 30SCook Sr36 TrR 10R 10MSPrelude*4/Line W (W3563) Sr37 TrS 30S 50STrident Sr38 Sr38 TrS 5S 10MR-MSTAM 107 Sr1RS-AM 20S 50S 50SCsSSrTmp SrTmp 10S 20S 30SBt/Wld SrWld-1 10S 20S 50SPavon 76 Sr2 complex 10MR 40MR 5MR-MSEnkorn Sr21 TrR 10MR 5MR-MSSeri 82 Sr31 5R 5MR 5MSPBW343=Attila with Sr31 Sr31 10R 30MR-MS 30MR-MS

Stem rust responses of differential genotypes in Egypt at adult stage over three cropping seasons (2012 - 2014) at Sakha Agric. Res. Stn. In Egypt

Stem rust symptoms on some entries of STEMRRSN and 10th ISRTN-15

Detection of Ug99 (TTKSK) race of stem rust fungus in Egypt

-Field observationsIdea of sending Sr31 infected samples from Egypt was generated in 2nd International wheat stripe rust symposium, Regional Cereal Rust Research Center, Izmir, Turkey 28 April - 1 May, 2014., discussion between David Hodson, Mogen Homveler and Atef Shahin.

-During April 2014, high levels of stem rust infection were observed on entries in wheat (Triticum aestivum) grown in a nursery at Sakha Agricultural Research Station in Egypt.

-Samples, collected from rusted stems of wheat cultivars, i.e. PBW343, and (Benno)/6*LMPG-6 DK42 which are known to carry stem rust resistance Sr31.

Code Host Varity Sample date Location Disease severity14EGY001 (Benno/)6*LMPG-6 DK42 08/05/2014 Sharqia MS

14EGY002 (Benno/)6*LMPG-6 DK42 08/05/2014 Sharqia MS

14EGY003 PBW343(8STEMRRSN) 03/05/2014 Sakha MR-MS

14EGY004 PBW343(GENETIC STOCK) 03/05/2014 Sakha MR-MS

14EGY005 PBW343(IBWSN) 03/05/2014 Sakha MS-S

14EGY006 (Benno/)6*LMPG-6 DK42 05/05/2014 Sharqia MS

Sr31 infected samples from Egypt

In the 2014 crop season in Egypt, the presence of virulence to Sr31 in Egypt was suspected based on preliminary field observations of Sr31 planted as international stem rust trap nursery at :

1) Sakha Agricultural Research Station in Kafrelsheikh (31.094059°N, 30.933899°E).2) Al- Sharqia (30.601400°N, 31.510383°E ).3) Nubaria (30.91464°N, 29.95543°E).

The presence of Sr31-virulence in Egypt in the 2014 growing season was supported by three independent observations: Stem rust scores of up to 30% stem/leaf coverage (30MR

to 30S). This was recorded by Dr. Atef Shahin, Sakha Agricultural Research Station, at the time of sampling on wheat cultivar PBW343, which is known to carry stem rust resistance Sr31.

Race typing of samples from Sr31-resistant wheat confirmed the presence of Sr31-virulence, suggesting the presence of Ug99. Tests were based on independent samples which were repeated 2-3 times. The presence of Sr31- virulence was confirmed beyond any doubt.

DNA/PCR test originally developed by the Cereal Disease

Lab, Minnesota, USA, was positive for the considered isolates carrying Sr31-virulence, which confirmed the presence of the Ug99 race group.

Mogens Støvring Hovmøller

Global Rust Reference Center www.wheatrust.org

Confirmation of stem rust Sr31-virulence (Ug99) in Egypt

Race identification by GRRC (Denmark)

*Summary results of race-typing 6 Egyptian samples done at CDL in the 2014 season

Total number of samples 6

Isolates typed as: TCMLC 1  TKKTF 9  TKTTF 4  TTKSK 2Total number confirmed isolates 16

*Source : Yue Jin, USDA-ARS and **Mogens Hovmøller, GRRC.

- ** Stem rust samples from the 2014 surveys of wheat fields in Egypt were sent to the Global Rust Reference Center (GRRC, Denmark) for race analysis The pathotyping was repeated in two or three independent experiments using single-pustule isolates. Isolates were also typed at the USDA-ARS Cereal Disease Laboratory (USA) for an independent confirmation.

- Three races in the Ug99 race group were detected; TTKST (four isolates) from Sharqia, TTKTK (13 isolates) from Sakha, and TTKSK (2 isolates) from Nubaria.

Confirmation of presence of Ug99 in Egypt

Two new Tmp-virulent races in the Ug99 race group

Conclusion

• Close cooperation between the wheat pathologists in Egypt and International research center and labs,

e.g. ICARDA, CIMMYT, GRRC, USDA, and University of Minnesota led to confirm the stem rust Sr31-

virulence (Ug99) in Egypt.

• Race analysis results indicated the presence three Ug99-races were detected in Egypt (2014), Ug99

(TTKSK) in addition to the new race TTKTK and TTKST. Also, Detection of non-Ug99 but Tmp-virulent race

(TKTTF) in Egypt.

• Sr25 virulence detected in two non-Ug99 races .

• Close cooperation has been established in Egypt between wheat breeders and wheat pathologist from

long time ago, generated a good system for rust diseases breeding.

• Despite of presence of TTKSK and TTKTK in Egypt, high level of stem rust resistance is observed on most

of the Egyptian wheat promising lines.

• Wheat genotypes having the combination of Sr2, Sr26 and Sr25 showed high stem rust resistance level

under the Egyptian condition.

Future Plan

• Studying role and the relationship between the present genetic makeup in both pathogen

and host should be continued to an overall breeding strategy to wheat rusts resistance.

• More cooperation between the wheat pathologists in Egypt and International research

center and labs, is very important.

• More cooperation is essential now between Egypt and countries having stem rust as a

common biotic stress i.e. Ethiopia, Kenya …etc. to exchange germplasm as well as test and

evaluate our materials.

• Studying the distribution of fungal pathotypes and Sr’s resistance genes of wheat rusts in

Egypt using molecular markers.

 

 

 

 

 

 

 

 

  

 

 

 

 

 

 

 

 

 

 

  

 

Acknowledgements• Agricultural Research Center,

ARC, Egypt• The Borlaug Global Rust Initiative

(BGRI)• GRRC, Denmark • USDA, ARS• Minnesota University• ICARDA• CIMMYT

Borlaug Global Rust Initiative (BGRI) Technical Workshop, Sydney, Australia, September 17-20, 2015

Thanks to all the people who have contributed to organizing this great event

 

 

 

 

 

 

 

 

  

 

 

 

 

 

 

 

 

 

 

  

 

If you want to go fast, go aloneIf you want to go far, go together

African proverb

Thank you for your attention

Contact : a.a.shahin@hotmail.comSeptember 18, 2015

Borlaug Global Rust Initiative (BGRI) Technical Workshop, Sydney, Australia, September 17-20, 2015