2nd iwsrs kang(2014-4-29)
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    2nd iwsrs kang(2014-4-29) 2nd iwsrs kang(2014-4-29) Presentation Transcript

    • Barberry plays an important role in natural infection of stripe rust on wheat in China Zhensheng Kang State Key Laboratory of Crop Stress Biology for Arid Areas, Northwest A & F University, Yangling, Shaanxi, China
    •  Diseased area: between 3-6 million ha.  Yield losses: 10-50%. Years Losses (million tonnes) 1950 6.0 1964 3.2 1990 2.6 2002 1.4 Wheat stripe rust is a destructive disease throughout all winter wheat regions and is considered the most important disease of wheat in China.
    • Virulence variation for rusts maybe due to  sexual hybridization  mutations  somatic hybridization However, the mechanism of sexual hybridization for Pst has been neglected since the sexual stage was presumed to be absent. Resistant wheat cultivars Rust virulence variation Huge yield losses
    • In 2010, some Berberis spp. were shown to serve as alternate hosts for the wheat stripe rust pathogen.  B. chinensis,  B. holstii,  B. koreana  B. vulgaris. Jin et al. Phytopathology, 2010
    • Question: Does the sexual stage of wheat stripe rust occur under natural conditions, particularly in China?  Field Surveys for Berberis spp.  Susceptible test of Berberis to stripe rust  Isolation of stripe rust from nature-infected Berberis  Virulence test for recovered isolates  Mock-inoculation on Berberis in fields
    • Surveys for Berberis spp.in China  215 of the ~500 described Berberis spp. in the world are endemic to China;  Many of China's Berberis spp. distribute in the western over-summering areas (hot-spot). Western China: Sichuan : 81; Chongqin : 30; Yunnan : 78; Tibet: 55; Guansu : 26; Shaanxi: 20; Guizhou : 19; Qinghai : 13; Xinjiang : 5; Ningxia : 3; Central China: Hubei : 24; Henan : 7; Shanxi : 10; Hunan : 9; Anhui : 2; Eastern China: Distribution of Berberis species in different regions of China
    • Surveys for Berberis spp.in China
    • No barberry plant Barberry plants distributedBarberry plants with rusts 35 barberry species were surveyed and identified. Over-summering area
    • Are these Berberis spp. susceptible to wheat stripe rust ?
    • Identification of Berberis spp. as alternate hosts of wheat stripe rust in green house Dew chamber We collected seeds and seedlings of Berberis spp. from the fields and inoculated using telia of Pst in the greenhouse.
    • Normally, we see pycnia on the leaves of susceptible Berberis spp. 10-13 days after inoculation. Infection of basidiospore and development of pycnia Pycnia on Berberis Identification of Berberis spp. as alternate hosts of wheat stripe rust in green house
    • About 20 days after inoculation, we see aecia develop on the leaves. Aeciospores can infect wheat through the stoma and produce typical rust symptoms (uredinia). Identification of Berberis spp. as alternate hosts of wheat stripe rust in green house
    • 23 Berberis species: alternate host for Pst Successful production of pycnia, and aecia on upper and back sides of barberry leaf after inoculation Zhao, J., et al. Phytopathology, 2013, 103(9):927-934
    • A B C D E F G H I J K L M N O P Q R Zhao, J., et al. Phytopathology, 2013, 103(9):927-934
    • No. Berberis spp. Orgin Distribution 1 B. aggregata Gansu, China Gansu, Sichuan, Hubei, Qinghai, Shanxi 2 B. brachypoda Gansu, China Gansu, Sichuan, Hubei, Qinghai, Shanxi,Henan, Shanxi 3 B. potaninii Gansu, China Gansu, Shaanxi, Sichuan 4 B. soulieana Gansu, China Gansu, Shaanxi, Sichuan,Hubei 5 B. dasystachya Shaanxi, China Gansu, Shaanxi,Hubei,Shanxi 6 B. shensiana Shaanxi, China Shaanxi,Gansu 7 B. atrocarpa Sichuan, China Sichuan, Yunnan, Hunan 8 B . ferdinandi-coburgii Yunnan, China Yunnan 9 B. phanera Yunnan, China Yunna, Sichuan 10 B. aggregate var. integrifolia Yunnan, China Gansu, Sichuan,Qinghai,Hubei,Shanxi 11 B. davidii Yunnan, China Yunnan 12 B. stenostachya Gansu, China Gansu, Shaanxi, Shanxi 13 B. wangii Yunnan, China Yunnan 14 B. circumserrata Shaanxi, China Shaanxi, Hubei, Gansu, Qinghai,Henan 15 B. poiretii Beijing, China Shaanxi, Qinghai, shanxi, Hebei, Jilin, Liaoning, 16 B. guizhouensis Guizhou, China Guizhou 17 B. wilsonae Yunnan, China Yunnan, Tibet, Shaanxi, Qinghai, Hubei, Guizhou 18 B. sp. (unidentified) Tibet, China Tibet 19 B. jamesiana Yunnan, China Yunnan , Qinghai, Chongqing, Tibet, Sichuan 20 B. coryi Yunnan, China Yunnan 21 B. wilsonae Yunnan, China Yunnan, Sichuan, Tibet, Gansu 22 B. sp. Tibet, China Tibet 23 B. sp. Tibet, China Tibet Species of barberry identified as alternate hosts for Pst by artificial inoculation using germinated teliospores in China
    •  Some species are evergreen, distributed in southwest regions.  Others are deciduous, distributed in northwest regions.  Some susceptible species (e.g., Berberis soulieana, B. brachypoda, and B. shensiana) are widely distributed in the western over- summering areas. Berberis soulieana B. shensiana
    • Can barberry species be infected by the stripe rust fungus (Pst) in nature? It is very common to see pycnia and aecia on infected Berberis leaves in fields in May.
    • Obtaining of Pst isolates from rust- infected barberry growing in nature 20 Pst isolates recovered from the infected barberry  4 Pst isolated from 3 barberry species in 2011  16 Pst isolates from 3 barberry species in 2013 collected the infected Berberis leaves Single aecium (SA) was used for inoculation Recovered SA isolate on wheat leaf
    • Berberis spp. No. of aecia No. of isolate Rate (%) IT0 IT1-2 Pst Pgt Pst Pgt B. brachypoda 1519 479 1032 2 6 0.13 0.39 B. shensiana 410 35 374 1 0 0.24 0.00 B. soulieana 384 367 16 1 0 0.26 0.00 B. potaninii 742 711 28 0 3 0.00 0.40 B. aggregata 648 633 13 0 2 0.00 0.31 Total 3703 2225 1463 4 11 0.11 0.30 Pst isolates recovered from rust-infected Berberis spp. in Gansu, and Shaanxi in 2011 *I T0 indicates no any necrosis, or/ and chlorosis on wheat after inoculation, I T1- 2 indicates necrosis, or/ and chlorosis; Pst=Puccinia st riif ormis f. sp. trit ici; Pgt =Puccinia graminis f. sp. t rit ici
    • Comparison of infection types on Chinese differential hosts of eight major races of Pst and the four Pst isolates recovered from three barberry species in 2011 Virulence tests demonstrated that the infection types of the four barberry-recovered cultures are different compared to the major Chinese races. Cultures and races Origin of cultures Differential hosts 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 B2011-1 Berberis brachypoda A V A V V V A V A VA V A A V A A A A A B2011-2 B. brachypoda A V A V V A A V A A V A A A A A A A A B2011-3 B. shensiana A V A V V A V V A AV V A A AV A A A A A B2011-4 B. soulieana A V A A V V A V A A A A A V A V A A A CYA33 Triticum aestivum V V V V V V V V V V V V V V A V A A A CYA32 T. aestivum V V V V V V V V V V V V V V A V V A A CYA31 T. aestivum V V V V V V V V V A V V A V A V V A A CYA30 T. aestivum V V V V V V V V V A V V A A A V V A A CYA29 T. aestivum V V V V V V V V V A V V A A A V A A A CYA28 T. aestivum V V V V V V V V V A V A A A A V A A A CYA23 T. aestivum V V V V A V V V V A V A A A A A A A A CYA17 T. aestivum VA V A VA A VA V A A A AV A A A A A A A A Zhao, J., et al. Phytopathology, 2013
    • Berberis spp. No. of aecia Number of isolate* Rate (%) IT 0 IT 1-2 Pst Pgt Pst Pgt B. shensiana 3677 3630 40 4 3 0.11 0.08 B. brachypoda 4477 4406 59 8 4 0.17 0.15 B. sp. (Tibet) 1163 1150 6 4 3 0.34 0.26 Total 9317 9174 105 16 12 0.17 0.13 *I T 0 indicates no any necrosis, or/ and chlorosis on wheat after inoculation, I T1- 2 indicates necrosis, or/ and chlorosis; Pst=Puccinia st riif ormis f. sp. trit ici; Pgt =Puccinia graminis f. sp. t rit ici Pst isolates obtained from the infected Berberis spp. in Shaanxi, Gansu and Tibet in 2013
    • Berberis (origin) Isolate No. Chinese differential hosts for Pst 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 (ck ) Race / race group   U1 A V V V V V V V V A V V A V VA A AV A A A V SuG   U2 A V V V V V V V A V A V A A V A A A A A V SuG    B.aggregata (Gansu)   U3 A V A AV V V V V A V A V A A V A A A A A V SuG U4 A V A V V A V V V V A V V V A V A V A A A V SuG U5 V V V V V V V V V V V V V V A V V A A V CYR32 U6 A V A V V V V V V A V V V A V A A A A A V SuG   U7 V V V V V V V V A V V A V A V A A A A A V SuG   U8 V V V V V V V V V V V V V V A V V A A V CYR32 B.shensiana (Shaanxi)       U 9 V V V V V V V V V V V V V V A V V A A V CYR32 U10 V V V V V V V V V V V V V V V V A A A V SuG U11 A V A V V V V V A A V A V A A V A A A A A V SuG U12 A V A AV V V V V A V A V A A V A A A A A V SuG U13 A V V V V A V V V A V V A A V A A A A A V SuG  3 recovered isolates were identified as race CYR 32.  Other 13 isolates have different virulence patterns as compared to the major Chinese races , and can be considered as new races. Chinese differential hosts for Pst A=avirulent ; V=virulent; AV/VA =segeration of resistant and susceptible; ck=Mingxian 169 susceptible to Pst. SuG=race group virulence for Suwon 11(YrSu); HyG=race group virulence for Hyrbrid 46 (Yr3b,Yr4b) CYR= chinese yellow rust Virulence patterns of 16 recovered isolates on Chinese differential host sets in 2013
    • Single Aecia from barberry Recovered isolate from single aecium Single uredium (Su) isolates Virulence test on NILs Does the sexual reproduction under natural conditions contribute to virulence variation for Pst ? • 120 single-uredium (SU) isolates were make from 16 recovered isolates from infected barberry. • Virulence test for each SU isolates on near-isogenic lines. • Remarkable variation in virulence patterns among the SU isolates.
    • Virulence patterns of single-uredium isolates from recovered Pst isolate on near-isogenic lines SU isolate Yr NILs 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 Yr2 Yr5 Yr6 Yr7 Yr8 Yr9 Yr10 Yr15 Yr17 Yr24 Yr26 Yr27 Yr44 YrSp YrTr1 YrExp2 Yr25 Yr28 CK U10* VA A V A A A A A A A AV A V A A A V V V U10-1 A A V A A V A A A A A A V A A A V V V U10-2 A A V A A A A A A A A A V A A A V V V U10-3 A A V A A A A A A A A A V A A A V V V U10-4 A A V A A V A A A A A A V V A A V V V U10-5 A A V A A V A A V A A A V V A A V V V U10-6 V A V AV A A A A V A A A V V A A V V V U10-7 A A V A A V A A V A A V V V A A V V V U10-8 A A V A A V A A A A A A V A A A V V V U10-9 A A V AV A V A A A A A A V A A A V V V • SU isolates U10-2 and U10-3 share the same virulence patterns, • Other 7 SU isolates have different virulence patterns. *U10 isolate recovered from Berberis shensiana in Shaanxi
    • SU isolate Yr NILs 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 Yr2 Yr5 Yr6 Yr7 Yr8 Yr9 Yr10 Yr15 Yr17 Yr24 Yr26 Yr27 Yr44 YrSp YrTr1 YrExp2 Yr25 Yr28 CK U2* A A A AV V A A A A A V A A A A V A A V U2-1 A A A AV V A A A A A V A A A A V A A V U2-2 A A A A V A A A AV A V A V A A V A A V U2-3 A A VA AV V A A A A A V A A A A A A A V U2-4 A A A A V A A A V A V A V A A V A A V U2-5 A A AV A V A A A A A V A A A A V A V V U2-6 A/V A V V V A A A A A V A V A A V A A V U2-7 A A A A V A A A A A V A A A A V A A V U2-8 VA A V A/V A V A A A V A V A VA A A V A A V U2-9 A A AV A V A A A A A V A V A A V A A V U2-10 A A A A V A A A A A V A A A A V A A V U2-11 A A AV VA V A A A A A V A A A A V A A V U2-12 A A AV A V A A A A A V A A A A V A A V U2-13 A/V A A A/V A V A A A V A V A A A A V A A V U2-14 A/V A A/V A/V A V A A A A A V A A/V A A V V VA V “/” indicates various infection type on first and second leaf of genotype wheat cultivar. AV, VA= segregation of resistant and susceptible. *U2 isolate recovered form Berberis aggregata in Gansu. • U2-7 and U2-10, share the same virulence patterns, • Other 12 SU isolates have different virulence patterns. Virulence patterns of single-uredium isolates from recovered Pst isolate on near-isogenic lines
    • Single Aecia from barberry Recovered isolate from single aecium Single uredium (Su) isolates Virulence test on NILs Does the sexual reproduction under natural conditions contribute to virulence variation for Pst? • The single-uredium isolates originally from aecia on naturally infected barberry plants consisted of diverse of virulence patterns, which should be due to sexual reproduction on the barberry plants.
    • Does infection of stripe rust on Barberry cause the disease in adjacent wheat?
    • Does infection of stripe rust on Barberry cause the disease in adjacent wheat?  Mock infection were performed in Gansu, China, by putting wheat tissues bearing telia on the top of barberry plants in Middle of April, 2013.  The inoculated barberry plants showed symptom in early May, 2013, and 70 Pst isolates were obtained from mock-inoculated barberry.
    •  Stripe rust appeared on wheat adjacent to the mock-inoculated barberry plants before the end of May, 2013.  No stripe rust on wheat in the same field far off mock- inoculated barberry plants, and in other fields at this time.  70 Pst isolates were obtained from nearby wheat. Does infection of stripe rust on Barberry cause the disease in adjacent wheat?
    • Comparison of pathotypes on Chinese differential hosts for Pst isolates from mock-inoculated barberry and from nearby wheat Origin Number of race Race type CYR32 CYR31 Guinong 22 Hy46-7 Other racesa Mock-inoculated barberry 70 11 1 20 1 37 Nearby wheat 70 8 0 19 8 35 a Infection type of other races were not matched with those of known Pst races. The results showed that the races derived from mock-inoculated barberry matched with those isolated from nearby wheat.
    •  23 of the 30 tested barberry species show susceptibility to Pst, indicating that there is a great diversity of potential alternate hosts for Pst in China.  20 Pst isolates obtained from different barberry species in the fields indicated that the sexual stage of Pst occurs under natural conditions.  Virulence tests for single-uredium isolates recovered from a single aecium demonstrate that the sexual reproduction contributes to virulence variation for Pst. Summary
    • Pst must now be considered a macrocyclic fungal pathogen.
    •  The earmarked fund for Modern Agro-industry Technology Research System in China  National Basic Research Program of China (973)  Nature Science Foundation of China  The 111 Project from the Chinese Ministry of Education Acknowledgement Dr. Jie Zhao Prof. Lili Huang Dr. Hongchang Zhang Dr. Gangming Zhan Dr. Qingmei Han Dr. Yonghong Zhang Dr. Dejun Han Dr. Xiaojie Wang Dr. Jun Guo Dr. Xueling Huang Dr. Wenming zheng Mrs. Guorong Wei All PhD and Master students in My Lab. Dr. X M. Chen, Dr. H. Buchenauer, Dr. Robert McIntosh , Dr. Colin Wellings, Dr. Scot H. Hulbert, Dr. Jin-Rong Xu, Dr. Shiping Wang, Dr. Hei Leung, Dr. J. Chong Dr. Yue Jin, Dr. Ravi Singh, Dr. Zacharias Pretorius
    • Thanks for your attention!