Establishment Of New Surveillance Stem For Chinese Raccs And Ug99of Puccinia Graminis F.sp. Tritici, Resistant Genes Defection In Commercial Wheat Varieties And Distinct Proteins Display Analysis Of Complementary Host Minn2761

Wheat stem rust caused by Puccinia graminis f.sp. tritici is a catastrophic wheat diseasebecause of its ability to cause complete annihilation of wheat crops over wide areas. China isin special wheat stem rust epidemic region and had suffered from several times of devastatingwheat losses due to the rust. For nearly four decades this disease has been under controlinternationally by widespread use of the1BL.1RS translocation with Sr31.Race variation isinternal reasons for wheat rust pandemic and resistance loss. Resurgence of raceUg99(TTKSK) is such a case. The race is highly virulent on Sr31and causes severe threat towheat production in China and the world. the urgent researches would be included in thefollowing aspects Race survey (a section of international wheat stem rust surveillance),Chinese resistant variety and germplasm screening, Chinese resistant variety gene detectionand new resistant gene mining, and Chinese important complementary host: Minn2761resistance related proteomics studies. The main progresses were as follows.1. The new sets of wheat stem rust differential single gene lines and five-letter racenomenclature system which could distinguish new races Ug99and its mutants were used toidentify75isolates in the year2009-2010. Eight pathotypes, namely,21C3CTHTM,34MKGRM,21C3CPHTM,21C3CTHTP,34MKGTM,21C3CFHTM,21C3CTHRM and34MKGRP, were identified with frequencies of42.7%,14.7%,13.3%,9.3%,6.7%,5.3%,4.0%and4.0%, respectively. Only a few races were found, without finding new pathotypeUg99. Race21C3still remained predominant. The virulence spectra of eight race pathotypeswere determined on47single gene lines. The virulence frequencies for Sr7a,7b,8a,9a,9b,9d,9f,9g,12,13,14,15,16,18,19,20,23,24,25,27,28,29,32,34,Tmp,Dp-2were100%; thosefor Sr6, Sr10,11,17, Wld-1, GT, between50%-80%; those for Sr5and Sr38, between10%-30%; and those for Sr9e，21，22，26，30，31，33，35，36，37，0. It was indicatedthat Sr5,9e,11,21,30and31were highly resistant to Chinese wheat stem rust races currentlybut susceptible to Ug99and only Sr22,26,33,35and37were resistant to both Chinese racesand Ug99and could be utilized in breeding for resistance.2. Using current main races of wheat stem rust in China, the resistance of57abroadUg99-resistant wheat cultivars and1418Chinese cultivars (lines) was appraised. It wasindicated that51wheat materials resistant to Chinese stem rust races and Ug99;52.0%of448cultivars from China also had good resistance, only1in105materials from Jiangsu was resistant to all races tested,64in236breeding lines from Shandong, resistant to all racestested;562in629wheat lines from Heilongjiang were highly resistant and442of them wereimmune.3. Molecular detection for stem rust resistant genes carried in448Chinese commercialcultivars was conducted by using validated closely linked stem rust resistant gene markers,such as those of available STS and SSR markers of Sr26(resistant to Ug99), Sr31and Sr36.The result was indicated that among those cultivars, Kenjiu10grown in HeilongjiangProvince and Baofeng104grown in Beijing were detected to contain Sr26, about70cultivarswere shown to carry the Sr31and no Sr36-carrying varieties were met with.4. For the first time in China, the mRNA differential display was used to study wheatstem rust resistance related protein of Chinese complementary differential: Minne2761.Twenty-four pairs of primers could amplify differential expression bands among Minn2761and Thatcher byDDRT-PCR analysis. Polymorphic bands were grouped into nine different types and four types of themwere supposed to correspond to disease-resistant genes. After sequencing cDNA differentialfragments and Blast homology analysis in the GenBank database it was indicated that therewas no known similar sequences found.5. A high-throughput, high-resolution and high-repeatability2-DE system was established for wheatleaf proteins. the proteins were extracted by using the TCA/acetones, the elements of lysis buffer werecomposed of urea:9mol/L, Thiourea:2mol/L,4%CHAPS，1%DTT，1%TBP and0.5%IPGbuffer. IEF condition was10000vh(17cm), SDS-PAGE was performed in12%polyacrylamide gels.6. The2-D maps of inoculated and un-inoculated Minn2761and Thatcher by of34MKGwere analyzed. The analysis result was shown that there were seven and six differentiallyexpressed protein spots, respectively, there were eleven differentially expressed protein spotsin inoculated Minne2761and Thatcher. Analysis of13interested protein spots was conductedwith MALDI-TOF-TOF. Among them,8proteins spots were obtained by Mascot databasesearching and inter-comparison preliminary identification. These protein spots could beclassified into three groups: group1was included in ascorbate peroxidase(66) and glutathionesynthetase (247), which is thought to relate to host defense; group2,ribulose-1,5-bisphosphate carboxylase/oxygenase large subunitRuBisCO large subunit(152), which isthought to relate to energy metabolism; group3,23kDa oxygen evolving protein ofphotosystem II(437,524) and group4was included in ADP-ribosylation factor(498),unknown(147,648).All of those up-or down-regulated proteins were probably parts of network of resistance of development-related proteins had close connection with plant defense responses,energy metabolism, cell signal transduction and transcriptional regulation.