A FIASCO-Based Approach for Detection and Diagnosis of Puccinia graminis f.sp.tritici in China  

作　　者：LIU Tai-guo WANG Xi GAO Li LIU Bo CHEN Wan-quan XIANG Wen-sheng 

机构地区：[1]State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences [2]College of Life Science, Northeast Agricultural University

出　　处：《Journal of Integrative Agriculture》2014年第11期2438-2444,共7页农业科学学报（英文版）

基　　金：Financial supports by the National 973 Program of China (2013CB127701 and 2011CB100403);the Ntional 863 Program of China (2012AA101501);the National Key Technologies Research and Development Program of China (2012BAD19BA04) from the Ministry of Science and Technology of China;the Special Fund for Agro-Scientific Research in the Public Interest, China (200903035);the China Agriculture Research System (CARS-03) from the Ministry of Agriculture of China;the National Natural Scientific Foundation of China (31371884)

摘　　要：Stem or black rust of wheat, caused by the fungus Puccinia graminis Pers. f. sp. tritici Eriks.＆E. Henn. （Pgt）, has historically caused severe losses to wheat （Triticum aestivum L.） production worldwide. In the Fujian and Guangdong provinces of China, six moderate-to-severe epidemics of wheat stem rust have occurred, which caused destructive losses of wheat between 1949 and 1966, although these were brought under control by integrated management. A rapid and reliable detection of the pathogen will contribute to the accurate forecast and seasonal control of this disease. The objective of this study was to develop a diagnostic molecular marker generated from simple sequence repeats （SSR） for the early rapid identiifcation of P. graminis. The genomic DNA of P. graminis, Puccinia striiformis, Puccinia triticina and seven other species was ampliifed by a pair of SSR primers generated by the FIASCO （fast isolation by AFLP sequences containing repeats） enrichment protocol. The primer set Pgtw （f）/Pgtw （r） generated a polymorphic pattern displaying a 330-bp DNA fragment speciifc for P. graminis whereas no DNA fragment was obtained from other non-target wheat fungal pathogens. The detection limit of the primer was 1 ng DNA in a 25-mL PCR reaction. The SSR markers of P. graminis can also be used to detect the presence of latent hyphae in Pgt-infected wheat leaves as early as 30 h post-inoculation. A rapid approach to distinguish P. graminis from similar pathogenic fungi would be anticipated in further study.Stem or black rust of wheat, caused by the fungus Puccinia graminis Pers. f. sp. tritici Eriks.＆E. Henn. （Pgt）, has historically caused severe losses to wheat （Triticum aestivum L.） production worldwide. In the Fujian and Guangdong provinces of China, six moderate-to-severe epidemics of wheat stem rust have occurred, which caused destructive losses of wheat between 1949 and 1966, although these were brought under control by integrated management. A rapid and reliable detection of the pathogen will contribute to the accurate forecast and seasonal control of this disease. The objective of this study was to develop a diagnostic molecular marker generated from simple sequence repeats （SSR） for the early rapid identiifcation of P. graminis. The genomic DNA of P. graminis, Puccinia striiformis, Puccinia triticina and seven other species was ampliifed by a pair of SSR primers generated by the FIASCO （fast isolation by AFLP sequences containing repeats） enrichment protocol. The primer set Pgtw （f）/Pgtw （r） generated a polymorphic pattern displaying a 330-bp DNA fragment speciifc for P. graminis whereas no DNA fragment was obtained from other non-target wheat fungal pathogens. The detection limit of the primer was 1 ng DNA in a 25-mL PCR reaction. The SSR markers of P. graminis can also be used to detect the presence of latent hyphae in Pgt-infected wheat leaves as early as 30 h post-inoculation. A rapid approach to distinguish P. graminis from similar pathogenic fungi would be anticipated in further study.

关 键 词：wheat stem rust Puccinia graminis f. sp. tritci MICROSATELLITE FIASCO PCR detection 

分 类 号：S435.121.4[农业科学—农业昆虫与害虫防治]