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| A FIASCO-Based Approach for Detection and Diagnosis of Puccinia graminis f. sp. tritici in China |
| LIU Tai-guo, WANG Xi, GAO Li, LIU Bo, CHEN Wan-quan , XIANG Wen-sheng |
1、State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences,
Beijing 100193, P.R.China
2、College of Life Science, Northeast Agricultural University, Harbin 150030, P.R.China |
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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 identification of P. graminis. The genomic DNA of P. graminis, Puccinia striiformis, Puccinia triticina and seven other species was amplified 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 specific 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.
Abstract 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 identification of P. graminis. The genomic DNA of P. graminis, Puccinia striiformis, Puccinia triticina and seven other species was amplified 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 specific 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.
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Received: 15 August 2013
Accepted:
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| Fund: Financial supports by the National 973 Program of China (2013CB127701 and 2011CB100403), the Ntional 863 Program of China (2012AA101501) and 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) and the China Agriculture Research System (CARS-03) from the Ministry of Agriculture of China, and the National Natural Scientific Foundation of China (31371884) are also gratefully acknowledged. The funding sources had no role in study design, data collection and analysis, decision to publish or the preparation of the manuscript. |
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Corresponding Authors:
CHEN Wan-quan, E-mail: wqchen@ippcaas.cn; XIANG Wen-sheng, E-mail: xiangwensheng@neau.edu.cn
E-mail: wqchen@ippcaas.cn;xiangwensheng@neau.edu.cn
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| About author: LIU Tai-guo, E-mail: tgliu@ippcaas.cn;* These authors contributed equally to this study. |
Cite this article:
LIU Tai-guo, WANG Xi, GAO Li, LIU Bo, CHEN Wan-quan , XIANG Wen-sheng.
2014.
A FIASCO-Based Approach for Detection and Diagnosis of Puccinia graminis f. sp. tritici in China. Journal of Integrative Agriculture, 13(11): 2438-2444.
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