An ISSR‐based Approach for the Molecular Detection and Diagnosis of Dwarf Bunt of Wheat,Caused by Tilletia controversa Kühn |
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| Authors: | Li Gao Wanquan Chen Taiguo Liu |
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| Institution: | Authors’ address: State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, CAAS, No 2, West Yuan Ming Yuan Road, Beijing 100193, China (correspondence to W. Chen. E‐mail: wqchen@ippcaas.cn) |
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| Abstract: | Dwarf bunt of wheat, caused by Tilletia controversa Kühn, is an important international quarantine disease in many countries. The objective of this investigation was to develop a diagnostic molecular marker generated from intersimple sequence repeat (ISSR) for rapid identification of T. controversa. A total of 60 primers were tested by ISSR to detect DNA polymorphisms between T. controversa and related species. The primer ISSR818 generated a polymorphic pattern displaying a 952‐ bp DNA fragment specific for T. controversa. The marker was converted into a sequence characterized amplified region (SCAR), and specific primers (TCKSF2/TCKSR2) were designed for use in a PCR detection assay. Its detection limit was 1 ng of DNA, which could be yielded by 1.1 μg of teliospores in a 25‐ μl PCR. Conclusively, a method to distinguish T. controversa from similar pathogenic fungi has been successfully developed based on the use of a SCAR marker. |
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| Keywords: | ISSR PCR SCAR Tilletia controversa Kü hn |
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