Journal
JOURNAL OF PHYTOPATHOLOGY
Volume 159, Issue 3, Pages 155-158Publisher
WILEY
DOI: 10.1111/j.1439-0434.2010.01735.x
Keywords
ISSR; PCR; SCAR; Tilletia controversa Kuhn
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Funding
- National Basic Research and Development Program of China [2009CB119200]
- National High Technology Research and Development Programme [2006AA10Z432]
- National Key Technologies Research and Development Programme [2006BAD08A14]
- Monitoring and Control of Plant Diseases and Insect Pests from Ministry of Agriculture, China [2130108]
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Dwarf bunt of wheat, caused by Tilletia controversa Kuhn, 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 mu g of teliospores in a 25- mu l 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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