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GrainGenes Reference Report: LAM-49-235

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Reference	LAM-49-235
Title	Development of a sequence-characterized amplified region marker for diagnosis of dwarf bunt of wheat and detection of Tilletia controversa Khn.
Journal	Letters in Applied Microbiology
Year	2009
Volume	49
Pages	235-240
Author	Liu JH Gao L Liu TG Chen WQ
Abstract	AIMS: Dwarf bunt of wheat, caused by Tilletia controversa Khn, is a destructive disease on wheat as well as an important international quarantined disease in many countries. The objective of this investigation was to develop a diagnostic molecular marker generated from amplified fragment length polymorphism (AFLP) for rapid identification of T. controversa. METHODS AND RESULTS: A total of 30 primer combinations were tested by AFLP to detect DNA polymorphisms between T. controversa and related species. The primer combination E08/M02 generated a polymorphic pattern displaying a 451-bp DNA fragment specific for T. controversa. The marker was converted into a sequence-characterized amplified region (SCAR), and specific primers (SC-01(49)/SC-02(415)), designed for use in PCR detection assays, amplified a unique DNA fragment in all isolates of T. controversa, but not in the related pathogens. The detection limit with the primer set SC-01(49)/SC-02(415) was 10 ng of DNA which could be obtained from 11 microg of teliospores in a 25-microl PCR reaction. CONCLUSIONS: An approach to distinguish T. controversa from similar pathogenic fungi has been developed based on the use of a SCAR marker. SIGNIFICANCE AND IMPACT OF THE STUDY: Development of the simple, high throughput assay kit for the rapid diagnosis of dwarf bunt of wheat and detection of T. controversa is anticipated in further studies.
External Databases	http://dx.doi.org/10.1111/j.1472-765X.2009.02645.x

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