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Development of a sequence‐characterized amplified region marker for diagnosis of dwarf bunt of wheat and detection of Tilletia controversa Kühn
Author(s) -
Liu J.H.,
Gao L.,
Liu T.G.,
Chen W.Q.
Publication year - 2009
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1111/j.1472-765x.2009.02645.x
Subject(s) - biology , amplified fragment length polymorphism , primer (cosmetics) , polymerase chain reaction , genetic marker , microbiology and biotechnology , molecular marker , genetics , gene , genetic diversity , population , chemistry , demography , organic chemistry , sociology
Aims: Dwarf bunt of wheat, caused by Tilletia controversa Kühn, 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 μ g of teliospores in a 25‐ μ l 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.