Development of a sequence-characterized amplified region marker for diagnosis of dwarf bunt of wheat and detection ofTilletia controversaKühn

Abstract

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‐0149/SC‐02415), 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‐0149/SC‐02415 was 10 ng of DNA which could be obtained from 11 μg of teliospores in a 25‐μl 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.

Detection of Tilletia controversa using immunofluorescent monoclonal antibodies

“…The immunologic assay described here for T. controversa and T. caries is significantly faster and simpler than conventional methods, such as morphologic and PCR‐based techniques (Liu et al . ; Gao et al . ,b, ).…”

“…Bakkeren et al (2000) reported that amplified fragment length polymorphism (AFLP) may provide better sequence-characterized amplified region (SCAR) markers for T. controversa. Liu et al (2009) identified a SCAR marker for T. controversa using the AFLP method. Gao et al (2010aGao et al ( , 2011 reported two SCAR markers using the ISSR method for detecting T. controversa.…”

Development of IRAP-SCAR marker for strain identification in Lentinula edodes

“…Therefore, it is very important to accurately identify right strains before cultivation. Strain-specific SCAR marker was proven to be a simple, reliable and effective method for strain identification and it can be derived from any PCR-based markers, such as RAPD (Li et al 2008), ISSR (Qin et al 2006;Su et al 2008a, b), SRAP (Wu et al 2010) and AFLP (Liu et al 2009). …”

Identification of a specific SCAR marker for detection of Tilletia foetida (Wall) Liro pathogen of wheat

“…Numerous specific SCAR markers have been developed from AFLP or RAPD markers in Tilletia caries [15] and Tilletia controversa [16,17].…”

“…3). Our previous successful con version from AFLP to SCAR investigation also showed that the corresponding fragment was 367 bp in size [16]. ln that investigation, a SCAR marker from the species specific AFLP fragment to distinguish T. controversa and other related pathogens has been coined.…”