Premium
Development of a sequence‐characterized amplified region marker using inter‐simple sequence repeats for detection of Puccinia striiformis f. sp . tritici
Author(s) -
Wang Shuhe,
Chu Bingyao,
Liu Qi,
Luo Yong,
Ma Zhanhong
Publication year - 2017
Publication title -
journal of phytopathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.53
H-Index - 60
eISSN - 1439-0434
pISSN - 0931-1785
DOI - 10.1111/jph.12578
Subject(s) - biology , polymerase chain reaction , genomic dna , puccinia striiformis , dna , genetics , pathogen , genetic marker , dna sequencing , sequence tagged site , microbiology and biotechnology , gene , chromosome , gene mapping
Abstract Stripe rust, caused by Puccinia striiformis f. sp . tritici ( Pst ), is the most devastating wheat disease in China. Early and accurate detection of the pathogens would facilitate effective control of the diseases. DNA ‐based methods now provide essential tools for accurate plant disease diagnosis. In this study, inter‐simple sequence repeats ( ISSR ) technique has been successfully applied to develop a sequence‐characterized amplified region ( SCAR ) marker for diagnosis of stripe rust of wheat and detection of Pst . In this study, one fragment unique to Pst was identified by ISSR and then sequenced. Based on the specific fragment, a pair of SCAR primers (616 AF /616 AR ) was designed to amplify a 299‐bp DNA fragment within the sequenced region. The primers can amplify a unique DNA fragment for all tested isolates of Pst but not for the other pathogens of wheat leaves and the uninfected leaves. The polymerase chain reaction ( PCR ) assay could detect as low as 0.1 ng of genomic DNA in a 25.0 μl PCR reaction mixture and detect the pathogen from asymptomatic wheat leaves inoculated with Pst under glasshouse conditions.