Development of a sequence-characterized amplified region marker using inter-simple sequence repeats for detection of Puccinia striiformis f.sp.tritici

作者：Wang, Shuhe; Chu, Bingyao; Liu, Qi; Luo, Yong; Ma, Zhanhong^*

来源：Journal of Phytopathology, 2017, 165(7-8): 442-447.

DOI：10.1111/jph.12578

• inter-simple sequence repeats
• Puccinia striiformis f.sp tritici
• sequence-characterized amplified region
• yellow rust of wheat

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 (616AF/616AR) 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 mu l PCR reaction mixture and detect the pathogen from asymptomatic wheat leaves inoculated with Pst under glasshouse conditions.

• 出版日期2017-8
• 单位中国农业大学; 新疆农业大学; 河南大学