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| Establishment and Application of Real-time PCR for Detecting SdhB-H278Y Mutation in The SdhB Gene Associated with Corynespora cassiicola in Cucumber |
| ZHU Guangxue1，2☆，YAN Yutao2☆，SUN Bingxue2，ZHOU Rongjia2，YUE Yuanyuan2，XIE Xuewen2，CHAI Ali2，LI Lei2，LI Baoju2*，SHI Yanxia2* |
| （1College of Horticulture，Qingdao Agricultural University，Qingdao 266109，Shandong，China；2Institute of Vegetables and Flowers，Chinese Academy of Agriculture Sciences，Beijing 100081，China） |
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Abstract Based on the differences of SdhB gene of Corynespora cassiicola registered in GenBank，specific primers were designed for SdhB-H278Y mutation，and the real-time PCR detection system was established for detecting SdhB-H278Y mutation．The results showed that C. cassiicola carried SdhB-H278Y and SdhB-I280V mutations．The SdhB-H278Y mutants have high resistance to boscalid，the EC50 value of SdhB-H278Y mutants was 21.47 μg·mL-1 or ＞ 30 μg·μL-1．The established standard curve of detection system existed a favorable linear correlation（R2 = 0.992 9）．The detection system can detect the SdhB-H278Y mutation specifically with sensitivity of 3.6 × 10-4 ng·μL-1，which was 10 times of AS-PCR．The expected values were highly correlated with the detected values by verifying genomic DNA carrying different mutation ratios of SdhB-H278Y（R2 = 0.999 7）．The system could also be used to detect the proportion of SdhB-H278Y mutant in the disease spot of corynespora leaf spot of cucumber，and the detection values were 0.12%-2.69% in Shandong．An efficient，sensitive and quantitative real-time PCR system was established for detecting SdhBH278Y mutation，providing technical support for corynespora leaf spot of cucumber controlling．
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Received: 16 September 2022
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