Ge Chenyang, Dong Daixing, Mao Chengxing, Zhang Qianqian, Zhang Chuanqing
College of Advanced Agricultural Sciences, Zhejiang Agriculture and Forest University, Hangzhou 311300, China.
Extension Centre of Agriculture Technology of Fuyang, Hangzhou 311400, China.
J Fungi (Basel). 2024 Aug 8;10(8):560. doi: 10.3390/jof10080560.
Rice bakanae disease (RBD) is a typical seed-borne fungal disease caused by . Prochloraz is a sterol demethylation inhibitor, which is among the most important classes of active ingredients for the management of RBD. In 2022, the total resistance frequency of to prochloraz in Zhejiang Province was 62.67%. The fitness of the prochloraz-resistant population was lower than that of the susceptible population, but its pathogenicity was slightly stronger. The S312T and F511S double mutations of were detected in the resistant isolates. Loop-mediated isothermal amplification (LAMP) technology based on S312T was established to rapidly determine prochloraz resistance in . LAMP primer mismatch design was performed based on the gene, and 100-300 bp sequences containing a mutation at codon 312 were amplified. In a 25 µL reaction tube, 1 pg/µL DNA of could be detected. The detection limit for the frequency of prochloraz resistance was 0.498% using this method. We performed LAMP detection on rice seedlings inoculated with prochloraz-sensitive and -resistant isolates and treated them with prochloraz. Prochloraz demonstrated good control in rice seedlings. A chromogenic reaction was observed in seedlings treated with prochloraz-resistant isolates, and the results were verified using electrophoresis. It has been demonstrated that LAMP technology based on the S312T genotype can quickly and specifically detect prochloraz-resistant isolates in rice seedlings.
水稻恶苗病（RBD）是一种由……引起的典型种传真菌病害。咪鲜胺是一种甾醇脱甲基抑制剂，是防治RBD最重要的活性成分类别之一。2022年，浙江省……对咪鲜胺的总抗性频率为62.67%。咪鲜胺抗性群体的适合度低于敏感群体，但其致病性略强。在抗性分离株中检测到……的S312T和F511S双突变。基于S312T建立了环介导等温扩增（LAMP）技术，以快速测定……对咪鲜胺的抗性。基于……基因进行LAMP引物错配设计，扩增包含第312位密码子突变的100 - 300 bp序列。在25 μL反应管中，可检测到1 pg/μL的……DNA。使用该方法检测咪鲜胺抗性频率的检测限为0.498%。我们对接种了咪鲜胺敏感和抗性分离株并用咪鲜胺处理的水稻幼苗进行了LAMP检测。咪鲜胺对水稻幼苗表现出良好的防治效果。在用咪鲜胺抗性分离株处理的幼苗中观察到显色反应，结果用电泳进行了验证。已经证明，基于S312T基因型的LAMP技术可以快速、特异性地检测水稻幼苗中的咪鲜胺抗性分离株。
