水稻稻曲病菌PCR快速检测技术及应用  

Rapid PCR detection technique and application for rice false smut

作　　者：曾蓉[1] 徐丽慧[1] 吴雁 高士刚[1] 刘欣 戴富明[1] ZENG Rong;XU Li-hui;WU Yan;GAO Shi-gang;LIU Xin;DAI Fu-ming(Shanghai Key Laboratory of Protection Horticultural Technology,Shanghai 201403,China)

机构地区：[1]上海市农业科学院生态环境保护研究所/上海市设施园艺重点实验室,上海201403

出　　处：《上海农业学报》2018年第6期23-26,共4页Acta Agriculturae Shanghai

基　　金：上海市科技兴农重点攻关项目[沪农科攻字(2015)第4-4号];上海市农业科学院卓越计划[2018(B-1)]

摘　　要：根据NCBI Gen Bank中已登记的水稻稻曲病菌(Ustilaginoidea virens)的全基因组序列,通过对生物信息学方法获得特异性基因,并根据核苷酸序列设计特异性引物。获得的检测引物UV-788F/UV-788R对所有水稻稻曲病菌具有很好的特异性,通过优化退火温度等反应体系,可稳定扩增出788 bp的目的片段,而其他参考菌株及健康种子均无条带。建立的检测方法对菌株基因组DNA的灵敏度为2 pg/μL。对测试样品基因组DNA扩增结果表明:受检的水稻种子检测到水稻稻曲病菌。研究建立的水稻稻曲病病菌的定性检测方法可实现对菌株的分子鉴定和种子的快速检测。According to the whole genome sequence of U.virens in GenBank,aspecific gene was obtained by bioinformatics analysis and used for primer design.The specific primers UV-788F and UV-788R showed good specificity and high sensitivity(2 pg genomic DNA),which being optimized by annealing,steadily generated the amplified target fragment of 788 bp.Meanwhile,the other strands or healthy seeds chosen for comparison turned out no polymorphic bands.The results of the experiment showed the seeds were infected with the disease.Based on the discovery,PRC technique is approved to be an effective method for the detection of the pathogen and assessment of rice seed healthiness.

关 键 词：水稻稻曲病菌 水稻种子 PCR检测 

分 类 号：S435.111.4[农业科学—农业昆虫与害虫防治]