摘要
【目的】研发简单、快速、准确的分子检测技术用于小麦秆锈菌的准确诊断和秆锈病的早期预警。【方法】采用FIASCO法构建秆锈菌基因组微卫星富集文库,分离微卫星DNA序列,设计合成小麦秆锈菌的特异性引物。【结果】根据小麦秆锈菌基因组微卫星富集文库,设计1对小麦秆锈菌特异性微卫星引物Pgtfssr1(f/r),可在来自中国不同麦区的20份小麦秆锈菌分离物基因组DNA中扩增出395 bp的特异性片段,而在小麦条锈菌、小麦叶锈菌和其它麦类病原真菌中未扩增出该特异性片段。病菌侵入寄主30 h后便可检测到特异性DNA片段的存在,灵敏度达到1 ng.μL-1模板DNA浓度水平。【结论】成功研发出小麦秆锈菌的特异性SSR分子标记,为小麦秆锈病早期诊断在生产上的应用奠定了基础。
【Objective】The objective of this study is to build a simple,quick and accurate molecular technique to detect Puccinia graminis f.sp.tritici and forecast this destructive disease.【Method】A microsatellite-enriched genomic library of Pgt was constructed by using the method of FIASCO.Based on the sequence of Pgt microsatellite,pairs of specific SSR primers were developed and screened.【Result】The primer Pgtfssr1（f/r） generated a polymorphic pattern displaying a 395 bp DNA fragment specific for Pgt,whereas no DNA fragment was obtained in other 24 non-target wheat fungal pathogens.The existence of specific DNA fragment was detected in the infected wheat tissues at 30 h post-inoculation,and the sensitivity of this molecular marker was Pgt DNA template of 1 ng·μL-1.【Conclusion】A specific SSR marker for the detection of wheat stem rust has been successfully developed,which could be used for the early diagnosis and forecast of wheat stem rust.
出处
《中国农业科学》
CAS
CSCD
北大核心
2011年第22期4593-4599,共7页
Scientia Agricultura Sinica
基金
国家重点基础研究发展计划“973”项目(2011CB100403,2010CB951503)
国家公益性行业(农业)科研专项(200903035)
国家小麦产业技术体系(CARS-03-04B)
关键词
小麦秆锈菌
微卫星标记
FIASCO
分子检测
Puccinia graminis f.sp.tritici
microsatellite marker
FIASCO
molecular detection
