摘要
尖孢镰刀菌从形态学和分子生物学上与属内多种菌难于区分,为准确鉴定豆科牧草上的尖孢镰刀菌,本研究基于尖孢镰刀菌与其他镰刀菌的核糖体DNA基因间间隔区IGS(intergenic spacer)序列间的差异,设计了一对特异性引物P_1/P_2,检测体系可以特异地从尖孢镰刀菌中扩增出一条1081bp的条带;引物P_1/P_2对尖孢镰刀菌基因组DNA的检测阈值为100pg,对土壤中尖孢镰刀菌分生孢子的检测灵敏度为100个孢子/g土;同时,引物P_1/P_2也可以对发病牧草根部组织中尖孢镰刀菌的存在进行特异性检测。检测体系可不经过室内病原菌的分离纯化即可有效的从土壤和病株中提取的DNA进行PCR检测鉴定,是一种快速有效的豆科牧草根腐病病原菌分子检测技术。
It is difficult to distinguish Fusarium o3cysporum from other Fusarium species using traditional mot phological observations or molecular analysis based on rDNA--ITS sequencing. In order to accurately identify F. oxysporumin legumes, a pair of primers named P~/P2 was designed based on differences in ribosomal DNA intergenic spacer （rDNA--IGS） sequences of the Fusarium genus, which can be used to amplify DNA from F. oacysporum by conventional PCR. More than 23 species of root rot pathogens were used to verify the specificity of the primers. P1/P2 could amplify a unique 1081 bp sequence from different biotypes of F. oxysporum while it could not amplify from other root rot pathogens. The sensitivity of P1/P2 was 100 pg for genomic DNA and 100 conidia/g soil for the root rot pathogens. Furthermore, this pair of primers could directly amplify se quences from the genomic DNA of F. oxysporum diseased plant samples without pathogen isolation, indicating that this is a rapid and effective legume root rot pathogen detection technology.
出处
《草业学报》
CSCD
北大核心
2016年第5期109-115,共7页
Acta Prataculturae Sinica
基金
农业公益性行业计划和草地病害防治技术研究与示范(201303057)资助
关键词
根腐病
尖孢镰刀菌
基因间隔区
分子检测
root rot
Fusarium oxysporum
ribosomal DNA intergenic spacer
molecular detection
