【摘要】 以卵孢小奥德蘑蛛网病菌（异形枝葡霉）为试材，以异形枝葡霉TEF1-α基因为靶标，采用聚合酶链式反应（PCR）方法，设计、合成并筛选出可特异检测异形枝葡霉的PCR引物，基于此建立并应用PCR检测方法，以期为科学防控卵孢小奥德蘑蛛网病提供分子基础。结果表明：利用基于T8F/R引物建立的PCR检测体系对16种枝葡霉属的22个菌株的基因组DNA进行扩增发现，该检测体系特异性强，仅以异形枝葡霉基因组DNA作为模板时，产物呈现出一条467 bp的条带，其它参照菌株及阴性对照均无条带。引物灵敏度验证结果表明，检测体系不受卵孢小奥德蘑组织液的干扰，可检测出最低90 pg·μL^-1的异形枝葡霉基因组DNA,土壤中最低检出限为230个孢子·g^-1。更多还原

【Abstract】 Cladobotryum varium,pathogen of cobweb disease on Oudemansiella raphanipes,was used as test material.Targeting TEF1-α gene of C.varium,design, synthesized and screened to established the system for detection C.varium using the method of polymerase chain reaction（PCR）,in order to provide molecular basis for scientific control of cobweb disease on O.raphanipes.The results showed that the genomic DNA of 22 strains of 16 Cladobotryum taxa were amplified with the PCR detection system established based on T8 F/R and found that the system showed strong specificity.Only when genomic DNA of C.varium was used as template, the PCR products showed a specific band of 467 bp.The sensitivity test results displayed that the detection system based on T8 F/R was not interfered by the tissue fluid of O.raphanipes,and could detect the lowest of 90 pg·μL^-1 C.varium genomic DNA.The minimum detection limit in soil was 230 spores·g^-1.更多还原