苏铁叶枯病菌检疫鉴定方法 Detection and identification of Didymella glomerata （Corda） Qian Chen & L. Cai  ICS  65.020.20 CCS  B 16   中华人民共和国国家标准 GB/T 44342—2024 2024-08-2 3发布 2025-03-0 1实施     国家市场监督管理总局 国家标准化管理委员会 发 布目　　次 前言   ·····································································································   Ⅲ 1     范围   ··································································································   1 2     规范性引用文件   ······················································································   1 3     术语和定义   ···························································································   1 4     苏铁叶枯病菌基本信息   ···············································································   1 5     方法原理   ······························································································   1 6     试剂、材料和培养基   ·················································································   1      6.1     试剂和材料   ······················································································   1      6.2     培养基   ···························································································   2 7     仪器和用具   ···························································································   2      7.1     仪器   ······························································································   2      7.2     用具   ······························································································   2 8     检疫鉴定方法   ·························································································   2      8.1     病害症状   ·························································································   2      8.2     病原菌分离培养和形态鉴定   ·····································································   2      8.3     分子生物学鉴定   ·················································································   3 9     结果判定   ······························································································   3 10     样品和档案保存   ·····················································································   3      10.1     样品保存与处理   ················································································   3      10.2     菌株保存与处理   ················································································   3      10.3     结果记录与资料保存   ···········································································   3 附录 A （资料性）　苏铁叶枯病菌的其他信息   ··························································   4      A.1     病害情况   ························································································   4      A.2     寄主范围   ························································································   4      A.3     分布范围   ························································································   4      A.4     传播方式   ························································································   4      A.5     病害症状   ························································································   4      A.6     病原菌形态特征   ·················································································   7 附录 B （资料性）　苏铁叶枯病菌培养基配制方法   ····················································   10      B.1     马铃薯葡萄糖琼脂（PDA）培养基   ····························································   10      B.2     燕麦片琼胶（OA）培养基   ·····································································   10 附录 C （资料性）　苏铁叶枯病菌DNA提取   ··························································   11      C.1     DNA提取方法（改良CTAB提取法）   ························································   11      C.2     DNA纯度与浓度的测定   ·······································································   11 GB/T 44342—2024     Ⅰ 附录 D （规范性）　苏铁叶枯病菌实时荧光PCR检测方法   ···········································   12      D.1     引物和探针序列   ···········