标准解读
《GB/T 44342-2024 苏铁叶枯病菌检疫鉴定方法》是一项由中国发布的国家标准,旨在为苏铁植物中叶枯病菌的检测与鉴定提供一套统一、科学的方法。该标准详细规定了从样本采集、预处理、病原菌分离培养到最终确诊的一系列操作流程和技术要求,以确保检验结果的准确性和可比性。以下是该标准的主要内容概述:
-
范围:明确了本标准适用于苏铁属植物中叶枯病症状的病原菌检疫鉴定工作,为防止该病害的传播和蔓延提供技术支撑。
-
规范性引用文件:列出了实施本标准时所依据或参考的其他相关国家标准或国际标准文档,确保了方法的标准化和兼容性。
-
术语和定义:对苏铁叶枯病菌、检疫、鉴定等相关专业术语进行了明确界定,便于读者理解并统一操作中的概念。
-
采样方法:详细说明了如何在苏铁植株上选择合适的病叶进行采样,包括采样时间、部位、数量以及采样工具的消毒处理等,确保样品具有代表性且不造成病害扩散。
-
样本预处理:描述了样本采集后如何进行初步处理,如清洗、表面消毒、切割等步骤,以减少污染并提高后续检测的准确性。
-
病原菌分离与培养:具体介绍了在无菌条件下,如何从样本中分离出疑似病原菌,并通过特定培养基进行培养观察其生长特征,包括培养条件(温度、湿度、光照)和培养时间等。
-
形态学鉴定:提供了病原菌的微观及宏观形态特征,如菌丝形态、孢子结构等的观察和描述方法,作为鉴定依据。
-
分子生物学鉴定:涉及利用PCR扩增、基因测序等现代生物技术手段,对分离得到的病原菌进行种属水平的确认,提高了鉴定的精确度。
-
结果判定与报告:制定了判断依据和标准,指导如何根据实验结果确定是否检出苏铁叶枯病菌,并规定了报告应包含的信息内容和格式。
-
质量控制:强调了实验过程中应采取的质量控制措施,如使用阳性对照和阴性对照、重复实验验证结果的可靠性等,以确保鉴定工作的科学性和准确性。
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安全与防护:提醒执行标准时应遵守的安全操作规程和采取必要的个人防护措施,以保护操作人员健康和实验室环境安全。
如需获取更多详尽信息,请直接参考下方经官方授权发布的权威标准文档。
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- 2025-03-01 实施
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文档简介
ICS
65.020.20
CCS
B16
中华人民共和国国家标准
GB/T44342—2024
苏铁叶枯病菌检疫鉴定方法
Detectionandidentificationof
Didymellaglomerata(Corda)QianChen&L.Cai
2024-08-23发布2025-03-01实施
国家市场监督管理总局发布
国家标准化管理委员会
GB/T44342—2024
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
Ⅰ
GB/T44342—2024
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
················································································
12
D.2
实时荧光PCR反应体系
·······································································
12
D.3
结果判定
·······················································································
12
附录E(规范性)苏铁叶枯病菌β微管蛋白tub2片段PCR扩增方法
·······························
13
E.1
引物序列
·······················································································
13
E.2
PCR反应体系及参数
···········································································
13
参考文献
································································································
15
Ⅱ
GB/T44342—2024
前言
本文件按照GB/T1.1—2020《标准化工作导则第1部分:标准化文件的结构和起草规则》的规
定起草。
请注意本文件的某些内容可能涉及专利。本文件的发布机构不承担识别专利的责任。
本文件由全国植物检疫标准化技术委员会(SAC/TC271)提出并归口。
本文件起草单位:中国检验检疫科学研究院、中国科学院微生物研究所、深圳仙湖植物园、宁波海
关技术中心、福建省热带作物科学研究所、西藏自治区高原生物研究所。
本文件主要起草人:刘芳、黄英、程颖慧、蔡磊、徐晗、段维军、蔡江桥、罗金水、王聪、严进、
普布多吉。
Ⅲ
GB/T44342—2024
苏铁叶枯病菌检疫鉴定方法
1范围
本文件描述了苏铁叶枯病菌(Didymellaglomerata)的分离培养、形态学鉴定、分子生物学鉴定
方法。
本文件适用于植物及其产品中苏铁叶枯病菌的筛查鉴定。
2规范性引用文件
本文件没有规范性引用文件。
3术语和定义
本文件没有需要界定的术语和定义。
4苏铁叶枯病菌基本信息
学名:Didymellaglomerata(Corda)QianChen&L.Cai
异名:ConiothyriumglomeratumCorda,Icon.Fungorum(Prague)4:39.1840.
Aposphaeriaglomerata(Corda)Sacc.,Syll.Fung.(Abellini)3:175.1884.
Phoma
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