標(biāo)準(zhǔn)解讀

《GB/T 44342-2024 蘇鐵葉枯病菌檢疫鑒定方法》是一項(xiàng)由中國發(fā)布的國家標(biāo)準(zhǔn)，旨在為蘇鐵植物中葉枯病菌的檢測與鑒定提供一套統(tǒng)一、科學(xué)的方法。該標(biāo)準(zhǔn)詳細(xì)規(guī)定了從樣本采集、預(yù)處理、病原菌分離培養(yǎng)到最終確診的一系列操作流程和技術(shù)要求，以確保檢驗(yàn)結(jié)果的準(zhǔn)確性和可比性。以下是該標(biāo)準(zhǔn)的主要內(nèi)容概述：

  1. 范圍：明確了本標(biāo)準(zhǔn)適用于蘇鐵屬植物中葉枯病癥狀的病原菌檢疫鑒定工作，為防止該病害的傳播和蔓延提供技術(shù)支撐。

  2. 規(guī)范性引用文件：列出了實(shí)施本標(biāo)準(zhǔn)時(shí)所依據(jù)或參考的其他相關(guān)國家標(biāo)準(zhǔn)或國際標(biāo)準(zhǔn)文檔，確保了方法的標(biāo)準(zhǔn)化和兼容性。

  3. 術(shù)語和定義：對蘇鐵葉枯病菌、檢疫、鑒定等相關(guān)專業(yè)術(shù)語進(jìn)行了明確界定，便于讀者理解并統(tǒng)一操作中的概念。

  4. 采樣方法：詳細(xì)說明了如何在蘇鐵植株上選擇合適的病葉進(jìn)行采樣，包括采樣時(shí)間、部位、數(shù)量以及采樣工具的消毒處理等，確保樣品具有代表性且不造成病害擴(kuò)散。

  5. 樣本預(yù)處理：描述了樣本采集后如何進(jìn)行初步處理，如清洗、表面消毒、切割等步驟，以減少污染并提高后續(xù)檢測的準(zhǔn)確性。

  6. 病原菌分離與培養(yǎng)：具體介紹了在無菌條件下，如何從樣本中分離出疑似病原菌，并通過特定培養(yǎng)基進(jìn)行培養(yǎng)觀察其生長特征，包括培養(yǎng)條件（溫度、濕度、光照）和培養(yǎng)時(shí)間等。

  7. 形態(tài)學(xué)鑒定：提供了病原菌的微觀及宏觀形態(tài)特征，如菌絲形態(tài)、孢子結(jié)構(gòu)等的觀察和描述方法，作為鑒定依據(jù)。

  8. 分子生物學(xué)鑒定：涉及利用PCR擴(kuò)增、基因測序等現(xiàn)代生物技術(shù)手段，對分離得到的病原菌進(jìn)行種屬水平的確認(rèn)，提高了鑒定的精確度。

  9. 結(jié)果判定與報(bào)告：制定了判斷依據(jù)和標(biāo)準(zhǔn)，指導(dǎo)如何根據(jù)實(shí)驗(yàn)結(jié)果確定是否檢出蘇鐵葉枯病菌，并規(guī)定了報(bào)告應(yīng)包含的信息內(nèi)容和格式。

  10. 質(zhì)量控制：強(qiáng)調(diào)了實(shí)驗(yàn)過程中應(yīng)采取的質(zhì)量控制措施，如使用陽性對照和陰性對照、重復(fù)實(shí)驗(yàn)驗(yàn)證結(jié)果的可靠性等，以確保鑒定工作的科學(xué)性和準(zhǔn)確性。

  11. 安全與防護(hù)：提醒執(zhí)行標(biāo)準(zhǔn)時(shí)應(yīng)遵守的安全操作規(guī)程和采取必要的個(gè)人防護(hù)措施，以保護(hù)操作人員健康和實(shí)驗(yàn)室環(huán)境安全。


如需獲取更多詳盡信息，請直接參考下方經(jīng)官方授權(quán)發(fā)布的權(quán)威標(biāo)準(zhǔn)文檔。

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文檔簡介

ICS

65.020.20

CCS

B16

中華人民共和國國家標(biāo)準(zhǔn)

GB/T44342—2024

蘇鐵葉枯病菌檢疫鑒定方法

Detectionandidentificationof

Didymellaglomerata（Corda）QianChen&L.Cai

2024-08-23發(fā)布2025-03-01實(shí)施

國家市場監(jiān)督管理總局發(fā)布

國家標(biāo)準(zhǔn)化管理委員會

GB/T44342—2024

目次

前言

·····································································································

Ⅲ

1

范圍

··································································································

1

2

規(guī)范性引用文件

······················································································

1

3

術(shù)語和定義

···························································································

1

4

蘇鐵葉枯病菌基本信息

···············································································

1

5

方法原理

······························································································

1

6

試劑、材料和培養(yǎng)基

·················································································

1

6.1

試劑和材料

······················································································

1

6.2

培養(yǎng)基

···························································································

2

7

儀器和用具

···························································································

2

7.1

儀器

······························································································

2

7.2

用具

······························································································

2

8

檢疫鑒定方法

·························································································

2

8.1

病害癥狀

·························································································

2

8.2

病原菌分離培養(yǎng)和形態(tài)鑒定

·····································································

2

8.3

分子生物學(xué)鑒定

·················································································

3

9

結(jié)果判定

······························································································

3

10

樣品和檔案保存

·····················································································

3

10.1

樣品保存與處理

················································································

3

10.2

菌株保存與處理

················································································

3

10.3

結(jié)果記錄與資料保存

···········································································

3

附錄A（資料性）蘇鐵葉枯病菌的其他信息

··························································

4

A.1

病害情況

························································································

4

A.2

寄主范圍

························································································

4

A.3

分布范圍

························································································

4

A.4

傳播方式

························································································

4

A.5

病害癥狀

························································································

4

A.6

病原菌形態(tài)特征

·················································································

7

附錄B（資料性）蘇鐵葉枯病菌培養(yǎng)基配制方法

····················································

10

B.1

馬鈴薯葡萄糖瓊脂（PDA）培養(yǎng)基

····························································

10

B.2

燕麥片瓊膠（OA）培養(yǎng)基

·····································································

10

附錄C（資料性）蘇鐵葉枯病菌DNA提取

··························································

11

C.1

DNA提取方法（改良CTAB提取法）

························································

11

C.2

DNA純度與濃度的測定

·······································································

11

Ⅰ

GB/T44342—2024

附錄D（規(guī)范性）蘇鐵葉枯病菌實(shí)時(shí)熒光PCR檢測方法

···········································

12

D.1

引物和探針序列

················································································

12

D.2

實(shí)時(shí)熒光PCR反應(yīng)體系

·······································································

12

D.3

結(jié)果判定

·······················································································

12

附錄E（規(guī)范性）蘇鐵葉枯病菌β微管蛋白tub2片段PCR擴(kuò)增方法

·······························

13

E.1

引物序列

·······················································································

13

E.2

PCR反應(yīng)體系及參數(shù)

···········································································

13

參考文獻(xiàn)

································································································

15

Ⅱ

GB/T44342—2024

前言

本文件按照GB/T1.1—2020《標(biāo)準(zhǔn)化工作導(dǎo)則第1部分：標(biāo)準(zhǔn)化文件的結(jié)構(gòu)和起草規(guī)則》的規(guī)

定起草。

請注意本文件的某些內(nèi)容可能涉及專利。本文件的發(fā)布機(jī)構(gòu)不承擔(dān)識別專利的責(zé)任。

本文件由全國植物檢疫標(biāo)準(zhǔn)化技術(shù)委員會（SAC/TC271）提出并歸口。

本文件起草單位：中國檢驗(yàn)檢疫科學(xué)研究院、中國科學(xué)院微生物研究所、深圳仙湖植物園、寧波海

關(guān)技術(shù)中心、福建省熱帶作物科學(xué)研究所、西藏自治區(qū)高原生物研究所。

本文件主要起草人：劉芳、黃英、程穎慧、蔡磊、徐晗、段維軍、蔡江橋、羅金水、王聰、嚴(yán)進(jìn)、

普布多吉。

Ⅲ

GB/T44342—2024

蘇鐵葉枯病菌檢疫鑒定方法

1范圍

本文件描述了蘇鐵葉枯病菌（Didymellaglomerata）的分離培養(yǎng)、形態(tài)學(xué)鑒定、分子生物學(xué)鑒定

方法。

本文件適用于植物及其產(chǎn)品中蘇鐵葉枯病菌的篩查鑒定。

2規(guī)范性引用文件

本文件沒有規(guī)范性引用文件。

3術(shù)語和定義

本文件沒有需要界定的術(shù)語和定義。

4蘇鐵葉枯病菌基本信息

學(xué)名：Didymellaglomerata（Corda）QianChen&L.Cai

異名：ConiothyriumglomeratumCorda,Icon.Fungorum（Prague）4:39.1840.

Aposphaeriaglomerata（Corda）Sacc.,Syll.Fung.（Abellini）3:175.1884.

Phoma

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