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GB/T44611—2024

目次

前言

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

1

范围

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

1

2

规范性引用文件

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

1

3

术语和定义

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

1

4

栽培介质基本信息

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

1

5

检测程序

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

1

6

检测对象及检测依据

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

2

6.1

检测对象

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

2

6.2

检测依据

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

2

7

现场检验

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

3

8

实验室检测

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

3

8.1

昆虫及软体动物检测

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

3

8.2

线虫检测

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

3

8.3

植物病原菌物检测

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

3

8.4

植物病原细菌检测

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

3

8.5

杂草检测

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

4

9

检测结果报告

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

4

10

综合判定

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

4

11

检测记录及样品保存

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

4

11.1

检测记录内容

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

4

11.2

样品及记录保存

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

4

附录A(资料性)

栽培介质中主要检疫性有害生物名单

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

6

A.1

线虫

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

6

A.2

菌物

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

6

A.3

细菌

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

7

附录B(规范性)

线虫分离方法

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

8

B.1

漂浮分离法

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

8

B.2

改良漏斗法

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

8

附录C(规范性)

菌物检测方法

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

9

C.1

洗涤检验法

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

9

C.2

稀释分离法

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

9

C.3

选择性培养基分离法

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

9

参考文献

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

10

GB/T44611—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、宁波海关技术中心、苏州晶睿生物科技有限公司、

满洲里海关、宁夏回族自治区农业技术推广总站、福州海关技术中心、山东大学。

本文件主要起草人:田茜、顾建锋、段维军、吴尧、刘玮琦、李秋实、徐晗、杜伟、于文涛、

张伟。

GB/T44611—2024

栽培介质有害生物检测规程

1范围

本文件给出了栽培介质的基本信息,确立了栽培介质有害生物检测程序,规定了检测对象的确定及

检测依据的选择、现场检验、实验室检测、出具检测结果报告以及综合判定的过程,描述了记录及保存

的证实方法。

本文件适用于进出境及国内生产流通的栽培介质中有害生物的检测鉴定。

2规范性引用文件

下列文件中的内容通过文中的规范性引用而构成本文件必不可少的条款。其中,注日期的引用文

件,仅该日期对应的版本适用于本文件;不注日期的引用文件,其最新版本(包括所有的修改单)适用

于本文件。

GB/T20478植物检疫术语

SN/T1848植物有害生物鉴定规范

3术语和定义

GB/T20478界定的以及下列术语和定义适用于本文件。

3.1

栽培介质growingmedium

植物的根系在其中生长,或能用于此目的的任何物质。

4栽培介质基本信息

栽培介质种类很多,一般分为两大类:无机介质和有机介质。无机介质包括砂、炉渣、矿渣、沸

石、煅烧黏土、陶粒、蛭石、珍珠岩、岩绵、浮石、片岩、火山岩、泡沫塑料等;来源为有机物的介质

称为有机介质,包括泥炭、树皮、木屑、稻壳、污泥、花生壳、甘蔗渣、垃圾堆肥、棉籽壳等。栽培介

质一般由以上一种或几种成分混合而成。

栽培介质的常用成分以及跨境运输中种植用植物与栽培介质通常组合见GB/T43164—2023中附录

A和附录C。

5检测程序

栽培介质有害生物检测程序流程图如图1所示。

1

GB/T44611—2024

图1栽培介质有害生物检测程序流程图

6检测对象及检测依据

6.1检测对象

本文件检测对象为栽培介质可能携带的检疫性有害生物和限定的非检疫性有害生物,主要包括昆虫

和软体动物、线虫、菌物、细菌、杂草等。栽培介质中主要检疫性有害生物名单见附录A,未列出种类

见《中华人民共和国进境植物检疫性有害生物名录》《全国农业植物检疫性有害生物名单》《全国林业

检疫性有害生物名单》等。

6.2检测依据

根据有害生物种类的不同,采用针对各类有害生物的标准,根据形态学特征、生物学特征、生化和

分子生物学特征等进行检疫鉴定。

植物有害生物检测依据可选择采用国际标准、地区性组织标准、国家标准、行业标准、实验室制定

的非标方法、国内外专著、资料及权威期刊公布的方法等。可采用一种或多种方法进行有害生物检测

鉴定。

其他要求按照SN/T1848的规定执行。

2

GB/T44611—2024

7现场检验

将扦取的原始样品放于白色塑料布上,翻开检查,观察有无昆虫,蜗牛、蛞蝓、螺及其他软体动

物,霉变,杂草(籽),植物残体等。如有发现,做初步识别,并收集送实验室做进一步鉴定。

8实验室检测

8.1昆虫及软体动物检测

8.1.1昆虫及软体动物检测步骤如下:

a)将待检栽培介质样品在样品袋内或瓷盘中混合均匀;

b)每批随机取50g~100g,根据其颗粒大小,选用合适孔径的样品筛,用回旋法过筛,回旋次数

不少于20次;

c)将筛上物和筛下物分别倒入白瓷盘或黑底玻璃盘中,用镊子或挑针进行挑选检查,对于不能确

定是否为昆虫的样品或昆虫残肢疑似物,将其放入培养皿在体视显微镜下镜检;

d)用形态特征、解剖、饲养等方法对检获的昆虫和软体动物种类进行形态鉴定,必要时进行分子

生物学鉴定。

8.1.2昆虫常规检测见SN/T2959,软体动物常规检测见SN/T3067。

8.1.3如检出疑似检疫性昆虫或软体动物,需进行进一步检测鉴定,具体检测步骤见相应检疫性昆虫检

疫鉴定标准。

8.2线虫检测

8.2.1线虫检测步骤如下:

a)将待检栽培介质样品在样品袋内或瓷盘中混合均匀;

b)每批随机取50g~100g,采用漂浮分离法和改良漏斗法分离线虫,具体分离步骤按附录B执行;

c)在显微镜下进行形态鉴定,必要时进行分子生物学鉴定。

8.2.2线虫常规检测见SN/T2757。

8.2.3如检出疑似检疫性线虫,需进行进一步检测鉴定,具体检测步骤见相应检疫性线虫检疫鉴定

标准。

8.3植物病原菌物检测

8.3.1植物病原菌物检测步骤如下:

a)将待检栽培介质样品在样品袋内或瓷盘中混合均匀;

b)采用洗涤检验法、稀释分离法和选择性培养基分离法分离菌物,具体分离培养步骤按附录C

执行;

c)用显微镜对分离或培养出的菌物进行形态检查,对可疑类群进行形态学特征观察或分子生物学

鉴定。

8.3.2植物病原菌物常规检测见SN/T2589。

8.3.3如检出疑似检疫性菌物,需进行进一步检测鉴定,具体检测步骤见相应检疫性菌物检疫鉴定

标准。

8.4植物病原细菌检测

8.4.1植物病原细菌检测步骤如下:

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GB/T44611—2024

a)将待检栽培介质样品在样品袋内或瓷盘中混合均匀;

b)每批随机取5g~10g,用无菌水浸泡,振荡10min充分混匀后浸泡4h;

c)浸出液简短离心,去除大的颗粒杂质,收集上清液离心浓缩制备成样品富集液后再进行梯度稀

释,涂布于营养琼脂培养基上,置于适温培养箱中培养;

d)每天检测细菌的生长情况,挑取不同菌落颜色或形态的单菌落进行纯化;

e)对可疑类群进行生理生化、血清学或分子生物学鉴定。

8.4.2植物病原细菌常规检测见SN/T2601。

8.4.3如检出疑似检疫性细菌,需进行进一步检测鉴定,具体检测步骤见相应检疫细菌检疫鉴定标准。

8.5杂草检测

8.5.1杂草检测步骤如下:

a)将待检栽培介质样品在样品袋内或瓷盘中混合均匀;

b)每批随机取50g~100g,平铺在试验台上,用白纸衬底,仔细检查介质样品中有无杂草籽、杂

草植株、带种子的杂草枯枝;

c)通过形态特征鉴定杂草籽,必要时进行分子生物学鉴定。

8.5.2杂草常规检测见SN/T3176。

8.5.3如检出疑似检疫性杂草,需进行进一步检测鉴定,具体检测步骤见相应检疫性杂草检疫鉴定

标准。

9检测结果报告

检测结束应出具检测结果报告。检测结果报告应包含样品的溯源性信息、检测方法、检测结果、检

测人签名、检测时间及检测机构负责人的签名或盖章。

10综合判定

根据现场检验情况和实验室检测结果,不携带有6.1中涉及的有害生物判定为合格,否则判定为不

合格。

11检测记录及样品保存

11.1检测记录内容

检测人员在进行检测时,应有检测过程的详细记录。记录内容包括:

a)所鉴定的植物有害生物学名和中文名;

b)样品的唯一性编码;

c)样品采集、检测和鉴定时间;

d)鉴定所使用的方法以及每种方法所获得的结果;

e)形态特征或测量值,绘图或照片;

f)对于生化和分子检测方法,试验结果应文件化,如鉴定结果所依据的电泳图片、酶联免疫吸附

(ELISA)读出结果的打印单、测序报告等。

11.2样品及记录保存

应保留检出有害生物的培养物、核酸、保存或封片的标本或试验材料、检测记录及结果证据。检测

4

GB/T44611—2024

记录所需保存时间根据鉴定结果而定。一般性有害生物检测记录及结果证据保存6个月,首次检出、重

大检出或当检测结果可能会对贸易方产生不利影响时,检测记录及结果证据保存1年以上,以备复验、

谈判和仲裁。

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GB/T44611—2024

附录A

(资料性)

栽培介质中主要检疫性有害生物名单

A.1线虫

草莓滑刃线虫

Aphelenchoidesfragariae(Ritzema﹘Bos)Christie

菊花滑刃线虫

Aphelenchoidesritzemabosi(Schwartz)Steiner&Buhrer

腐烂茎线虫

DitylenchusdestructorThorne

鳞球茎茎线虫

Ditylenchusdipsaci(Kühn)Filip’

ev

长针线虫属(传毒种类)LongidorusMicoletzky(Filipjev)(Thespeciestransmittingviruses)

根结线虫属(非中国种)MeloidogyneGoeldi(non﹘Chinesespecies)

拟毛刺线虫属(传毒种类)ParatrichodorusSiddiqi(Thespeciestransmittingviruses)

短体线虫(非中国种)PratylenchusFilipjev(non﹘Chinesespecies)

香蕉穿孔线虫

Radopholussimilis(Cobb)Thorne,1949

毛刺线虫属(传毒种类)TrichodorusCobb(Thespeciestransmittingviruses)

剑线虫属(传毒种类)XiphinemaCobb(Thespeciestransmittingviruses)

A.2菌物

葡萄藤猝倒病菌Eutypalata(Pers.)Tul.&C.Tul.

南美大豆猝死综合症病菌FusariumtucumaniaeT.Aoki,O’Donnell,Yos.Homma&Lattanzi

北美大豆猝死综合症病菌FusariumvirguliformeO’Donnell&T.Aoki

松树脂溃疡病菌FusariumcircinatumNirenberg&O’Donnell

芹菜枯萎病菌Fusariumoxysporumf.sp.apii(Nelson&Sherbakoff)Snyder&Hansen

芦笋枯菱病菌Fusariumoxysporumf.sp.asparagiCohen&Heald

香蕉枯菱病菌(4号小种和非中国小种)Fusariumoxysporumf.sp.cubense(E.F.Sm.)W.C.

Snyder&H.N.Hansen(Race4non﹘Chineseraces)

油棕枯萎病菌Fusariumoxysporumf.sp.elaeidisToovey

草莓枯菱病菌Fusariumoxysporumf.sp.fragariaeWinks&Williams

木层孔褐根腐病菌Phellinusnoxius(Corner)G.Cunn.

马铃薯坏疽病菌Phomaexiguaf.sp.foveata(Foister)Boerema

葡萄茎枯病菌Phomaglomerata(Corda)Wollenw.&Hochapfel

栗疫霉黑水病菌Phytophthoracambivora(Petri)Buisman

马铃薯疫霉绯腐病菌PhytophthoraerythrosepticaPethybridge

草莓疫霉红心病菌PhytophthorafragariaeHickman

树莓疫霉根腐病菌Phytophthorafragariaevar.rubiWilcox&Duncan

柑橘冬生疫霉褐腐病菌PhytophthorahibernalisCarne

雪松疫霉根腐病菌PhytophthoralateralisTucker&Milbrath

苜蓿疫霉根腐病菌PhytophthoramedicaginisE.M.Hansen&D.P.Maxwell

菜豆疫霉病菌PhytophthoraphaseoliThaxter

栎树猝死病菌PhytophthoraramorumWerres,DeCock&Manin’tVeld

大豆疫霉病菌PhytophthorasojaeKaufm.&Gerd.

6

GB/T44611—2024

丁香疫霉病菌PhytophthorasyringaeKleb.

小麦基腐病菌Pseudocercosporellaherpotrichoides(Fron)Deighton

油棕猝倒病菌PythiumsplendensBraun

草莓花枯病菌RhizoctoniafragariaeHusain&McKeen

小麦矮腥黑穗病菌TilletiacontroversaJ.G.Kühn

小麦印度腥黑穗病菌TilletiaindicaMitra

苜蓿黄萎病菌Verticilliumalbo﹘atrumReinke&Berthold

棉花黄萎病菌VerticilliumdahliaeKleb.

A.3细菌

魔芋细菌性叶斑病菌Acidovoraxkonjaci(Goto)Willemsetal.

洋葱腐烂病菌Burkholderiagladiolipv.alliicola(Burkholder)comb.nov.

番茄溃疡病菌Clavibactermichiganensissubsp.michiganensis(Smith)Davisetal.

菜豆细菌性萎蔫病菌Curtobacteriumflaccumfacienspv.flaccumfaciens(Hedges)CollinsetJones

菊基腐病菌Dickeyachrysanthemi(Burkholderetal.)Samsonetal.

十字花科蔬菜细菌性黑斑病菌Pseudomonassyringaepv.maculicola(McCulloch)Youngetal.

番茄细菌性叶斑病菌Pseudomonassyringaepv.tomato(Okabe)Young,Dye&Wilkie

香蕉细菌性枯萎病菌Ralstoniasolanacearumrace2(Smith)Yabuuchietal.

7

GB/T44611—2024

附录B

(规范性)

线虫分离方法

B.1漂浮分离法

将待检介质样品放入1000mL烧杯中,加入约800mL水,轻轻搅拌,静置1min~2min。较轻的

孢囊和一些有机杂物陆续向上漂浮,集中于水面。将漂浮物倒入20目(筛网孔径0.85mm)和60目

(筛网孔径0.25mm)的套筛,淋洗60目下筛的漂浮物。用滤纸过滤漂浮物,收集漂浮物在滤纸上,

在解剖镜下检查孢囊。

B.2改良漏斗法

改良漏斗分离装置由一只较大并类似于贝曼漏斗的玻璃或有机玻璃作为外壳,上部装置则与浅盘装

置相似。将线虫滤纸或双层纸巾平放在筛盘筛网上,用水淋湿滤纸边缘与筛盘结合部分,将待分离线虫

的样品放置其上,从筛盘下部的空隙中将水注入分离器中,以淹没供分离的材料为止,在

20

℃~25℃

下放置24h~48h后,打开止水夹,用离心管接取约5mL的水样,静置20min左右或1500r/min离心

3min,吸去离心管内上层清液,即获得浓度较高的线虫水悬浮液。

8

GB/T44611—2024

附录C

(规范性)

菌物检测方法

C.1洗涤检验法

随机取50g待检介质样品,倒入灭菌处理的250mL三角瓶内,加蒸馏水100mL,表面活性剂

吐温﹘201滴~2滴。用铝箔纸或Parafilm膜将三角瓶封口,置于振荡器上振荡10min。振荡结束立即取

下三角瓶,将洗涤悬浮液倒入孔径分别为上层120µm、中层50µm、下层15µm的混合套筛。将

15µm套筛筛上物加水约10mL收集,注

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