青海省海东市设施番茄根结线虫病原鉴定

doi:10.3969/j.issn.1004-1524.20241094

浙江农业学报 ›› 2025, Vol. 37 ›› Issue (12): 2545-2553.DOI: 10.3969/j.issn.1004-1524.20241094

青海省海东市设施番茄根结线虫病原鉴定

鲍梦楠¹(), 李劲斌¹, 张宪², 闫佳会¹^,^*()

1.青海大学农业病虫害综合治理重点实验室,农业农村部西宁农作物病虫害科学观测实验站,青海西宁 810016
2.海东市平安区蔬菜技术服务中心,青海海东 810699

收稿日期:2024-12-17 出版日期:2025-12-25 发布日期:2026-01-09
作者简介:*闫佳会,E-mail:15209787170@163.com
鲍梦楠(2000—),女,河南洛阳人,硕士研究生,研究方向为植物病理学。E-mail:15737982286@163.com
通讯作者: 闫佳会
基金资助:
青海省中央引导地方科技发展资金项目(2024ZY025)

Identification of Meloidogyne spp. infecting tomato in Haidong City, Qinghai Province, China

BAO Mengnan¹(), LI Jinbin¹, ZHANG Xian², YAN Jiahui¹^,^*()

1. Key Laboratory of Agricultural Integrated Pest Management of Qinghai Province, Xining Crop Diseases and Insect Pests Scientific Observation Experimental Station of Ministry of Agriculture and Rural Affairs, Qinghai University, Xining 810016, China
2. Vegetable Technology Service Center, Ping’an District, Haidong City, Haidong 810699, Qinghai, China

Received:2024-12-17 Online:2025-12-25 Published:2026-01-09
Contact: YAN Jiahui

摘要/Abstract

摘要： 为明确青海省海东市平安区农鑫种植专业合作社番茄大棚中根结线虫(Meloidogyne spp.)的种类,本研究采用形态学与分子生物学方法对该线虫群体进行种类鉴定,并通过室内盆栽接种试验测定其对番茄的致病性。结果表明,该群体雌虫的体值与会阴花纹特征均与花生根结线虫(Meloidogyne arenaria)基本一致;利用花生根结线虫特异序列扩增标记(SCAR)引物Far/Rar进行PCR扩增,获得长度为420 bp的特异性条带。基于rDNA ITS区与28S rRNA D2A/D3B区序列构建的系统发育树显示,该线虫群体与其他花生根结线虫序列聚为一支,同源性高于95%。综合形态学与分子生物学鉴定结果,确定引起该地区番茄根结线虫病的病原为花生根结线虫(M. arenaria)。盆栽接种试验表明,接种该线虫后番茄植株表现为植株矮小、茎秆细弱、叶片黄化;接种30 d后根部可见明显根结,经分离鉴定确认为花生根结线虫。本研究首次在青海省发现并报道了花生根结线虫,为该地区番茄根结线虫病的科学防控提供了依据。

关键词: 番茄, 根结线虫, 花生根结线虫, 病原, 致病性

Abstract:

To identify the root-knot nematode species (Meloidogyne spp.) infecting tomato in greenhouses of the Nongxin Planting Cooperative in Ping’an District, Haidong City, Qinghai Province, this study combined morphological and molecular biological methods for species identification and evaluated its pathogenicity on tomato using indoor pot inoculation tests. The results showed that the morphometric values and perineal patterns of the female adults were consistent with those of Meloidogyne arenaria. PCR amplification with the species-specific SCAR markers Far/Rar produced a specific band of 420 bp. Phylogenetic analysis based on the rDNA ITS and 28S rRNA D2A/D3B regions revealed that the nematode population clustered in a clade with other M. arenaria sequences, showing more than 95% homology. Based on the integrated morphological and molecular data, the pathogen was identified as M. arenaria. Inoculated tomato plants exhibited stunted growth, thin stems, and yellowing leaves. Thirty days after inoculation, distinct root galls were observed, and the re-isolated nematodes were confirmed to be M. arenaria. This study reports the first identification and documentation of M. arenaria in Qinghai Province, providing a scientific basis for the management of tomato root-knot nematode disease in this region.

Key words: tomato, Meloidogyne spp., Meloidogyne arenaria, pathogen, pathogenicity

中图分类号:

S436.412

鲍梦楠, 李劲斌, 张宪, 闫佳会. 青海省海东市设施番茄根结线虫病原鉴定[J]. 浙江农业学报, 2025, 37(12): 2545-2553.

BAO Mengnan, LI Jinbin, ZHANG Xian, YAN Jiahui. Identification of Meloidogyne spp. infecting tomato in Haidong City, Qinghai Province, China[J]. Acta Agriculturae Zhejiangensis, 2025, 37(12): 2545-2553.

图/表 8

表1 本研究所用引物信息

Table 1 Primers used in this study

目标片段 Target region	引物名称 Primer name	序列(5'→3') Sequence(5'→3')	扩增长度 Amplification size/bp	退火温度 Annealing temperature/℃	退火时间 Annealing time/s
rDNA ITS	18S	TTGATTACGTCCCTGCCCTTT	771	55	45
	26S	TTTCACTCGCCGTTACTACGG
28S rDNA	D2A	ACAAGTACCGTGAGGGAAAGTTG	754	56	45
	D3B	TCGGAAGGAACCAGCTACTA
SCAR标记	Far	TCGGCGATAGAGGTAAATGAC	420	61	27
SCAR marker	Rar	TCGGCGATAGACACTACAACT

图1 番茄根结线虫病发病症状 a,发病前期地上部表现缺素;b,发病前期根部出现根结;c,健康番茄植株根部; d,发病后期根部变为褐色并腐烂。

Fig.1 Symptoms of tomato plants infected with root-knot nematodes a, Aboveground symptoms of nutrient deficiency at the early stage of disease; b, Root knots in the root system at the early stage; c, Roots of healthy tomato plant; d, Roots turning brown and rotting at the late stage of disease.

图2 番茄根结线虫群体形态学特征 a,二龄幼虫头部;b,二龄幼虫尾部;c,二龄幼虫整体;d,雌虫;e,雌虫体前端;f和g,会阴花纹。比例尺:a、b、e、f、g为10 μm,c、d为100 μm。

Fig.2 Morphological characteristics of the root-knot nematode population in tomato a, Anterior region of J2; b, Tail region of J2; c, Entire body of J2; d, Female; e, Anterior region of female; f and g, Perineal pattern. Scale bars: a, b, e, f, g=10 μm, c and d=100 μm.

表2 根结线虫群体雌虫与二龄幼虫的体值

Table 2 Morphometric parameters of female and second-stage juveniles (J2) of root-knot nematode population

样本 Sample	体长 Body length/μm	体宽 Body width/μm	口针长 Stylet length/μm	背食道腺开口到口针基部球的距离 Distance from dorsal esophageal gland opening to the stylet knot/μm	尾长 Tail length/ μm	体长/体宽 Body length/ Body width	体长/尾长 Body length/ Tail length	尾长/肛门处虫体宽 Tail length/ Anal body diameter	中食道球宽 Metacorpus width/μm
雌虫 Female (n=20)	571.33± 78.61 (455.51~ 696.03)	403.45± 59.21 (302.02~ 492.74)	12.48± 1.50 (10.09~ 14.80)	3.98±0.57 (3.09~4.98)	—	—	—	—	30.23± 6.33 (21.48~ 39.42)
二龄幼虫 Second-stage juveniles(J2) (n=20)	413.53± 32.64 (372.52~ 486.10)	—	11.33± 1.35 (8.90~ 13.12)	9.23±2.64 (5.04~14.42)	31.19± 2.97 (22.50~ 40.38)	31.30± 2.63	16.34± 1.68	5.10± 1.03	—

图3 番茄群体根结线虫分子生物学鉴定结果 a,根结线虫rDNA ITS与28S rDNA D2/D3区扩增结果;b,花生根结线虫SCAR标记引物(Far/Rar)扩增结果。

Fig.3 Molecular biological identification results of root-knot nematodes in tomato population a, Amplification of rDNA ITS and 28S rDNA D2/D3 regions in root-knot nematodes; b, Amplification with the M. arenaria-specific SCAR primers Far/Rar.

图4 基于rDNA ITS序列构建的根结线虫系统发育树

Fig.4 Phylogenetic tree of root-knot nematodes based on rDNA ITS sequences

图5 基于28S rDNA序列构建的根结线虫系统发育树

Fig.5 Phylogenetic tree of root-knot nematodes based on 28S rDNA sequences

图6 盆栽番茄接种根结线虫后的根部症状与染色结果 a,接种40 d后番茄根部症状;b,接种70 d后番茄根部症状;c,侵染后期番茄根部症状;d,健康植株根部;e~h,根系染色结果。

Fig.6 Root symptoms and staining results of potted tomato plants following root-knot nematode inoculation a, Tomato roots on 40 days post-inoculation; b, Tomato roots on 70 days post-inoculation; c, Roots at late infection stage; d, Roots of healthy plants; e-h, Root staining results.

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青海省海东市设施番茄根结线虫病原鉴定

Identification of Meloidogyne spp. infecting tomato in Haidong City, Qinghai Province, China

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