作物杂志, 2026, 42(4): 95-100 doi: 10.16035/j.issn.1001-7283.2026.04.011

遗传育种·种质资源·生物技术

中俄大豆种质资源对大豆胞囊线虫3号生理小种的抗性评价

项鹏,1, 魏然1, 杨树1, 张武1, 韩德志1, 尤佳2, 栗铭徽3, 刘大伟,4

1 黑龙江省农业科学院黑河分院/农业农村部黑河野外综合科学观测研究站164300黑龙江黑河

2 黑龙江省农业科学院大豆研究所150086黑龙江哈尔滨

3 北安市大龙种业有限责任公司164021黑龙江北安

4 东北农业大学植物保护学院150030黑龙江哈尔滨

Resistance Evaluation of Chinese and Russian Soybean Germplasm Resources against Soybean Cyst Nematode Race 3

Xiang Peng,1, Wei Ran1, Yang Shu1, Zhang Wu1, Han Dezhi1, You Jia2, Li Minghui3, Liu Dawei,4

1 Heihe Branch of Heilongjiang Academy of Agricultural Sciences / Heihe Comprehensive Field Scientific Observation and Research Station, Ministry of Agriculture and Rural Affairs, Heihe 164300, Heilongjiang, China

2 Soybean Research Institute, Heilongjiang Academy of Agricultural Sciences, Harbin 150086, Heilongjiang, China

3 Limited Liability Company of Dalong Seed Industry, Beian 164021, Heilongjiang, China

4 College of Plant Protection, Northeast Agricultural University, Harbin 150030, Heilongjiang, China

通讯作者: 刘大伟,主要从事农作物病虫害研究,E-mail:liudawei353@163.com

收稿日期: 2026-01-4   修回日期: 2026-06-3   网络出版日期: 2026-06-09

基金资助: 黑龙江省自然科学基金资助项目(PL2024C032)
黑龙江省重点研发计划项目(2024ZXDXB34)
国家大豆产业技术体系线虫防控岗(CARS-04-PS27)

Received: 2026-01-4   Revised: 2026-06-3   Online: 2026-06-09

作者简介 About authors

项鹏,主要从事植物线虫研究,E-mail:xp_303@126.com

摘要

在自然条件下利用田间和盆栽相结合的传统方法,从537份中俄大豆种质中鉴定出19份对大豆胞囊线虫3号生理小种具有良好抗性的资源,占鉴定总数的3.54%,其中中国抗性资源11份,俄罗斯抗性资源8份。通过KASP标记技术,对控制该性状的主效位点Rhg1Rhg4进行了等位基因检测,其中9份抗性材料在Rhg1Rhg4有抗性位点,4份材料在Rhg4有抗性位点。这些抗性资源的农艺性状优良,不仅在生产中为大豆胞囊线虫3号生理小种高发区提供了抗性品种,也可以直接作为抗性亲本应用于育种,推进大豆胞囊线虫抗病品种选育的进程。

关键词: 大豆种质资源; 3号生理小种; 抗性评价; KASP标记

Abstract

The resistance of 537 soybean germplasm materials originating from China and Russia to soybean cyst nematode (SCN) race 3 was evaluated using a combination of field screening under natural infestation and pot experiments with infested soil. A total of 19 resistant materials were identified, accounting for 3.54% of the tested germplasm, including 11 accessions from China and eight from Russia. Kompetitive Allele-Specific PCR (KASP) markers were used to genotype the major resistance loci, Rhg1 and Rhg4. Among the resistant materials, nine carried resistance-associated alleles at both Rhg1 and Rhg4, while four possessed resistance alleles only at the Rhg4 locus. These resistant germplasm resources exhibited favorable agronomic traits and can serve not only as resistant cultivars for areas with a high incidence of soybean cyst nematode race 3 but also as valuable parental materials for resistance breeding programs. The identified resources provide important genetic materials for accelerating the development of soybean cyst nematode-resistant soybean cultivars.

Keywords: Soybean germplasm resources; Cyst nematode race 3; Resistance evaluation; KASP marker

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项鹏, 魏然, 杨树, 张武, 韩德志, 尤佳, 栗铭徽, 刘大伟. 中俄大豆种质资源对大豆胞囊线虫3号生理小种的抗性评价. 作物杂志, 2026, 42(4): 95-100 doi:10.16035/j.issn.1001-7283.2026.04.011

Xiang Peng, Wei Ran, Yang Shu, Zhang Wu, Han Dezhi, You Jia, Li Minghui, Liu Dawei. Resistance Evaluation of Chinese and Russian Soybean Germplasm Resources against Soybean Cyst Nematode Race 3. Crops, 2026, 42(4): 95-100 doi:10.16035/j.issn.1001-7283.2026.04.011

大豆胞囊线虫病(soybean cyst nematode,SCN)是大豆生产中的重要病害,一般可造成大豆减产5%~10%,危害严重的地块甚至绝收,该病害很大程度上制约着我国大豆经济及产量的增长[1]。防治大豆胞囊线虫方法有很多,而应用抗病品种是最经济有效的解决途径。3号生理小种是黑龙江地区的优势病原群体[2],因此筛选针对该小种的抗源对于保障当地大豆抗病育种的成效具有至关重要的意义。

我国大豆抗胞囊线虫病育种的早期工作为后续相关研究奠定了重要基础。全国大豆种质抗胞囊线虫鉴定研究协作组研究[3]表明,1986-1990年间,通过对10 000余份大豆种质开展系统鉴定,成功筛选出对3号生理小种免疫材料3份、抗性材料18份以及抗4号小种材料2份。周长军等[4]在2020-2021年,通过田间病圃与室内盆栽接种试验,对620份东北地区常用大豆亲本进行了3号生理小种抗性鉴定,结果表明达到中抗及以上水平的品种(系)共30份,占鉴定总数的4.84%。李明姝等[5]于2014年在温室利用盆栽鉴定的方法对3号生理小种进行了抗病性鉴定,从779份大豆种质资源中筛选出19份抗病种质,占鉴定总数的2.44%。此后,大豆种质资源对大豆胞囊线虫不同生理小种的抗性鉴定逐渐成为一个活跃的研究方向,相关研究得以不断推进[6-8]

随着科技的发展,Kadam等[9]基于Rhg1Rhg4位点的基因组信息,开发了相应的KASP标记,实现了大豆抗SCN相关位点的高通量检测。练云等[10]利用KASP标记技术在487份材料中鉴定出22份携带Rhg1Rhg4抗性位点的材料,通过接种鉴定发现,仅5份材料对2号生理小种表现出实际抗性。魏荷等[11]从333份大豆种质中鉴定出19份对2号生理小种具有抗性的材料,经KASP标记证实这些抗性种质均含有Rhg1Rhg4主效位点。KASP标记的应用提高了对抗病材料的选择效率,因此,开展抗大豆胞囊线虫病评价时,需要分子标记和抗性鉴定相结合。

本研究以中国黑龙江省与俄罗斯阿穆尔州大豆主产区的537份核心种质为试验材料,采用表型鉴定与KASP分子标记检测相结合的技术体系,系统评价核心种质对大豆胞囊线虫3号生理小种的抗性,明确抗性位点基因型与农艺品质特征,旨在为黑龙江省大豆抗胞囊线虫育种提供多元化抗源与科学依据。

1 材料与方法

1.1 试验材料

本研究共收集了中俄大豆种质资源537份,其中中国大豆种质资源278份,主要来源于我国黑龙江大豆主产区,俄罗斯大豆种质资源259份,主要来源于俄罗斯阿穆尔州大豆主产区,具体材料信息详见网络增强出版附加材料附表1。大豆胞囊线虫采自黑龙江省农业科学院黑河分院大豆胞囊线虫病圃。

表1   抗性鉴定分级标准

Table 1  Grading standard for resistance identification

抗性级别
Resistance
class
分级标准
Grading standard
抗性评价
Resistance
evaluation
1单株平均胞囊数为0个,植株生长正常免疫(I)
3
单株平均胞囊数在0.1~3.0个,植株生长正常高抗(R)
5
单株平均胞囊数在3.1~10.0个,植株生长基本正常或部分矮黄中抗(MR)
7
单株平均胞囊数在10.1~30.0个,植株矮小,叶片发黄,结实少感(S)
9
单株平均胞囊数在30.1个以上,植株不结实,干枯死亡高感(HS)

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1.2 试验设计

1.2.1 生理小种鉴定

采用Golden等[12]建立的一套鉴别寄主,鉴定试验病圃中的生理小种,以监测生理小种的动态变化。鉴定方法采用Riggs等[13]的鉴定模式,以雌虫指数(female index,FI)为抗性级别,FI≥10%为感病(S);FI<10%为抗病(R)。

1.2.2 田间鉴定

试验在黑龙江省农业科学院黑河分院大豆胞囊线虫病圃进行,地块自然感染3号生理小种。田间鉴定采用顺序排列,每份材料播种3行,行长2 m,株距5.00 cm,行距0.65 m。大豆出苗30 d后监测感病品种根部SCN的发育情况,当SCN白色雌虫完全突出表皮时,记录根上的白色雌虫数,每份材料调查10株。参照中国大豆资源抗胞囊线虫鉴定分级标准[3],对大豆种质资源进行抗性评价(表1)。

1.2.3 盆栽抗性鉴定

对田间表现抗性的材料进行盆栽抗性鉴定。将平均胞囊量40~52个/100 g风干土的病土装入塑料盆(直径20 cm)中,每个供试材料和鉴别寄主各播种3盆,每盆留苗5株,播后30 d左右第1代胞囊显囊期,扣盆洗根,对所有鉴定材料逐株调查,记录根上着生胞囊数,并统计雌虫指数以评价抗性级别。按Schmitt等[14]提出的鉴定大豆抗病性标准进行分级(表2)。FI(%)=鉴别材料单株平均着生胞囊数/Lee(CKS)单株平均着生胞囊数×100。

表2   抗性鉴定分级标准

Table 2  Grading standards for resistance identification

FI (%)抗性级别Resistance class
0.0~10.0高抗(R)
10.1~30.0中抗(MR)
30.1~60.0中感(MS)
> 60.0感(S)

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1.2.4 KASP标记的分析

采用CTAB法从抗性材料叶片中提取基因组DNA,并利用Nanodrop 2000测定其浓度与纯度。参照Kadam等[9]报道的KASP标记设计合成引物,根据LGC Genomics KASPTM基因分型试剂盒(KBS-1016-002)说明书配制PCR反应体系,反应程序参照魏荷等[11]的方法。将扩增产物置于Pherastar SNP基因分型仪上进行荧光检测,依据荧光信号比值判定SNP基因型。以抗病品种PI90763(CKR)和感病品种Lee(CKS)的KASP基因型为参照[15-16]

1.2.5 抗性种质资源农艺及品质性状调查

将田间鉴定和盆栽鉴定筛选出来的19份抗性材料随机区组排列播种在黑河分院试验基地,在大豆生长期调查抗性材料的农艺性状,参照邱丽娟等[17]的方法调查指标。在收获期调查百粒重,并测定蛋白质和脂肪含量。

1.3 数据处理

采用WPS软件进行数据处理与图表制作,使用WPS函数计算每组数据的平均值。

2 结果与分析

2.1 3号生理小种的鉴定

根据生理小种鉴定结果(表3),确定试验病圃中的线虫种群为大豆胞囊线虫3号生理小种,其生理小种类型未发生变异。

表3   大豆胞囊线虫3号生理小种鉴定结果

Table 3  Identification results of soybean cyst nematode race 3

鉴定材料
Identify
material
单株胞囊平均数
Average number
of cysts per plant
FI
(%)
抗性评价
Resistance
evaluation
Pickett1.000.68R
Peking4.001.90R
PI887883.002.04R
PI907630.670.46R
Lee(CKS147.00100.00S

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2.2 大豆种质资源对3号生理小种的田间抗性鉴定评价

537份中俄大豆种质资源对3号生理小种抗性鉴定结果(表4)未发现表现免疫的品种。其中,19个大豆种质资源表现为抗病,占鉴定材料总数的3.54%,其中6个材料表现为高抗,13个材料表现为中抗。有518个大豆种质资源对3号生理小种表现为高感或感病,占鉴定材料总数的96.46%。

表4   537份大豆种质资源对3号生理小种抗性评价结果

Table 4  Evaluation results of resistance to cyst nematode race 3 in 537 soybean germplasm resources

抗性评价
Resistance evaluation
材料数
Number of materials
所占比例
Percentage (%)
R61.12
MR132.42
S19836.87
HS32059.59

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2.3 大豆种质资源对3号生理小种的盆栽鉴定评价

经鉴定(表5),19份材料对大豆胞囊线虫3号生理小种均表现抗病,其中表现高抗的材料有安2-318、庆豆13、黑农531、抗线虫12、октябрь70和соната,表现中抗的材料有嫩丰18、农庆豆24、黑河52、黑河46、鹏豆158、齐农5、齐农12、Вецауцес、Киевская、Малета、Лондон、Росинка和Сентябрика。19份抗性材料中11份来自中国,8份来自俄罗斯。

表5   19份抗病材料的盆栽鉴定结果

Table 5  Results of pot identification for 19 resistant materials

编号
Number
材料
Material
平均雌虫数
Average number
of females
FI
(%)
抗性评价
Resistance
evaluation
来源
Origin
1安2-3188.476.26R中国
2庆豆139.176.78R中国
3黑农5316.895.10R中国
4抗线虫124.553.37R中国
5октябрь705.894.36R俄罗斯
6соната6.224.60R俄罗斯
7嫩丰1815.4111.40MR中国
8农庆豆2421.3615.80MR中国
9黑河5241.2229.75MR中国
10黑河4638.2428.28MR中国
11鹏豆15817.2412.75MR中国
12齐农522.1816.40MR中国
13齐农1222.6416.74MR中国
14Вецауцес33.4624.74MR俄罗斯
15Киевская15.3411.35MR俄罗斯
16Малета20.1414.90MR俄罗斯
17Лондон33.1524.52MR俄罗斯
18Росинка32.5724.09MR俄罗斯
19Сентябрика28.2920.92MR俄罗斯
20Lee(CKS135.21100.00

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2.4 抗性位点分析

利用KASP标记对19份材料进行检测(表6),其中有9份材料基因型与抗病品种PI90763一致,在Rhg1Rhg4位点都有SCN抗性位点,属于Peking抗性类型。有4份材料在Rhg4位点检测到有SCN抗性位点,属于Rhg4抗性类型。6份材料基因型与感病品种Lee一致,没有检测到Rhg1Rhg4位点,这些材料的抗性可能源于其他未知的抗性位点。

表6   19份抗性材料的KASP标记分析

Table 6  KASP marker analysis of 19 resistant materials

编号
Number
材料
Material
Rhg1位点标记
Rhg1 locus marker
Rhg4位点标记
Rhg4 locus marker
Rhg1-2Rhg1-5Rhg4-3Rhg4-5
1安2-318++++
2庆豆13++++
3黑农531++++
4抗线虫12++++
5октябрь70--++
6соната--++
7嫩丰18++++
8农庆豆24++++
9黑河52----
10黑河46----
11鹏豆158++++
12齐农5++++
13齐农12--++
14Вецауцес----
15Киевская----
16Малета----
17Лондон----
18Росинка--++
19Сентябрика++++
20Lee(CKSCCGGAACC
21PI90763(CKRGGCCTTGG

“+”表示携带抗性等位基因;“-”表示未携带抗性等位基因。

“+”indicates carrying the resistance allele;“-”indicates no carrying the resistance allele.

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2.5 抗性种质资源农艺及品质性状分析

对19份抗病材料的农艺及品质性状调查结果(表7)显示,其中6份高抗材料种皮均为黄色,可在抗病育种中作为抗性亲本直接应用。oктябрь70和соната俄罗斯高抗材料种皮呈黄色,种脐呈淡黄色,且蛋白质和脂肪总含量都超过了60.00%,也是较好的抗性亲本材料。黑农531对3号生理小种表现出稳定抗性,可在大豆胞囊线虫高发区推广应用。

表7   19份抗性材料的农艺及品质性状分析

Table 7  Analysis of agronomic and quality traits of 19 resistant materials

编号
Number
材料
Material
株高
Plant
height (cm)
生育期
Growth
period (d)
花色
Flower
color
叶形
Leaf
shape
脐色
Hilum
color
种皮颜色
Seed coat
color
蛋白质含量
Protein
content (%)
脂肪含量
Fat content
(%)
百粒重
100-seed
weight (g)
1安2-31880123白色圆叶黄色黄色42.0720.3020.0
2庆豆1390123紫色圆叶黑色黄色41.0621.0919.0
3黑农53185123白色尖叶黄色黄色38.1722.3421.3
4抗线虫1290123紫色圆叶淡褐色黄色39.7720.8919.4
5октябрь7080105紫色尖叶淡褐色黄色40.3021.6018.0
6соната7598紫色尖叶淡黄色黄色40.9020.7015.6
7嫩丰1885116紫色圆叶淡褐色黄色40.2819.9719.5
8农庆豆2478123白色圆叶褐色黄色38.5321.0219.0
9黑河5290110紫色尖叶淡黄色黄色40.5521.5220.6
10黑河4675112紫色尖叶淡黄色黄色39.7420.1117.9
11鹏豆15880115白色尖叶淡褐色黄色39.0822.1621.8
12齐农5100123白色尖叶淡褐色黄色41.5021.9020.0
13齐农1293120紫色圆叶黄色黄色39.7120.7621.0
14Вецауцес75115紫色尖叶淡黄色黄色41.8418.9820.0
15Киевская85117紫色卵圆叶褐色黄色41.3921.8521.0
16Малета78108白色尖叶黑色黑色39.0521.9119.4
17Лондон90117白色圆叶淡褐色黄色38.5621.0920.5
18Росинка80100紫色尖叶黄色黄色41.4121.5016.0
19Сентябрика7395白色卵圆叶褐色黑色41.4217.9615.5

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3 讨论

虽然我国大豆品种资源丰富,在抗病资源发掘与利用方面具有独特优势,但是这些抗线品种的遗传基础非常狭窄,抗病基因来源单一[18]。大豆胞囊线虫具有毒力多样性,在田间以混合群体形式存在,存在多个生理小种。长期依赖单一抗线品种进行防治,会对线虫群体产生强大的定向选择压力,从而导致毒力小种迅速富集,最终致使该品种丧失抗性。研究[2]表明,在黑龙江省,大豆胞囊线虫以毒力较弱的3号生理小种为主,6号、4号和14号生理小种偶有发生。黑龙江地区大豆生产中大多数抗性品种的抗性单一,仅有的抗线品种对3号生理小种的抗性已出现减弱或丧失[19]。因此,利用新的抗源或新型抗病基因是拓宽抗病品种遗传基础的重要途径。国内外研究[20-21]主要集中在抗病基因鉴定与利用、新型抗病基因发掘2个方面。

大豆胞囊线虫的抗性是受多个位点控制的数量性状遗传[22],研究[23-24]表明,Rhg1Rhg4是提供大豆胞囊线虫抗性的重要位点。Rhg1位于第18号染色体上,控制着大豆对多个生理小种的抗性[25],此位点内的GmAAT、a-SNAP以及WIP蛋白3个基因参与了对大豆胞囊线虫的抗性,在感病材料中将这3个基因都进行超表达后,发现感病材料的抗性明显提升[26]Rhg4位于第8号染色体上[27],能够编码丝氨酸羟甲基转移酶(Shmt),Shmt的活性被认为是决定抗性程度的关键因子,在不同抗性品种中存在显著差异[28]。对试验材料接种3号生理小种后进行功能验证,证明定位在Rhg4位点的GmSHMT基因可正向调控大豆种质对胞囊线虫病的抗性[29]。除Rhg1Rhg4外,基因组中还分布多个尚未广泛利用的主效QTL、微效位点及新型抗性基因,可独立或协同介导抗性。研究[30-32]发现,10号染色体上的位点可独立介导广谱抗性,过表达候选基因GmTGA1-10GmSCT-10可显著降低胞囊数[30];14号染色体上的GmSNAP14通过感病基因缺失突变赋予广谱抗性,与Rhg1Rhg4机制完全不同[31];此外在7、9、11、14、19和20号染色体上均鉴定到可稳定表达的新型抗性QTL,部分位点在Rhg1Rhg4感病背景下仍能发挥作用[32]。本研究利用的KASP分子标记技术在抗胞囊线虫基因型筛选鉴定中已得到广泛应用[33],检测显示有6份材料在Rhg1Rhg4的2个主效位点均未检测到抗性等位基因,但其田间与盆栽鉴定均表现出稳定的抗性,明确证明这些材料不依赖已知主效位点,而携带新型抗性遗传位点。这一结果对破解当前抗大豆胞囊线虫育种遗传基础狭窄和抗性易失效的关键问题具有重要理论与应用价值。

研究[34-36]报道,在黑龙江省的63个大豆种植区都检测到大豆胞囊线虫,个别地区发病尤为严重。应用抗病品种是最经济有效的防治手段,能有效挽回因病害造成的产量损失,直接提升单产和总产,可保障大豆产业安全,增加农民收入,同时还可以显著减少化学杀线虫剂的使用,符合“双减”和绿色可持续发展的国家战略。在重病区种植抗病品种,能够有效降低土壤中的线虫基数,逐步修复被大豆胞囊线虫严重侵染的耕地,恢复其生产能力,实现藏粮于地的战略目标。

4 结论

本研究首次对中俄大豆种质资源开展大豆胞囊线虫3号生理小种抗性系统评价,田间自然病圃与盆栽人工接种鉴定结果完全一致,其中获得11份中国资源、8份俄罗斯种质资源,为黑龙江省抗性资源的丰富及应用奠定了基础。本研究发现了6份不依赖Rhg1Rhg4主效位点的抗性材料,是挖掘新抗病基因和解析新型抗性机制的核心材料,将这些核心材料的新型位点导入骨干亲本,可构建多基因聚合抗病材料,从根本上提升黑龙江省大豆抗大豆胞囊线虫育种的遗传宽度与长久抗性,为绿色防控与粮食安全提供关键支撑。

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大豆在我国国民经济中扮演着重要角色,目前我国是全球最大的大豆消费国、进口国,且进口大豆数量逐年递增。大豆孢囊线虫病是威胁全球主要大豆产地的重要病害,每年全球范围内造成超过数十亿美元经济损失,防控形势严峻。抗性品种的种植是防控大豆孢囊线虫病最经济有效的措施。然而,单一抗性品种的过度使用及大豆孢囊线虫生理小种不断演化,导致抗性降低,威胁大豆产业安全。随着生物技术的发展,大豆孢囊线虫抗性机制研究不断深入,在遗传学、转录组学、蛋白功能等相关方面的研究取得了长足进展。本文综述了已知的大豆主要抗性位点(Rhg1和Rhg4)的抗性机制及囊泡运输、植物激素通路与大豆孢囊线虫抗性产生的关系,讨论了相关功能蛋白对大豆抗性的意义以及研究方向上可能存在的问题,最后展望了该领域的后续研究。相关的研究将有利于充分发掘大豆优良抗性基因,为抗大豆孢囊线虫转基因大豆新种质创制奠定理论基础,服务于我国大豆产业的长久安全发展。

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