Genetic analysis and identification of two soybean mosaic virus resistance genes in soybean [Glycine max (L.) Merr]

Soybean mosaic virus (SMV) commonly affects soybean production worldwide, and the SC18 strain has been widespread in China. This study aimed to characterize and map the SC18 resistance genes present in soybean cultivars ‘Kefeng No. 1’ and ‘Qihuang 22’. Inheritance analysis revealed that two independent single dominant genes in Kefeng No. 1 and Qihuang 22 confer resistance to SC18. Using simple sequence repeat (SSR) markers and bulked segregant analysis, the Kefeng No. 1 and Qihuang 22 resistance genes were located on soybean chromosomes 2 and 13, respectively. We further screened two populations of recombinant inbred lines with 32 SSR markers in the target region, where the resistance gene in Kefeng No. 1 was fine mapped to an 80‐kb region containing six putative genes. Sequence and expression analyses of these genes revealed that SMV resistance in Kefeng No. 1 was probably attributable to three of the candidate genes (i.e. Glyma.02G127800, Glyma.02G128200 and Glyma.02G128300). Collectively, the results of this study will greatly facilitate the cloning of SC18 resistance genes and marker‐assisted breeding of SMV‐resistant soybean cultivars.     

Characterization of broad‐spectrum resistance to Soybean mosaic virus in soybean [Glycine max (L.) Merr.] cultivar ‘RN‐9’

Soybean mosaic virus (SMV) can cause serious yield losses in soybean. Soybean cultivar ‘RN‐9’ is resistant to 15 of 21 SMV strains. To well‐characterize this invaluable broad‐spectrum SMV‐resistance, populations (F₁, F₂ and F_2:3) derived from resistant (R) × susceptible (S) and R × R crosses were tested for SMV‐SC18 resistance. Genetic analysis revealed that SC18 resistance in ‘RN‐9’ plus two elite SMV‐resistant genotypes (‘Qihuang No.1’ and ‘Kefeng No.1’) are controlled by independently single dominant genes. Linkage analysis showed that the resistance of ‘RN‐9’ to SMV strains SC10, SC14, SC15 and SC18 is controlled by more than one gene(s). Moreover, Rsc10‐r and Rsc18‐r were both positioned between the two simple sequence repeats markers Satt286 and Satt277, while Rsc14‐r was fine‐mapped in 136.8‐kb genomic region containing sixteen genes, flanked by BARCSOYSSR_06_0786 and BARCSOYSSR_06_0790 at genetic distances of 3.79 and 4.14 cM, respectively. Allelic sequence comparison showed that Cytochrome P450‐encoding genes (Glyma.06g176000 and Glyma.06g176100) likely confer the resistance to SC14 in ‘RN‐9’. Our results would facilitate the breeding of broad‐spectrum and durable SMV resistance in soybeans.     

大豆对大豆花叶病毒株系SC6和SC17抗病基因的精细定位

针对我国北方和长江流域大豆产区广泛分布的SMV株系SC6和SC17,利用2个抗病大豆品种Q0926和中豆35分别与感病品种南农1138-2和南农菜豆5号配制2个抗感杂交组合Q0926×南农1138-2和中豆35×南农菜豆5号以及一个抗抗组合Q0926×中豆35,研究3个组合的F₁、F₂、F_2:3抗性遗传规律,探讨Q0926对SC6和中豆35对SC17及2个抗病品种对同一SMV株系抗性基因的等位关系,并对大豆对2个株系的抗病基因进行了标记定位。结果显示,Q0926×南农1138-2和中豆35×南农菜豆5号2个抗感杂交组合在分别接种SC6和SC17后,F₁表现抗病,F₂呈3抗∶1感分离比例,F_2:3家系呈1抗∶2分离∶1感病的分离比率,表明Q0926对SC6和中豆35对SC17的抗病性分别由1对显性基因控制;抗抗组合Q0926×中豆35的F₁和F₂在接种2个株系后均未发现感病单株,表明Q0926与中豆35对SC6和SC17株系的抗病基因分别是等位或紧密连锁的。分别利用2个抗感组合的F₂和F_2:3群体对2个抗病基因的定位结果显示,第2染色体上的25个SSR标记与抗SC6的基因R_SC6连锁,最近的2个标记与抗性基因R_SC6的排列次序和遗传距离为BARCSOYSSR_02_0617(0.775 cM)-R_SC6-BARCSOYSSR_02_0621(0.519 cM);第2染色体上的38个SSR标记与抗SC17的基因R_SC17连锁。最近的2个标记与抗性基因R_SC17的排列次序和遗传距离为BARCSOYSSR_02_0622(0.264 cM)-R_SC17-BARCSOYSSR_02_0627(0.262 cM),其对应的物理区间分别为52 kb和60 kb。抗性遗传研究为抗大豆花叶病毒育种的亲本选配、后代选择提供了理论指导,抗性基因的标记定位研究为抗性基因的分子标记辅助选择和抗病基因的图位克隆奠定了基础。     

大豆花叶病毒(SMV)株系SC4和SC8的抗性遗传分析

选用我国黄淮和长江流域大豆产区发生频繁的SMV株系SC4和SC8,利用大豆抗病材料和感病材料配制抗感和抗抗杂交组合,研究抗病材料对SC4和SC8株系的遗传方式以及不同大豆材料对SMV抗性基因位点间的等位性关系。结果表明, 接种SC4株系后,由冀LD42、徐豆1号、跃进4号、科丰1号、PI96983、晋大74、汾豆56、大白麻和齐黄22为抗源配制的9个抗感组合的F₁均表现抗病,经卡方测验, F₂抗感分离比例3∶1,F_2:3家系分离比例为1(抗)∶2(分离)∶1(感),表明这些抗源均有1对基因控制对SC4株系的抗性,且抗病表现为显性;5个抗抗组合的F₁均表现抗病,F₂群体分离比例15(抗)∶1(感),表明大白麻与汾豆56、科丰1号和齐黄1号携带抗SC4的基因是不等位的,冀LD42与汾豆56,晋大74与中作229是不等位的。接种SC8株系后,用齐黄1号、中作229、NY58、科丰1号、PI96983、晋大74、汾豆56、大白麻和齐黄22为抗源配制的抗感组合杂交后代分离符合1对基因的分离比例且F₁均表现抗病,说明这些品种对SC8株系的抗性也均由1对显性基因控制。抗抗组合晋大74×汾豆56接种SC8株系后的F₂群体全部表现抗病,F_2:3家系没有抗感分离,表明抗病品种晋大74与汾豆56携带的抗病基因是等位的;齐黄1号×科丰1号、大白麻×汾豆56的F₂群体分离比例15(抗)∶1(感),表明抗病亲本齐黄1号与科丰1号、大白麻与汾豆56携带抗SC8的基因是不等位的,而且独立遗传。     

Characterization and allelism of soybean mosaic virus strain SC3 resistance in ‘Qihuang-1’ derived soybean cultivars

Soybean mosaic virus is a severe constraint of soybean production in China. A total of country-wide 22 SMV strains (SC1-SC22) were identified. Of these, SC3 is a major strain widely distributed in Huanghuai and Yangtze River Valley region of China. Soybean cultivar ‘Qihuang-1’ contains R_SC3Q locus conditioning the resistance to SC3 and is an important parental line extensively used to breed the soybean cultivars in China. The objective of this study was to elucidate the genetic pattern of SC3 resistance genes in cultivars developed from ‘Qihuang-1’ or its derivative lines. Hence, we have evaluated the SC3 resistance in 91 cultivars developed from ‘Qihuang-1’ or its derivative lines. The results showed that a total of 43 cultivars exhibited resistance to the SC3 strain. Among them, 37 cultivars were derived from ‘Qihuang-1’. Then, we have detected the R_SC3Q loci in these cultivars using four SSR markers (Satt334, Sct_033, BARCSOYSSR_13_1114 and BARCSOYSSR_13_1136). It revealed that, among the 37 resistant cultivars derived from ‘Qihuang-1’, there are 20 cultivars containing R_SC3Q loci. Moreover, the allelic relationship of resistance genes was analysed using the crosses from resistance × resistance between ‘Qihuang-1’ and its resistant derived cultivars. The results showed that the resistance genes of ‘Qihuang-1’ and its 20 cultivars were allelic. But it is not allelic with those of the other 17 cultivars, different from ‘Qihuang-1’, and also, R_SC3Q does not condition the resistance. These results will be beneficial to exploring the transmission of resistance genes of ‘Qihuang-1’ and will be useful to the disease resistance breeding of soybean.     

大豆对SMV SC-7株系群的抗性遗传与基因定位

科丰1号×南农1138-2的P₁、P₂、F₁和180个重组自交家系接种SC-7株系群的鉴定表明,P₁与F₁全抗,P₂全感,说明抗性为显性;重组自交家系抗、感按1∶1分离,说明抗性由一对基因控制。利用王永军等的遗传连锁图对SC-7株系群的抗性基因进行连锁分析,将抗病基因Rsc-7定位于N8-D1b＋W连锁群上,并与已定位的5个抗性基因中的3个连锁,还有一个与之相连锁的标记LC5T,其排列顺序和遗传距离为Rsa (30.6 cM) Rsc-7 (22.1 cM) Rn3 (10.3 cM) Rn1 (15.8 cM) LC5T。     

Identification and characterization of a RAPD/SCAR marker linked to a resistance gene for soybean mosaic virus in soybean

Soybean line `ICGR95-5383' [Glycinemax (L.) Merr.] is a newly releasedgermplasm from China and is resistant (R)to soybean mosaic virus (SMV). ICGR95-5383was crossed to the susceptible (S)cultivars `HB1', `Tiefeng21', `Amsoy', and`Williams' to investigate the inheritanceof SMV resistance. The F<sub>1</sub> and F<sub>2</sub>plants were inoculated with SMV-3 (the mostvirulent) strain from Northeast China. Theresults showed that F<sub>1</sub> plants from thefour R × S crosses were necrotic (N) andall F<sub>2</sub> populations segregated in a3(R+N):1S ratio, indicating thatICGR95-5383 carries a single gene withincomplete dominance for resistance to SMV. In a bulked segregant analysis (BSA) of theF<sub>2</sub>population from ICGR95-5383 × HB1, a codominant RAPD marker,OPN11<sub>980/1070</sub>, was found to be linkedto the resistance gene in ICGR95-5383. The980-base pair (bp) fragment OPN11<sub>980</sub>was amplified in the R parent ICGR95-5383,R bulk, and resistant F<sub>2</sub> plants. Theother 1070-bp fragment OPN11<sub>1070</sub> wasamplified in the S parent HB1, S bulk, andsusceptible F<sub>2</sub>plants.OPN11<sub>980/1070</sub> was amplified in theF<sub>1</sub> plants and the necroticF<sub>2</sub> plants from the R×S cross.Segregation analysis of the RAPD marker inthe F<sub>2</sub> population revealed that themarker OPN11<sub>980/1070</sub> is closely linkedto the resistance gene with a map distanceof 3.03 cM. OPN11<sub>980/1070</sub> was clonedand sequenced, and specific PCR primerswere designed to convertOPN11<sub>980/1070</sub> into sequencecharacterized amplified region (SCAR) makerSCN11<sub>980/1070</sub>. SCAR analysis of theF<sub>2</sub> population confirmed thatOPN11<sub>980/1070</sub> and SCN11<sub>980/1070</sub> areat the same locus linked to the SMVresistance gene. The RAPD markerOPN11<sub>980</sub> was used as RFLP probefor southern hybridization to soybeangenomic DNA. Southern analysis showed thatsoybean genome contains low-copy sequenceof OPN11<sub>980</sub>. Using a recombinant inbredmapping population of `Kefeng No.1' (R) ×Nannong1138-2'(S), OPN11_980/1070 was mapped to thesoybean molecular linkage group (MLG) Fbetween the restriction fragment lengthpolymorphism (RFLP) markers B212 (0.7 cM) and K07 (6.7 cM) and 3.03 cM apart from theSMV resistance gene.     

Grouping patterns of rainfed winter wheat test locations and the role of climatic variables

Soybean mosaic virus (SMV) is a member of genus Potyvirus, which causes worldwide soybean [Glycine max (L.) Merr.] yield loss and seed quality deterioration. It is of great significance to find new resistance loci and genes for cultivation of soybean variety. In the present study, a recombinant inbred line (RIL) population and a genome-wide association study (GWAS) panel, which contained 193 lines and 379 germplasms, respectively, were used for QTL mapping of resistance to SMV. Linkage mapping identified a major QTL, qSMV13, on chromosome 13, conferring resistance to SMV SC3 and SC7 strains, explaining phenotypic variations 71.21 and 76.59?%, respectively. The QTL qSMV13 was located close to the known SMV resistance loci Rsv1-h. GWAS analysis revealed five single nucleotide polymorphisms (SNPs) significantly associated with resistance to SC3 on chromosomes 2, 11, 13, 14 and 16. One of the SNP markers, ss715614844, was the right flanking marker of qSMV13. Combining linkage mapping and GWAS analysis enabled us to delimit qSMV13 in a 97.2-kb genomic region containing seven genes. A LRR-RLK protein was proposed as the candidate gene of qSMV13. These results provided selection markers and candidate genes for SMV resistance in soybean molecular breeding programs.

     

农杆菌介导的RNAi CP基因在大豆中的转化

花叶病毒(soybean mosaic virus, SMV)病是大豆主要病害之一,生产上常采用种植抗性品种方法来防治。本研究以RNA干扰花叶病毒衣壳蛋白(coat protein, CP)基因为表达载体,Bar基因作为筛选标记基因,成熟子叶节为外植体,采用农杆菌介导法获得了22株T₀代转基因大豆生根苗,经草丁膦涂抹、Bar试纸条和PCR法鉴定,获得RNAi CP转基因植株18株;对转基因植株T₁代的遗传分析表明,外源基因能够稳定遗传到下一代且符合孟德尔遗传规律;T₁代Southern杂交表明,导入的干扰片段为单拷贝;花叶病毒摩擦接种表明RNAi CP转基因大豆植株具有抗花叶病毒特性;摩擦接种后3周,DAS-ELISA检测进一步表明,RNAi CP转基因植株花叶病毒检出率仅为7.69%,而非转基因植株为100%。这表明RNAi花叶病毒CP基因可用于抗大豆花叶病毒的研究。     

Molecular tagging of Asiatic soybean rust resistance in exotic genotype EC 241780 reveals complementation of two genes

Asian soybean rust (ASR) caused by Phakopsora pachyrhizi severely reduces seed yield in soybean. Molecular tagging of ASR resistance can help in the process of resistance breeding. In this study, an F₂ population of cross (susceptible cultivar ‘NRC 7’ × resistant exotic genotype EC 241780) was used for bulked segregant analysis (BSA) with 25 SSR (simple sequence repeat) primers linked with six Rpp genes. Among them, five polymorphic SSR markers, viz., Sct 187, SSR 1859, Satt 191 (Rpp1b like loci) and Satt 215, Sat_361 (Rpp2 loci) distinguished the ASR resistant and susceptible bulks and individuals. In combined marker analysis, the markers Satt 191 (Rpp1b like loci) and Satt 215 (Rpp2 loci) were linked with ASR severity score and were also confirmed in individual 110 F₂ segregants. Hence, these markers could be utilized in the marker assisted rust resistance breeding of Rpp1b like and Rpp2 genes. In silico candidate gene analysis for hypersensitive response revealed that Satt 191 linked region was rich in genes encoding apoptotic ATPase having leucine‐rich repeat (LRR) domain.     

Stripe rust resistance gene Yr18 and its suppressor gene in Chinese wheat landraces

The slow‐rusting and mildewing gene Yr18/Lr34/Pm38/Sr57 confers partial, durable resistance to multiple fungal pathogens and has its origins in China. A number of diagnostic markers were developed for this gene based on the gene sequence, but these markers do not always predict the presence of the resistant phenotype as some wheat varieties with the gene are susceptible to stripe rust in China. We hypothesized that these varieties have a suppressor of Yr18. This study was undertaken to determine the presence of Yr18, the suppressor and/or another resistance gene in 144 Chinese wheat landraces using molecular markers and stripe rust field data. Forty‐three landraces were predicted to have Yr18 based on the presence of the markers, but had final disease severities higher than 70%, indicating that this gene may be under the influence of a suppressor. Four of these landraces, ‘Sichuanyonggang 2’, ‘Baikemai’, ‘Youmai’ and ‘Zhangsihuang’, were chosen for genetic studies. Crosses were made between the lines and ‘Avocet S’, with further crosses of Sichuanyonggang 2 ×  ‘Huixianhong’ and Sichuanyonggang 2 ×  ‘Chinese Spring’. The F₁ plants of Sichuanyonggang 2/Chinese Spring was susceptible indicating the presence of a dominant suppressor gene. The results of genetic analyses of F_2:3 and BC₁F₂ families derived from these crosses indicated the presence of Yr18, a Yr18 suppressor and another additive resistance gene. The Yr18 region in Sichuanyonggang 2 was sequenced to ensure that it contained the functional allele. This is the first report of a suppressor of Yr18/Lr34/Pm38/Sr57 gene with respect to stripe rust response.     

Marker-assisted identification of resistance genes to soybean mosaic virus in soybean lines

Soybean plants react differentially to soybean mosaic virus (SMV) strains because of interactions among different resistant genes in the soybean genome. Three independent genes resistant to SMV have been identified by inheritance studies and linkage analyses. To develop durable SMV-resistant soybean cultivars, it is necessary to determine which soybean SMV resistance genes can be readily transferred from resistant to susceptible cultivars in a breeding system. Here, we report the type and number of resistance gene(s) in four Korean elite soybean cultivars using a combination of disease reaction symptoms, inheritance studies, and molecular marker mappings. The disease reactions of Sowonkong and Keunolkong soybean varietals in response to infection with SMV strains suggested that both cultivars most likely harbor the Rsv1 gene similar to that in York. Subsequent inheritance studies confirmed that Sowonkong has the Rsv1 gene. The inheritance studies suggested that Sinpaldalkong harbored the Rsv1 gene, which was then confirmed by molecular marker mapping. The inheritance studies also suggested that Jinpumkong 2, which is the most resistant to SMV infection among the four cultivars, contained the Rsv1 and Rsv3 genes; this was confirmed by molecular marker mapping. Our approach, which combined inheritance studies and molecular linkage analyses, allowed the efficient identification of resistance gene(s) in four Korean soybean cultivars.     

大豆引进种质抗胞囊线虫病、抗花叶病毒病和耐盐基因型鉴定及优异等位基因聚合种质筛选

我国从美国、俄罗斯、日本等26个国家或地区共引进大豆种质3218份, 仅对部分种质进行了大豆胞囊线虫病(Soybean cyst nematode, SCN)、大豆花叶病毒病(Soybean mosaic virus, SMV)和盐敏感性的抗性鉴定, 但基因型的系统分析尚未见报道。本研究针对大豆抗胞囊线虫病3个基因(rhg1、Rhg4、SCN3-11)和耐盐基因(GmSALT3)开发KASP标记5个, 结合与大豆花叶病毒抗性相关的1个SCAR标记(SCN11), 对1489份大豆引进种质进行基因型鉴定。结果表明, 具有优异等位基因的种质共1084份; 携带3个位点优异等位基因的种质19份, 包括抗胞囊线虫病3个位点(rhg1、Rhg4、SCN3-11)叠加(Peking型)种质3份, 聚合抗胞囊线虫病基因和抗花叶病毒病标记7份, 聚合抗胞囊线虫病和耐盐基因2份, 聚合抗胞囊线虫病、抗花叶病毒病和耐盐基因7份; 携带4个位点优异等位基因的种质9份, 包括聚合抗胞囊线虫病基因和抗花叶病毒病标记6份, 聚合抗胞囊线虫病和耐盐基因2份, 聚合抗胞囊线虫病、抗花叶病毒病和耐盐7份; 携带5个位点优异等位基因8份, 聚合了抗胞囊线虫病、抗花叶病毒病和耐盐优异等位变异。在这些携带优异等位变异的种质中, 44份已由前人证明具有相应的抗性。携带3个或3个以上优异等位基因的36份种质中, 有52.78%种质的一种或两种特性已被报道。在不携带抗性优异等位变异的种质中, 93份已证明有耐盐性或对SMV3号株系抗性, 这些种质可能存在新的抗性(等位)基因。本研究利用高通量分子标记筛选出的携带抗病、抗逆优异等位基因的种质为我国大豆资源表型鉴定、抗源的快速筛选及利用提供理论依据和新思路。     

Molecular mapping of two recessive genes controlling resistance to bacterial leaf pustule disease in soybean (Glycine max)

Bacterial leaf pustule (BLP) caused by Xanthomonas axonopodis pv. glycines (Xag) is a serious soybean disease. A BLP resistant genotype ‘TS-3’ was crossed with a BLP susceptible genotype ‘PK472’, and a segregating F₂ mapping population was developed for genetic analysis and mapping. The F₂ population segregation pattern in 15:1 susceptible/resistance ratio against Xag inoculum indicated that the resistance to BLP in ‘TS-3’ was governed by two recessive genes. A total of 12 SSR markers, five SSR markers located on chromosome 2 and seven SSR markers located on chromosome 6 were identified as linked to BLP resistance. One of the resistance loci (r1) was mapped with flanking SSR markers Sat_183 and BARCSOYSSR_02_1613 at a distance of 0.9 and 2.1 cM, respectively. Similarly, SSR markers BARCSOYSSR_06_0024 and BARCSOYSSR_06_0013 flanked the second locus (r2) at distances of 1.5 and 2.1 cM, respectively. The identified two recessive genes imparting resistance to BLP disease and the SSR markers tightly linked to these loci would serve as important genetic and molecular resources to develop BLP resistant genotypes in soybean.     

The transition time from the juvenile phase to the adult phase differs in soybean cultivars ‘Enrei’ and ‘Peking’

Plants develop juvenile phase to adult phase in vegetative stage. Although soybean is a very important crop worldwide, there has been only one study of the juvenile–adult phase change. In this study, we determined that the juvenile–adult phase change occurred at different stages in two soybean cultivars that differ in their photosensitivity. Cultivar ‘Enrei’ (E1e2e3E4) is weakly photosensitive and cultivar ‘Peking’ (E1E2E3E4) is strongly photosensitive. In ‘Enrei’, the leaf size gradually increased at a constant leaf position regardless of the difference in day length. In ‘Peking’ plants transferred to short‐day conditions at several leaf development stages, leaf size gradually increased at different leaf positions. Expression of miR156 by ‘Enrei’ transferred to short‐day conditions had nearly the same pattern as that of ‘Enrei’ grown under long‐day conditions. In ‘Peking’, the expression of miR156 had different patterns in younger leaves of plants subjected to either a short‐day treatment or long‐day conditions. These results indicate that the E2 and E3 loci that regulate photosensitivity also regulate the expression of miR156 and the juvenile–adult phase change in soybean.     

大豆重组自交系群体NJRIKY遗传图谱的加密及其应用效果

作物基因组研究,包括基因或数量性状位点(QTL)定位、图位克隆以及物理图谱构建等,首先必须建立具有丰富标记信息的高密度遗传连锁图谱。由科丰1号和南农1138-2杂交组合衍生的重组自交系群体NJRIKY已经构建了4张大豆遗传连锁图谱,但由于遗传信息和标记数目不够充分,在基因和QTL作图时仍然存在精确度和准确度问题。为增加NJRIKY图谱密度,本研究在967对SSR引物中获得了401个多态性SSR标记。结合其他分子数据,使用作图软件Mapmaker/Exp3.0b,获得一张含有553个遗传标记,25个连锁群,总长2071.6cM,平均图距3.70cM的新遗传连锁图谱,其中SSR标记316个,RFLP标记197个,EST标记39个,形态标记1个。连锁群上大于20cM的标记间隔由原来42个减少到2个。原图谱的3个SMV抗性基因定位于D1b连锁群末端的开放区间上且仅与一个RFLP标记连锁,利用加密图谱对Rsc-3、Rsc-7、Rsc-9、Rsc-13、Rsa、Rn1和Rn3等7个SMV抗性基因重定位,全部位于D1b连锁群,与相邻分子标记距离均小于6cM,其中Rsc-9、Rn1、Rsa的距离小于1cM,Rsc-13与EST标记GMKF168a共分离。对本群体农艺性状进行QTL重定位,获得8个性状相关的42个主效QTL,其中20个QTL遗传贡献率大于10%,与原图谱比较,新定位的各QTL的标记区间明显缩短,与相邻标记的连锁更加紧密。     

QTL mapping and gene mining to identify genes on soybean (Glycine max) associated with NopB of Sinorhizobium fredii HH103

Soybean is a special crop that can utilize N₂ in the air via symbioses with Rhizobium spp. The formation of effective nodules is a complex process in which nodulation outer proteins (Nops) are determinants of establishment of a symbiotic relationship. We constructed a Sinorhizobium fredii HH103ΩnopB mutant. A nodulation test showed that the mutant had a negative effect on the Suinong14, ZYD00006, Dongnong594 and Charleston soybean lines. Recombinant inbred soybean lines were independently inoculated with the mutant and wild‐type strains, and five and four quantitative trait loci (QTLs) were identified by analysing the nodule number (NN) and nodule dry weight (NDW), respectively. We chose one QTL that overlapped with other studies and a novel QTL identified in our study and selected six candidate genes for further analysis. The qRT‐PCR analysis showed that only changes in Glyma.17G166200 expression depended on NopB. Further analysis showed that Glyma.17G166200 encoded a protein with a D‐glucose‐binding domain and a serine‐threonine/tyrosine protein kinase catalytic domain that was involved in the abscisic acid (ABA) pathway.