Identification and fine mapping of qSW2 for leaf slow wilting in soybean

Drought is one of the abiotic stresses limiting the production of soybean(Glycine max).Elucidation of the genetic and molecular basis of the slow-wilting(SW)trait of this crop offers the prospect of its genetic improvement.A panel of 188 accessions and a set of recombinant inbred lines produced from a cross between cultivars Liaodou 14 and Liaodou 21 were used to identify quantitative-trait loci(QTL)associated with SW.Plants were genotyped by Specific-locus amplified fragment sequencing and seedling leaf wilting was assessed under three water-stress treatments.A genome-wide association study identified 26 SW-associated single-nucleotide polymorphisms(SNPs),including three located in a 248-kb linkage-disequilibrium(LD)block on chromosome 2.Linkage mapping revealed a major-effect QTL,qSW2,associated with all three treatments and adjacent to the LD block.Fine mapping in a BC_(2)F_(3) population derived from a backcross between Liaodou 21 and R26 confined qSW2 to a 60-kb interval.Gene expression and sequence variation analysis identified the gene Glyma.02 g218100,encoding an auxin transcription factor,as a candidate gene for qSW2.Our results will contribute significantly to improving drought-resistant soybean cultivars by providing genetic information and resources.

Genome-wide association with transcriptomics reveals a shade-tolerance gene network in soybean

Shade tolerance is essential for soybeans in inter/relay cropping systems.A genome-wide association study(GWAS)integrated with transcriptome sequencing was performed to identify genes and construct a genetic network governing the trait in a set of recombinant inbred lines derived from two soybean parents with contrasting shade tolerance.An improved GWAS procedure,restricted two-stage multi-locus genome-wide association study based on gene/allele sequence markers(GASM-RTM-GWAS),identified 140 genes and their alleles associated with shade-tolerance index(STI),146 with relative pith cell length(RCL),and nine with both.Annotation of these genes by biological categories allowed the construction of a protein–protein interaction network by 187 genes,of which half were differentially expressed under shading and non-shading conditions as well as at different growth stages.From the identified genes,three ones jointly identified for both traits by both GWAS and transcriptome and two genes with maximum links were chosen as beginners for entrance into the network.Altogether,both STI and RCL gene systems worked for shade-tolerance with genes interacted each other,this confirmed that shadetolerance is regulated by more than single group of interacted genes,involving multiple biological functions as a gene network.

Natural variations and geographical distributions of seed carotenoids and chlorophylls in 1167 Chinese soybean accessions

Understanding the composition and contents of carotenoids in various soybean seed accessions is important for their nutritional assessment.This study investigated the variability in the concentrations of carotenoids and chlorophylls and revealed their associations with other nutritional quality traits in a genetically diverse set of Chinese soybean accessions comprised of cultivars and landraces.Genotype,planting year,accession type,seed cotyledon color,and ecoregion of origin significantly influenced the accumulation of carotenoids and chlorophylls.The mean total carotenoid content was in the range of 8.15–14.72μg g–1 across the ecoregions.The total carotenoid content was 1.2-fold higher in the landraces than in the cultivars.Soybeans with green cotyledons had higher contents of carotenoids and chlorophylls than those with yellow cotyledons.Remarkably,lutein was the most abundant carotenoid in all the germplasms,ranging from 1.35–37.44μg g–1.Carotenoids and chlorophylls showed significant correlations with other quality traits,which will help to set breeding strategies for enhancing soybean carotenoids without affecting the other components.Collectively,our results demonstrate that carotenoids are adequately accumulated in soybean seeds,however,they are strongly influenced by genetic factors,accession type,and germplasm origin.We identified novel germplasms with the highest total carotenoid contents across the various ecoregions of China that could serve as the genetic materials for soybean carotenoid breeding programs,and thereby as the raw materials for food sectors,pharmaceuticals,and the cosmetic industry.

A novel procedure for identifying a hybrid QTL-allele system for hybrid-vigor improvement, with a case study in soybean(Glycine max)yield

“Breeding by design” for pure lines may be achieved by construction of an additive QTL-allele matrix in a germplasm panel or breeding population, but this option is not available for hybrids, where both additive and dominance QTL-allele matrices must be constructed. In this study, a hybrid-QTL identification approach, designated PLSRGA, using partial least squares regression(PLSR) for model fitting integrated with a genetic algorithm(GA) for variable selection based on a multi-locus, multi-allele model is described for additive and dominance QTL-allele detection in a diallel hybrid population(DHP). The PLSRGA was shown by simulation experiments to be superior to single-marker analysis and was then used for QTL-allele identification in a soybean DPH yield experiment with eight parents. Twenty-eight main-effect QTL with 138 alleles and nine QTL × environment QTL with 46 alleles were identified, with respective contributions of 61.8% and 23.5% of phenotypic variation. Main-effect additive and dominance QTL-allele matrices were established as a compact form of the DHP genetic structure. The mechanism of heterosis superior-to-parents(or superior-to-parents heterosis, SPH) was explored and might be explained by a complementary locus-set composed of OD+(showing positive over-dominance, most often), PD+(showing positive partial-to-complete dominance, less often) and HA+(showing positive homozygous additivity, occasionally) loci, depending on the parental materials. Any locus-type, whether OD+, PD + and HA+, could be the best genotype of a locus. All hybrids showed various numbers of better or best genotypes at many but not necessarily all loci, indicating further SPH improvement. Based on the additive/dominance QTL-allele matrices, the best hybrid genotype was predicted, and a hybrid improvement approach is suggested. PLSRGA is powerful for hybrid QTL-allele detection and cross-SPH improvement.

垄作和覆膜下盐碱地水盐运移和大豆产量的变化

选择河北省黄骅市中度盐碱土区布置夏播大豆[Glycine max(L.)Merr.]田间试验,共设平播(对照)、平播覆膜、垄作及垄作覆膜4个处理,研究垄作和覆膜对土壤水盐运移和大豆产量的影响。结果表明,与传统平播方式相比,平播覆膜处理对土壤水盐运移和大豆产量的影响不显著;垄作处理能显著改变土壤水盐分布,特别是大豆生长中后期降雨量开始减少后,垄作定植沟会产生叠加集雨效果,显著提高大豆生长中期0~20 cm和20~40 cm土壤含水量,减缓作物生长后期向表层返盐进程,有效降低土壤表层盐分含量,对保障大豆植株生长和产量均有一定的促进效果,可增产27.2%;垄作覆膜处理效果与垄作处理类似,覆膜措施影响不显著。垄作技术可以作为滨海盐碱地夏播大豆种植的农耕措施。

大豆次生代谢产物结构、生物活性及其作用机制研究进展

大豆为豆科大豆属一年生草本植物,不仅含有优质的蛋白质和油脂,还含有大量的活性次生代谢成分,如大豆异黄酮、大豆皂苷等。其次生代谢产物具有独特的结构和多种生物活性,受到广泛关注。本文通过Web of Science、PubMed等数据库,综述了近年来大豆中主要的次生代谢产物大豆异黄酮、大豆皂苷的结构及活性化合物的功能及作用机制。鉴定得到的大豆异黄酮类共12个,通过丝裂原活化蛋白激酶(mitogen activated protein kinase,MAPK)、核因子κB(nuclear factor kappa B,NF-κB)、磷脂酰肌醇3-激酶/蛋白激酶B、核因子-红细胞2相关因子2(nuclear factor erythroid 2-related factor 2,Nrf2)等发挥预防心脑血管疾病、抗骨质疏松症、抗氧化、抗炎、降血糖、抗肿瘤、抗中风、神经保护、保肝等作用。鉴定得到的皂苷类化合物共30个,具有抗炎、抗氧化、预防心血管疾病、抗肥胖、预防糖尿病以及抗肿瘤等作用,涉及的信号通路包括Toll样受体4/髓样分化因子88/NF-κB、MAPK、非受体型蛋白酪氨酸激酶家族和Nrf2等,这些次生代谢产物间因结构存在差异,活性侧重点不同,大豆异黄酮主要体现为植物雌激素活性,而大豆皂苷则以抗肿瘤、抗炎等为主。旨在为大豆次生代谢产物的深入挖掘提供依据,为大豆资源的进一步开发奠定基础。

大豆膨胀素基因GmEXPB5和GmEXPB7生物信息学及表达分析

膨胀素(expansin,EXP)通过调控细胞壁的松弛在植物应对环境胁迫过程中起着重要作用。为研究EXP基因在大豆应对非生物胁迫过程中的作用,该文对大豆中的两个EXP基因(GmEXPB5和GmEXPB7)及其蛋白序列进行生物信息学分析,通过实时荧光定量PCR(qRT-PCR)检测基因表达量。结果表明:(1)GmEXPB5和GmEXPB7分别位于大豆第10号和第12号染色体上,编码的蛋白序列长度分别为272和267个氨基酸。GmEXPB5蛋白分子量为29.07 kD,理论等电点为7.51;GmEXPB7蛋白分子量为29.09 kD,理论等电点为8.66。GmEXPB5和GmEXPB7均为稳定的亲水蛋白且定位于细胞壁中。GmEXPB5和GmEXPB7蛋白均含有一段信号肽序列和一个保守的DPBB_1结构域。(2)GmEXPB5蛋白与鹰嘴豆CaEXPB15蛋白亲缘关系最近,GmEXPB7蛋白与密花豆、赤豆和豇豆的EXPB3蛋白有着较近的亲缘关系。(3)GmEXPB5和GmEXPB7在大豆根、茎和叶中均有表达且它们在根和叶中的表达量均显著高于茎中的表达量。(4)GmEXPB5和GmEXPB7在大豆幼苗中可以响应盐、干旱和低温胁迫。(5)GmEXPB5启动子区域含有2种与逆境相关的顺式作用元件(ABRE和ARE);GmEXPB7启动子区域含有5种与逆境相关的顺式作用元件(ABRE、ARE、CGTCA-motif、TC-rich repeats和MBS)。综上所述,GmEXPB5和GmEXPB7能够参与大豆对非生物胁迫的应答。

Cd Distribution and Subcellular Localization in Leaf and Its Effects on Growth of Soybean(Glycine max) Seedlings

In order to investigate Cd accumulation, subcellular distribution, and local-ization in soybean seedlings leaves, soybean seedlings were cultivated in solution containing different concentrations of Cd. The results showed that most Cd associ-ated with the cellwal s and soluble fractions, and a minor part of Cd presented in mitochondria fractions, nuclear and chloroplast fractions, especial y exposure to high Cd concentrations. Under 20.00 mg/L Cd stress, Cd subcellular distribution fol owed a sequence as： soluble fractions （55.00%）＆gt;cellwal s （30.0%）＆gt;mitochondria fractions （8.21%）＆gt;nuclear and chloroplast fractions （6.79%）. Deposited Cd black particles were observed in cellwal s, chloroplasts, nuclei, and vacuoles through electrical microscope slice. This fact indicated that the cellwal s of soybean leaves were the first protecting organel es from Cd toxicity, and the cellwal s and soluble fractions were the main place for Cd storage. Due to Cd accumulated in the organel es, the intercellular space was enlarged and the subcellular structure was damaged, especial y for the chloroplasts.

大豆(G.max)种子蛋白SBTi位点新类型的发现及甘肃省大豆SBTi位点各等位基因频率的研究

1991年,用PAGE技术检测了甘肃省243份大豆(G.max)品种种子蛋白中SBTi位点的等位基因型。检测结果:Ti^a型品种22份,占品种总数的90.9％,Ti^b21份,占8.6％,未发现Ti^c及ti型。发现一份材料的电泳图谱在与标样上Ti^a、Ti^b、Ti^c对应位置上没有谱带,而在Ti^b位置紧上方,有一条新谱带(Rf=0.79,Ti^b=0.82)。经加胰蛋白酶等措施多次验证,初步认为这很可能是在Ti位点上的一个新的等位基因控制下形成的蛋白谱带。

干旱响应的大豆G-box结合因子GmDIBL1的克隆和功能鉴定

干旱是典型的非生物逆境之一,严重影响作物的产量品质.鉴定作物抗/耐旱关键基因功能,解析其调控机制,对于培育抗/耐旱作物新种质、保障粮食安全具有重要意义.大豆是对干旱逆境较敏感的作物,挖掘寻找大豆抗/耐旱基因、鉴定其功能、明确其调控机制是大豆抗逆研究领域的热点.通过分析干旱处理的大豆根转录组数据,获得一个受干旱诱导的G-box结合蛋白基因GmDIBL1(Drought Induced B12D Like 1),克隆的cDNA序列包含1个长261 bp的开放阅读框,编码86个氨基酸残基,推导的分子量为10.1 kD,等电点为9.647.GmDIBL1蛋白质序列与其他物种的G-box结合蛋白之间具有较高的保守性,包含B12D结构域(6~73个氨基酸残基)和跨膜螺旋,N-端具有信号肽,C-端构成非胞质区域(34~86氨基酸残基),该蛋白定位于细胞核.GmDIBL1基因在根瘤中的表达水平最高,根中次之.干旱处理12 h后,其表达水平显著升高.在耐旱性较强的‘晋豆21’根中的表达水平显著高于干旱敏感的‘绥农26’中的表达水平.进一步通过在大豆毛状根中超量表达和敲除GmDIBL1基因,鉴定了转基因材料对干旱的响应,证明超量表达GmDIBL1基因显著提高毛状根复合体的抗旱性.这些结果说明GmDIBL1基因是大豆中一个响应干旱胁迫的基因,在大豆抗/耐干旱逆境中具有重要作用.

Rapid HPLC Method for Determination of 12 Isoflavone Components in Soybean Seeds

It is important to determine the isoflavone components by high-performance liquid chromatography （HPLC） for the molecular assistant selection of isoflavone in soybean. Based on the standard samples of 12 isoflavone components, the isoflavone components were analyzed using the determination of absorbance peaks method by HPLC. The results showed that there were different maximum ultraviolet （UV） absorbance for the aglycones of daidzein, glycitein, and genistein, which were at 250, 257, and 260 nm, respectively. A linear gradient elution of acetonitrile （13-30%） containing 0. 1% acetic acid as a mobile phase was applied on a YMC-C18 column at 35℃. The 12 isoflavone components were determined using the UV detector by HPLC. We concluded that this is a rapid and precise method which adapted to determine the large numbers of samples with microanalysis.

Mapping and validation of a dominant salt tolerance gene in the cultivated soybean(Glycine max) variety Tiefeng 8

Salt is an abiotic stress factor that strongly affects soybean growth and production. A single dominant gene has been shown to confer salt tolerance in the soybean cultivar Tiefeng 8.The objective of the present study was to genetically map the salt-tolerance gene in an F2:3population and a recombinant inbred line(RIL) population derived from a cross between two cultivated soybeans, Tiefeng 8(tolerant) and 85-140(sensitive). The F2:3families and RILs were treated with 200 mmol L-1Na Cl to evaluate salt tolerance. The F2:3population showed 1(42 tolerant): 2(132 segregating): 1(65 sensitive) segregation, indicating a single dominant gene for salt tolerance in Tiefeng 8. A sequence-characterized amplified region(SCAR) marker from a previously identified random amplified polymorphic DNA(RAPD)marker and four insertion/deletion polymorphism(In Del) markers were developed within the mapping region. Using these markers along with SSR markers, the salt-tolerance gene was mapped within 209 kb flanked by SCAR marker QS08064 and SSR marker Barcsoyssr_3_1301 on chromosome 3. Three markers that cosegregated with the salt tolerance gene and SCAR marker QS08064 were used to genotype 35 tolerant and 23 sensitive soybean accessions. These markers showed selection efficiencies of 76.2% to94.2%. The results indicate that these markers will be useful for marker-assisted breeding and facilitating map-based cloning of the salt tolerance gene in soybean.

Effect of Phosphorus Fertilization to P Uptake and Dry Matter Accumulation in Soybean with Different P Efficiencies

Phosphorus （P） is an essential element for plant growth and yield. Improving phosphorus use efficiency of crops could potentially reduce the application of chemical fertilizer and alleviate environmental damage. Soybean （Glycine max （L.） Merr.） is sensitive to phosphorus （P） in the whole life history. Soybean cultivars with different P efficiencies were used to study P uptake and dry matter accumulation under different P levels. Under low P conditions, the P contents of leaf in high P efficiency cultivars were greater than those in low P efficiency cultivars at the branching stage. The P accumulation in stems of high P efficiency cultivars and in leaves of low P efficiency cultivars increased with increasing P concentration at the branching stage. At the late podding stage, the P accumulation of seeds in high and low P efficiency cultivars were 22.5 and 26.0%, respectively; and at the mature stage were 69.8 and 74.2%, respectively. In average, the P accumulation in whole plants and each organ was improved by 24.4% in high P efficiency cultivars compared to low P efficiency cultivars. The biomass between high and low P efficiency cultivars were the same under extended P condition, while a significant difference was observed at late pod filling stage. At the pod setting stage, the biomass of high P efficiency cultivars were significant greater （17.4%） than those of low P efficiency cultivars under high P condition. Meanwhile, under optimum growth conditions, there was little difference ofbiomass between the two types of cultivars, however, the P agronomic efficiency and P harvest index were significant higher in high P efficiency cultivars than those in low P efficiency cultivars.

Identification of a novel seed size associated locus SW9-1 in soybean

Seed size is one of the vital traits determining seed appearance, quality, and yield. Untangling the genetic mechanisms regulating soybean 100-seed weight (100-SW), seed length and seed width across environments may provide a theoretical basis for improving seed yield. However, there are few reports related to QTL mapping of 100-SW across multiple ecological regions. In this study, 21 loci associated with seed size traits were identified using a genome-wide association of 5361 single nucleotide polymorphisms (SNPs) across three ecoregions in China, which could explain 8.12%–14.25% of the phenotypic variance respectively. A new locus, named as SW9-1 on chromosome 9 that explained 10.05%–10.93% of the seed weight variance was found significantly related to seed size traits, and was not previously reported. The selection effect analysis showed that SW9-1 locus has a relatively high phenotypic effect (13.67) on 100-SW, with a greater contribution by the accessions with bigger seeds (3.69) than the accessions with small seeds (1.66). Increases in seed weight were accompanied by increases in the frequency of SW9-1T allele, with >90% of the bred varieties with a 100-SW >30 g carrying SW9-1T. Analysis of SW9-1 allelic variation in additional soybean accessions showed that SW9-1T allele accounting for 13.83% of the wild accessions, while in 46.55% and 51.57% of the landraces and bred accessions, respectively, this results indicating that the SW9-1 locus has been subjected to artificial selection during the early stages of soybean breeding, especially the utilization of SW9-1T in edamame for big seed. These results suggest that SW9-1 is a novel and reliable locus associated with seed size traits, and might have an important implication for increasing soybean seed weight in molecular design breeding. Cloning this locus in future may provide new insights into the genetic mechanisms underlying soybean seed size traits.

Analysis of QTLs for the Trichome Density on the Upper and Downer Surface of Leaf Blade in Soybean [Glycine max (L.) Merr.]

Trichomes （plant hairs） are present on nearly all land plants and are known to play important roles in plant protection, specifically against insect herbivory, drought, and UV radiation. The identification of quantitative trait loci （QTL） associated with trichome density should help to interpret the molecular genetic mechanism of soybean trichome density. 184 recombinant inbred lines （RILs）, derived from a cross between soybean cultivars Kefeng 1 and Nannong 1138-2 were used as segregating population for evaluation of TDU （trichome density on the upper surface of leaf blade） and TDD （trichome density on the downer surface of leaf blade）. A total of 15 QTL were detected on molecular linkage groups （MLG） A2, Dla, Dlb, E and H by composite interval mapping （CIM） and among all the QTL, qtuA2-1, qtuD 1 a-1, qtuD lb-2, qtuH-2 qtuE-1, qtdDlb-2, and qtdH- 2 were affirmed by multiple interval mapping （MIM）. The contribution ofphenotypic variance of qtuH-2 was 31.81 and 29.4% by CIM and MIM, respectively, suggesting it might be major gene Ps loci. Only 10 pairs of main QTL interactions for TDU were detected, explained a range of 0.2-5.1% of phenotypic variations for each pair for a total of 22.8%. The QTL on MLG Dlb affecting trichome density were mapped near to Rsc-7 conditioning resistance to SMV （soybean mosaic virus）. This study showed that the genetic mechanism of trichome density was the mixed major gene and polygene inheritance, and also suggested that the causal nature between trichome density and other agronomic traits.

Overexpression of G10-EPSPS in soybean provides high glyphosate tolerance

Glyphosate is a highly efficient, broad-spectrum nonspecific herbicide that inhibits the 5-enolpyruvylshikimate-3-phosphate synthase(EPSPS)-mediated pathway of shikimic acid. The screening of glyphosate-resistant EPSPS gene is a major means for the development of new genetically modified glyphosate-resistant transgenic crop. Currently, the main commercialized glyphosate-resistant soybean contains glyphosate-resistant gene CP4-EPSPS. In this study, a G10-EPSPS gene was reported providing glyphosate resistance in Zhongdou 32. Here, G10-EPSPS gene was introduced into soybeans through Agrobacterium-mediated soybean cotyledon node. PCR, Southern blotting, semi-quantitative RT-PCR, qRT-PCR, and Western blotting were used, and the results revealed that G10-EPSPS had been integrated into the soybean genome and could be expressed steadily at both mRNA and protein levels. In addition, glyphosate resistance analysis showed that the growth of transgenic soybean had not been affected by concentrations of 900 and 2 700 g a.e. ha–1 of glyphosate. All the results indicated that G10-EPSPS could provide high glyphosate resistance in soybeans and be applied in production of glyphosate-resistant soybean.

Genetic Analysis on Isoflavone Content in Soybean Seeds

Fifteen combinations with six soybean cultivars of different isoflavone content were formulated and planted in a randomized complete-block design model; genetic factors of isoflavone quantity were analyzed. Results indicated that genetic factors of isoflavone contents in F2 population inherited quantitatively. Isoflavone content of F1 F2 seeds normally trended. There were heterosis in F1, F2 of most combinations, and also heterobeltiosis in part of the crosses. The broad sense heritability of F2 was higher in parts of the crosses. It predicted the selection might be carried out preliminarily in F2 hybrids. There was significant positive correlation between hybrids and mid-parent.

Identification of novel QTL associated with soybean isoflavone content

Soybean isoflavones are essential secondary metabolites synthesized in the phenylpropanoid pathway and benefit human health. In the present study, highresolution QTL mapping for isoflavone components was performed using specific-locus amplified fragment sequencing(SLAF-seq) with a recombinant inbred line(RIL) population(F5:7) derived from a cross between two cultivated soybean varieties, Luheidou 2(LHD2) and Nanhuizao(NHZ). Using a high-density genetic map comprising 3541 SLAF markers and the isoflavone contents of soybean seeds in the 200 lines in four environments, 24 stable QTL were identified for isoflavone components, explaining 4.2%–21.2% of phenotypic variation.Of these QTL, four novel stable QTL(qG8, qMD19, qMG18, and qTIF19) were identified for genistin, malonyldaidzin, malonylgenistin, and total isoflavones, respectively. Gene annotation revealed three genes involved in isoflavone biosynthesis(Gm4CL, GmIFR, and GmCHR) and 13 MYB-like genes within genomic regions corresponding to stable QTL intervals, suggesting candidate genes underlying these loci. Nine epistatic QTL were identified for isoflavone components, explaining 4.7%–15.6% of phenotypic variation. These results will facilitate understanding the genetic basis of isoflavone accumulation in soybean seeds. The stable QTL and tightly linked SLAF markers may be used for markerassisted selection in soybean breeding programs.

Sampling Survey and Identification of Races of Soybean Cyst Nematode (Heterodera glycines Ichinohe) in Huang-Huai Valleys

Soybean cyst nematode （SCN Heterodera glycines Ichinohe） is one of the most important nationwide soybean diseases in China. A total of 38 soil specimens or locations in the area was sampled and tested for SCN races during 2001-2003 for the inspection of race distribution in Huang-Huai Valleys. A map of race distribution was constructed according to the data from both the present study and the published reports cited. Three areas, namely, the area of southeast to Jinan in Shangdong Province; the area of northern Henan Province and its border region to south of Hebei Province; and the area of Luohe, Zhoukou of Henan Province and Fuyang of Anhui Province mainly infested with Race 1 were identified. Race 4 was predominant in Shanxi Province, Beijing and the adjacent area of Henan, Shandong, and Anhui provinces, and the delta of Huanghe River in Shandong Province. Race 2 was mainly found in Liaocheng, Dezhou of Shangdong Province and Shijiazhuang of Hebei Province, and Jiaozuo and Huojia of Henan Province. Race 7 was distributed in the west part of Jiaodong Peninsula of Shandong Province and Kaifeng, Huaxian, Wenxian of Henan Province. Race 5 was found and scattered in Hebei and Henan Province. Race 9 was found in Shangqiu of Henan Province, which was reported for the first time in China. It can be seen that Race 1 and Race 4 were the two predominant races in Huang-Huai Valleys, and that research should focus on developing resistant cultivars of these races. There might exist other races in an area with some predominant races. The race substitution in the past decade was not obviously found, therefore, the results should be meaningful to future breeding for resistance to SCN in Huang-Huai Valleys.

Constitutive expression of feedback-insensitive cystathionine γ-synthase increases methionine levels in soybean leaves and seeds

Soybean （Glycine max （L.） Merr.） is a major crop that provides plant-origin protein and oil for humans and livestock. Al- though the soybean vegetative tissues and seeds provide a major source of high-quality protein, they suffer from low con- centration of an essential sulfur-containing amino acid, methionine, which significantly limits their nutritional quality. The level of methionine is mainly controlled by the first unique enzyme of methionine synthesis, cystathione y-synthase （CGS）. Aiming to elevate methionine level in vegetative tissues and seeds, we constitutively over-expressed a feedback-insensitive Arabidopsis CGS （AtD-CGS） in soybean cultivars, Zigongdongdou （ZD） and Jilinxiaoli 1 （JX）. The levels of soluble methionine increased remarkably in leaves of transgenic soybeans compared to wild-type plants （6.6- and 7.3-fold in two transgenic ZD lines, and 3.7-fold in one transgenic JX line）. Furthermore, the total methionine contents were significantly increased in seeds of the transgenic ZD lines （1.5- to 4.8-fold increase） and the transgenic JX lines （1.3- to 2.3-fold increase） than in the wild type. The protein contents of the transgenic soybean seeds were significantly elevated compared to the wild type, suggesting that the scarcity of methionine in soybeans may limit protein accumulation in soybean seeds. The increased protein content did not alter the profile of major storage proteins in the seeds. Generally, this study provides a promising strategy to increase the levels of methionine and protein in soybean through the breeding programs.