The male sterile line of soybean is crucial for hybrid seed production, and has allowed significant advancements in soybean germplasm innovation and yield increase in China. In this study, we created two bulks of sterile plants and collected a natural population consisting of 100 elite soybean germplasms. There were significant phenotypic differences between the sterile and natural populations resulting from flowers and pods. The sterile plants exhibited fleshy spherical pods and large black-green leaves in the maturity stage, while the leaves of the fertile plants fell off. After I2-KI staining, the pollen of the sterile line turned light brown and yellow, while the pollen of the fertile line turned black. On the basis of the SNP sequencing results, the sterility genes were located on eight chromosomes. Additionally, they were fine-mapped to 13 regions on six chromosomes using 72 pairs of SSR markers. Five genes involved in auxin response and pollen development were predicted as candidate genes underlying soybean sterility. These candidate genes for soybean sterility will help with gene cloning and functional analysis and accelerate the widespread use of hybrid seed production and yield increase in soybean grown in cool regions.

Soybean [Glycine max (L.) Merr.], an essential staple food and oil crop worldwide, boasts abundant vegetable proteins and fats beneficial for both human and animal consumption. However, the soybean pod borer (Leguminivora glycinivorella) (SPB) stands as the most destructive soybean insect pest in northeast China and other northeastern Asian regions, leading to significant annual losses in soybean yield and economic burden. Therefore, this study aims to investigate the introduction of a previously tested codon-optimized cry1c gene, cry1c*, into the soybean genome and assess its effect on the SPB infestation by generating and characterizing stable transgenic soybeans overexpressing cry1c*. The transgenic soybean lines that constitutively overexpressed cry1c* exhibited a significant reduction in the percentage of damaged seeds, reaching as low as 5% in plants under field conditions. Additionally, feeding transgenic leaves to the larvae of S. exigua, S. litura, and M. separta resulted in inhibited larval growth, decreased larval body weight, and lower survival rates compared to larvae fed on wild-type leaves. These findings showed that the transgenic lines maintained their resistance to SPB and other lepidopteran pests, especially the transgenic line KC1. Southern blotting and genome-wide resequencing analysis revealed that T-DNA integration occurred as a single copy between loci 50,868,122 and 50,868,123 of chromosome 10 in the transgenic line KC1. Therefore, the transgenic line KC1, overexpressing high levels of cry1c* in leaves and seeds, holds strong potential for commercial use in the integrated management of SPB and other lepidopteran pests.

Summary High light stress is an important factor limiting crop yield. Light receptors play an important role in the response to high light stress, but their mechanisms are still poorly understood. Here, we found that the abundance of GmPLP1, a positive blue light receptor protein, was significantly inhibited by high light stress and mainly responded to high blue light. GmPLP1 RNA‐interference soybean lines exhibited higher light energy utilization ability and less light damage and reactive oxygen species (ROS) accumulation in leaves under high light stress, while the phenotype of GmPLP1:GmPLP1‐Flag overexpression soybean showed the opposite characteristics. Then, we identified a protein–protein interaction between GmPLP1 and GmVTC2, and the intensity of this interaction was primarily affected by sensing the intensity of blue light. More importantly, overexpression of GmVTC2b improved soybean tolerance to high light stress by enhancing the ROS scavenging capability through increasing the biosynthesis of ascorbic acid. This regulation was significantly enhanced after interfering with a GmPLP1 ‐interference fragment in GmVTC2b‐ox soybean leaves, but was weakened when GmPLP1 was transiently overexpressed. These findings demonstrate that GmPLP1 regulates the photosynthetic capacity and ROS accumulation of soybean to adapt to changes in light intensity by sensing blue light. In summary, this study discovered a new mechanism through which GmPLP1 participates in high light stress in soybean, which has great significance for improving soybean yield and the adaptability of soybean to high light.

为了解大豆节间在不同温度和外源赤霉素(gibberellic acid,GA)诱导条件下的表型变化规律,分析GA合成重要途径和挖掘调控节间的关键候选基因,本研究将大豆品种Charleston在培养箱条件和室外盆栽条件种植,进行不同温度处理和外源GA涂抹处理,利用徒手切片配合显微照相方法,分析大豆节间长度和细胞形态变化;利用液相色谱-质谱联机结合转录组测序方法分析大豆节间GA合成主要通路并挖掘调控节间生长的关键候选基因.结果表明在25℃和30℃条件下,外源涂抹不同浓度GA可以诱导大豆生长节间伸长,随着伸长量的增加,大豆节间都变得纤细.外源GA对细胞作用效果主要为促进伸长,对宽度影响不明显.高温处理对节间的伸长效果高于低温处理.本研究鉴定到GA2氧化酶基因在大豆生长节间表达和较高含量的GA19和GA53,及这2种GA下游的GA20(活性GA前体),以及这条合成途径的活性GA产物GA3也被检测到都存在于细胞伸长区组织,说明从GA前体物质到GA53,再到GA19,通过GA20最终合成GA3是大豆节间生长的一条重要GA合成通路,进一步说明GA2氧化酶在大豆节间生长过程中有重要作用.挖掘到某些DELLA、GA和PIF基因家族成员具有组织表达特异性,为调节大豆节间和株高提供了候选基因.

IntroductionGlyceraldehyde-3-phosphate dehydrogenase (GAPDH) is an essential key enzyme in the glycolytic pathway and plays an important role in stress responses. Although GAPDH family genes have been found in different plant species, the determination of their gene family analysis and their functional roles in soybean are still unknown. MethodsIn this study, gene sequence and expression data were obtained using online tools, and systematic evolution, expression profile analysis, and qRT-PCR analysis were conducted.Results and DiscussionHere a total of 16 GmGAPDH genes were identified on nine chromosomes, which were classified into three clusters. Additionally, all GmGAPDH genes harbor two highly conserved domains, including Gp_dh_N (PF00044) and Gp_dh_C (PF02800). The qRTPCR analysis also showed that most GmGAPDH genes significantly responded to multiple abiotic stresses, including NaHCO3, polyethylene glycol, cold, and salt. Among them, GmGAPDH14 was extraordinarily induced by salt stress. The GmGAPDH14 gene was cloned and overexpressed through soybean hair roots. The overexpressed transgenic soybean plants of the GmGAPDH14 gene have also shown better growth than that of control plants. Moreover, the overexpressed transgenic plants of GmGAPDH14 gene had higher activities of superoxide dismutase but lower malonaldehyde (MDA) content than those of control plants under salt stress. Meanwhile, a total of four haplotypes were found for the GmGAPDH14 gene, and haplotypes 2, 3, and 4 were beneficial for the tolerance of soybean to salt stress. These results suggest that the GmGAPDH14 gene might be involved in the process of soybean tolerance to salt stress. The results of this study will be valuable in understanding the role of GAPDH genes in the abiotic stress response of soybean.

The maturity group (MG) system is a widely recognized and effective approach for assessing the photothermal sensitivity of soybean cultivars and determining their optimal adaptation zones. In China, soybean have been cultivated extensively for thousands of years, evolving into various ecotypes through natural and artificial selection. In this study, the relative maturity groups (RMGs) of a total of 766 soybean cultivars collected from the Northeast Spring Planting Sub-region (NE), Huang-Huai-Hai Summer Planting Region (HH), Northwest Spring Planting Sub-region (NW,), and South Multiple Cropping Region (SC) of China were evaluated using linear regression models at 36 sites nationwide. The results show that the RMGs of Chinese soybean cultivars range from MG m1.6 to MG 9.5. Among all the identified soybean cultivars, MG III cultivars account for the largest proportion of 22.19 %, followed by MG II (21.67 %) and MG I (18.54 %). Conversely, MG IX has the fewest number of cultivars, representing only 0.26 % of all cultivars. The MG ranges for spring- and summer-sowing cultivars are MG 000-V and MG II-IX, respectively. The adaptive soybean MG zones across China were mapped using the method of Kriging interpolation based on the soybean RMGs data from 816 sites of 29 soybean-producing provinces. Cultivars of MG 0 and earlier groups are primarily distributed in the NE, and MG III and MG IV are the major MGs in HH and NW. The cultivars in MG V and later groups mainly distribute in SC. This study realizes the unification and normalization of the MG system between China and other major soybean- producing countries worldwide. Such unification and standardization will facilitate global germplasm exchanges and assist soybean producers in making more informed decisions when selecting cultivars.

In this study, 255 soybean germplasm were used as experimental materials, the mixed alkali was used to simulate the alkali tolerance environment of soda saline-alkali land in Daqing City.The alkali tolerance related traits were investigated at germination and seedling stages of soybean germplasm. BLINK and Farm CPU were used for genome-wide association analysis. The results showed that a total of 64 associated SNP loci were detected by the two methods, of which seven loci such as 3:37513903, 3:37490361 and 9:37033917 were relatively stable SNP loci with high reliability. A total of 26genes with functional annotation were screened out. According to functional annotation and gene expression level, three genes related to alkali tolerance were speculated, which were Glyma.03G160200,Glyma.09G149900 and Glyma.19G067400.

Lodging is an important agronomic trait related to crop yield and is easily susceptible to environmental influences. In this study, a recombinant inbred line population from soybean (Glycine max (L.) Merr.) Hefeng 25 × Dongnong L28 including 109 lines was used to identify quantitative trait loci (QTLs) related to soybean lodging. Seven QTLs were identified in the three environments (Harbin in 2017, 2018 and 2019), and these could explain 2.21–20.17% of the phenotypic variation. Among these QTLs, qLDG-I-1 (Chr20_24146101–Chr20_24297321) was stable for multiple environments. A residue heterozygous line, which was heterozygous at the qLDG-I-1 locus, was used to verify qLDG-I-1, and the results showed that this QTL could significantly improve lodging resistance of soybean. Meanwhile, 13 pairs of epistatic QTLs were detected, which could explain 3.26–18.24% of the phenotypic variation. QTL × environment interaction mapping was also used, and it detected 31 QTLs, which could explain 1.61–7.94% of the phenotypic variation. In total, 122 pairs of epistatic QTLs were detected, and they could explain 5.39–27.81% of the phenotypic variation. Additionally, candidate genes related to soybean lodging in the qLDG-I-1 interval were predicted, and Glyma.20g068000 was mined as a candidate gene based on quantitative real-time PCR analysis. The QTLs and candidate genes identified in this study are of great significance to position cloning, and could accelerate the progress of breeding resistance to lodging in soybean.

研究采用芽期耐盐性状差异显著的东农46和L-100杂交衍生的包含127个家系的F2:12和F2:13重组自交系群体为试验材料,调查耐盐性状:吸胀率(IR)、发芽指数(GI)和发芽率(GR),计算3个相对耐盐指数:相对吸胀率(STIR)、相对发芽指数(STGI)和相对发芽率(STGR),利用QTL IciMapping 4.1.0.0软件对3个相对耐盐指数作QTL分析.结果表明,共检测到12个加性QTL,贡献率为4.88％～15.91％,2个位点有重叠区间,3个性状表型贡献率最高QTL分别为qSTIR-9-1(15.91％)、qSTGI-2-1(10.21％)和qSTGR-20-1(12.23％),共检测到108个上位性QTL,贡献率为0.55％～26.59％.该耐盐性状QTL为分子辅助培育抗盐大豆品种奠定基础.

Soybean is sensitive to drought stress, and increasing tolerance to drought stresses is an important target for improving the performance of soybean in the field. The genetic mechanisms underlying soybean’s drought tolerance remain largely unknown. Via a genome-wide association study (GWAS) combined with linkage analysis, we identified 11 single-nucleotide polymorphisms (SNPs) and 22 quantitative trait locus (QTLs) that are significantly associated with soybean drought tolerance. One of these loci, namely qGI10-1, was co-located by GWAS and linkage mapping. The two intervals of qGI10-1 were differentiated between wild and cultivated soybean. A nuclear factor Y transcription factor, GmNFYB17, was located in one of the differentiated regions of qGI10-1 and thus selected as a candidate gene for further analyses. The analysis of 29 homologous genes of GmNFYB17 in soybean showed that most of the genes from this family were involved in drought stress. The over-expression of GmNFYB17 in soybean enhanced drought resistance and yield accumulation. The transgenic plants grew better than control under limited water conditions and showed a lower degree of leaf damage and MDA content but higher RWC, SOD activity and proline content compared with control. Moreover, the transgenic plants showed a fast-growing root system, especially regarding a higher root–top ratio and more branching roots and lateral roots. The better agronomic traits of yield were also found in GmNFYB17 transgenic plants. Thus, the GmNFYB17 gene was proven to positively regulate drought stress resistance and modulate root growth in soybean. These results provide important insights into the molecular mechanisms underlying drought tolerance in soybean.

WRINKLED1 (WRI1), an APETALA2/ethylene-responsive-element-binding protein (AP2/EREBP) subfamily transcription factor, plays a crucial role in the transcriptional regulation of plant fatty acid biosynthesis. In this study, GmWRI1a was overexpressed in the soybean cultivar ‘Dongnong 50’ using Agrobacterium-mediated transformation to generate three transgenic lines with high seed oil contents. PCR and Southern blotting analysis showed that the T-DNA was inserted into the genome at precise insertion sites and was stably inherited by the progeny. Expression analysis using qRT-PCR and Western blotting indicated that GmWRI1a and bar driven by the CaMV 35S promoter were significantly upregulated in the transgenic plants at different developmental stages. Transcriptome sequencing results showed there were obvious differences in gene expression between transgenic line and transgenic receptor during seed developmental stages. KEGG analysis found that the differentially expressed genes mainly annotated to metabolic pathways, such as carbohydrated metabolism and lipid metabolism. A 2-year single-location field trial revealed that three transgenic lines overexpressing GmWRI1a (GmWRI1a-OE) showed a stable increase in seed oil content of 4.97–10.35%. Importantly, no significant effect on protein content and yield was observed. Overexpression of GmWRI1a changed the fatty acid composition by increasing the linoleic acid (C18:2) content and decreasing the palmitic acid (C16:0) content in the seed. The three GmWRI1a-OE lines showed no significant changes in agronomic traits. The results demonstrated that the three GmWRI1a overexpression lines exhibited consistent increases in seed oil content compared with that of the wild type and did not significantly affect the seed yield and agronomic traits. The genetic engineering of GmWRI1a will be an effective strategy for the improvement of seed oil content and value in soybean.

Isoflavones, one of the most important secondary metabolites produced by soybeans (Glycine max (L.) Merr.), are important for a variety of biological processes, and are beneficial for human health. To identify genetic loci underlying soybean isoflavone content, a mapping population containing 119 F5:18 recombinant inbred lines, derived by crossing soybean cultivar "Zhongdou27" with "Dongong8004," was used. We identified 15 QTLs associated with isoflavone contents. A novel loci, qISO19-1, was mapped onto soybean chromosome 19 and was fine-mapped to a 62.8 kb region using a BC2F2 population. We considered GmMT1 as a candidate gene for the qISO19-1 locus due to the significant positive correlation recovered between its expression level and isoflavone content in the seeds of 43 soybean germplasms. Overexpression of GmMT1 in Arabidopsis and soybean cultivars increased isoflavone contents. Transgenic soybeans overexpressing GmMT1 also exhibited improved resistance to pathogenic infection, while transgenic Arabidopsis resisted salt and drought stress.

Key message Genetic resources contributes to the sustainable protein production in soybean. Abstract Soybean is an important crop for food, oil, and forage and is the main source of edible vegetable oil and vegetable protein. It plays an important role in maintaining balanced dietary nutrients for human health. The soybean protein content is a quantitative trait mainly controlled by gene additive effects and is usually negatively correlated with agronomic traits such as the oil content and yield. The selection of soybean varieties with high protein content and high yield to secure sustainable protein production is one of the difficulties in soybean breeding. The abundant genetic variation of soybean germplasm resources is the basis for overcoming the obstacles in breeding for soybean varieties with high yield and high protein content. Soybean has been cultivated for more than 5000 years and has spread from China to other parts of the world. The rich genetic resources play an important role in promoting the sustainable production of soybean protein worldwide. In this paper, the origin and spread of soybean and the current status of soybean production are reviewed; the genetic characteristics of soybean protein and the distribution of resources are expounded based on phenotypes; the discovery of soybean seed protein-related genes as well as transcriptomic, metabolomic, and proteomic studies in soybean are elaborated; the creation and utilization of high-protein germplasm resources are introduced; and the prospect of high-protein soybean breeding is described.

Plant height is an important component of plant architecture, and significantly affects crop quality and yield. A soybean GmRAV (Related to ABI3/VP1) transcription factor containing both AP2 and B3 domains is a growth repressor. Three GmRAV-overexpressing (GmRAV-ox) transgenic lines displayed extremely shorter height and shortened internodes compared with control plants, whereas transgenic inhibition of GmRAV expression resulted in increased plant height. GmRAV-ox soybean plants showed a low active gibberellin level and the dwarf phenotype could be rescued by treatment with exogenous GA3 treatment. ChIP (Chromatin immunoprecipitation)-qPCR assay showed that GmRAV could directly regulate the expression of the GA4 biosynthetic genes GA3-oxidase (GmGA3ox) by binding two CAACA motifs in the GmGA3ox promoter. The GmGA3ox promoter was bound by GmRAV, whose expression levels in leaves were both elevated in GmRAV-i-3 and decreased in GmRAV-ox-7 soybean plants. Transient expression assay in N. benthamiana also showed that the proGmRAV:GmRAV-3F6H effector strongly repressed the expression of LUC reporter gene driven by GmGA3ox promoter containing two CAACA motifs. Together, our results suggested that GmRAV protein repressed the expression of GmGA3ox by directly binding to the two CAACA motifs in the promoter to limit soybean plant height.

大豆是重要的粮油作物,而我国大豆主要依靠进口,提高大豆产量对保障国家粮油安全意义重大.为定位大豆产量相关性状,本研究以产量差异显著的东农42和东农50作为杂交亲本,构建了包含168个家系的重组自交系(recombination inbred lines,RILs)群体,对其进行全基因组重测序,构建高密度遗传图谱,并利用R/qtl软件的复合区间作图法(composite interval mapping,CIM)结合两年六点的大豆产量相关性状表型数据,进行QTL定位.结果表明:利用测序获得的660316个SNP标记构建了一张分布在20个连锁群的包含6227个bin标记的大豆高密度遗传图谱,总图距和平均图距分别为2739.15 cM,0.44 cM.在12个染色体上定位到22个大豆产量相关性状QTL,四粒荚数、单株荚数、单株粒重和百粒重性状定位到的QTL分别为5、4、5和8个.在3号和19号染色体上各有一段基因组区域在两年间重复定位,涉及6个主效QTL,分别为qNFSP-19-1(22.976％)、qNFSP-19-2(11.977％)、qNFSP-19-3(17.203％)、qHSW-3-1(11.346％)、qHSW-3-2(11.346％)和qHSW-3-3(11.175％),加性效应值均为负值.在3、7、11、12和20号染色体上新定位到7个产量相关性状QTL,其中表型贡献率最高的为qHSW-3-3(14.276％),包含具有重复定位区间的qHSW-3-2和qHSW-3-3.与前人定位的结果相比,更多QTL极大地缩短了定位区间,表明本文报道的高密度遗传图谱更准确,可以为大豆产量相关性状的精细定位、候选基因的挖掘及分子标记辅助育种奠定基础.

Soybean frogeye leaf spot (FLS) is a worldwide fungal disease. Its higher occurrence frequency and wider distribution range always led to severe yield losses of soybean, therefore, breeding new cultivars with FLS resistance has been an important breeding goal for soybean breeders. In this study, an association panel of 183 representative soybean accessions was used to evaluate their resistance to FLS race 1, and to identify quantitative trait nucleotides (QTNs) and candidate genes based on genome-wide association study (GWAS) and high-throughput single-nucleotide polymorphisms (SNPs). A total of 23,156 high-quality SNPs were developed using the specific locus-amplified fragment sequencing (SLAF-seq) approach. Finally, 13 novel association signals associated with FLS race 1 resistance were identified by the compressed mixed linear model (CMLM). In addition, 119 candidate genes were found within the 200-kb flanking genomic region of these 13 peak SNPs. Based on the gene-based association analysis, haplotype analysis, expression pattern analysis, and virus-induced gene silencing (VIGS) systems, four genes (Glyma.05G121100, Glyma.17G228300, Glyma.19G006900, and Glyma.19G008700) were preliminarily proved to play an important role in the soybean resistance to FLS race 1.

为明确大豆节间木质素积累规律,深入理解木质素积累与大豆植株抗倒伏间的关系,进而为调控大豆植株表型和抗倒伏大豆新品种选育提供参考依据,选取具有代表性的有限生长习性品种Charleston,按照20,35株/m2的2种密度种植桶栽大豆,在2种不同密度下测定主茎的木质素含量,分析Charleston主茎第3节间的细胞伸长区(EZ)和次生细胞壁成熟区(MZ)木质素含量、在木质素合成过程中的4-香豆酸:辅酶A连接酶(4CL)、苯丙氨酸转氨酶(PAL)、肉桂酸脱氢酶(CAD)等关键酶活性以及对应基因的表达量.手工切片及分光光度计测定的结果表明,大豆主茎木质素含量由形态学上端向下端逐渐增加.低密度处理主茎中的木质素含量高于高密度.无论是高密度还是低密度处理,单一节间(茎3)木质素含量均为MZ>EZ;低密度处理茎中4CL、PAL、CAD的活性高于高密度处理.2个密度处理下茎中4CL、PAL、CAD活性变化规律较为一致,与基因表达结果相符合.大豆植株主茎中木质素含量受到木质素合成途径中的多种酶调控.木质素合成的关键酶基因多以基因家族形式存在,同一基因家族中的酶基因在同一物种植株的相同部位表达存在差异,同一基因在同一物种的不同组织和器官中的表达情况也不尽相同.

采用共沉淀法合成了MnCo2O4催化剂,并采用水热法合成了氧化锰八面体分子筛(OMS-2)催化剂,与NaY分子筛催化纤维素热解效果对比后对NaY进一步改性,采用浸渍法制备了Co/NaY、Sn/NaY以及Co-Sn/NaY,并研究了所制备催化剂对纤维素热解气催化重整制备呋喃类化合物的影响.采用氮气吸附-脱附、X射线衍射(XRD)仪、扫描电镜显微镜(SEM)和氨气吸附-脱附法(NH3-TPD)对催化剂进行表征.实验结果表明:金属Co、Sn分散在NaY表面,且没有改变NaY的晶体结构和形貌;负载后催化剂的比表面积和孔容减小,平均孔径增大,且比表面积远大于MnCo2O4、OMS-2;负载Sn几乎不改变NaY分子筛的酸性位点,而Co会减少酸性位点.几种催化剂的加入均促进了醇类、酸类、醛类的生成,抑制了糖类的生成.在纤维素与催化剂的质量比为1:14的条件下,NaY和Co/NaY均能够显著提高呋喃类化合物质量分数,从未添加催化剂的15.34％分别提高到了48.16％和61.88％,金属负载NaY催化剂均促进了呋喃酮的生成,抑制了呋喃的产生.

Soybean mosaic virus (SMV) is one of the most widespread and devastating viral diseases worldwide. The genetic architecture of qualitative resistance to SMV in soybean remains unclear. Here, the Rsvg2 locus was identified as underlying soybean resistance to SMV by genome-wide association and linkage analyses. Fine mapping results showed that soybean resistance to SMV strains G2 and G3 was controlled by a single dominant gene, GmST1, on chromosome 13, encoding a sulfotransferase (SOT). A key variation at position 506 in the coding region of GmST1 associated with the structure of the encoded SOT and changed SOT activity levels between RSVG2-S and RSVG2-R alleles. In RSVG2-S allele carrier "Hefeng25", the overexpression of GmST1 carrying the RSVG2-R allele from the SMV-resistant line "Dongnong93-046" conferred resistance to SMV strains G2 and G3. Compared to Hefeng25, the accumulation of SMV was decreased in transgenic plants carrying the RSVG2-R allele. SMV infection differentiated both the accumulation of jasmonates and expression patterns of genes involved in jasmonic acid (JA) signalling, biosynthesis and catabolism in RSVG2-R and RSVG2-S allele carriers. This characterization of GmST1 suggests a new scenario explaining soybean resistance to SMV.

研究以150份大豆种质为试验材料开展2年3点试验,在收获期测定抗倒伏相关性状,并利用RTM-GWAS方法作关联分析,共检测到99个显著关联SNP位点,其中48个效应显著位点,解释0.28％～3.38％表型变异;89个与环境互作显著位点,解释0.53％～7.04％表型变异;其中,21个位点主效表型变异解释率高于1％;与6个倒伏相关性状显著关联SNP位点中,与茎粗显著关联SNP位点最多,与抗倒指数显著关联SNP位点最少;获得22个有功能注释基因,根据基因表达量和基因功能注释,推测3个基因(Glyma.18G149700、Glyma.04G244100、Glyma.18G011400)可作为与大豆抗倒伏相关候选基因.