Excessive rainfall provides a favorable condition for field mold infection of plants, which triggers field mold (FM) stress. If FM stress occurs during the late maturation stage of soybean seed, it negatively affects seed yield and quality. To investigate the responses of soybean seed against FM stress and identify the underlying biochemical pathways involved, a greenhouse was equipped with an artificial rain producing system to allow the induction of mold growth on soybean seed. The induced quality changes and stress responses were revealed on the levels of both transcriptome and metabolome. The results showed that soybean seeds produced under FM stress conditions had an abnormal and inferior appearance, and also contained less storage reserves, such as protein and polysaccharide. Transcriptional analysis demonstrated that genes involved in amino acid metabolism, glycolysis, tricarboxylic acid, β-oxidation of fatty acids, and isoflavone biosynthesis were induced by FM stress. These results were supported by a multiple metabolic analysis which exhibited increases in the concentrations of a variety of amino acids, sugars, organic acids, and isoflavones, as well as reductions of several fatty acids. Reprogramming of these metabolic pathways mobilized and consumed stored protein, sugar and fatty acid reserves in the soybean seed in order to meet the energy and substrate demand on the defense system, but led to deterioration of seed quality. In general, FM stress induced catabolism of storage reserves and diminished the quality of soybean seed in the field. This study provides a more profound insight into seed deterioration caused by FM stress.

Black soybean seed (BSS) is a traditional edible and medical food. The cotyledons of BSS are either yellow or green, but that difference's effect on functional and bioactivities are not clear. In the present study, the concentrations of the main functional components, including anthocyanins, isoflavones, fatty acids, protein, and polysaccharides, and in vitro antioxidant activities of yellow-cotyledon and green-cotyledon BSS (YBS and GBS, respectively) were compared. The anti-fatigue activities of the ethanol extract of BSS (EEB) were evaluated using mouse swimming endurance tests. Additionally, a metabolomics comparison between YBS and GBS was done using ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC/Q-TOF MS) on high doses of EEB-treated mouse plasma. The results indicated that YBS showed significantly higher contents of anthocyanin and protein, and stronger antioxidant activity and anti-fatigue activity than GBS; while GBS showed higher levels of isoflavone, stearic acid, and polysaccharide. By infusing the feed with EEB, the swimming time of mice increased in a dose-dependent manner. The metabolomics results suggested that the anti-fatigue effect of EEB correlated well with serotonin and melatonin biosynthesis in vivo. All of the above results showed that BSS has anti-fatigue potential and might be used as a potential source of functional food ingredients.

通过本草文献综述,梳理了黑豆作为药材使用的历史简况;并结合药化分析试验比较了不同籽重黑豆的化学成分和生物活性,对其化学鉴别、质量评价方法提出了新的商榷建议.推荐临床以百粒重小于18 g的小黑豆入药,建议新版《中国药典》明确以矢车菊素葡萄糖苷为对照品鉴别其真伪;采用高效液相色谱法检测其含量,并以此作为黑豆质量评价标准.

采用高效液相色谱质谱联用技术(HPLC-MS),建立黑豆异黄酮和花色苷含量测定方法.应用WatersXselect HSS T3色谱柱,以乙腈-0.1％乙酸水溶液为流动相梯度洗脱黑豆异黄酮;应用X-Terra MS C18色谱柱,以10％甲酸水溶液-甲醇/乙腈/水/甲酸(22.5/22.5/40/10,v/v)混合液为流动相梯度洗脱黑豆花色苷;以电喷雾离子源(ESI)选择离子监控模式(SIM)检测目标化合物,并通过标准品对照比较,实现黑豆12个异黄酮及8个花色苷单体化合物的定量测定.该方法操作简便、稳定、耗时少,具有良好的灵敏度、重现性和回收率,可用于黑豆种质资源的化学评价及质量控制.

Targeted metabolomics analysis was conducted to quantify fatty acids in soybean seeds using GC-MS. PLS-DA helped to determine the appropriate row spacing in the intercropping system.

Maize-soybean relay strip intercropping systems is an important planting mode in Southwest China. However,shade-tolerant germplasms of black soybean are scarce and the mechanism of shade tolerance of black soybean is still unclear, which significantly affects further promotion of the mode. The objective of this study was to establish a mathematical model to evaluate shade-tolerance ability of black soybean seedlings, screen shade-tolerance appraisal indicators and discuss the diverse responses of the root structures of different black soybean germplasms seedlings to shade stress.The study was also intended to lay the foundation for breeding varieties and for explaining the mechanism of shade tolerance of black soybean. Thus in the study, shade tolerance of 23 black soybean germplasms were evaluated in a pot culture experiment with normal light and shade treatments. The morphological characteristics (plant height, stem diameter, hypocotyl length, leaf dry weight, stem dry weight, main stem count, total root length, root surface area, total root volume, etc.) and physiological parameters were measured at the 3rd trifoliolate leaf stage (V3). An evaluation mathematical model was established using multivariate statistical analysis. Additionally, based on the evaluation results, correlation analysis was conducted to determine differential responses of roots to shade stress. The 23 black soybean germplasms were divided into three groups, including tolerant, moderately tolerant and sensitive germplasms. A total of 5 evaluation indexes closely related to shade tolerance were screened out. The study showed that the response of root structure to shade stress varied with shade-tolerance ability of black soybean germplasms. Root pa-rameters such as total root length, root surface area, total root volume and root dry weight all had a similar order, which was shade tolerant germplasms > moderate shade tolerant germplasms > sensitive germplasms. The results indicated that shade tolerant germplasms of black soybean had a stronger root system.

[目的]通过分离、纯化和鉴定黑豆种皮中的主要化学成分,对其田间霉菌抑制活性进行系统评价.[方法]以抑菌活性为指导,采用液相萃取、硅胶柱层析等方法,分离纯化黑豆种皮醇提物中的化学成分,结合核磁共振波谱(NMR)数据鉴定抑菌化合物的化学结构;并采用高效液相色谱质谱联用技术(HPLC-MS)对黑豆种皮中花色苷组分进行定量分析.[结果]黑豆种皮抑菌活性成分主要集中在乙酸乙酯萃取层,从中纯化得到主要活性成分表儿茶素,其对黄曲霉菌(Aspergillus flavus)、黑曲霉菌(Aspergillus nige)、串珠镰刀菌(Fusarium moniliforme)和产黄青霉菌(Penicillium chrysogenum)具有明显的抑制作用.定量分析结果表明,黑豆种皮中富含矢车菊素葡萄糖苷等花色苷类物质,其在提取过程中可转化形成表儿茶素.[结论]表儿茶素作为黑豆种皮中多酚类物质的重要组成成分,具有较强的抑菌活性,是黑豆抗田间霉菌的主要活性物质.

The isoflavone profiles of seeds of various soybean genotypes with different levels of shade tolerance at the seedling stage were investigated. High-performance liquid chromatography (HPLC) was used to quantify 12 isoflavones, and the data were analyzed using a multivariate statistical analysis. Combined with field experimental data and an orthogonal partial least-squares discriminant analysis (OPLS-DA), several aglycones (genistein (GE), daidzein (DE), and glycitein (GLE)) were selected and identified as key compounds involved in the shade tolerance of soybean seedlings. Additional correlation analysis and laboratory shading stress experiments with soybean seedlings also confirmed the function of these selected isoflavones, especially GE, in the discrimination of soybean seedlings with different levels of shade tolerance. Furthermore, the structure–antioxidant activity relationships between a range of isoflavones and the plant shade-tolerance mechanism are discussed. Targeted metabolomic analyses of isoflavones could reveal the diversity of shade tolerance in soybean seedlings, thus contributing to the breeding of excellent soybean varieties.

Twelve isoflavones and eight anthocyanins were quantified in black soybean seeds by HPLC-MS analyses; the coupled OPLS-DA multivariate statistical analysis helped us to determine their geographical origin.

The reduction in the ratio of red light (R) to far-red light (FR) is an important signal of shade stress.In the present study,an experiment was conducted to study the effect of normal and low R/FR irradiation on the isoflavone contents of two soybean varieties:high shade-tolerance species'ND12'and low shade-tolerance species'C103'.Relative expression levels of key genes,which encoded isoflavone synthetic enzymes in leaves,hypocotyls and roots were also studied.The results indicated that compared with seedlings under normal light,low R/FR increased total contents of isoflavone and genistein significantly in the leaves of'ND12',but decreased in hypocotyls;while the opposite result was obtained in'C103'.Expression of isoflavone synthesis key genes,including IFS2,4CL,C4H and CHI correlated with isoflavone accumulation under low R/FR irradiation.Shading signal induced isoflavone accumulation particularly in leave of shade-tolerance varieties'.This study preliminary clarified the shading signal (low R/FR) that how to control the rules of isoflavone accumulation in soybean seedlings.This study laid a foundation to research regulation mechanism of soybean isoflavone response to shading signal.

In this study 17 soybean genotypes were introduced from Japan to the Sichuan Basin with an aim to assess their adaptability.Comparison of quality traits showed that variation in isoflavone content was highest;the coefficient of variation values were generally greater than 30% (apart from aglycone),while the coefficient variation for other seed compounds was around 10%.The soluble protein contents of the tested soybean seeds were relatively high while fatty acids,soluble polysaccharides and isoflavone were low.Principal component a-nalysis and cluster analysis divided the 17 germplasms into five groups.Individually,Group-Ⅰ comprised high-oil varieties,their linoleic acid content were>55‰,suggesting that they could be used to produce high-oil hy-brid varieties.Group-Ⅲ comprised high-protein varieties,potentially useful for developing varieties to produce soy raw materials;Group-Ⅴ comprised high-isoflavone varieties,which could potentially be used to produce varieties useful for functional food or fodder.

以大豆下胚轴为试验材料,研究了绿原酸(CGA)及其与植物生长调节剂(IAA、IBA)共同作用对大豆下胚轴不定根发生的影响.结果表明,当CGA施用量达到200 μmol/L及400μmol/L时,大豆下胚轴生根率分别显著降低了17.8％和26.7％,平均根长分别显著降低了18.7％和35.4％;当植物生长调节剂施用量为2μmol/L时,可显著缓解CGA(100 μmol/L)对大豆不定根的抑制作用,使其生根率分别提高12.5％和20.0％,随着生长素施用量的进一步增加(10～50 u mol/L),其生根率显著下降;大豆不定根培养24h内,对照下胚轴基部木质素相对含量降低,CGA处理大豆的木质素相对含量升高.高浓度CGA对大豆不定根形成具有抑制作用,其通过与木质素、生长素的共同作用,实现对大豆不定根发生的调控.

Seedpods are the outermost barrier of legume plants encountered by pests and pathogens, but research on this tissue, especially regarding their chemical constituents, is limited. In the present study, a mildew-index-model-based cluster analysis was used to evaluate and identify groups of soybean genotypes with different organ-specific resistance against field mold. The constituents of soybean pods, including proteins, carbohydrates, fatty acids, and isoflavones, were analyzed. Linear regression and correlation analyses were also conducted between these main pod constituents and the organ-specific mildew indexes of seed (MIS) and pod (MIP). With increases in the contents of infection constituents, such as proteins, carbohydrates, and fatty acids, the MIP increased and the MIS decreased. The MIS decreased with increases in the contents of glycitein (GLE)-type isoflavonoids, which act as antibiotic constituents. Although the infection constituents in the soybean pods caused pod mildew, they also helped mitigate the corresponding seed mildew to a certain extent.

The effects of storage duration on the seed germination and metabolite profiling of soybean seeds with five different coloured coats were studied. Their germination, constituents and transcript expressions of isoflavones and free fatty acids (FFAs) were compared using chromatographic metabolomic profiling and transcriptome sequencing. The seed water content was characterized using nuclear magnetic resonance (NMR) relaxometry. Results showed that dark-coloured seeds were less inactivated than light-coloured seeds. The aglycone and β-glucoside concentrations of upstream constituents increased significantly, whereas the acetylglucosides and malonylglucosides of downstream constituents decreased with an increase in the storage period. FFAs increased considerably in the soybean seeds as a result of storage. These results indicate that dark-coloured soybean seeds have better storability than light-coloured seeds, and seed water content plays a role in seed inactivation. It was concluded that there are certain metabolic regularities that are associated with different coloured seed coats of soybeans under storage conditions.

以西南地区不同来源的15个大豆种质资源为研究对象,研究大豆主要化学成分与其田间霉变抗性间的相关性.采用高效液相色谱法测定大豆中12种异黄酮含量,气质联用法测定大豆中19种脂肪酸含量,分光光度法测定大豆皂苷、可溶性多糖、蛋白含量,并进行聚类及相关性分析.结果表明,15个大豆种质资源依据化学成分含量被分为5个类别,C103等材料的品质性状优良,具有较好的田间霉变抗性,可作为优质大豆种质资源进一步开发利用.棕榈酸、反亚油酸和芥酸等脂肪酸组分含量高的大豆材料其霉变程度更高,A环C-6位甲氧基化的GL型异黄酮(黄豆黄素、黄豆黄苷、乙酰黄豆黄苷、丙二酰黄豆黄苷)及异黄酮苷元含量高的大豆具有更大的抗霉变潜力.

In this present study, two soybean cultivars with different drought tolerance in serial number of JP-6 (high drought-tolerant species) and JP-16 (low drought-tolerant species) were researched. The HPLC and real-time PCR analyses were used to determine the isoflavone contents and relative expression levels of key genes, which encoded isoflavone synthesis relative enzymes in lea-ves and roots under different drought stress levels, respectively. The results indicated that the isoflavone contents in roots were significantly higher than that in leaves, whereas the relative expression of isoflavone synthetic enzyme related genes in leaves was significantly higher than that in roots. Analysis of isoflavone accumulation by comparing two different drought tolerance soybean cultivars found that the isoflavone accumulation in roots of JP-6 was greater than that in others. With increa-sing levels of the drought stress, there were significant differences in both isoflavone synthesis and accumulation between JP-6 and JP-16 soybean cultivars. In JP-6, the isoflavone accumulation in root and leaf increased after slight decreasing, while the opposite result was obtained in JP-16, in which the isoflavone accumulation in different parts were decreased after slight increasing. The expression of isoflavone synthesis relative enzyme genes presented a trend that decreased and then gradually increased with the increasing level of drought stress, except C4H, 4CL and IFS2 which were synthesis genes upstream of isoflavones in leaves of JP-6. Soybean isoflavones were mainly synthesized in leaves, little was synthesized in roots. The isoflavone synthesis and accumulation of low drought-tolerant species were scare, while those of high drought-resistant variety were relatively higher. The variety with high isoflavone accumulation in the root was more drought-resistant.

The profiles of isoflavone and fatty acids constitute important quality traits in soybean seeds, for making soy-based functional food products, due to their important contributions to the flavor and nutritional value of these products. In general, the composition of these constituents in raw soybeans is affected by cultivation factors, such as sunlight; however, the relationship of the isoflavone and fatty acid profiles with cultivation factors is not well understood. This study evaluated the isoflavone and fatty acid profiles in soybeans grown under a maize–soybean relay strip intercropping system with different row spacings, and with changes in the photosynthetic active radiation (PAR) transmittance. The effects of PAR on the isoflavone and fatty acid contents were found to be quadratic. Appropriate intercropping shading may reduce the bitterness of soybeans caused by soy aglycone and could improve their fatty acid composition.