利用中国秋大豆(Glycine max(L.) Merr)筛选SSR核心位点的研究

选择中国秋大豆为试验材料 ,对基因组DNA进行SSR标记筛选和鉴定。经过 2 0 0个位点在琼脂糖胶上初筛和 96个位点在变性聚丙烯酰胺胶上复鉴 ,选出 6 0个位点 ,这些位点具有以下特点 :(1)分布在大豆 2 0个整合遗传连锁群 ,相邻位点间平均遗传距离在 5 0cM左右。除连锁群C2 、O上分别有 5个位点 ,G、K、M上分别有 2个位点外 ,其余 15个连锁群均分布有 3个位点 ;(2 )与 96个位点在 80份秋大豆种质检测到种质间遗传关系达到极显著相关 (r =0 .910 ) ;(3)在 80份秋大豆初选核心种质中表现出较高多态性 ,平均每个位点等位变异数为 9.3,多态性信息含量 (PIC)值为 0 .773;(4)在检测的秋大豆绝大多数种质基因组中 ,均为单一拷贝的位点 ,具有较高特异性 ;(5 )在相同的PCR扩增条件下 ,同一位点不同等位变异间易于识别且扩增强度较为一致。这套SSR核心位点的确定为中国大豆核心种质的构建奠定了基础。

大豆(Glycine max(L.) Merr.)叶片抗氧化酶对干旱及复水的响应机制

采用10%PEG-6000模拟干旱胁迫,研究了抗旱品种晋大74与不抗旱品种晋大75两种大豆幼苗在水分胁迫及复水后叶片MDA含量、O2-·含量、SOD、CAT、APX及POD等的活性变化,旨在揭示抗氧化酶对干旱及复水的响应机制。结果表明,在干旱胁迫下,两种大豆叶片MDA及O2-·,含量都高于对照,且晋大75增加幅度显著大于晋大74;复水后晋大74中MDA及O2-·接近于对照水平,表明旱后复水产生了补偿效应。干旱胁迫下,晋大74通过保持较高的SOD活性或较低的APX、CAT降幅以减弱活性氧伤害,而旱后复水过程中晋大74通过维持较高的SOD、APX、CAT酶活性,利用其协同作用有效清除活性氧,避免膜伤害。总之,耐旱品种在干旱及复水过程中具有更强的抗氧化修复能力。

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.

Genetic Analysis of Embryo, Cytoplasm and Maternal Effects and Their Environment Interactions for Isoflavone Content in Soybean [Glycine max (L.) Merr.]

Soybean seed products contain isoflavones （genistein, daidzein, and glycitein） that display biological effects when ingested by humans and animals. These effects are species, dose and age dependent. Therefore, the content and quality of isoflavones in soybeans is a key factor to the biological effect. Our objective was to identify the genetic effects that underlie the isoflavone content in soybean seeds. A genetic model for quantitative traits of seeds in diploid plants was applied to estimate the genetic main effects and genotype x environment （GE） interaction effects for the isoflavone content （IC） of soybean seeds by using two years experimental data with an incomplete diallel mating design of six parents. Results showed that the IC of soybean seeds was simultaneously controlled by the genetic effects of maternal, embryo, and cytoplasm, of which maternal genetic effects were most important, followed by embryo and cytoplasmic genetic effects. The main effects of different genetic systems on IC trait were more important than environment interaction effects. The strong dominance effects on isoflavone from residual was made easily by environment conditions. Therefore, the improvement of the IC of soybean seeds would be more efficient when selection is based on maternal plants than that on the single seed. Maternal heritability （65.73%） was most important for IC, followed by embryo heritability （25.87%） and cytoplasmic heritability （8.39%）. Based on predicated genetic effects, Yudou 29 and Zheng 90007 were better than other parents for increasing IC in the progeny and improving the quality of soybean, The significant effects of maternal and embryo dominance effects in variance show that the embryo heterosis and maternal heterosis are existent and uninfluenced by environment interaction effects.

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.

Genetic Analysis of Combining Abilities and Heterosis for the Contents of Soybean Isoflavone and Its Components Among the Soybean Varieties [Glycine max (L.) Merr.]

The genetic analysis of soybean isoflavone content and its components were carried out based on the NC Ⅱ matingdesign in eight soybean varieties. The results showed that the isoflavone contents and its components of soybean seedare quite differences among the tested materials, the contents of isoflavone and daidzein are controlled not only byadditive effects and but also by non-additive effects, while the content of genistin is dominated by non-additive effects,and genistein, glycitin and daidzin are mainly controlled by additive effects. There are significant differences in thecontents of isoflavone and its components among the combinations derived from different parents. Results also indicatedthat the tested traits are negatively heterosis except for the contens of daidzein and daidzin are positively heterosis basedon the data of the GCA and SCA in average heterosis values. In this research we have a suggestion that soybean varietywith high isoflavone should be used as one of the parents in the breeding program, and it is the best choice that thecombinations crossed between two high isoflavone varieties or a high variety and a low one.

Proteomic Studies of Petal-specific Proteins in Soybean [Glycine Max(L.)Merr.] Florets

A survey of petal-specific proteomes of soybean(Glycine max(L.) Merr[Non-italic].) was conducted comparing protein expression profiles in different petals. Two-dimensional polyacrylamide gel electrophoresis reference maps of protein extracts from standard petals(SP), lateral wings(LW), keel petals(KP), and reproductive organs(RO)(a mixture of stamen and carpel) were obtained. Protein expression in the three petal types was compared using Image Master TM 2 D platinum 6.0 software. This indicated that the proportion of homologous proteins between SP and LW was 59.27%, between SP and KP was 61.48%, and between LW and KP was 60.05%. Within a mass range of 6.5-200.0 ku and pH 4.0-7.0, approximately 590, 646, 544, and 700 protein spots were detected in SP, LW, KP, and RO, respectively. A total of 82 differentially expressed proteins were detected. Sixty-four of these detected spots were differentially expressed and showed more than 2-fold changes in abundance; of these 64 proteins, 26 showed increased expression and 38 showed decreased expression. Among these spots, single organ-specific proteins were also identified.They were ID 49(60.9 ku), ID 45(50.0 ku), and ID 46(40.5 ku) in RO, ID 98(42.0 ku) in SP, and ID 05(29.0 ku) in KP. A total of 14 protein spots from 82 differentially expressed proteins were identified with LC-MS/MS. Further protein identification was conducted using the SwissProt and NCBInr databases. The identified proteins and their putative functions were discussed further. This was the first study reporting the comparison of petal protein profiles of soybean florets using proteomics tools.

野生大豆(Glycine soja)YD63和栽培大豆(G. max)ZD19茎秆解剖结构比较

以野生大豆YD63和栽培大豆ZD19为研究对象,通过组织化学方法观察茎秆解剖结构的差异,分析和阐述这些解剖结构与功能和环境适应性间的关系,旨在为大豆抗逆性研究提供解剖学依据。结果表明:1.野生大豆表皮毛和腺毛多于栽培大豆,且角质层厚度、表皮厚度和表皮比例均大于栽培大豆,表皮和外皮层细胞的木质化和木栓化程度也高于栽培大豆;2.野生大豆皮层、韧皮部、木薄壁组织和髓的比例均大于栽培大豆,茎秆机械强度降低,可塑性升高,抗逆性增强;3.栽培大豆木质部、木纤维和总纤维比例均大于野生大豆,并且表皮细胞壁、韧皮纤维壁、木纤维壁和导管壁厚度均大于野生大豆。栽培大豆组织木质化的比例大于野生大豆,茎秆的机械强度升高,可以更好地维持直立生长和形态构建;4.栽培大豆微管形成层的细胞层数和厚度均大于野生大豆。栽培大豆木质部的比例大于韧皮部的比例,而野生大豆两者比例基本相同;5.野生大豆韧皮部厚壁组织几乎是连续分布,仅在髓射线处中断,而栽培大豆是不连续的,呈片状分布,野生大豆韧皮部厚壁组织的比例大于栽培大豆;6.野生大豆导管壁强度(t/b)2和小导管比例大于栽培大豆,水分运输的安全性较高,但野生大豆木质部的连通性和水分运输的效率低于栽培大豆。

Effects of salinity on activities of H^+-ATPase, H^+-PPase and membrane lipid composition in plasma membrane and tonoplast vesicles isolated from soybean(Glycine max L.) seedlings

The effects of NaCl stress on the H +-ATPase, H +-PPase activity and lipid composition of plasma membrane(PM) and tonoplast(TP) vesicles isolated from roots and leaves of two soybean cultivars(Glycine max L.) differing in salt tolerance(Wenfeng7, salt-tolerant; Union, salt-sensitive) were investigated. When Wenfeng7 was treated with 0.3%(W/V) NaCl for 3 d, the H +-ATPase activities in PM and TP from roots and leaves exhibited a reduction and an enhancement, respectively. The H +-PPase activity in TP from roots also increased. Similar effects were not observed in roots of Union. In addition, the increases of phospholipid content and ratios of phospholipid to galactolipid in PM and TP from roots and leaves of Wenfeng7 may also change membrane permeability and hence affect salt tolerance.

大豆对胞囊线虫(Heterodera glycines Ichinohe)1号和4号生理小种抗性的遗传分析

大豆胞囊线虫(Heterodera glycinesIchinohe)是我国大豆的全国性主要病害之一。1号和4号生理小种是黄淮地区的优势小种。以Essex×ZDD2315、Peking×ZDD2315、PI88788×ZDD2226、Peking×ZDD2226的P1、P2、F1、BC1F2为材料,用主基因+多基因混合遗传模型分析大豆对胞囊线虫1号和4号生理小种抗性的遗传机制。结果表明,ZDD2315、ZDD2226对1号生理小种的抗性受主效基因控制,未发现多基因效应,且与Peking存在相同的抗病基因;抗性遗传表现组合特异性,Essex×ZDD2315组合为3对加性主基因遗传模型,主基因遗传率72.02%,PI88788×ZDD2226组合为2对显性上位主基因遗传模型,主基因遗传率62.33%。对4号生理小种的抗性为主基因+多基因混合遗传模型,Essex×ZDD2315、Peking×ZDD2315、PI88788×ZDD2226等3个组合为3对主基因+多基因遗传模型,主基因遗传率分别为67.76%7、2.46%和53.25%,多基因遗传率分别为24.48%、21.31%和35.77%;Peking×ZDD2226表现为2对主基因遗传模型,主基因遗传率45.40%。抗性基因表现为隐性,育种上可以在早代选择。培育多抗品种应以抗4号生理小种为主要目标进行基因聚合。

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.

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.

外源茉莉酸对大豆中异戊烯黄酮Glyceollins及其前体Glycinol累积的影响

以中黄13号大豆种子为材料,取生长8天的健康子叶,在其表面挖出约2×2mm的伤口,将50μl不同浓度的茉莉酸溶液和1mM的硝酸银溶液滴在伤口处,黑暗中(25℃)保温不同时间后取样,经甲醇提取,用高压液相色谱(HPLC)测定其中异戊烯化异黄酮glyceollins及其前体glycinol 的累积情况.结果显示,健康子叶中没有glyceollins和glycinol的累积,1mM硝酸银比茉莉酸具有更强的诱导glyceollins和glycinol累积的能力,glyceollins的累积与茉莉酸浓度呈负相关.在各种处理中,glyceollinⅠ、Ⅱ、Ⅲ所占比例分别为:50%～80%、10%～40%和10%～20%,但是glyceollin三种异构体的比例随保温时间的延长而有所变化,glyceollin Ⅰ比例逐渐下降,glyceollin Ⅱ所占比例逐渐升高,在1mM硝酸银处理的子叶中表现更为明显.

大豆中Glycinol的分离与鉴定

Glycinol是大豆中主要的植物抗毒素和植物雌激素glyceollins的直接前体,是研究大豆中异戊烯转移酶必需的重要底物。该研究以新鲜大豆为原料,采用硝酸银诱导大豆子叶使glycinol在其中累积,再用甲醇提取,经制备高效液相色谱分离获得单一成分;经液相色谱/四极杆-飞行时间串联质谱及紫外光谱鉴定,并与标准品对照,确定所获得的单一成分为glycinol。采用该分离方法可从390g鲜重的大豆中分离得到100mg高纯度的glycinol。该研究提供了一种分离纯化和较大量制备glycinol的方法。

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.

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。

Science Letters:Optimization of soybean (Glycine max (L.) Merrill) in planta ovary transformation using a linear minimal gus gene cassette

Soybean transformation by ovary-drip was improved by optimizing the length of the transformation pathway by cutting the styles. These modifications facilitated soybean transformation manipulation and improved transformation reproducibility and efficiency. Using a linear minimal gus gene cassette as the foreign DNA, a maximum transformation frequency of 11% was obtained in flowers of the soybean cultivar ‘Liaodou 14’ with their styles mostly removed, whereas removal of only the stigma, partial style cutting and partial ovary cutting gave transformation frequencies of 0%, 1%, and 2%, respectively. An average transformation frequency of 8.2% was obtained when 619 flowers from three soybean cultivars (‘Liaodou 14’, ‘Liaodou 13’, and ‘Tiefeng 29’) were transformed by this optimized method. Southern blotting analysis showed that the gus reporter gene (encoding β-glucuronidase) was stably inherited with a simple pattern. Reverse transcription-polymerase chain reaction (RT-PCR) and GUS staining confirmed the expression of the gus gene in transgenic plants.

多环境下野生大豆染色体片段代换系群体农艺性状相关QTL/片段的鉴定

【目的】改进染色体片段代换系群体,挖掘野生大豆(Glycine soja Sieb.et Zucc.)中蕴藏的农艺性状优异等位变异,为拓宽栽培大豆(Glycine max(L.)Merr.)的遗传基础提供材料和依据。【方法】通过标记加密和剔除部分单标记型片段的方法,改进以野生大豆N24852为供体,栽培大豆NN1138-2为受体的染色体片段代换系(CSSL)群体Soja CSSLP1;对改进后的群体(Soja CSSLP2)进行3年2点田间试验,通过单标记分析、区间作图、完备复合区间作图和基于混合线性模型的复合区间作图等4种定位方法,结合与轮回亲本有显著差异的染色体片段代换系间相互比对,检测与大豆开花期、株高、主茎节数、单株荚数、百粒重和单株粒重相关的野生片段。【结果】改进后的群体(Soja CSSLP2)由150个CSSL构成,其中,有130个家系与Soja CSSLP1相同;在原遗传图谱上,新增40个SSR标记,相邻标记间平均遗传距离由16.15 c M变为12.91 c M,大于20 c M的区段由32个减少至17个,标记覆盖遗传距离总长度较原图谱(2 063.04 c M)增加103.52 c M;群体NN1138-2背景回复率变幅为79.45%—99.70%,平均为94.62%。利用Soja CSSLP2群体,分别鉴定到与开花期、株高、主茎节数、单株荚数、百粒重和单株粒重相关的4、5、5、7、14和3个工作QTL(working QTL)/片段,其中有15个工作QTL/片段能在多个环境下检测到,属共性工作QTL(joint working QTL);除片段Sct_190—Sat_293上的主茎节数位点外,野生等位变异具有的加性效应方向与双亲表型差异方向一致;单个位点分别能解释5%—64%的表型变异;同时,分别检测到3、2和2个与地点存在互作的株高、主茎节数和单株荚数QTL/片段,其中与凤阳环境的互作均具有增加表型的效应,这可能与凤阳较南京所处纬度高有关;这些位点/片段分布在26个染色体片段上,其中有7个片段与2个及以上性状相关,可能是性状相关的遗传基础;与前人结果比较,有3个开花期、3个株高、2个主茎节数、2个单株荚数、8个百粒重、2个单株粒重位点能在其他遗传背景栽培大豆中检测到,说明在这些位点上野生大豆和栽培大豆间及栽培大豆间均存在遗传差异;另外18个位点(片段)为本研究利用野生大豆的新发现。【结论】大豆开花期、株高和主茎节数的遗传基础较百粒重简单,前者均存在效应较大位点/片段,后者多由小效应位点控制,遗传基础极为复杂;野生大豆中蕴藏着新的等位变异,能拓宽栽培大豆遗传基础。

快速检测大豆籽粒中十二种异黄酮组分的HPLC方法

【目的】利用高效液相色谱(HPLC)技术快速检测大豆异黄酮组分,为大豆的异黄酮标记辅助育种提供重要依据。【方法】以大豆异黄酮12种组分为标准样品,利用HPLC技术结合吸收波峰鉴定的方法进行异黄酮组分的分析。【结果】不同异黄酮苷元组分黄豆苷元(daidzein)、大豆黄素(glycitein)和染料木素(genistein)分别具有不同的最大紫外光谱吸收波长,分别位于250、257和260nm左右。采用YMC-C18反相色谱柱,以含0.1%(v/v)乙酸13%～30%(v/v)的乙腈为流动相,在35℃柱温条件下,经梯度洗脱,结合紫外吸收检测,可以准确地定性和定量鉴定出12种不同异黄酮组分。【结论】该方法样品用量少,检测异黄酮含量准确、快速,适合大量样本分析。