磷素对不同大豆品种籽粒异黄酮含量的影响

为探索磷素对不同大豆品种籽粒异黄酮含量的影响,寻找不同基因型大豆品种最佳施磷水平,以期提高大豆籽粒异黄酮的含量,改善其品质。选用‘黑农48’（高蛋白品种）、‘黑农37’（中间型品种）、‘黑农44’（高油品种）3个大豆品种作为试验材料。采用盆栽,在每kg土壤施N和K2O各为0.033 g基础上,设P1、P2、P3、P4 4个P处理（即每kg土壤分别施P2O5 0、0.033、0.067、0.100 g）。采用紫外分光光度法测定不同大豆品种籽粒总异黄酮的含量。结果表明：同一品种P2处理大豆总异黄酮含量显著高于P1、P3、P4处理,‘黑农37’、‘黑农44’、‘黑农48’3个品种大豆P2处理大豆总异黄酮含量分别比对照组增加3.7%、4.3%、3.8%;不同品种同一处理都是‘黑农44’总异黄酮含量最高;在12个处理组合中‘黑农44’P2处理总异黄酮含量最高,3个大豆品种异黄酮含量在品种间和施磷处理间差异显著。施磷对3个大豆品种异黄酮含量有影响,适宜的施磷量有利于提高大豆籽粒异黄酮的含量。     

蚜虫取食大豆诱导大豆异黄酮变化的规律

采用高效液相色谱分析方法研究蚜虫取食大豆诱导大豆异黄酮含量和组分的变化规律。结果表明,蚜虫取食处理不仅能诱导虫害叶内异黄酮含量的显著变化,而且能诱导同株未被害叶内异黄酮含量发生类似的变化,说明蚜虫取食诱导大豆叶片植株合成3种异黄酮苷元组分及总异黄酮的反应是一种系统诱导抗性;接蚜数量不同的大豆植株间,异黄酮含量差异显著,而且在一定范围的受害程度内,大豆叶片内异黄酮含量基本一致。蚜虫取食对大豆叶片异黄酮的诱导反应与光照无关,只与植株是否被害有关。     

大豆异黄酮含量相关SSR分子标记的研究

大豆异黄酮是大豆生长过程中形成的一类次级代谢产物,是一种重要的生物活性物质。本研究以异黄酮含量差异显著的‘中豆27’(4 447 ug/g)和‘台湾75’(2 062 ug/g)为亲本,构建了包括160个家系F2:6高代重组自交系群体。采用高效液相色谱方法测定双亲及群体各单株的不同异黄酮组分的含量。异黄酮不同组别中,大豆苷类含量最高,黄豆黄素苷类含量最低。异黄酮不同存在形式中,丙二酰基葡萄糖苷型含量最多,苷元型含量最少。使用197对在双亲间表现出多态性的SSR引物对RIL群体进行扩增。而后使用SAS程序的Proc GLM命令进行异黄酮含量和标记间的方差分析,在p0.01显著水平上共检测到包括21个SSR标记的41个显著性互作。其中部分标记同时与多个不同的异黄酮成分相关;4个SSR标记与总异黄酮含量相关。本研究为高异黄酮含量育种的分子标记辅助选择奠定了基础。     

大豆异黄酮及其组分含量的遗传分析与QTL检测

以栽培大豆晋豆23为母本,以山西农家品种大豆灰布支黑豆为父本杂交衍生的447个RIL作为供试群体构建遗传图谱,利用高效液相色谱法定性、定量测定样品中的异黄酮及其组分含量。采用主基因+多基因混合遗传分离分析法和Win QTLCart 2.5复合区间作图法,对大豆异黄酮及其组分含量进行混合遗传分析和QTL定位。结果表明,大豆苷、黄豆苷元、染料木素、染料木苷、大豆苷元和异黄酮总含量分别受4、4、2、3、2和2对主基因控制,并有多基因修饰。检测到44个与大豆异黄酮及其组分含量相关的QTL,与大豆苷、染料木素、黄豆苷元、大豆苷元、染料木苷和异黄酮总含量相关的QTL分别有10、9、4、7、8和6个。连续2年分别检测到与大豆苷、染料木苷、黄豆苷元和异黄酮关联,分别位于标记区间satt430~satt359、satt038~satt570、satt197~sat_128和satt249~satt285的稳定表达QTL,可尝试用于分子标记辅助育种。     

硫素对不同基因型大豆脂肪酸含量的影响

为了提高大豆脂肪酸含量,改善大豆品质,通过施用硫素,揭示其对不同基因型大豆脂肪酸含量的影响,寻找3个基因型品种最佳施硫水平。选用‘黑农37’(HN37)、‘黑农44’(HN44)‘黑农48’(HN48)3个大豆品种作为试验材料,采用盆栽,每个品种设S1、S2、S3、S4 4个硫处理（即每千克土壤施用单质硫0、0.02、0.04、0.06 g）。采用高效气相色谱法,对成熟大豆籽粒脂肪酸的含量进行测定。结果表明：3个基因型大豆籽粒中脂肪酸含量均有所增加。同一处理不同品种NH44含量增加较为明显,同一品种不同处理S3条件下各脂肪酸含量增加较为明显。并且在S3条件下,HN37和HN48籽粒中出现了微量的豆蔻酸;HN44籽粒中亚油酸和硬脂酸2种脂肪酸含量均远高于其他品种;HN48籽粒中油酸的含量最高;HN44、HN48和HN37三个基因型大豆籽粒中α-亚麻酸和棕榈酸含量差异不明显。施硫对3个大豆品种脂肪酸含量有影响,适宜的施硫量有利于提高大豆脂肪酸的含量。     

异黄酮含量与大豆对大豆蚜虫抗性之间的关系

利用高效液相色谱分析方法测定了10个抗感大豆品种蚜虫取食处理叶片和茎的异黄酮含量。结果表明,蚜虫取食诱导抗虫品种叶片大豆苷、染料木苷和异黄酮总含量增加,而感蚜品种异黄酮含量无明显变化。相关分析表明,大豆品种受害程度与叶片大豆苷、染料木苷和异黄酮总含量呈负相关。但茎的异黄酮含量与大豆品种的抗蚜性无明显相关。     

大豆异黄酮组分HPLC快速分析技术及其在豆腐加工中的应用

大豆异黄酮育种与大豆制品加工研究需要进行大批量样品12种异黄酮组分的快速定量分析。在前人研究基础上利用Agilent 1100高效液相色谱(HPLC)系统,以大豆品种南农95C-13为材料,研究大豆苷元,大豆苷,乙酰基大豆苷,丙二酰基大豆苷,染料木苷元,染料木苷,乙酰基染料木苷,丙二酰基染料木苷,黄豆苷元,黄豆苷,乙酰基黄豆苷,丙二酰基黄豆苷等12种标准品外标法快速定量技术。从样品制备与色谱条件对分析12种异黄酮组分的准确度和分离度入手,确定分析流程,以(科丰1号×南农1138-2)的184个重组自交系(NJRIKY)为材料,研究豆腐加工中总量和各组分的变化特点。(1)样品以80%甲醇水溶液50℃超声1 h提取;色谱条件为,检测波长254 nm, 柱温36℃,流速2.0 mL min^–1, 进样量 10 μL, 流动相0.1%(V/V)乙酸水溶液(A)和100%甲醇(B),0~2 min,27% B (V/V)→2~3 min,27%~38% B→3~10 min, 38% B→10~12 min,38%~39% B→12~14 min, 39% B→14~15 min, 39~27% B梯度洗脱;在15 min内将12个组分良好分离,各组分峰面积与其相应浓度均呈良好线性关系(R²为0.9976~0.9999);加标回收率均大于99%,变异系数低于2%。(2)NJRIKY群体籽粒、豆乳、豆腐异黄酮总量和组分的大量分析验证了HPLC快速技术的效果。籽粒异黄酮总含量(3 695.00 μg g^–1)在豆乳加工中平均85.15%转入豆乳 (3 146.12 μg g^–1),14.85% (548.88 μg g^–1)进入豆渣。传统豆腐加工通过硫酸钙絮凝,只有17.32% (639.89 μg g^–1)转入豆腐,67.83% (2 506.23 μg g^–1)留在黄浆水中。豆乳中12种组分含量比籽粒稍低,均以丙酰基染料木苷含量最高;而豆腐中乙酰基染料木苷和乙酰基黄豆苷缺失,以染料木苷元与大豆苷元含量较高。大豆科丰1号与南农1138-2杂交重组后家系间的异黄酮遗传变异增大,增加了对其遗传改良的潜力。     

硫素对不同基因型大豆球蛋白组成及含量的影响

为了探讨施硫量对不同基因型大豆7S和11S球蛋白亚基组成及含量的影响,寻找不同基因型大豆品种最佳施硫水平,以期提高大豆籽粒球蛋白的含量,改善期品质。选用在黑龙江省种植面积较大的‘黑农48’（高蛋白）、‘黑农37’（中间型）和‘黑农44’（高油）3种大豆作为试验材料。采用盆栽种植,每个品种设S1、S2、S3、S4 4个S处理（即每千克土壤分别施单质硫0、0.02、0.04、0.06 g）。采用SDS-PAGE凝胶电泳方法,对成熟大豆籽粒球蛋白亚基组成和含量2个指标进行研究。结果表明：在3个大豆品种中,7S球蛋白由α′、α、γ、β 4个亚基组成;11S球蛋白由酸性亚基和碱性亚基组成;各种亚基分子量在品种间的差异不显著。同一品种不同处理中均以S3水平下蛋白含量最高,对不同品种施用硫肥时,以高油品种提高球蛋白含量效果显著。施用硫肥对3个大豆品种的7S和11S球蛋白各种亚基组成都没有影响,对分子量的大小影响很小。施用硫肥对3个大豆品种中的7S和11S球蛋白和亚基含量有显著影响,适宜的施用硫肥有利于提高各品种球蛋白和亚基的含量。     

硫素对不同大豆品种膳食纤维含量的影响

硫是生物体重要组成成分,与大豆膳食纤维代谢密切相关,确定最佳施硫量,对改善大豆品质,提高大豆营养价值具有重要意义。为研究不同施硫量对不同基因型大豆膳食纤维含量的影响,以黑龙江省3个主栽大豆品种,‘黑农37’（HN37,中间型品种）、‘黑农44’（HN44,高油品种）、‘黑农48’（HN48,高蛋白品种）为试验材料,采用盆栽,每个品种设4个处理组（即每Kg土壤分别施单质硫0、0.02、0.04、0.06,即S1、S2、S3、S4）。采用酶-重量法测定不同大豆品种籽粒总膳食纤维的含量。结果表明：同一品种不同处理大豆总膳食纤维含量, HN48和HN37在S3处理含量最高,HN44在S2处理含量最高;同一处理不同品种间大豆总膳食纤维含量,在4个施硫条件下均表现出HN48含量最高,其次是HN37和HN44;在不同品种不同处理中,大豆总膳食纤维含量峰值出现在HN37的S3水平下;施硫对3个大豆品种膳食纤维含量有影响,合理适量的施硫可有效提高大豆籽粒膳食纤维的含量。     

小麦腺苷二磷酸葡萄糖焦磷酸化酶同工酶基因型与酶活性及淀粉含量的关系

采用非变性聚丙烯酰胺凝胶电泳鉴定了我国60个代表性小麦品种的腺苷二磷酸葡萄糖焦磷酸化酶(AGP)同工酶基因型, 并测定了AGP活性及总淀粉含量, 以明确小麦籽粒AGP同工酶基因型组成及其与AGP活性和淀粉含量的关系。结果表明, AGP有AGPa、AGPb、AGPc和AGPd 4个等位基因位点, 其出现频率分别为96.7%、80.0%、86.7%和16.7%; 共检测到5种基因型, 其中基因型AGPabc出现频率最高, 为46.7%。不同基因型的品种间AGP活性和总淀粉含量差异显著(P<0.05)。其中具有基因型AGPabcd的品种酶活性及总淀粉含量最高, 表明小麦籽粒中不同AGP同工酶基因型对酶活性及淀粉含量有不同遗传效应。     

环境因素对大豆异黄酮含量的影响研究进展

影响大豆异黄酮含量的因素主要有遗传因素（基因）和环境因素。异黄酮是受主-微效基因共同控制的;不同地理位置、不同栽培贮藏条件、不同品种、不同年份、不同籽粒的特征特性等,其异黄酮含量也有差异;另外野生大豆异黄酮的含量,因分布区域的不同而有变化。     

Genetic parameters relating isoflavone and protein content in soybean seeds

Isoflavones are a class of compounds present in high amounts in soybean seeds, which can be used for prevention and treatment of several chronic diseases. Proteins present in soybean seeds are the basis for the high nutritional value and versatility of this leguminous species in animal and human feeding. The main goals of this work were to estimate heritabilities for isoflavone contents in soybean seeds and the correlation between isoflavone and protein contents. Commercial variety IAC-100 (high isoflavone and normal protein contents) and the line BARC-8 (low isoflavone and high protein contents) were crossed, and one single F₁ plant derived 97 F₂ seeds, which were used to obtain F₃ seeds. A sample of F₃ seeds from each F₂ plant was used for isoflavone determination by HPLC and protein by the Kjeldahl method. Six isoflavone forms were detected: daidzin, genistin, glycitin, malonyldaidzin, malonylgenistin and malonylglycitin. Total isoflavone contents ranged from 427.92 to 965.89 μg per gram of dry seed and the protein content ranged from 45.17 to 34.95% in BARC-8 and IAC-100, respectively. Our results indicate that it is possible to select for high isoflavone content in early breeding generations because the broad sense heritabilities for the contents of the various isoflavone forms were higher than 90%. In addition, high correlation values among the contents of the individual isoflavone forms were observed (between 0.80 and 0.98). However, negative correlation values were obtained between isoflavone and protein contents, ranging from −0.51 to −0.37 for the different isoflavone forms. The correlation value of −0.47 between total isoflavone and protein contents confirmed the negative correlation between these two parameters, as reported by other authors. This revised version was published online in August 2006 with corrections to the Cover Date.     

Dynamic quantitative trait loci underlies isoflavone accumulation in soybean seed

Most of quantitative trait loci (QTL) underlying soybean seed isoflavone contents were derived from the harvest stage of plant development, which uncover the genetic effects that were expressed in earlier seed developmental stages. The aim of this study was to detect conditional QTL associated with isoflavone accumulation during the entire seed development. A total of 112 recombinant inbred lines developed from the cross between ‘Zhongdou 27’ (higher seed isoflavone content) and ‘Jiunong 20’ (lower seed isoflavone content) were used for the conditional QTL analysis of daidzein (DZ), genistein (GT), glycitein (GC) and total isoflavone (TI) accumulations through composite interval mapping with mixed genetic model. The results indicated that the number and type of QTL and their additive effects for individual and total isoflavone accumulations were different among R3 to R8 developmental stages. Three unconditional QTL and six conditional QTL for DZ, four unconditional QTL and five conditional QTL for GT, six unconditional QTL and five conditional QTL for GC, six unconditional QTL and seven conditional QTL for TI were identified at different developmental stages, respectively. Unconditional and conditional QTL that affect individual and total isoflavone accumulations exhibited multiple expression patterns, implying that some QTL are active for long period and others are transient. Two genomic regions, Satt144‐Satt569 in linkage group F (LG F; chromosome 13, chr 13) for DZ, GC, GT and TI accumulations and Satt540‐Sat_240 in LG M (chr 07) for TI and GC accumulations, were found to significantly affect individual and total isoflavone accumulations in multiple developmental stages, suggesting that the accumulation of soybean seed isoflavones is governed by time‐dependent gene expression.     

Changes in anthocyanin and isoflavone concentrations in black seed-coated soybean at different planting locations

This study assessed the altitudinal variations in the anthocyanin and isoflavone contents of six black seed coated soybean [Glycine max (L.) Merrill] cultivars. The black soybean cultivars Heugcheong, Seonheuk, Geomjeong 1, Geomjeong 2, Cheongja 2, and Cheongja 3 were planted at Milyang (12 m above mean sea level — low altitude) and Muju (600 m — high altitude), Korea on 10 June 2005 and 2006. The total anthocyanin and isoflavone contents and individual components were investigated by reverse-phase high performance liquid chromatography (HPLC). All black soybean cultivars cultivated in high altitude possessed significantly higher total anthocyanin (p < 0.01) and isoflavone (p < 0.01) contents than those grown in low altitude. For anthocyanin composition, cyanidin-3-O-glucoside, cyanidin-3-O-galactoside, and peonidin-3-O-glucoside contents were significantly higher while delphinidin-3-O-glucoside contents was significantly lower at high altitude. The composition of individual isoflavones, 6″-O-malonyldaidzin, and 6″-O-malonylgenistin contents significantly increased at high altitude.     

Inheritance of isoflavone contents in soybean seeds

Summary We studied the genetic basis of isoflavone content inheritance in soybean seeds. The progenitors BARC-8 (low isoflavone content), IAC-100 (high isoflavone content), the F₁ and F₂ populations derived from reciprocal crosses, and backcross populations were analyzed for isoflavone content and composition. Six isoflavones were detected: daidzin (DZ), genistin (GT), glycitin (GC), malonyldaidzin (MDZ), malonylgenistin (MGT) and malonylglycitin (MGC). DZ, GT, MDZ and MGT contents were influenced by the cytoplasm and the nuclear genes of the maternal parent. For this reason, a genetic model was considered that included the cytoplasmic effect and epistasis between nuclear and cytoplasmic genes. Except for GT, the additive effect was the most important one. For GT content the cytoplasmic effect was the most important. Except for MDZ, the epistatic effects were significant for all the isoflavone forms. Our data indicate that genetic improvement for these traits should explore the additive genetic variances in superior lines or the cytoplasmic effect and the epistatic interactions between cytoplasmic and nuclear genes to obtain the largest selection gains.     

R2R3-MYB转录因子GmMYB184调节大豆异黄酮合成

异黄酮是一类主要含在豆科植物中的次生代谢物, 在植物防御体系中发挥重要作用, 并与人类健康密切相关。大豆异黄酮含量受多基因和复杂代谢网络控制, 调控代谢途径上的结构基因不能显著改变大豆异黄酮含量, 与异黄酮代谢途径相关的转录因子的鉴定和应用可能会有效解决这个问题。本研究克隆了一个与大豆异黄酮合成相关的R2R3类型MYB转录因子GmMYB184, 并进行了初步的功能验证。亚细胞定位研究结果表明GmMYB184转录因子定位于细胞核。组织表达分析结果表明该转录因子基因与IFS2 (异黄酮合酶2编码基因)的表达模式相同。同时, GmMYB184和IFS2的表达模式与异黄酮的积累模式相似。谷胱甘肽诱导表达分析表明该转录因子基因与IFS2共同被诱导, 说明这两个基因可能参与同一或相似的生物过程。采用双荧光素酶报告系统分析其对异黄酮合成途径关键基因的转录激活活性影响, 发现GmMYB184能够促进IFS2和CHS8启动子表达活性分别提高5倍和7倍。最后, 通过发根农杆菌介导的遗传转化系统, 找到该转录因子在异黄酮合成调控中的直接作用证据。沉默GmMYB184导致大豆毛状根异黄酮含量的显著下降。但是, 过表达GmMYB184不足以显著提高毛状根中异黄酮的含量。总之, 本研究为大豆异黄酮合成分子机制探索及大豆异黄酮品质改良提供了理论依据。     

Molecular loci associated with seed isoflavone content may underlie resistance to soybean pod borer (Leguminivora glycinivorella)

Soybean pod borer (SPB) (Leguminivora glycinivorella (Mats.) Obraztsov) causes severe loss of soybean (Glycine max L. Merr.) seed yield and quality in some regions of the world, especially in north‐eastern China, Japan and Russia. Isoflavones in soybean seed play a crucial role in plant resistance to diseases and pests. The aim of this study was to find whether SPB resistance QTL are associated with soybean seed isoflavone content. A cross was made between ‘Zhongdou 27’ (higher isoflavone content) and ‘Jiunong 20’ (lower isoflavone content). One hundred and twelve F_5:10 recombinant inbred lines were derived through single‐seed descent. A plastic‐net cabinet was used to cover the plants in early August, and thirty SPB moths per square metre were put in to infest the soybean green pods. The results indicated that the percentage of seeds damaged by SPB was positively correlated with glycitein content (GC), whereas it was negatively correlated with genistein (GT), daidzein (DZ) and total isoflavone content (TI). Four QTL underlying SPB damage to seeds were identified and the phenotypic variation for SPB resistance explained by the four QTL ranged from 2% to 14% on chromosomes Gm7, 10, 13 and 17. Moreover, eleven QTL underlying isoflavone content were identified, and ten of them were encompassed within the same four marker intervals as the SPB QTL (BARC‐Satt208‐Sat292, Satt144‐Sat074, Satt540‐Sat244 and Satt345‐Satt592). These QTL could be useful in marker‐assisted selection for breeding soybean cultivars with both SPB resistance and high seed isoflavone content.     

Quantitative trait loci underlying soybean seed tocopherol content with main additive,epistatic and QTL × environment effects

Soybean (Glycine max [L.] Merrill) seeds are a major source of tocopherols (Toc), which could significantly improve immune system health of human and prevent or treat many serious diseases. Selection for higher Toc contents of seeds could increase nutritional value of soybean‐derived food, laying on an important breeding goal for many soybean breeders. The present objectives of the work were to evaluate various genetic effects of QTL associated with individual and total Toc content based on a RIL population (“Beifeng 9” × “Freeborn”) in six environments to improve the efficiency of molecular marker‐assisted selection (MAS) for high‐Toc breeding. The results described that eighteen, thirteen, eleven and thirteen QTL were associated with α‐Toc, γ‐Toc, δ‐Toc and total Toc content, respectively, and have additive main effects (a) and/or additive × environment interaction effects (ae) in certain environments. Among them, four QTL for α‐Toc, two QTL for γ‐Toc, one QTL for δ‐Toc and four QTL for total Toc could increase α‐Toc, γ‐Toc, δ‐Toc and total Toc content via significant a effect, respectively, which have stronger stability in different years and locations. It implied a value for MAS. Additionally, twenty‐five, fifteen, eleven and twenty epistatic pairwise QTL associated with α‐Toc, γ‐Toc, δ‐Toc and total Toc contents, respectively, were detected. The genetic information of the QTL effects obtained here would be beneficial for breeding soybean variety with high‐Toc content by MAS.     

大豆不同环境下脂肪酸组分含量的QTL分析

大豆油的品质取决于脂肪酸各组分在大豆中的比例, 为发掘控制大豆5种脂肪酸含量的数量性状位点(QTL), 利用冀豆12和黑豆重组自交系群体构建遗传图谱, 采用Windows QTL Cartographer 2.5和QTL Network-2.0软件的CIM和MCIM法对大豆5种脂肪酸组分进行数量性状定位。结果表明,在石家庄和三亚各环境下共检测到16个QTL, 位于连锁群A2、B2、C2、F、G、I、L上。对2个环境联合分析, 检测到13个QTL, 其中9个用2种方法被检测到, 但这13个位点与环境互作的贡献率明显小于加性效应。其中在B2连锁群Satt168~Satt556控制硬脂酸的QTL Ste-1在河北石家庄和海南三亚均能被检测到, 贡献率均为12%, 在双尾群体和间隔挑选群体中也能检测到控制硬脂酸的QTL Ste-1, 说明这一QTL稳定存在于本组合群体中, 为今后大豆硬脂酸的QTL精细定位奠定了基础。