大豆脂肪酸主要组分含量QTL定位

以中黄13×中黄20的100个BC₂F₂家系为作图群体,构建了一张包含131个SSR分子标记的遗传连锁图谱,图谱总长为2157.3 cM,平均遗传距离为16.5 cM,涵盖了大豆的20个连锁群。利用气相色谱技术测定BC₂F₂、BC₂F₃和BC₂F₄回交群体的脂肪酸主要组分含量,采用IciMapping 3.3完备区间作图法定位QTL,共检测到5种脂肪酸组分相关的QTL 26个,与棕榈酸、硬脂酸、油酸、亚油酸和亚麻酸相关的QTL分别为5、5、7、5和4个;3个区间在不同年份被检测到与同一脂肪酸组分相关,sat_294~satt228连续3年被检测到与棕榈酸含量相关,sat_253~satt323和sat_292~satt397连续2年被检测到与油酸含量相关;4个区间被检测到与2种脂肪酸组分相关,其中sat_294~satt228与棕榈酸、油酸相关,satt308~sat_422与硬脂酸、亚油酸相关,sat_292~satt397与油酸、亚油酸相关,satt374~satt269与油酸、亚麻酸相关。     

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。     

大豆苗期耐淹性的遗传与QTL分析

洪涝灾害是大豆生产的主要逆境之一,培育耐涝品种是抗灾保收的重要措施。大豆耐涝性育种方案的设计必须以耐涝性遗传为前提。以苏88-M21(淹水不敏感)×新沂小黑豆(淹水敏感)衍生的175个重组自交系(NJRISX)为材料,在盆栽V2期土壤表层保持5~7 cm水层20 d的淹水条件下,研究大豆苗期耐淹性的遗传和QTL定位。通过对8个耐淹性有关性状的相关分析和主成份分析,确定以处理前后株高变化量、处理终叶龄和成熟期株高3个性状的平均耐淹指数为评价指标。NJRISX家系间耐淹性差异极显著,存在超亲分离。主基因+多基因分离分析表明该群体的耐淹性为2对连锁主基因+多基因遗传,主基因遗传率为62.83%,多基因的遗传率为8.90%。WinQTLCart2.5复合区间及多区间QTL定位分析均检测到2个QTL,位于连锁群L2上的satt229~satt527和satt527~satt286区间,对表型的解释率分别为11.76%~25.20%和10.10%~12.34%。大豆NJRISX群体苗期耐淹性遗传分离分析与QTL定位结果相对一致。     

黑龙江省大豆籽粒异黄酮含量生态差异

大豆异黄酮由于其特有的生理保健功能而受到日益广泛的关注。为了明确黑龙江省大豆籽粒异黄酮含量的生态差异规律, 2005选定5个生态条件差异明显的试验点及4个大豆品种进行试验。结果表明, 年份、地点、基因型及基因型×环境互作对大豆籽粒异黄酮总含量及3种酸解处理后所得苷元的含量具有显著效应。2006年的大豆异黄酮总含量及3种苷元组分含量的平均值显著高于其他2年;大豆籽粒异黄酮含量不同基因型间及不同地点间均差异显著,大豆籽粒异黄酮总含量及其3种酸解后所得苷元含量与纬度呈极显著正相关。据此认为,大豆籽粒异黄酮含量在黑龙江省存在优势生产区, 对大豆籽粒异黄酮含量进行品质区划是可行的。     

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。     

六种大豆异黄酮溶液的稳定性

用制备液相色谱仪(Shimadzu LC-20AP)制备了大豆苷、黄豆黄苷、染料木苷、大豆素、黄豆黄素、染料木素6种大豆异黄酮单体溶液,对其进行了1个月的短期稳定性和6个月的长期稳定性研究,特性量值为溶液的浓度值和纯度值。对在4个不同温度(–20℃、4℃、25℃、60℃)下保存一个月后的测定值与初始值进行了方差分析,短期稳定性试验结果表明,除染料木苷外,其余5种色谱纯大豆异黄酮溶液的浓度在4℃、25℃条件下保存无显著变化,6种大豆异黄酮溶液的纯度在–20℃、4℃、25℃条件下保存无显著变化,因此认为4℃、25℃是大豆异黄酮溶液短期储存的最适温度。在4℃条件下,对经过0、1、2、3、4、6、8、12、16和24周的大豆异黄酮测定值与初始值进行t检验,长期稳定性结果表明,6个月的保存期内6种色谱纯大豆异黄酮溶液的纯度无显著性变化,但浓度均显著增加,分析认为分装容器的密封性是导致浓度增加的原因。本研究结果为大豆异黄酮单体溶液标准溶液产业化提供了技术支撑。     

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[Glycine max（L.）]成熟期近等基因系导入片段分析

选择243个多态性SSR标记, 分析轮回亲本Harosoy及23个携带大豆4个不同成熟期基因的近等基因系(near isogenic line, NIL), 共检测出导入片段266个, 平均每个NIL有导入片段11.6个。其中由携带E1基因的NIL检测出导入片段150个, 主要集中在第6号染色体;由携带E2、e3和E5基因的NIL各检测出导入片段55个、49个和73个, 分别集中在第20号、第12号和第20号染色体, 根据NIL的SSR分析结果聚类,具有相同熟期基因的NILs趋向聚在一起。通过对导入片段进行分析, 推测E1基因与第6号染色体的satt643~sat_312区间及第11号染色体的sat_095位点相关, E2和E5基因与第20号染色体的satt587~satt496区间相关, e3与第12号染色体的satt317~satt181区间相关。结果表明, 利用近等基因系不仅验证了已知E1基因所在的染色体区间, 发现了一个新的标记位点与E1相关, 还鉴定出与E2、e3和E5基因相关的标记, 明确了轮回亲本中成熟期基因所在导入片段大小及其位置, 为成熟期基因的精细定位和克隆提供了信息。     

HPLC法同时测定安徽产野葛、粉葛中6种异黄酮成分

以安徽葛根主产区收集的6种样品(太湖野葛、岳西野葛、金寨野葛、宣城野葛、六安粉葛、亳州葛根)为试材,建立高效液相色谱法同时测定安徽野葛、粉葛中葛根素、大豆苷、染料木苷、大豆素、染料木素、刺芒柄花素6种异黄酮类成分的含量,样品前处理采用30%乙醇超声波提取,采用Phenomenex Column luna C18(2)100A(250 mm×4.6 mm,5μm)色谱柱,以甲醇(A)-水(B)为流动相,梯度洗脱,流速为1 m L/min,柱温35℃,检测波长为250 nm。标准品在范围内呈良好线性关系,相关系数R2分别为0.999 3、0.999 4、0.999 4、0.997 8、0.997 4、0.998 6,最低检出限分别为0.562 7、0.453 4、0.283 0、0.234 0、0.256 0、0.261 0μg/m L,精密度、稳定性和重现性良好,平均加标回收率分别为100.5%、99.3%、104.2%、98.6%、97.1%、104.5%。安徽产区的野葛与粉葛中葛根素等6种异黄酮含量差别显著。该方法准确、快速,操作简单,分离效果好,可用于野葛、粉葛中葛根素、大豆苷、染料木苷、大豆素、染料木素、刺芒柄花素6种异黄酮含量的同时测定。     

大豆脂肪及脂肪酸组分含量的QTL定位

脂肪及脂肪酸组分的改良是大豆油脂品质育种的主要方面。本研究旨在构建遗传图谱,定位大豆脂肪及脂肪酸组分的QTL,为大豆油脂品质育种提供参考。以Essex×ZDD2315的114个BC₁F₁单株为作图群体,构建了250个SSR标记和1个形态标记,具有25个连锁群的遗传图谱,覆盖大豆基因组2 963.5 cM,平均每个连锁群上10.0个标记,标记平均间距11.8 cM。用BC₁F₃家系3个重复的表型平均值代表相对应的BC₁F₁单株表型值,采用Win QTL Cartographer 2.5复合区间作图法（CIM）检测到18个控制脂肪及脂肪酸组分含量的QTL,位于9个不同的连锁群上,表型贡献率为9.6%~34.5%;多区间作图法（MIM）检测到与CIM区间相同的7个QTL（fat-1, pal-1, st-1, ole-1, lin-1, lin-4和lio-2）,区间相近的2个QTL（ole-4和lin-5）,位于6个不同的连锁群上,表型贡献率为8.2%~39.3%。CIM法检测到的其他9个QTL有待进一步验证。大豆脂肪及脂肪酸组分含量的主效QTL数量不多,效应大的不多,可能还受许多未能检测出来的微效基因控制,育种中既要注意主效QTL的利用,又要考虑微效多基因的积聚。     

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.     

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.     

Genistein and Daidzein Concentrations and Contents in Seedling Roots of Three Soybean Cultivars Grown under Three Root Zone Temperatures

Daidzein and genistein are plant-to-bacterium signal compounds involved in soybean nodule formation. They can induce nod gens expression in Bradyrhizobium japonicum. The objective of this study was to determine whether the production of signal molecules was affected by low root zone temperatures (RZTs) in a manner that varied among soybean cultivars. Daidzein and genistein concentrations of soybean seedling roots were measured at three RZTs by high performance liquid chromatography (HPLC). The results indicated that daidzein content and concentration per plant were higher at 15 and 17.5°C than those at 25°C. AC Bravor had higher daidzein contents and concentrations than did Maple Glen and KG20. At 17.5°C. KG20 had higher genistein content and concentration levels than Maple Glen, and no difference existed for the two cultivars at 15 and 25 C. Daidzein contents and concentrations of Maple Glen and AC Bravor increased with harvest time. However, for cultivar KG20, the content and concentration decreased at 19 days after inoculation. Genistein contents and concentrations of the three cultivars increased under each RZT up to the last harvest. There was an interaction between soybean cultivar and RZT for root genistein and daidzein contents and concentrations. The content and concentration of daidzein in soybean seedling roots were much higher (more than five times) than those of genistein.     

Parental effects on nutritional and antioxidants constituents in seeds of peanut cv. Boreom 1

In spite of the importance of parental inheritance effects, many aspects remain inadequately explained. The objective of this study was to explore parental effects on their progeny by estimating the variability of nutritional content in peanut seeds. The peanut cv. Boreom 1 was derived from Suwon 108 (female) and Milyang 4 (male). Protein and amino acid analyses revealed that male and female parent influences on protein, Asp, Thr, Ser, Glu, Pro, Ala, Cys, Val, Met, Ile, Leu, Tyr, Phe, His, and Arg content in their progeny. The oil and fatty acid composition (C16:0, C18:0, C18:1, C18:2, C20:0, C20:1, and C22:0) of the progeny were controlled by both parental types. The synergistic interaction of female and male parental genes was also shown on sucrose and stachiyose content, while paternal effect was found on raffinose in their progeny. The antioxidant activity, daidzin, glycitin, genistin, mal-glycitin, maldaidzin, mal-genistin, and daidzein content of Boreom 1 were significantly correlated with those secondary metabolites of female parent. Our results documented that both parental genes differentially influenced on nutritional composition of their progeny, Boreom 1. This biochemical evidence of parental effects would be support to identify the suitable cultivars to improve the crop breeding.     

大豆光合气体交换参数的QTL分析

光合气体交换参数是用来表示植物光合能力的常用指标。利用来自大豆品种科丰1号和南农1138-2的重组自交系群体NJRIKY(184个家系)及其分子遗传图谱,通过两年盆栽试验定位与光合速率、气孔导度、胞间CO₂浓度和蒸腾速率有关的QTL。结果表明,4个参数的遗传力中等偏低,在0.48~0.60之间;两两间存在极显著正相关关系,相关系数在0.192~0.686之间;两年共检测到15个QTL,分别位于C1、C2、D2、E、H、I和O连锁群上,LOD值在2.25~6.31之间,贡献率为4.80%~12.30%,;有6个QTL在不同环境下稳定表达,它们分别是控制光合速率的qPnC1.1、控制气孔导度的qSCD2.1和qSCI.1、控制胞间CO₂浓度的qCiI.1和qCiO.1,以及控制蒸腾速率的qTrO.1;检测到4个同时控制两个或两个以上参数的标记区间,它们分别是C1连锁群上控制光合速率和气孔导度的sat_311~sct_191区间,E连锁群上控制光合速率、气孔导度和胞间CO₂浓度的sat_172~satt268区间,I连锁群上控制气孔导度和胞间CO₂浓度的satt726~satt330区间,以及D2连锁群上控制气孔导度和胞间CO₂浓度的sat_296~sat_277区间。     

大豆叶片性状和叶绿素含量QTL间的上位性和环境互作效应

以丰产性好、抗旱力强的栽培大豆晋豆23为母本,山西农家品种半野生大豆灰布支黑豆为父本杂交衍生的447个RIL作为供试群体。将亲本及447个家系分别于2011、2012和2013年采用随机试验种植,按照标准测量叶长、叶宽和叶柄长3个性状,并于2012年8月1日和8月8日和2013年8月2日和8月9日各测量1次叶绿素含量。采用QTLNETwork 2.0混合线性模型分析方法和主基因+多基因混合遗传分离分析法,对大豆叶片性状和叶绿素含量进行遗传分析和QTL间的上位性和环境互作效应研究。结果表明,叶长受2对加性-加性×加性上位性混合主基因控制,叶宽受3对等效主基因控制,叶柄长受4对加性-加性×加性上位性主基因控制,叶绿素含量受4对加性主基因控制;检测到10个与叶长、叶宽、叶柄长和叶绿素含量相关的QTL,分别位于A1、A2、C2、H_1、L和O染色体。其中2个叶长QTL分别位于C2和L染色体,是2对加性×加性上位互作效应及环境互作效应QTL;3个叶宽加性与环境互作QTL分别位于A2、C2和O染色体;2个叶柄长QTL分别位于L和O染色体;3个叶绿素含量QTL分别位于A1、C2和H_1染色体。叶片性状和叶绿素含量的遗传机制较复杂,加性效应、加性×加性上位互作效应及环境互作效应是大豆叶片性状和叶绿素含量的重要遗传基础。建议大豆分子标记辅助育种中,一方面要考虑起主要作用的QTL,另一方面要注重上位性QTL的影响,这对于性状的遗传和稳定表达具有积极的意义。