大豆7S球蛋白α-亚基缺失型种质创新

采用常规杂交育种方法,人工去雄、授粉,以10份综合农艺性状优良的黑龙江省主栽品种(或优良品系)为母本、亚基组成为(α'+α+11S酸性亚基)-缺失型育种材料日B为父本进行有性杂交。将F1杂交种南繁,单粒点播、单株收获得到F2代分离群体。聚丙烯酰胺凝胶电泳(SDS-PAGE)法分析表明,F1代杂交种子7S球蛋白α'-与α-亚基的表现型全部为正常型。在杂交F2代种子中获得了具有中国大豆遗传背景的α-缺失、(α+A1aA1bA2)-缺失、A3-缺失、(α'+A4)-缺失和(α'+α)-缺失的贮藏蛋白亚基组成新类型种质,为我国大豆蛋白组分育种选择提供了重要的中间材料。     

具有中国大豆遗传背景的7S与11S多亚基缺失型大豆新品系的创制

利用我国高油大豆品种东农47与日本引进多亚基缺失型育种材料日B,采用回交、三交的育种方法,综合系谱选择,通过SDS-PAGE技术分析亚基组成,在BC₁、BC₃及三交种F₈群体内,选育到(α+11S groupⅡa)-缺失型、(α′+11S groupⅡa)-缺失型、[(α′+α)+11S groupⅡa、Ⅱb]-缺失型、[(α′+α)+11S groupⅡa]-缺失型、[(α′+α)+11S groupⅡb+X₁X₂]-缺失型、[(α′+α)+11S groupⅡb]-缺失型和(α′+11S groupⅠ、Ⅱa)-缺失型共7种具有中国大豆遗传背景的7S球蛋白α′、α亚基与11S球蛋白groupⅠ(A_1aB_1b,A₂B_1a,A_1bB₂)、groupⅡa(A₄A₅B₃)和groupⅡb(A₃B₄)不同亚基缺失组合新种质。测定优良品系的综合农艺性状及氨基酸组成、含量,结果表明,与对照相比,各种缺失突变体的各种氨基酸组分含量普遍提高,蛋白总量普遍高于轮回亲本,精氨酸含量特别是游离精氨酸的含量大幅提高。其中亚基组成为(α+11S groupⅡa)-缺失型品系G2-2-3的17种氨基酸含量、氨基酸总量、蛋氨酸含量均显著高于轮回亲本东农47,特别是游离精氨酸含量高出7.27mg/g。以上结果表明,7S与11S多亚基缺失型优良品系在有效去除致敏蛋白的同时,可以提高大豆蛋白氨基酸含量, 改善大豆蛋白氨基酸组分配比。各种致敏蛋白缺失型大豆优良新品系的获得,大大丰富了我国蛋白质组分改良育种的种质基础。     

大豆7S球蛋白（α＋β）-亚基双缺失体遗传特征分析

采用聚丙烯酰胺凝胶电泳法（SDS-PAGE）对大豆7S球蛋白（α＋β）-亚基双缺失体后代的亚基组成表现进行分析,为明晰该缺失体的遗传规律提供理论依据。结果筛选到β-低含量型、（α-缺失＋β-低含量）型、β-缺失型、（α＋11S group Ⅰ＋llSA4A5B3）-缺失型和（α′＋α）-亚基缺失型5种新的7S球蛋白亚基缺失变异类型。首次在（α＋β）-亚基缺失型材料系统内检测到（α-缺失＋-低含量）型和β-缺失型重组个体,说明该系统内α-亚基缺失与β-亚基缺失特性的独立遗传是可能的。     

小麦高分子量谷蛋白亚基缺失品质效应研究进展

高分子量谷蛋白亚基（high-molecular-weight glutenin subunits,HMW-GS）是小麦籽粒贮藏蛋白的重要组成成分,其数量和质量与品质密切相关。聚合优质HMW-GS可改良强筋小麦制品面包品质,相反HMW-GS缺失可能在弱筋小麦制品饼干、糕点或其他特色食品品质改良上具有重要应用价值。本文从小麦HMW-GS缺失的类型和机制、贮藏蛋白组分和蛋白体发育、面粉和面团品质效应以及食品加工品质改良等方面进行了综述,分析了当前小麦HMW-GS缺失研究利用中存在的问题,并对未来研究方向进行了展望。     

大豆子粒蛋白亚基变异种质的鉴定与筛选

本研究以大豆优良品种中品661的298份甲基磺酸乙酯（EMS）诱变株系和610份大豆品种（系）为试验材料,利用聚丙烯酰胺凝胶电泳（SDS-PAGE）技术,分离大豆种子蛋白并计算各蛋白亚基的相对含量及11S/7S值。研究结果表明,突变群体与自然群体中不同亚基的变异系数差异极显著,但均以β亚基变异范围最大。另外,突变群体的α、β和酸性蛋白亚基的变异范围均大于自然群体。相关性分析显示：11S球蛋白与7S球蛋白含量呈极显著负相关;11S/7S值与蛋白含量相关性不显著,11S/7S值与11S和7S球蛋白各组成亚基均呈显著相关。此外,本研究还筛选出亚基明显变异材料6份,其中Ax亚基突变体尚未见报道,11S/7S值大于3的材料10份,蛋白含量大于48%的材料7份。本研究鉴定和筛选的种子蛋白变异种质为大豆品质相关基因发掘和品种改良提供了材料基础。     

部分山东小麦高分子量谷蛋白1Bx14亚基特异PCR标记分析

在1Bx14亚基的基因序列的基础上,选取不同位点设计特异引物,从中筛选出了一对引物,对HMW-GS在Glu-Bx1位点已知的10个小麦品种进行了PCR扩增。结果表明:凡具有1Bx14亚基的4个品种都能扩增出一条1256bp左右的特异带,而不具该亚基的品种则未能扩增出这一特异带。用这一特异标记对山东省种植面积较大的40个品种进行PCR检测,发现仅有5个品种携带1Bx14亚基。该标记可用于检测小麦品种在这一位点的亚基组成,与SDS-PAGE相比,可显著提高检测的准确性和效率。     

栽培大豆蛋白亚基11S/7S组成及过敏蛋白缺失分析

以175份中国大豆品种资源为材料,利用SDS-PAGE电泳和Western杂交技术,分析蛋白亚基11S和7S的含量,以及Gly m Bd 28K和 Gly m Bd 30K两种过敏蛋白的缺失情况。结果表明,参试品种的11S和7S的含量呈极显著负相关（r＝－0.95）,11S/7S比值范围为0.77～4.67,并鉴定出1份自然缺失β亚基的材料。在175个品种中没有检测到缺失30     

大豆立枯丝核菌G蛋白β亚基基因的克隆与分析

由立枯丝核菌[Rhizoctonia solani Kuhn,有性世代:Thanatephorus cucumeris(Frank)Donk]引起的大豆纹枯病(soybean sharp eyespot)是一种重要病害.G蛋白β亚基(guanine nucleotide binding protein beta-subunit)作为重要的信号传导因子,在植物病原菌致病分子机制中起着重要作用.为了解G蛋白β亚基基因的结构与功能,根据同源物种G蛋白β亚基相关序列设计引物,利用PCR和RT-PCR技术克隆了大豆立枯丝核菌G蛋白β亚基的基因序列和开放阅读框(G-protein beta-subunit of soybean Rhizoctonia solani,简写为gbsrs1,GenBank登录号为EU663628).该片段全长1 864 bp,含有4个内含子和5个外显子;开放阅读框(ORF)长1 047 bp,编码348氨基酸残基,与多种真菌G蛋白β亚基的氨基酸序列相似程度较高,达79.72%-99-43%;该蛋白质具有2个α-螺旋和7个β-折叠的二级结构,形成无规则卷曲连接的桶形三级结构.将gbsrs1的ORF连接于原核融合表达载体pGEX-4T-2中,经IPTG诱导,获得了相应蛋白的表达.gbsrs1的克隆和特性研究为了解大豆立枯丝核菌的致病机理、有效防治纹枯病奠定了基础.     

甘肃小麦部分种质高分子量麦谷蛋白亚基(HMWGS)遗传变异分析

应用十二烷基磺酸钠——聚丙烯酰胺凝胶电泳（SDS-PAGE）方法,分析了372份甘肃小麦种质的高分子量麦谷蛋白亚基（high molecular weight glutenin subunit,HMWGS）组成,以了解甘肃省小麦种质的遗传变异。研究表明,供试的甘肃省小麦种质可检测到20种HMW亚基变异类型和36种HMW亚基组合类型。地方品种的HMW亚基组合类型较少,而且以Null、7＋8和2＋12模式为主,育成品种的HMW亚基变异类型比地方品种丰富,以1、7＋8和2＋12三种变异类型的出现频率为最高。供试材料HMW-GS的品质评分在4～12分之间,平均得分为7.0,所筛选出的一批含优质亚基组合的种质可供育种工作进一步利用。     

山西省不同时期小麦品种麦谷蛋白亚基组成分析

为深入了解山西省不同时期大面积推广品种的HMW—GS组成情况,为该区品质育种提供依据,利用SDS-PAGE电泳技术对152份山西省不同时期大面积推广品种HMW—GS组成、变异及出现频率进行了分析,并计算了参试品种的品质得分。结果表明,历年来主栽品种在Glu-Al位点具有3种亚其类型（NULL,1,2＋）,Glu—B1位,具有4种亚基类型（7＋8,7＋9,14＋15,17＋18）,Glu-D1位,具有2种亚基类型（2＋12,5＋10）。总体而言,参试小麦品种的HMW—GS构成欠佳,优质亚基组合类型单一,2＋、17＋18、5＋10、14＋15亚基的频率很低。具有较高品质得分的亚基组合“1,7＋8,5＋10”、“1,14＋15,5＋10”和“2＊,17＋18,5＋10”的品种较少。     

大豆蛋白质有关性状的QTL定位

以科丰1号×南农1138-2组合衍生的184个重组自交家系(简称RIKY)和(Essex×ZDD2315)×ZDD2315衍生的114个BC₁F₂家系(简称BIEX)为材料,对蛋白质含量、蛋油总量与油脂含量,11S、7S、11S/7S,11S-1~11S-4, 7S-1~7S-6等4组16个性状利用WinQTL Cartographer Ver.2.5的复合区间作图法(CIM)、多区间作图法(MIM)和IciMapping Ver.2.0的完备区间作图法(ICIM)进行QTL分析, 结果表明：(1) 在RIKY和BIEX群体分别定位到17⁺个和21⁺个QTL,合计38⁺个QTL;在RIKY有蛋白、油脂、蛋油总量QTL11个,在11S和7S亚基组上分别只有1⁺和3⁺个;在BIEX有前性状QTL2⁺个,有后性状QTL分别9⁺和6⁺个;(2) 两群体16个性状上均没有检测到共享的QTL,说明两群体的蛋白质有关性状具有完全不同的遗传基础;RIKY的两个亲本间蛋白、油脂和蛋油总量有明显遗传差异,但在亚基组上遗传差异不大,而BIEX则反之;(3) 4组总、分性状中,两群体一致表现出蛋白、油脂和蛋油总量和11S、7S和11S/7S比值两组在总、分性状间共享QTL(共同遗传基础), 而11S亚基组和7S亚基组两组性状在总、分性状间无共享的QTL;(4) 蛋白质有关性状QTL定位结果和分离分析结果共同表明这类性状主效基因和微效基因均占较大比重,要考虑两者兼用的育种方法。     

Effect of Salinity Stress on Growth and Nutrient Composition of Three Soybean (Glycine max L. Merrill) Cultivars

Soil salinity is a major limitation to legume production in many areas of the world. The salinity sensitivity of soybean was studied to determine the effect of salinity on seed germination, shoot and root dry weights, and leaf mineral contents. Three soybean cultivars, Lee, Coquitt, and Clark 63, were planted in soils of different salinity levels. The electrical conductivity (EC) of the soils used in this experiment was 0.5 dS m^?1. The soil salinity treatments were 0.5, 2.5 4.5, 6.5 and 8.5 dS m^?1. Saline drainage water from a drainage canal with an EC of 15 dS m^?1 was used to treat the soil samples in order to obtain the desired salinity levels. Germination percentages were recorded 10 days after planting. Shoot and root dry weights of 45‐day‐old plants were measured. Nutrient concentrations for Na⁺, K⁺, Ca²⁺, Mg²⁺ and Cl^? were determined. Germination percentages were significantly reduced with increasing salinity levels. The cultivar Lee was less affected by salinity stress than Coquitt and Clark 63. At 8.5 dS m^?1 a significant reduction in plant height was found in all three cultivars. However, Lee plants were taller than plants of the other two cultivars. Salinity stress induced a significant increase in leaf sodium (Na⁺) and chloride (Cl^?) in all cultivars. However, the cultivar Lee maintained lower Na⁺ and Cl⁺ concentrations, a higher potassium (K⁺) concentration and a higher K⁺/Na⁺ ratio at higher salinity levels than Coquitt and Clark 63. Saline stress reduced the accumulation of K⁺, calcium (Ca²⁺) and magnesium (Mg²⁺) in the leaves of the cultivars studied. This study suggests that Lee is the most tolerant cultivar, and that there is a relationship between the salt tolerance of the cultivar and macronutrient accumulation in the leaves.     

基因枪转化大豆胚尖草胺膦抗性浓度的确定

为了探讨基因枪法遗传转化时草胺膦的适宜浓度,以‘荷豆12’成熟种子胚尖为材料,将经6-BA诱导过的大豆胚尖转移至含不同草胺磷浓度的培养基上,观察生长及芽伸长情况。结果表明,随着草胺磷浓度增加,大豆胚尖死亡率增加,丛生芽伸长受抑制,再生能力减弱。添加草胺膦的浓度高于0.5 mg/L时,生长受严重抑制甚至死亡,移至基本培养基上也不能再生;在低于0.5 mg/L时,不能有效抑制其生长。因此,0.5 mg/L可作为基因枪法转化外源基因时草胺膦的筛选标准。     

大豆蛋白质有关性状遗传的分离分析

采用科丰1号×南农1138-2衍生的重组自交家系群体(RIKY)和(Essex×兴县灰布支)×兴县灰布支回交自交衍生群体(BIEX)为材料,应用SDS-PAGE电泳测定11S、7S、11S/7S比值及其10个亚基组的相对含量,用凯氏(Kjeltec)自动定氮仪测定蛋白质含量、自动索氏(Soxtec)抽提仪测定油脂含量。结果表明: (1) 两群体蛋白质组分及其亚基组有关性状以及油脂与蛋油总量等均有不同程度超亲分离,双亲间存在不同程度的位点互补。(2) 蛋白质含量遗传,两群体均为1对主基因+多基因模型,主基因和多基因遗传率分别为31.3%~40.9%和37.2%~53.7%。蛋油总量均为3对主基因+多基因模型,主、多基因遗传率分别为59.9%~66.6%和23.2%~27.9%。油脂含量为2~3对主基因+多基因模型,两类遗传率分别为48.6%~71.7%和4.2%~29.7%。(3) 11S组分均为3对主基因+多基因模型,两类遗传率分别为14.3%~60.7%和17.0%~50.7%。7S组分均为3对主基因+多基因模型,两类遗传率分别为34.5%~44.1%和21.5%~45.1%。11S/7S比值遗传均为2对主基因+多基因模型,两类遗传率分别为56.6%~74.8%和10.1%~20.1%。(4) 11S的4个亚基组依次分别为2~3对主基因+多基因、2对主基因+多基因、2对主基因+多基因、2~3对主基因+多基因模型。(5) 7S的6个亚基组依次分别为1~2对主基因+多基因、2对主基因+多基因、2~3对主基因+多基因、3对主基因+多基因、1~2对主基因+多基因和2对主基因+多基因模型。所有16个性状都由主基因和多基因控制,其中有10个性状两群体具相同的主基因数,其他6个性状两群体间相差1对主基因,两群体间遗传模型大同小异。这些相关性状的育种既要利用主基因还必须利用微效多基因,要考虑兼用两者的育种方法。     

Transgenic fertile plants of soybean [Glycine max (L.) Merrill] obtained from bombarded embryogenic tissue

This work describes the production of transgenic, fertile plants of soybean [Glycine max (L.) Merrill]. The transformation method combines the advantages of somatic embryo genesis with the efficiency of particle bombardment of tissues that have a great capacity for in vitro proliferation and regeneration. The results described here represent the first report of transformation of soybean cultivars recommended for commercial growing in South Brazil using somatic embryogenesis, and may open the field for the improvement of this crop in this country by genetic engineering. This revised version was published online in August 2006 with corrections to the Cover Date.     

Insect-mediated cross-pollination in soybean [Glycine max (L.) Merrill]: II. Phenotypic recurrent selection

Recurrent selection is a method for population improvement which has been used in soybean [Glycine max (L.) Merrill] to modify traits such as grain yield, seed-protein content, seed-oil content, tolerance to iron-deficiency chlorosis, and seed size. Nuclear male-sterility with insect-mediated cross-pollination has been successfully used in recurrent selection schemes in soybean. However, little attention has been given to selection to increase the agronomic performance of male-sterile plants per se. The objective of this study was to evaluate the response of male-sterile lines segregating for male-sterile alleles ms2, ms3, ms6, ms8, and ms9 to phenotypic recurrent selection for increased seed-set after 3 cycles, using a selected group of male parents. Bees halictidae, anthophoridae, andrenidae, and megachilidae were utilized as the pollinator vector. The results indicated that recurrent selection in a favorable environment was successful to increase the number of seeds per male-sterile plant. Although a differential response was observed among populations, the seed-set observed would justify the use of some specific male-sterile selections as female parents in a hybrid soybean seed production system.     

Growth and Development of Soybean (Glycine max [L.] Merr.) Cultivars under Shade in Coconut Garden

The growth and development of seven soybean cultivars were studied under shade in a coconut garden. Higher shoot height, internodal elongation and lower leaf area index were the most significant growth changes noticed under shade. Leaf net photosynthesis, crop growth rate and seed yield were also reduced under shade. The cultivars CO 1, UGM 30 and UGM 37 recorded higher yield under shade when compared to other cultivars tested.     

Interaction Amongst Soybean [Glycine max (L.) Merrill] Genotype, Soil Type and Inoculant Strain with Regard to N2 Fixation

The nitrogen (N₂)‐fixing bacterial inoculant strain for soybean [Glycine max (L.) Merrill] is not indigenous to South African soils. The interaction between soybean genotype, soil type and inoculant strain, however, has a definite influence on soybean production and compatibility should be optimized. This paper reports a growth chamber study using three different soybean genotypes (Barc‐9, Avuturda and Talana), three Bradyrhizobium japonicum inoculant strains (WB108, WB112 and WB1) and three soil types (Avalon, Arcadia and sand) to evaluate the effectiveness of N₂ fixation by different genotype × soil type × inoculant strain combinations, using different measuring parameters. These parameters included nodule fresh mass (NFM), amount of N₂ fixed (P_fix), as determined by the ureide method, seed protein content (SPC), average seed mass per plant (SMP) and average foliar N content (FNC). The comparison amongst the three‐way interactions, genotype × soil type × inoculant strain, did not differ significantly for the parameters used. Significant two‐way interactions were soil × inoculant for FNC, P_fix and SMP; soil × genotype for FNC and SMP, and inoculant × genotype for FNC (P < 0.05). The soil × inoculant strain interaction was significant for P_fix (P < 0.05). NFM, P_fix, FNC, SMP and SPC correlated positively with soil pH and negatively with soil clay content and soil NO₃^– and NH₄⁺ content (P < 0.05). SPC was significantly different (P < 0.05) for soil type, genotype and inoculant strain. P_fix and NFM did not reflect the protein content of the seeds, indicating that nodule evaluation should be used with caution as a N₂ fixation parameter. Low soil pH and high mineral N content inhibited N₂ fixation. NFM correlated negatively with the clay content of the soil. This finding confirms that soybean production in South Africa can be improved by appropriate selection of genotypes and inoculant strains for their compatibility in different soils.     

河南栽培大豆的RAPD品种鉴定和聚类分析

用CTAB法提取了10个大豆品种总DNA,使用RAPD技术对其进行了品种鉴定和聚类分析。从73条随机引物中筛选出12条扩增出较稳定DNA条带的引物扩增这些品种总DNA,共扩增出67条带,大小0．2-5 kb,每种引物扩增出4-9条带;多态性条带共51条,多态性比例为76．12％。利用SPSS11．0软件所做的统计分析和聚类分析结果表明,10个品种间的相似系数在0．528-0．776之间,平均相似系数为0．641 6;可聚成3类:豫豆24号、周豆11号、周豆 12号、豫豆11号、周豆13号、豫豆6号,可聚为A类;中作98-3和豫豆15号可聚为B类;豫豆26号和豫豆22号聚为 C类。同一类品种间体现了一定的相关性。