中国野生和栽培大豆11S及7S蛋白质相对含量的比较分析

大豆种质蛋白质11S和7S及其亚基组相对含量的遗传变异是专用型品种选育的基础.以全国各生态区的野生豆138份和地方品种409份,国内育成品种148份、国外育成品种83份,合计778份大豆种质为材料.采用SDS-PAGE电泳技术测定蛋白质11S和7S组分及其亚基组相对含量,研究其遗传变异.在南京同一条件下的结果表明:全国野生豆、地方品种和育成品种11S相对含量平均分别为54.7%、64.8%和71.7%,变幅28.8%～82.6%、38.8%～79.4%和48.2%～88.9%;7S相对含量平均分别为44.7%、34.9%和27.9%,变幅20.6%～71.2%、20.6%～61.1%和15.7%～47.8%;11S/7S比值平均分别为1.4、2.0和2.7,变幅0.4～3.9、0.6～3.9和0.9～4.0.野生豆驯化为栽培豆并经选育后11S相对含量和11S/7S比值上升,7S相对含量下降,变幅均减小;亚基组11S-2和11S-3相对含量增加;7S的6个亚基组,尤其7S-1和7S-6,相对含量下降.11S、7S、11S/7S以及各亚基组在各群体各生态区内均有较大变异,但与来源地纬度、蛋白质和油脂含量均无显著相关.从中优选到11S/7S比值大于3.7、11S相对含量为78.9%～88.9%的8份种质,发现有11S的4个亚基组相对含量分别大于37%、7S的6个亚基组相对含量分别大于24%、以及11S-1和7S的6个亚基组缺失的种质,这些特异种质可供蛋白质组分育种利用.     

小麦种质资源宁春4号的研究和利用

小麦品种宁春4号不仅是中国的一个优良春小麦品种,也是一个优异的种质资源.它含有来自美洲、欧洲、亚洲、澳洲优良小麦品种的遗传物质,含有Rht2等基因,具有1、17+18、5+10优质HMW-GS组合,以及Glu-A3b、Glu-B3f优质LMW-GS组合,属于非1BL/1RS易位系小麦品种.与其他宁夏春小麦品种相比,宁春4号在不同生育期功能叶片的叶绿素含量、缓降期内的平均叶绿素含量均最高;光饱和点最小;从孕穗期至开花期旗叶光合速率日平均值、最高值最大.宁春4号与部分小麦近缘属种杂交表现出较高的可杂交性,利用其做亲本与其他小麦杂交已选育出一批优良品种.由于长期坚持对宁春4号小麦品种提纯复壮,使其在生产上28年不衰退.宁春4号是一种兼性类型的小麦品种,现已筛选出冬性宁春4号,品质有较大的改善.     

Chilling Stress Accelerates Degradation of Seed Storage Protein and Photosynthetic Protein during Cotton Seed Germination

Low temperature is one of the major abiotic stresses limiting the productivity and geographical distribution of many important crops. To gain a better understanding of chilling stress responses in cotton ( Gossypium hirsutum L . ), we carried out a comparative proteomic analysis. Cotyledon proteins of 1-week-old cotton seedlings treated with or without chilling treatment at 4 °C were extracted, separated by a two-dimensional gel electrophoresis and compared. Among 1500 protein spots reproducibly detected on each gel, 25 protein spots were down-regulated and 29 were up-regulated. Seven cotton proteins were identified by mass spectrometry analysis as beta-globulin A precursor fragments. One was identified as a Rubisco large subunit fragment, providing evidence of both seed storage protein and photosynthetic protein destruction by chilling stress. The related physiological and biochemical analysis showed that there was a significant increase in endopeptidase activity and activated oxygen generation rate, and an obvious decrease in carboxylation efficiency and maximum net photosynthetic rate of cotton seedlings under chilling stress. Based on the above results, the degradation mechanisms of beta-globulin and Rubisco under chilling stress were discussed. In conclusion, our study provides new insights into chilling-stress responses in cotton and demonstrates the advantages of proteomic analysis.     

硬粒小麦Glu-B3基因家族新成员的克隆及其分子标记的开发

获得小麦低分子量谷蛋白基因家族新成员, 为深入研究该基因家族的组成及其在染色体上的分布与进化奠定基础。本研究根据已发表的小麦低分子量麦谷蛋白基因序列设计了1对简并引物, 以从硬粒小麦品种Langdon BAC文库中筛选的携带小麦低分子量谷蛋白基因的BAC为模板, 利用同源序列克隆技术克隆了1个MET类型的小麦低分子量谷蛋白基因LMWLDN-M, 该基因编码350个氨基酸。利用LMWLDN-M与本实验室克隆的Langdon品种其他类型的小麦低分子量谷蛋白基因之间的SNP, 设计了该类型基因特异的引物。以Langdon代换系和中国春小麦缺体-四体为模板进行PCR扩增, 将该基因定位到小麦B基因组上。序列分析表明, LMWLDN-M为Glu-B3基因家族的一个新成员。     

Development of two multiplex PCR assays targeting improvement of bread-making and noodle qualities in common wheat

Wheat quality properties are genetically determined by the compositions of high and low molecular weight glutenin subunits, grain hardness, polyphenol oxidase (PPO) activity and starch viscosity. Two multiplex PCR assays were developed and validated using 70 cultivars and advanced lines from Chinese autumn‐sown wheat regions. Multiplex PCR I includes molecular markers for genes/loci ω‐secalin, Glu‐B1‐2a (By8), Glu‐D1‐1d (Dx5), Glu‐A3d, Glu‐B3 (for non‐1B·1R type) and Pinb‐D1b targeting improved gluten parameters and pan bread quality. Multiplex PCR II comprises markers for genes/loci Ppo‐A1, Ppo‐D1 and Wx‐B1b targeting improved noodle quality. The results were consistent with those achieved by SDS‐PAGE and RP‐HPLC, indicating that the two multiplex assays were highly effective, with good repeatability and low costs enabling their use in wheat breeding programmes. In total, nine alleles (subunits) at locus Glu‐B1, four at Glu‐D1 and five at Glu‐A3 locus were identified, and the alleles (subunits) Glu‐B1b (7 + 8), Glu‐B1c (7 + 9), Glu‐D1a (2 + 12), Glu‐D1d (5 + 10), Glu‐A3a, Glu‐A3c and Glu‐A3d were most frequently present in the cultivars and lines tested. The 1B·1R translocation was present in 28 (40.0%) lines, whereas the Wx‐B1 null allele for better noodle quality was present in only seven (10.0%) cultivars and advanced lines, and 37 (52.9%) lines had Pinb‐D1b associated with hard grains. The allele Ppo‐A1b on chromosome 2AL associated with lower PPO activity was present in 38 (54.3%) genotypes, whereas the less effective allele Ppo‐D1a on chromosome 2DL, also associated with low PPO activity was present in 45 (64.3%) of genotypes. These two multiplex PCR assays should be effective in marker assisted selection targeting improved pan bread‐making and noodle qualities.     

棉蚜ATP合成酶基因AgoATPb的克隆与表达

为确定棉蚜(Aphis gossypii)ATP合成酶B亚基基因在棉蚜不同组织和日龄,以及取食不同植物的表达情况,以棉蚜为研究对象,采用RT-PCR和RACE技术获得AgoATPb的全长cDNA序列,通过Expasy、Sig-nalP-4.0 Server等在线工具对其进行了生物信息学分析,同时利用实时荧光定量PCR技...     

香蕉RuBPCase小亚基基因全长cDNA的克隆和分析

利用RACE和RT-PCR技术,从香蕉中克隆出编码RuBPCase小亚基的全长cDNA,并与GenBank中已经报道的其他部分香蕉bcS基因的核苷酸和推导的氨基酸序列进行同源性分析,为进一步研究香蕉RUBPCase 的功能与结构提供依据。     

小麦高分子量麦谷蛋白亚基与品质性状的关系

对2002年国家黄淮南片区试参试小麦品系的蛋白质、湿面筋、沉降值和稳定时间等品质性状进行了测定,并运用SDS-PAGE分析了参试品系的高分子量(HMW)麦谷蛋白亚基组成及其与品质性状的关系。结果表明,在26个参试品系中,仅内乡188、郑农16达到强筋小麦标准;5+10亚基频率有所提高(53.8%),但具5+10亚基的品种间品质具有高度不稳定性,在品质改良中仅靠转育5+10亚基是不够的,要注重亚基组合的选育,其中以1,7+8,5+10和1,7+9,5+10亚基组合对品质的贡献最大,以14+15,5+10亚基组合对品质的贡献最小。     

椰麸组分蛋白的氨基酸组成、结构与乳化性分析

为提高椰子加工副产物的利用,本研究从椰麸中依次提取4种组分蛋白,包括清蛋白、球蛋白、谷蛋白-1和谷蛋白-2.分析这4种组分蛋白的氨基酸组成,并利用SDS-PAGE技术分析其亚基组成和分子量.同时分析其表面疏水性,并研究不同离子强度对椰麸各组分蛋白的乳化性和乳化稳定性的影响.结果表明,椰麸蛋白质中清单白、球蛋白、谷蛋白-...     

江苏省小麦品种的谷蛋白亚基组成分析

为了解江苏省小麦品种的谷蛋白亚基组成现状,为品质育种提供理论指导,选用77份来自江苏省淮南和淮北主要小麦育种单位的育成品种或高代品系,利用聚丙烯酰胺凝胶电泳法(SDS-PAGE)对Glu-1和Glu-3位点进行了鉴定。结果表明,参试品种(系)Glu-1和Glu-3位点的高分子量麦谷蛋白亚基(HMW-GS)和低分子量麦谷蛋白亚基(LMW-GS)变异丰富。共检测到12个HMW-GS等位变异,其中Glu-A1位点有3个,Glu-B1位点有5个,Glu-D1位点有4个。LMW-GS的Glu-A3和Glu-B3位点则分别检测到5个和6个等位变异。淮北小麦的HMW-GS多态性高于淮南小麦。Glu-1和Glu-3位点等位变异的分布频率差异较大,且淮北和淮南表现不同,优质亚基比例较低。Glu-1位点主要亚基类型为0(46.8%)、1(49.4%)、7+8(63.6%)和2+12(63.6%),Glu-A3位点主要亚基类型为a(23.4%)、c(57.1%)和d(15.6%),Glu-B3位点主要亚基类型为b(22.1%)、f(49.4%)和j(1B/1R易位)(16.9%)。淮北小麦的高分子量谷蛋白优质亚基及亚基组合较少,Glu-B3j分布频率高(54.5%),亚基质量有待提高;淮南小麦的优质亚基及组成亦较少,需根据中筋和弱筋小麦育种目标并结合蛋白质含量和亚基选择进行改良。