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利用贮藏蛋白PAGE电泳鉴定花生远缘杂种的研究 总被引：1，自引：1，他引：1
以花生属二倍体野生种A.cardenasii为父本,地栽培种铁岭四粒红杂交,获得远缘杂种F1。应用花生种子贮藏蛋白SDS-PAGE电泳技术,对交本,母本及F1进行遗传鉴定,并结合形态性状进行分析。电泳结果表明,F1代出现了父本相似带,母本相似带及父母本都没有的新带,按迁移率计,父母本相似系数为0.50,遗传距离为0.50,杂种F1与父本相似系数为0.59,遗传距离为0.41,与母本相似系数为0.65,遗传距离为0.35,杂种F1的形态性状则表现为显性遗传,共显性遗传及超显性遗传。从杂种F1的形态性状观察,表明花生种子贮藏蛋白SDS-PAGE电泳技术鉴定远缘杂种的真实性是有效的。 相似文献
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为构建Waxy蛋白亚基的快速、高效、高通量毛细管电泳分离鉴定体系,为该亚基理化特性的分析、功能的发掘及含Waxy蛋白亚基种质的快速筛选提供参考依据,以小麦中国春为材料,采用逐一优化毛细管电泳体系参数的方法,系统研究了毛细管电泳中缓冲液各成分以及pH值对中国春中Waxy蛋白亚基出峰效果的影响,初步构建了普通小麦Waxy蛋白亚基的最佳分离体系。结果表明,在检测波长为214 nm、以0.05 mol·L-1硼砂+8% 乙腈+0.8% SDS 溶液（pH 9.5）为缓冲液、分离电压20 kV、温度25 ℃、进样压力0.5 psi×5 s时,在6~9 min处出现峰高约0.015 au的三个主峰,且图形清晰,基线水平,重复性好。本研究构建的毛细管电泳系统分离体系可有效分离普通小麦Waxy蛋白亚基。 相似文献
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为了进一步研究茸毛偃毛草基因向小麦中的导入情况,采用种子贮藏蛋白电泳技术对新创制的小麦-茸毛偃麦草部分双二倍体TE-3的醇溶蛋白和谷蛋白亚基组成进行分析,发现源自茸毛偃麦草的部分醇溶蛋白特征带在双二倍体中得到表达。利用不含1RS/1BL染色体的四川小麦品种对小麦与茸毛偃麦草的杂种F1进行连续回交,并结合染色体逐代观察,现已获得一批变异类型丰富的种质材料Y176,对其中农艺性状优良的部分株系,如Y176-12等进行的蛋白电泳分析,发现了茸毛偃麦草的特异醇溶蛋白带的导入。 相似文献
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SRAP-cDNA方法在植物基因差异表达分析中的应用 总被引：3，自引：0，他引：3
分离差异表达的基因一直是近些年的热点问题，国内外科学家发展了许多技术来研究植物基因的差异表达，如SSH、RAD、SAGE、DDRT-PCR、AFLP-cDNA、cDNA-microarray等。SRAP是近些年发展起来的一种新的分子标记，目前SRAP主要应用于植物基因组DNA水平上的研究，对于RNA即基因表达上的研究还比较少。本文对SRAP技术的原理方法、优点进行了介绍，并阐述了SRAP-cDNA方法的实验条件优化、应用现状及前景，表明SRAP-cDNA方法具有简便、重复性好、实验成本低等优点，可以在研究植物基因差异表达领域广泛应用，并成为作物育种、杂种优势分析等工作的重要工具。 相似文献
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国内外巴西橡胶树替代作物及技术研发现状 总被引：2，自引：0，他引：2
天然橡胶是重要的工业原料和战略物资.近年来其供需日趋紧张,而目前大面积商业化种植生产天然橡胶的巴西橡胶树,只能种植在亚洲和非洲等地的热带地区.为此,美国、欧盟的某些机构相继开展了巴西橡胶树替代作物的研究,并在近期取得了重要进展.这对中国及世界其它国家以巴西橡胶树种植为主的天然橡胶产业构成了潜在威胁.为了对目前国内外非巴西橡胶树生产天然橡胶的研究和开发现状进行较为深入系统地了解,本文对国内外巴西橡胶树替代产胶作物及技术开发利用现状进行概括分析,并建议迅速确立专项开展银胶菊及转基因橡胶草等替代巴西橡胶树产胶作物及技术调研研究. 相似文献
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建立同时测定槟榔壳中6种酚类物质的高效毛细管电泳方法,研究分析不同浓度和不同pH值缓冲液对10种标准品(即:表儿茶素、儿茶素、柚皮素、芦丁、山奈素、阿魏酸、绿原酸、香豆酸、咖啡酸和没食子酸)的分离效果。结果显示,最佳电解缓冲液为0.1 mol/L、pH 9.0硼酸缓冲液,紫外检测波长为280 nm,分离电压为20 kV。该方法简便快速,能在20 min之内将10种酚类物质完全分离开,具有良好的精密度、回收率和线性关系。检测限范围为0.587 5～4.415 2μg/mL。 相似文献
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以精选表型和基因型丰富的29份代表性玉米自交系以及三联体验证材料,基于叶绿体基因组中11个InDel位点进行引物设计、筛选评估,确定2对玉米S型不育鉴定特异引物。两对引物CPMIDP01CE和CPMIDP02CE的多态分别为83个和5个碱基的插入缺失变异,属于trnS-trnG和ndhJ-ndhK基因间区。CPMIDP01CE和CP?MIDP02CE引物的扩增产物如果含有83 bp插入序列(片段长度为339 bp),不含有5 bp的插入序列(片段长度为239 bp),所分析样品即为S型胞质不育材料。两对特异鉴定引物基于变性荧光毛细管、非变性凝胶毛细管、琼脂糖等不同电泳平台均建立了检测方法,并且具有兼容更高效的KASP、TaqMan等技术体系的潜力。 相似文献
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Lab-on-a-chip capillary electrophoresis has been used for identification of wheat variety and quality type. Analysis of each chip takes 30 minutes for 10 samples, and distinction can be made between members of a set of 40 commonly grown Australian wheat varieties. Quality type could be predicted by analysis of the HMW and LMW glutenin subunits. The technique has also been applied to the separation of proteins from other grains and legumes, and may also be useful for identifying variety and/or quality type in these crops. 相似文献
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During the malting process, storage proteins are degraded by proteolytic enzymes into small peptides and amino acids. The activity of these enzymes was measured during malting of oats and was found to be increased. To quantify proteolytic degradation, proteins of unmalted, germinating and malted grains were fractionated. After extracting the oat proteins (Osborne fractionation), protein fractions were analysed using a Lab-on-a-Chip technique, which separates the proteins – based on their molecular weight – by capillary electrophoresis. This new technique for the analysis of proteins was supported by using two-dimensional gel electrophoresis. In addition, amino acid analysis was carried out. In general a degradation of proteins to small peptides and amino acids could be observed in the globulin, prolamin and glutelin fractions. In the albumin fraction a protein increase was observed, which is due to the fact that this fraction contains the majority of the metabolically active proteins. Amino acid analysis supported the observation of increased protein amount in the albumin fraction and decreased protein amounts in the other fractions. Some proteins, which have not been described in the literature, were detected in the albumin and glutelin fraction, since Lab-on-a-Chip technique allows detection of proteins with low molecular weights of 4.5 kDa. 相似文献
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为了给种质资源和育种研究、种子管理及新品种保护提供理论依据,用优质面包小麦中优9507和中优9701的4个杂交组合CA9614/中优9507、CA9614/中优9701、中优16/津麦2号//中优9701和中优9507/CA9640的46份高代品系,研究了毛细管电泳在鉴别亲缘关系很近的小麦品种中的应用潜力及醇溶蛋白组成与加工品质的关系.结果表明,4个杂交组合醇溶蛋白毛细管电泳分离峰的聚类分析结果与其选择历史一致,说明毛细管电泳是小麦品种(系)鉴定的有效方法.来自于同一F3或F4株系的亲缘关系很近的杂交后代在6～8 min或10～12 min的分离峰中存在较大差异,说明毛细管电泳能够用于鉴定亲缘关系很近的品种.品质表现一致的材料其醇溶蛋白组成无明显共性,说明醇溶蛋白组成对小麦蛋白质含量、SDS沉淀值和揉面特性等品质性状影响不大. 相似文献
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《Journal of Cereal Science》1998,27(1):27-36
Gliadin and glutenin are characterized by specific ultrastructures, which depend on variety and separation conditions. Gliadins, extracted with 80% ethanol, of Israeli spring wheat ‘Ariel’ appeared as spherical bodies within an amorphous perforated matrix. The gliadins of a commercial sample of U.S. hard red winter wheat were deposited in bundles of bodies of concentric membrane-like units during water dialysis and they tended to separate while heating at 120 °C. Acetic-acid-extracted glutenins heated at 120 °C appeared as amorphous compact agglomerated particles beside dispersed aggregates, fibril-like patterns and oil-like bodies. The extracted glutenins, which were dialysed vs water, appeared as dispersed or aggregated particles beside oil-like bodies embedded in and/or encapsulated by coagulated protein. Differences were found in high-performance capillary electrophoresis patterns of both gliadins and glutenins between the Israeli spring wheat ‘Ariel’ and the good baking quality U.S. hard red winter wheat ‘Karl 92’; indicating that the structural differences are a result of different proteins and differences in the physicochemical properties of the proteins. 相似文献
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《Journal of Cereal Science》1995,22(2):147-155
The water-insoluble storage proteins of barley seeds reside in the starchy endosperm tissue. This tissue, when expressed from germinating barley, has a pH of 4·8. The hydrolysis of storage proteins during germination (malting) occurs mainly in the endosperm, so proteinases that are located in endosperm and are active at pH 4·8 are probably important to the storage protein hydrolytic process. This study reports our continued investigations of the endoproteinases of germinating barley (Hordeum vulgare L., cv. Morex) with a two-dimensional gel separation method that uses isoelectric focusing (IEF) and non-denaturing polyacrylamide gel electrophoresis (PAGE) in gels containing incorporated substrate protein. We identified the endoproteinases that were active at pH 4·8 and determined when they appeared during germination and where they were located in 4-day germinated barley (green malt). A total of nine cysteine, four aspartic, and two serine class proteolytic activities that were active at pH 4·8 were extracted from the endosperm tissue of green malt. It seems probable that some or all of these endosperm endoproteinases, especially proteinases C7, C8, C11, D3, E3 and E4, are the ones most intimately involved in hydrolyzing the storage proteins during malting. 相似文献
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《Journal of Cereal Science》1995,22(3):211-224
The polypeptide subunits present in SDS-unextractable glutenin, the glutenin macropolymer (GMP) and the 70% (v/v) ethanol unextractable protein, the Osborne glutenin fraction, of various cultivars were separated by RP–HPLC and capillary electrophoresis (CE) under denaturing (urea and SDS, respectively) and reducing conditions. In addition, the SDS-extractable protein was separated by CE. HighMrglutenin subunits were well separated by CE, while the separation of lowMrglutenin subunits was better by RP–HPLC. HighMrglutenin subunits separated by RP–HPLC were collected and separated by CE. The subunits were identified unequivocally using the combined information from these two techniques and from SDS–PAGE patterns using the cvs. Spring and Troy Spring. By both RP–HPLC and CE it could be demonstrated for flour from three wheat cvs. (Camp Remy, Obelisk and Rektor) and a blend of flour from two of those cvs. (Camp Remy/Obelisk) that the highMrglutenin subunit content of the GMP was 29–31%. In contrast, the SDS-extractable protein consisted of 4–6% highMrglutenin subunits, which accounted for 14–23% of the highMrglutenin subunits in flour. Interestingly, the SDS-extractable highMrglutenin subunits consisted mainly (90–96%) of x-type subunits whereas, in the GMP, only 70–75% of the highMrsubunits were x-type subunits. Although the SDS extractable protein was not separated by RP–HPLC, results similar to those obtained by CE could be inferred from the subtraction of the contents of glutenin subunits of the GMP from the contents in the Osborne glutenin fraction. The results suggest that x- and y-type highMrglutenin subunits may have a different role in the structure (size and composition) of glutenin polymers. 相似文献