利用A-PAGE研究谷子籽粒蛋白质多态性

研究谷子贮藏蛋白多态性对谷子品种选育和鉴定评价具有重要的意义。本文利用A-PAGE方法对来自不同生态区的24份谷子品种籽粒的水溶蛋白、盐溶蛋白、酸溶蛋白和醇溶蛋白的多态性进行了研究。结果发现,不同谷子品种之间籽粒水溶蛋白、酸溶蛋白谱带相似,基本没有差异;每个谷子品种的盐溶蛋白谱带有20多条,仅有4个品种出现多态性谱带,多态性也较低。说明谷子品种水溶蛋白、盐溶蛋白和酸溶蛋白A-PAGE谱带不适用于谷子鉴定研究。谷子醇溶蛋白谱带存在一定的异质性,每个品种有7~10条醇溶蛋白谱带,其中5条为公共条带,2~5条为多态性谱带,醇溶蛋白A-PAGE谱带可以作为谷子品种鉴定评价的依据。结果说明,和玉米、小麦等禾谷类作物相比,不同生态区谷子栽培品种蛋白质变异较小,遗传背景变化较小,因此需要不断丰富谷子品种的遗传基础,为谷子品种选育提供更加丰富的材料。     

杂交玉米及亲本种子碱溶蛋白电泳鉴定

用乳酸-聚丙烯酰胺凝胶电泳对玉米不同杂交种及其亲本种子碱溶蛋白进行分析,结果表明：玉米碱溶蛋白在蛋白质组成上存在丰富的多态性,各杂交种和自交系的电泳谱带差异明显,可视为各自的“特征”指纹,通过指纹分析能有效地将各供试品种区分开来。因此,碱溶蛋白可作为玉米品种鉴定的依据。     

利用SDS-PAGE鉴定不同地区谷子籽粒醇溶蛋白差异

醇溶蛋白是谷子籽粒主要的贮藏蛋白,研究谷子醇溶蛋白的差异对于谷子品种选育及种质鉴定评价有重要的意义。本研究利用SDS-PAGE方法对来自不同省份的46份谷子材料进行醇溶蛋白谱带分析,结果表明,单个品种可分离出14~24条醇溶蛋白谱带,平均每个品种18.74条带,46份品种共分离出32条醇溶蛋白谱带,其中7条为共有带,25条为多态性带,多态性带数占分离出总带数的78.1%。根据迁移率大小将醇溶蛋白谱带分为A、B、C、D四个区段,四个区段内分别有6、16、15、4种带型。在B、C区段醇溶蛋白有较丰富的多态性,A、B区段弱带较多,C、D区段强带较多。醇溶蛋白分子量在13.0~55.0 kDa之间,在29,25 kDa(第23条带)、22.5 kDa(第26条带)、17.5 kDa(第28条带)位点附近醇溶蛋白表达丰富且品种之间存在较大差异。品种之间醇溶蛋白谱带存在较大的差异,可作为品种鉴定与评价的重要依据。聚类分析表明,在GD值为0.45时,46个品种可以分为5类,其中34个品种聚为一类,其他四类所包含的材料较少,这表明大部分材料醇溶蛋白相似度较高。     

棉花种子的品种鉴定方法探讨

对棉花三个新品种潞棉11号、潞棉147、潞棉150脱脂处理后.分步提取棉花种子中的水溶、盐溶、醇溶三种主要贮藏蛋白,用十二烷基磺酸钠聚丙烯酰胺凝胶电泳、染色、脱色、拍照,并用德国GDS-9000凝胶成像系统对电泳图谱扫描分析.结果显示,同一品种间蛋白谱带稍有差异,不同品种以不同类型的贮藏蛋白为特征,潞棉11号的水溶蛋白带数多,与潞棉147、潞棉150差异较大;醇溶蛋白在路棉147号样品中电泳带数多,与潞棉11号、潞棉150差异显著;潞棉150盐溶蛋白与潞棉11号、潞棉147相比有自己的特征.采用这样的方法进行种子鉴定较单一蛋白更全面、可信.     

大麦籽粒总蛋白含量及醇溶蛋白含量的差异性研究

为研究大麦籽粒总蛋白含量及醇溶蛋白的差异性。以来源不同的38份大麦品种为材料,采用FOSS凯氏定氮仪测定其籽粒总蛋白和醇溶蛋白含量,利用SDS-PAGE凝胶电泳技术分析参试材料醇溶蛋白带型。对参试材料的籽粒总蛋白含量、醇溶蛋白含量及醇溶蛋白带型的差异性和遗传多样性分析表明:38份大麦品种的总蛋白含量和醇溶蛋白含量的遗传差异显著,醇溶蛋白含量与总蛋白含量呈极显著正相关。根据醇溶蛋白含量将参试材料聚为3类;38个大麦品种SDS-PAGE凝胶电泳共分离出12条迁移率不同的条带,根据电泳谱带将共参试材料聚为构成18种类型,不同类型大麦品种数量差异较大。本研究为适宜蛋白质含量与醇溶蛋白含量大麦品种的选育提供了依据。     

不同种子蛋白在杂交油葵品种纯度定中应用的可行性研究

本文旨在通过对杂交油葵品种不同种子蛋白电泳图谱比较分析,筛选出一种用于杂交油葵品种纯度鉴定的蛋白。研究表明,与其他四种蛋白相比,水溶蛋白提取最为简便,提取效率最高,而且电泳谱带质量高,分离效果好,多态性最为丰富。因此,水溶蛋白电泳技术适宜应用于杂交油葵品种纯度鉴定和亲本纯度鉴定。     

中国北方冬麦区主栽品种醇溶蛋白组成的遗传演变分析

用改良的pH3.2A-PAGE技术分析了我国北方冬麦区建国后不同时期的51个主栽品种和21个骨干亲本的醇溶蛋白组成及其遗传演化规律.结果表明,供试主栽品种具有丰富的醇溶蛋白变异类型,是今后小麦育种的重要物质基础.本实验材料共分离出72种醇溶蛋白组分,供试主栽品种含有其全部组分,其单品种谱带数目在21～41之间,品种间各类蛋白组     

芸豆种子蛋白组分及其在种子萌发过程中的变化

以4个具有明显差异的芸豆品种为材料,采用SDS-PAGE方法研究芸豆种子萌发过程中水溶、盐溶和总蛋白组分变化及种子胚和子叶变化过程中蛋白组分变化.试验结果表明,芸豆种子的子叶、胚含有丰富的蛋白亚基,其中,同一品种子叶和胚贮藏蛋白亚基差异明显,子叶、胚的水溶蛋白、盐溶蛋白和总蛋白相差不大、条带丰富;不同品种之间条带存在差异;在发芽过程中,高分子量蛋白亚基的降解速度快于低分子量蛋白亚基,子叶亚基的降解速度明显慢于胚的亚基降解速度;不同品种间子叶、胚条带差异明显,种子活力高的品种(Y05,Y06)降解速度快于种子活力低的品种(Y09,Y1 1),而蛋白含量高的品种(Y1 1,Y05)与蛋白含量低的品种(Y06,Y09)差异不明显.研究结果为芸豆品种鉴定、品种改良、优质栽培及发芽机理提供理论依据.     

普通小麦醇溶蛋白组份的分布及其与HMW——麦谷蛋白亚基对品质的组合效应

本研究利用SDS-PAGE和改良的APAGE技术分析了1124份普通小麦材料的HMW—麦谷蛋白亚基和麦醇溶蛋白谱带的组成,共分离出18种主要的HMW麦谷蛋白的等位变异和67条不同迁移率的醇溶蛋白潜带。根据多元回归分析,筛选出gli42.3,62.7,39.6(2),11.4,23,Glu 5 10等蛋白谱带对以沉降值为代表的面包烘烤品质起增效作用。品种间品质性状变异的28％～42％可以由两种蛋白组份的差异所决定,而一种蛋白组份只能决定12.3％～31.0％。国内品种中优质谱带少,非优质谱带比例高,这可能是造成我国小麦面包烘烤品质差的原因之一。提出在后代选育中要立足于优中选优,避免非优质谱带组合的出现。按高效谱带做第一次分类效果最好。此外,对这两项生化技术在品质育种中的应用进行了讨论。     

饲用燕麦(Avena sativa L.)品种的种子醇溶蛋白超薄层等电聚焦电泳快速鉴定

利用超薄层等电聚焦电泳（UTLIEF）技术对青藏高原牧区普遍种植的6个饲用燕麦品种及1个国外品种进行种子醇溶蛋白鉴定分析.结果表明,参试品种种子醇溶蛋白UTLIIEF电泳图谱分辨率高,重复性好,可作为燕麦品种鉴定的蛋白质指纹图谱.电泳共分离出34条不同迁移率的谱带,其中85%的谱带具有多态性.根据Rf=0.469～0.781这一区域中不同品种所具有的A带数目,将参试品种分为4组：（1）丹麦444,锋利（具有2条A带）;（2）加拿大,473（3条A带）;（3）林纳,001（4条A带）;（4）甜燕麦（5条A带）.第1组中,444两条A带的迁移率为0.547和0.617,锋利两条A带的迁移率为0.469和0.594;第2组中,加拿大和473的差异仅仅在2条迁移率不同的B带上,加拿大具有Rf=0.594的B带,473具有0.742的B带;第3组中,001比林纳多了1条Rf=0.781的B带.据此,可将7个品种区分开.同时对参试7个品种进行了纯度检验,每个品种设3个重复,每个重复随机取50粒种子,进行单粒种子醇溶蛋白UTLIEF电泳,以此分析品种纯度.以丹麦444和林纳燕麦为例,根据品种纯度公式得到这2个品种的品种纯度,分别为81.3%和100%.其他5个品种的纯度值分别为锋利：96.7%;加拿大：99.3%;473：91.3%;001：93.3%;甜燕麦：99.3%.     

玉米杂交种及其亲本自交系的生化指纹鉴定

本试验以浙单9号等五个玉米杂交组合及其亲本自交系为材料,进行种子盐溶蛋白聚丙烯酰胺凝胶电泳等多种电泳鉴定方法的研究,以揭示玉米杂交种及其亲本自交系的“生化指纹”(biochenucal fingerprint),以及筛选出适合于玉米杂交种及其亲本自交系真实性和纯度鉴定的方法。结果表明,各供试玉米杂交种及其亲本自交系都具有相应的、唯一的种子盐溶蛋白聚丙烯酰胺凝胶电泳所显现的生化指纹。对于有些组合。玉米芽鞘和叶片绿色组织过氧化物酶同工酶电泳图谱存在阴极第4、第5酶带差异,因这两条酶带的差异稳定,并且重现性好,故能用过氧化物酶同工酶技术对其进行有效地鉴定。上述两种方法,尤其是前者,因技术要求不高,费用低,快速及重现性好等特点,能满足我国目前种子检验室日常玉米品种纯度快速测定工作的要求,具育良好的应用前景。     

苦瓜品种资源聚类分析

本试验对14个不同绿苦瓜、白苦瓜和广西野生苦瓜品种的种子可溶性蛋白、盐溶性蛋白、醇溶性蛋白和碱溶性蛋白进行聚丙烯酰胺凝胶电泳,运用遗传距离系数对14个苦瓜品种进行聚类分析方法;同时利用8个数量性状计算主成分遗传距离,对14个苦瓜品种进行聚类分析。研究结果表明,广西野生苦瓜与其他13个栽培品种之间遗传距离系数大,数量遗传距离也大,亲缘关系远,它们分别归为不同的品种类群,聚类分析将广西野生苦瓜确定为不同的亚种;栽培品种间差异小,很难进行类群的清晰划分。     

西瓜同工酶及可溶性蛋白分析

对京欣一号西瓜的亲本及杂种一代的几种同工酶系统及可溶性蛋白进行了分析。结果表明，在种子萌发过程中，过氧化物酶、酯酶和可溶性蛋白随发育阶段或营养状况的改变而改变；而在相同萌发时期，过氧物酶工酶、酯酶同工酶、过氧化氢酶同工酶和淀粉酶同工酶在亲本及杂种间未见差异，杂种的可溶性蛋白图谱表现与母本相同，而与父本有差异。此外，对６个品种的西瓜、非洲西瓜和瓠瓜的分析结果表明，非洲西瓜和瓠瓜的过氧化物酶酶谱、酯酶     

Symbiotic Interactions between Rhizobium leguminosarum Strains and Elite Cultivars of Pisum sativum L.

Pea ( Pisum sativum L.) is an important world grain legume and vegetable in the south of Europe where it is grown in small farms and gardens during the cool season. Most of the pea production of this area occurs in high nitrogen (N) soils. Inoculation is required to increase yields through N fixation and reduce the external inputs. Three pea elite cultivars representing human food market classes (green, sugar and dry peas) were studied in order to characterize the cultivars' variability in symbiotic characters using two Rhizobium leguminosarum biovar viceae commercial strains and the indigenous soil strain, and three N-fertilization rates. Significant differences were observed among pea elite cultivars for the weight and length of plant parts, and the traits associated with N fixation. Pea cultivars showed a significant cultivar–strain interaction for shoot fresh weight and significant differences among strains were found for nodule fresh weight. Green and sugar pea cultivars had good N fixation with a particular commercial strain, while the dry pea cultivar did not show an effective symbiotic combination. The differences between the pea cultivars could be exploited in breeding programs for enhanced nodulation and N fixation in each pea market class.     

Biochemical characterization of white onion landraces (Allium cepa L.) through HPLC analysis of endosperm seed proteins

Water-, salt-, alcohol- and alkali-soluble seed storage proteins, extracted from 21 cipolla bianca di Pompei landraces (Allium cepa L.), were analyzed by anionic exchange-high performance liquid chromatography (AE-HPLC). Chromatographic elution profiles (time range 0–40 min) at 280 nm of water-soluble seed proteins evidenced the presence of 21 peaks, which allowed all the landraces studied to be distinguished from each other. The differences detected were both qualitative (presence/absence of one or more peaks) and quantitative; the water-soluble proteins were useful in differentiating landraces and cultivars while the other seed protein fractions only showed a weak polymorphism. The cluster analysis, based on HPLC data, showed that the landraces clustered with a genetic similarity degree ranging between 69% and 94%. The possibility of discriminating among closely related onion landraces during the course of breeding programmes could allow the identification of biochemical markers linked to useful agronomical traits. As observed by chromatographic analysis, the globulin composition of onion water-soluble seed protein appears to be independent of environmental growth conditions. The biochemical characterization of the available typical onion germplasm may contribute to obtain a community recognition and denomination, such as Denomination of Protected Origin (D.O.P.), Indication of Protected Origin (I.G.P.) or Specificity Attestation (A.S.). The biochemical method here developed resulted of high resolution, cost-effective and time-saving for characterization and genetic purity assessment of the landraces studied.     

Relationships between Seed Yield and Chemical Composition of Field Peas Grown under Semi-arid Mediterranean Conditions

Relationships between seed protein, lipid, starch and sugar concentrations and seed yield of 14 pea (Pisum sativum L.) cultivars were examined under rain-fed conditions in northern Jordan. Significant differences amongst cultivars for each character were noted. The cultivar-by-environment interaction was significant for each character except lipid and starch concentrations. A negative correlation was noted between seed yield and seed protein and fructose concentrations. Positive non-significant correlations between seed yield and seed lipid and starch concentrations were noted. The cultivars DMR-8, P 350-1 and 88P022-6 had similar seed yields, but varied seed chemical components. Seeds of the Praire No. 11 cultivar had the highest protein concentration amongst cultivars, although it produced the lowest seed yield. These results indicate that factors other than seed yield are important for determining the chemical composition of seeds, and that one-sided selection for high seed yield tends to reduce the nutritional quality of the pea seed.     

辣椒种子贮藏蛋白的含量测定和电泳分析

对12个辣椒品种种子内4种贮藏蛋白清蛋白、球蛋白、醇溶蛋白和谷蛋白进行了含量测定和SDS，PAGE电泳分析。结果表明：辣椒种子贮藏蛋白含量以清蛋白和谷蛋白含量最高，其次是球蛋白，醇溶蛋白的含量最低。品种间蛋白质含量差异不明显。各品种都能通过清蛋白、球蛋白和谷蛋白的蛋白质亚基组成信息区别出其差异性。     

Seed proteins of the wild and the cultivated Amaranthus species

The 13–21% variation in seed protein content was observed in wild and cultivated forms of amaranth. Seed proteins of amaranth are highly nutritive and composed presumably of easily digestable albumins and globulins (over 50% of total protein); of 20.8% alkali-soluble proteins-glutelins with similar nutritive value and only of 12% alcohol-soluble proteins-prolamins which are lacking in essential amino acids. The results of polyacrylamide gel electrophoresis (buffer pH being 3.2) indicate that seed proteins of amaranth are heterogeneous and compose of 38 bands which, by their electrophoretic mobility, can be tentatively assigned to four zones: A, B, C and D. By their protein patterns all Amaranthus species were assigned to 7 biotypes. The cytogenetic and electrophoretic comparison enabled us to determine the degree of diversity among amaranth forms studied. The phylogenetic relationship between A. paniculatus L. and A. hybridus L. was confirmed. A relationship was also supposed between these two species and A. lividus L. and between A. powellis L. and A. deflexus L. which by their electrophoretic patterns belong to the same biotype. A taxonomic classification of 5 samples whose classification has not been done yet was performed, as well as of cultivars Elbrus and Progress which were assigned to specious A. edulis L. It was confirmed that the method of electrophoretic analysis of seed storage proteins is very promising for detecting the phylogenetic relationship between Amaranthus species. This revised version was published online in July 2006 with corrections to the Cover Date.