中国小麦地方品种内和品种间醇溶蛋白遗传多样性分析

为了揭示中国小麦地方品种内遗传异质性和品种间的遗传多样性,采用A-PAGE方法,对72份来自不同生态区的地方品种进行醇溶蛋白构成分析。结果发现,全部供试地方品种共观察到101条迁移率不同的务带,构成229种醇溶蛋白构型,每个品种醇溶蛋白条带数目为14—24。63份（87．5％）地方品种在品种内具有2种以上醇溶蛋白变异类型,其中,变异类型最多的品种二红皮小麦（ZM004659）30个子粒中有14种之多,多数品种具有2—3种变异类型。品种内醇溶蛋白构型一致的品种共有9个,占12．5％。这表明供试的大多数小麦地方品种内个体间在醇溶蛋白构成上具有遗传异质性。聚类分析表明,相同生态区的地方品种没有整齐地聚为一类。     

醇溶蛋白水平上甘、青两省春小麦品种间的遗传多样性现状及演变趋势

用APAGE技术检测了43个春小麦品种在醇溶蛋白水平上的遗传变异,在计算Jaccard相似系数的基础上用UPGMA方法进行聚类分析。共得到42种不同的带型,电泳共分离出37条带,其中33条具有多态性。品种间在醇溶蛋白水平上的遗传距离变异范围很大(0．0000～0．8148),平均遗传距离GD=0．5073。说明43个品种在醇溶蛋白编码位点上存在较大变异。聚类结果显示,除佛手麦白成一类外(GS=0．28),地方品种和引进品种与大多数育成品种(GS=0．46)分属不同的亚类,说明育成品种同地方品种和引进品种在醇溶蛋白编码基因上存在较大变异。育成品种间在醇溶蛋白水平上的遗传多样性程度不高,这一结果提示在这一地区进行与醇溶蛋白相关的品质育种很难在现有的育成品种间杂交的基础上取得成功。从遗传多样性的演变趋势来看,历史上以地方品种间醇溶蛋白的遗传变异程度最大(GD=0．5455),50年代引进品种变异程度最小(GD=0．3310)。从60～90年代,育成品种醇溶蛋白水平上的遗传多样性有一先增后减的过程,但总体上变异程度要低于地方品种,其原因与亲本单一和育种目标由高产到优质变化条件下人工选择压由弱到强有关。     

基于醇溶蛋白的20份小麦种质遗传完整性分析

采用醇溶蛋白电泳技术对同一品种不同繁殖年份的20份小麦种质进行遗传完整性分析。结果表明：供试种质中有10份具有一种醇溶蛋白谱带带型的同质性种质;另外10份具有2～4种醇溶蛋白谱带带型的异质性种质,其中6份为地方品种。表明地方品种具有较高的遗传多样性。在10份异质性种质中,两个繁殖年份种质之间的醇溶蛋白带型频率变化差异不显著的有5份,其第一繁殖年份的种质发芽率均高于75％,而另外5份存在显著差异的种质,第一年份的发芽率都低于66％。进一步分析表明,这10份异质性种质在两个繁殖年份之间,其发芽率差值与带型频率差值之间呈极显著正相关,相关系数为0．8665。上速结果表明,小麦更新时较高的发芽率是维持异质性种质遗传完整性的关键因素。     

野生大麦与栽培大麦醇溶蛋白遗传多样性比较

利用A-PAGE(acid-polyacrylamide gel electrophoresis)法对采自以色列的野生大麦的一个野生自然群体的15个系和来自世界不同国家的14份栽培大麦品种醇溶蛋白的遗传多样性进行了分析.结果表明:在所有的29份供试材料中,共发现52条相对迁移率不同的谱带.52条谱带的出现频率为3.44%～93.1%,多样性指数为0.066～0.368;以中国春醇溶蛋白为标准,ω区大麦醇溶蛋白的谱带数最多,其次是β区;野生大麦Shannon多样性指数依次为β区>ω区>α区>γ区,而栽培大麦Shannon多样性指数依次为ω区β>区>γ区>a区;野生大麦自然群体和栽培大麦品种间的遗传相似系数变幅相当,且聚类分析结果显示,野生大麦自然群体和来自全球不同区域栽培大麦品种间的醇溶蛋白遗传多样性同样丰富.以上结果说明,野生大麦中保存了较栽培大麦更为丰富的基因资源,今后栽培大麦的品质改良应该重视野生大麦资源的合理利用.     

人工合成六倍体小麦醇溶蛋白遗传多样性分析

运用酸性聚丙烯酰胺凝胶电泳(A-PAGE)技术,对96份人工合成六倍体小麦的醇溶蛋白多样性进行了分析。结果显示,96份人工合成小麦中,共分离出65条不同的醇溶蛋白谱带,其中ω区22条,β和γ区各17条,α区9条,但各醇溶蛋白在电泳图谱中出现的频率差异较大,其变化范围为1.04%~91.67%。醇溶蛋白遗传多样性指数(H′)及多态性信息含量(PIC)分析结果显示,β、ω两个谱带区醇溶蛋白组成最为丰富,而α区最低;聚类分析结果显示,材料间的平均遗传距离为0.86,在遗传距离为0.83水平上,96份材料被划分为4个主要类群,类群间的关系基本反映了合成双二倍体的亲缘关系。研究结果表明,人工合成六倍体小麦醇溶蛋白基因位点表现出广泛的遗传变异,具有丰富的遗传多样性。     

鹅观草种质资源醇溶蛋白遗传多样性研究

以8个地理类群的90份鹅观草野生种质材料为研究对象,采用A-PAGE(酸性聚丙烯酰胺)凝胶电泳技术进行蛋白质水平遗传多样性检测.研究结果表明,来源于不同居群的鹅观草共分离出26条谱带,每个材料可以分离出5～26条迁移率不同的谱带, 平均谱带数为16.39条,其中平均多态性谱带为12.65条,多态性比例为77.22%;基于供试材料醇溶蛋白每个位点谱带出现的频率,分别计算了地理类群内多样性指数(0.345)和总多样性指数(0.471),类群间的遗传分化系数为26.8%,表明鹅观草变异的73.2% 来源于类群内;90份供试材料醇溶蛋白的Jaccard遗传相似系数变异范围为0.133 3～1.000,平均遗传相似系数为0.395 7;利用种子醇溶蛋白可将90份材料分为12类,鹅观草种质资源之间的亲缘关系呈现出一定的地域性规律;不同地理类群间的遗传多样性指数从高到低的排列顺序依次为云南＞四川＞内蒙古＞新疆＞山西＞甘肃＞宁夏＞河北.因此在进行鹅观草种质资源收集和原地保护时,建议对云南和四川地区的鹅观草种质资源应给予极大关注.     

偃麦草属植物醇溶蛋白和谷蛋白多态性及系统学研究

运用酸性聚丙烯酰胺凝胶电泳(A-PAGE)和十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)对偃麦草属(Elytrigia)24个物种的醇溶蛋白和谷蛋白进行了研究。以中国春为参照,按醇溶蛋白和谷蛋白电泳图谱条带迁移距离大小和条带多态性对供试材料进行聚类分析。结果显示,24份供试材料均呈现不同的醇溶蛋白和谷蛋白电泳图谱,共分离出醇溶蛋白带纹83条和谷蛋白带纹53条,多态性均达100%,并且在相同染色体组组成的物种中,染色体数目越多,其蛋白带纹越多;偃麦草属24个物种的醇溶蛋白和谷蛋白图谱均有明显差异,蛋白图谱可作为鉴定偃麦草属物种的指纹图谱。聚类分析结果显示,在材料间的醇溶蛋白遗传相似性系数为0.66时,24份材料被划分为6个主要类群,含E和St基因组的物种具有较近的亲缘关系;在谷蛋白遗传相似性系数为0.62时,24份材料被聚为4大类。聚类结果表明,染色体组成未知的物种Et.pachynera可能为异源多倍体物种。     

119份小麦种质资源醇溶蛋白遗传多样性分析

利用A-PAGE对119份小麦种质资源进行了醇溶蛋白遗传多样性分析。结果显示,共分离出蛋白带1 938条,迁移率不同的谱带类型116种,其中编号为10和14的2种谱带出现频率最高,分别为54.62%和96.44%,其余114种谱带类型具有较强的多态性;每个种质材料的醇溶蛋白谱带数为10~24条,大部分为13~19条;供试种质间遗传距离在0.24~0.83,平均为0.54;聚类分析将所测材料分为6大类,与种质资源所反映的系统关系类似,表明醇蛋白在一定程度上能反映种质间的亲缘关系。     

东方小麦(Triticum turanicum Jakubz.) 醇溶蛋白遗传多样性分析

为了进一步利用东方小麦(Triticum turanicum Jakubz.)遗传资源,对来自埃塞俄比亚、伊拉克、伊朗、阿塞拜疆、阿富汗、摩洛哥等国家共87份东方小麦醇溶蛋白位点的遗传多样性进行了分析.结果发现,供试材料存在丰富的遗传变异,87份材料共产生72种谱带类型,分离出33条带纹,每份材料可电泳11～22条带,平均16条;在α、β、γ和ω四个区均差异较大,分别有16、11、20和20种谱带变异类型.聚类分析发现,醇溶蛋白揭示的材料间遗传多样性与其地理来源有一定关系.     

波斯小麦醇溶蛋白遗传多样性分析

应用APAGE技术对来源于16个国家(地区)96份波斯小麦材料进行醇溶蛋白遗传多样性分析,结果表明,波斯小麦具有丰富的醇溶蛋白等位变异,共分离出55条迁移率不同的带纹。每份材料可电泳出13～26条,平均18.6条,带纹多态性为100%。96份材料共出现了75种电泳图谱类型,其中31份材料共同拥有10种图谱,剩余的65份材料的电泳图谱各不相同。供试材料间GS值变异范围为0.176～1.000,平均值为0.538。聚类分析表明,在GS值0.538水平上供试材料可聚为五大类群,且遗传聚类关系与材料的地理来源有一定的相关性。     

野生垂穗鹅观草醇溶蛋白遗传多样性研究

利用酸性聚丙烯酰胺凝胶电泳(Acid-polyacrylamidegel electrophoresis)法对来源于内蒙古、山西、甘肃、新疆、云南、四川和西藏的25份垂穗鹅观草材料进行醇溶蛋白检测。研究结果表明,供试材料可分离出30条相对迁移率不同的谱带,每份材料可电泳出11～22条谱带,其中只有1条(3.45%)带纹是25份材料共有的,其余29条谱带均具有不同程度的多态性,多态性谱带数目占分离出总条带的96.67%,说明野生垂穗鹅观草具有较丰富的遗传多样性。聚类分析发现,材料的遗传相似系数(GS)变异范围为0.111～0.9286,平均值为0.4821。在GS值为0.4821的水平上供试材料可聚为6个类群,某些具有相同地理来源的材料聚成一类或亚类,即醇溶蛋白图谱类型与材料的生态地理环境具有相关性。     

赖草属植物醇溶蛋白的遗传多态性

用酸性聚丙烯酰胺凝胶电泳 (A PAGE)对小麦族赖草属 2 0种和 1亚种的 45份材料进行醇溶蛋白遗传分析 ,结果表明 :(1 ) 4 5份材料共出现 43种不同的醇溶蛋白图谱 ,从中分离出的 38条带纹多态性高达 1 0 0 % ;(2 )赖草属植物具有丰富的醇溶蛋白遗传多态性 ,其种间和种内不同居群间均存在明显的醇溶蛋白遗传变异 ,其种间变异大于种内变异 ,醇溶蛋白图谱可以作为鉴定赖草属植物的指纹图谱 ;(3)醇溶蛋白图谱的聚类结果与形态学结果基本一致 ,表明醇溶蛋白资料可以运用于赖草属植物种间、种内遗传差异及亲缘关系研究     

鹅观草属三个物种及其居群间的醇溶蛋白分析

利用酸性聚丙烯酰胺凝胶电泳技术(APAGE)对小麦族鹅观草属3个物种:毛叶鹅观草、纤毛鹅观草和竖立鹅观草进行了醇溶蛋白电泳分析,23份材料电泳分离出22条相对迁移率不同的谱带,其中3条(16.6%)共同带,19条(83.4%)具多态性,每个材料可分离出9～16条谱带。结果表明:(1)三个物种具有相似的醇溶蛋白带型,但存在明显的种间差异;(2)同种不同居群间也存在遗传差异;(3)种间差异大于种内差异。     

山羊草Aegilops kotschyi及其供体种Ae.longissima和Ae.umbellulata的醇溶蛋白研究

采用酸性聚丙烯酰胺凝胶电泳(APAGE)法对11份A担心Aegilops kotschyi及其S^1染色体组供体种Ae.longissima2份和U染色体组供体种Ae.umbellulata6份进行了醇溶蛋白位点的研究。结果表明：11份Ae.kotschyi共分离出32条带,31条具有多态性,占96．88％,每份材料可以分离出10-17条谱带,其中仅1条(3．12％)是共有带;11份Ae.kotschyi的遗传距离的变异范围在0-0.704之间,平均为0．409;11份Ae.kotschyi分离出的多数醇溶蛋白谱带均与其染色体组供体种Ae.longissi-ma及Ae.umbellulata相同,但仍有8条谱带未在两供体种中找到;11份Ae.kotschyi的醇溶蛋白多态性(96．88％)明显高于Ae.longissima(52．94％)与Ae.umbellulata(88．89％)11份Ae.kotschyi中有4份表现出了一定的特征带,分析知可能在γ区发生了较大的变异。     

Genetic diversity of French common wheat germplasm based on gliadin alleles

 Analysis of gliadin electrophoretic (APAGE) patterns made it possible to identify 79 alleles at six Gli-1 and Gli-2 loci (from 9 to 18 per locus) and 173 gliadin genotypes in the 187 French common wheat cultivars considered. Six new alleles were registered in the catalogue of gliadin alleles. The genetic diversity of French common wheats was found to be high (H=0.714) and had not changed much during the last 25–50 years. Analysis of genetic distances showed some gradual changes in French wheat germplasm over the course of time. Genetic distances between French and several European wheat germplasm were analysed; genotypes of European wheats were found to relate very distantly to Canadian genotypes. The considerable differentiation of wheat genotypes from different countries and cereal companies might be caused by breeders’ personal preferences and by hidden natural selection specific to each local environment. In French cultivars, genetic variation in earliness, and in the North/South habit of the cultivars studied, correlated significantly with allelic variation at Gli-B1, Gli-A2 and Gli-D2 for earliness, and at Gli-D2 for the North/ South habit. Early and late cultivars are grown mainly in Southern and Northern France, respectively (r ²=0.30). Cultivars having either the 1B/1R translocation or allele Gli-D2g are, on average, later and more resistant to cold; they hence are grown in the North of France. Alternatively, cultivars with the allele Gli-D2m are earlier and cold-sensitive, and are grown in the South of France. Received: 5 February 1997 / Accepted: 19 September 1997     

Comparison of RAPD,ISSR, and AFLP Molecular Markers to Reveal and Classify Orchardgrass (Dactylis glomerata L.) Germplasm Variations

Three different DNA-based techniques, Random Amplified Polymorphic DNA (RAPD), Inter Simple Sequence Repeat (ISSR) and Amplified Fragment Length Polymorphism (AFLP) markers, were used for fingerprinting Dactylis glomerata genotypes and for detecting genetic variation between the three different subspecies. In this study, RAPD assays produced 97 bands, of which 40 were polymorphic (41.2%). The ISSR primers amplified 91 bands, and 54 showed polymorphism (59.3%). Finally, the AFLP showed 100 bands, of which 92 were polymorphic (92%). The fragments were scored as present (1) or absent (0), and those readings were entered in a computer file as a binary matrix (one for each marker). Three cluster analyses were performed to express–in the form of dendrograms–the relationships among the genotypes and the genetic variability detected. All DNA-based techniques used were able to amplify all of the genotypes. There were highly significant correlation coefficients between cophenetic matrices based on the genetic distance for the RAPD, ISSR, AFLP, and combined RAPD-ISSR-AFLP data (0.68, 0.78, 0.70, and 0.70, respectively). Two hypotheses were formulated to explain these results; both of them are in agreement with the results obtained using these three types of molecular markers. We conclude that when we study genotypes close related, the analysis of variability could require more than one DNA-based technique; in fact, the genetic variation present in different sources could interfere or combine with the more or less polymorphic ability, as our results showed for RAPD, ISSR and AFLP markers. Our results indicate that AFLP seemed to be the best-suited molecular assay for fingerprinting and assessing genetic relationship among genotypes of Dactylis glomerata.     

Genetic control of endosperm proteins in wheat

Summary Total endosperm proteins extracted from both several common wheat cultivars and some intervarietal substitution lines derived from them were fractionated according to their molecular weight in a high resolution one-dimensional gel electrophoresis. The four donor cultivars and the recipient one — Chinese Spring, possessed differentially migrating protein bands in the fractions of high molecular weight (HMW) glutenins and gliadins. Several of these bands were identified for the first time in this study. By utilizing intervarietal substitution lines the control of the HMW glutenins and gliadins by chromosomes of homoeologous group 1 was either reaffirmed or, for the new bands, established. Several HMW gliadin subunits showed a considerable variation in their staining intensity in the intervarietal substitution lines indicating that their expression was dependent on the genetic background.This paper is based on a portion of a dissertation to be submitted by G. Galili in partial fulfilment of the Ph.D. requirements of the Feinberg Graduate School, The Weizmann Institute of Science, RehovotThe Marshall and Edith Korshak Professor of Plant Cytogenetics     

AFLP analysis of genetic variability in New Guinea impatiens

New Guinea impatiens ( Impatiens hawkeri) is an economically important floral crop, however, little work has been conducted to further our understanding of the genetics of this crop. In this study, we used amplified fragment length polymorphism (AFLP) technology to investigate the level of polymorphism present among 41 commercial cultivars of New Guinea impatiens, study their genetic relatedness, and assess the genetic diversity in this material. An efficient DNA extraction protocol was developed, and a total of 48 EcoRI and MseI primer combinations were used for PCR amplification. Amplification products were then subjected to polyacrylamide gel electrophoresis. The AFLP analysis showed that all 41 cultivars generated between 73 and 130 scoreable polymorphic bands per primer combination. Gower's Genetic Dissimilarity estimates for the entire set of cultivars ranged between 0.940 and 0.488. A dendogram was generated from these dissimilarity data that revealed four groupings among these 41 cultivars. The implications of these results on genotypic variation, genetic relationships, and genetic diversity in New Guinea impatiens will be discussed.