基于ISSR标记分析豌豆农家品种遗传多样性

为掌握内蒙古部分地区种植的农家豌豆品种的亲缘关系,本研究通过ISSR分子标记技术,对来自内蒙古自治区部分地区及甘肃省张掖市的29份豌豆农家品种的遗传多样性进行分析。结果显示,从19个ISSR引物中筛选出多态性明显、条带清晰、反应稳定的引物共7个。29份DNA材料共扩增出118条条带,其中116条为多态性条带,平均每个引物扩增出的条带数为16.9条,多态性比率为98.31%。Shannon多样性指数平均为0.505 9,每个位点的有效等位基因数为1.569 6,品种间遗传相似系数变幅为0.370 4～0.928 6,表明本研究的29个豌豆农家品种具有丰富的遗传多样性。利用UPGMA聚类分析,以遗传相似系数0.69为界限,将29份材料划分为5类,聚类结果与豌豆种群地域性分布规律关联性较高,表明ISSR分子标记技术的遗传多样性分析能够将地理来源相近的居群进行聚类划分,其研究结果可运用于豌豆种质资源保护及育种工作中。     

东北地区野豌豆属四个种间遗传关系的ISSR分析

对野豌豆属的山野豌豆、狭叶山野豌豆、广布野豌豆和东方野豌豆4份材料,利用ISSR分子标记进行遗传关系分析。6个引物共扩增出52条带,其中多态性条带总数为46条,平均多态性比率为88.46%。Shannon多样性指数分析结果表明,种内平均遗传多态度为1.72,遗传多态度总量为4.39;种内变异值在0.15~0.72之间,种内的遗传变异均值为0.42;种间的遗传变异均值为0.58。UPGMA法遗传关系聚类显示,山野豌豆和狭叶山野豌豆亲缘关系最近,与东方野豌豆亲缘关系最远。利用ISSR标记技术可较准确分析野豌豆属材料间的亲缘及遗传多样性。     

基于ISSR标记的福建茶树品种(系)遗传多样性分析

本研究通过对所收集的55份福建茶树品种(系)进行遗传多样性和亲缘关系分析,采用ISSR分子标记技术进行扩增,并通过NTSYS-pc2.10e软件进行数据分析。以7份福建茶树品种为材料,对50条通用引物进行筛选,得到多态性好、扩增条带清晰的引物15条。15条引物共扩增出213条条带,208条为多态性条带,多态性所占比率为97.65%。55份茶树资源遗传相似系数介于0.54~0.91之间,表明这55份茶树遗传背景差异较大。并根据ISSR扩增图谱构建了遗传聚类图,明确了各茶树品系与品种间的亲缘关系,为茶树培育新品种奠定科学基础、提供理论依据。     

贵州金荞麦种质资源遗传多样性ISSR研究

应用ISSR分子标记方法,以来源于11个不同地域的金荞麦种质为材料,进行遗传多样性与亲缘关系分析。从40条通用引物中筛选出条带清晰、多态性丰富、重复性好的11个引物PCR扩增共获得103条稳定的条带,其中75个位点具有多态性,其多态性位点百分率(PPB)为72.8%。应用Nei-Li相似系数法对11份不同来源的金荞麦品种间遗传相似系数(GS)进行了估算,其GS为0.48~0.90,遗传多样性较丰富,11种金荞麦品种间遗传距离GD在0.10~0.52,平均为0.27,种间遗传差异较小。聚类分析表明:11个品种被分为2大类,在第一大类中,大方和水城最先聚在一起,二者亲缘关系较近,而贵阳与另外5个地区金荞麦亲缘关系较远则单独聚类;第二大类中,罗甸和独山最先聚为一类,且遗传距离短,二者亲缘关系近。本研究可为金荞麦的亲本利用及开展金荞麦遗传连锁图的构建和核心种质指纹图谱的绘制提供参考。     

人工合成六倍体小麦的ISSR位点遗传多样性分析

为了解人工合成六倍体小麦品种的遗传多样性,选用了100条ISSR引物对11份人工合成小麦和3个普通小麦材料(TP、HTS-1、CS)进行了遗传多样性分析。筛选出了42条引物可用于遗传多样性分析,其中有24条引物对供试材料具有较强的鉴别能力。42条引物共扩增出了273个条带,每条引物扩增出的条带数是3~12,平均每条为6.5,同时检测到在14份小麦试材中有170(占62.3%)个等位变异位点,引物等位变异数为1~11,平均每个位点出现4.5个等位变异。42条引物扩增产物的多态性信息量(PIC)为0.499~0.879,平均为0.767,多数引物扩增的多态性信息量在0.800左右。14份材料间的遗传相似系数为0.663~0.934,平均为0.790。根据材料间的遗传相似系数聚类,14个材料被聚为3类,其中聚为第2类的试材最多。本研究表明,14份小麦材料遗传相似性较大,亲缘关系较近,但参试材料总体遗传多样性水平较高。聚类分析结果与试材系谱及亲本来源相吻合。结果表明ISSR标记技术可应用于物种的鉴别、遗传多样性研究和遗传图谱构建。     

28份茉莉花种质基于TRAP技术的遗传多样性分析

本研究利用TRAP分子标记对茉莉花的遗传多样性进行分析,从36对TRAP候选引物中筛选出25对多态性丰富的引物,以28份茉莉花基因组DNA为模板进行PCR分析,共获得148条带,其中136条具有明显的多态性,多态率为91.9%;每对引物扩增出3~8个多态性带,平均5.44个条带。28份茉莉花种质材料间遗传相似系数为0.37~0.92,平均值为0.65;UPGMA聚类分析表明在遗传相似系数0.64时,可将28份材料划分为五个聚类组。研究结果表明,TRAP标记在茉莉花材料中具有丰富的多态性,可以进一步应用该技术进行茉莉花遗传多样性及亲缘关系的研究。     

青麻叶大白菜核基因雄性不育性遗传模式的研究

以稳定遗传的青麻叶类型大白菜甲型两用系2个、乙型可育株系4个及可育品系26个为材料，经过杂交、自交、测交及育性鉴定等手段，对青麻叶类型大白菜核基因雄性不育性的遗传模式进行了验证，并对青麻叶类型材料的育性基因型进行了测定。结果表明，青麻叶类型大白菜核基因雄性不育性的遗传模式符合复等位基因遗传模式；在经过多代纯化的青麻叶材料中，基因型为msms的材料所占比例最大，为65．4％，基因型为Ms^fms的材料所占比例最小，为7．7％，基因型为Ms^fMs^f的材料所占比例居中，为26．9％。     

基于SSR标记的冬瓜种质资源遗传多样性分析

为了加强冬瓜种质资源的保护和评价,明确资源间的亲缘关系,本研究利用SSR标记在分子水平上对111份冬瓜种质资源进行遗传多样性分析。结果表明,19对SSR引物共扩增出182条条带,其中多态性条带169条,多态率92.86%;通过POPGENE 1.31软件计算每对引物的平均有效等位基因数(Ne)为1.393 2,平均Nei's遗传多样性指数(He)为0.260 1,平均Shannon's信息指数(I)为0.418 7。采用Jaccard相似系数进行种质间的聚类分析,在遗传距离为0.70的位置可将全部种质分为6个类群。通过聚类结果可知,种质材料未按照地理来源划分,但相同果型、籽型的种质资源间亲缘关系较近。本研究基于SSR标记分析冬瓜种质资源遗传多样性并进行亲缘关系的聚类,将为冬瓜种质的保存和利用提供理论依据。     

8份剑麻种质亲缘关系的ISSR和RAPD分析

为了揭示剑麻栽培品种的遗传多样性,利用ISSR和RAPD分子标记技术对8份剑麻种质的亲缘关系进行分析。结果表明,筛选后选用的8条ISSR引物和8条RAPD引物,分别产生了53条和66条扩增条带,其中多态性条带分别为44条和61条,多态性条带百分率分别为83.02%和92.42%。根据2种标记的扩增结果,用UPGMA法对8份剑麻种质进行聚类分析,供试材料之间具有较高的遗传多样性,其品种间遗传相似系数分别为0.59~0.80和0.52~0.76。2个标记的聚类结果基本一致,但有点差异,可将供试的8份剑麻种质划分为2类群,而且2个标记聚类结果呈显著相关性,相关系数为0.70。可见,剑麻种质资源的遗传多样性丰富。     

基于SSR标记的芍药种质资源遗传多样性研究

基于SSR分子标记技术对来自河南、湖北、安徽3省100种芍药种质资源间的遗传关系进行分析。结果表明:8对多态性SSR分子标记共扩增获得54条条带,平均每对引物可扩增6条条带,平均每对引物的有效等位基因数为6.75,Shannon信息指数范围为0.443 3~2.132 6;使用UPGMA法构建所有试材的系统聚类树,供试材料共分为3个组及3个亚组,聚类结果与试材地域来源关系密切。进一步说明SSR是研究芍药种质资源遗传多样性及亲缘关系的有效手段,为芍药亲本组合选配、种质创制和遗传育种奠定基础。     

AFLP分析小豆种(Vigna angularis)内遗传多样性

利用12对AFLP引物,对来自于世界6个国家的146份小豆(Vigna angularis)栽培变种(var. angularis)和野生变种(var. nipponensis)种质的基因组DNA进行扩增,得到580条清晰的显带,其中313(53.93%)条呈多态性,平均每对AFLP引物得到26.08条多态性带;平均遗传距离0.35,变异幅度为0.00～0.87.利用AFLP多态性数据进行的Jaccard''s遗传距     

采用AFLP和SSR标记对扁蓿豆遗传多样性进行比较分析

为了筛选出优异的扁蓿豆育种新材料,本研究采用AFLP和SSR分子标记技术对来自于中国7个省市自治区的15份扁蓿豆种质资源进行遗传多样性的比较分析,结果表明:18对SSR引物扩增出109个多态位点,8个AFLP引物组合扩增出640条带,其中472条多态带.AFLP标记的平均Nei′s遗传多样性指数、Shannon多样性指数和遗传分化系数均高于SSR标记.15份扁蓿豆种质的遗传距离和遗传相似系数与地理类群很接近.AFLP和SSR数据的聚类分析显示:15份扁蓿豆种质分为4大类,但是聚类结果与地理类群不完全相符,主成分结果与聚类结果相似,Mantel 检测表明:AFLP和SSR数据有较高的显著相关性,AFLP和SSR标记能够有效地对扁蓿豆进行遗传多样性分析,其结果为扁蓿豆育种和资源保护具有指导意义.     

Bulked AFLP analysis for the assessment of genetic diversity in white clover (Trifolium repens L.)

The use of bulked leaf samples from individual plants for amplified fragment length polymorphism (AFLP) analysis was evaluated as a tool for assessment of genetic diversity in white clover (Trifolium repens L.). Bulking of leaf samples produced slightly simpler AFLP profiles compared to the combined profiles of individual plants from the same cultivar. Approximately 90% of bands which were present in individual plants were present in bulked samples of the same cultivar. The majority of those absent were rare bands, shared by less than 25% of individual plants. Replicate bulk samples gave almost identical banding patterns, demonstrating the robustness of the bulked AFLP technique. Cluster analysis of AFLP data derived from individual plants resulted in a phenogram similar to that produced from data derived from bulked samples of the same plants. AFLP analysis of bulked samples detected significant amounts of genetic variability among 52 cultivars and accessions with genetic similarity values ranging from 0.42 to 0.92. However, cluster analysis of AFLP data only partially reflected the geographic origin of cultivars and accessions and was not congruent with cluster analysis based on variation for morphophysiological characters. Bulked AFLP analysis provides a powerful tool for rapid assessment of genetic variability in white clover and may also be used for cultivar identification. This revised version was published online in August 2006 with corrections to the Cover Date.     

裸燕麦品质资源AFLP标记遗传多样性分析

用20对AFLP引物组合对281份栽培裸燕麦(Avena nuda)进行遗传多样性分析,共得到1 137条带,其中260条为多态性带,引物的平均多态性百分率为22.96%,平均多样性信息指数(PIC)为0.0326。以地理来源分组,不同来源的组群Simpson指数在1.235~1.495之间,Shannon指数范围为0.1558~0.4437,组群内变异贡献率为83.45%,组群间变异占16.55%。组群大小与多态性位点数、组群内变异贡献率、Simpson指数及Shannon指数显著相关。内蒙古和山西资源多样性丰富,东北地区资源独特,西部地区资源遗传结构单一,东欧组群与内蒙古组群遗传关系最近。国内组群的遗传多样性水平高于国外组群。地方品种与育成品种相比,组群内变异贡献率较高。建议在遗传多样性丰富地区进一步收集裸燕麦资源,并加强对材料少、代表性较差的地区,如西北和西南地区的裸燕麦地方品种的收集,以丰富我国的裸燕麦基因源。     

裸燕麦种质资源AFLP标记遗传多样性分析

用20对AFLP引物组合对281份栽培裸燕麦(Avena nuda)进行遗传多样性分析,共得到1137条带,其中260条为多态性带,引物的平均多态性百分率为22.96%,平均多样性信息指数(PIC)为0.0326.以地理来源分组,不同来源的组群Simpson指数在1.235～1.495之间,Shannon指数范围为0.1558～0.4437,组群内变异贡献率为83.45%,组群间变异占16.55%.组群大小与多态性位点数、组群内变异贡献率、Simpson指数及Shannon指数显著相关.内蒙古和山西资源多样性丰富,东北地区资源独特,西部地区资源遗传结构单一,东欧组群与中国内蒙古组群遗传关系最近.国内组群的遗传多样性水平高于国外组群.地方品种与育成品种相比,组群内变异贡献率较高.建议在遗传多样性丰富地区进一步收集裸燕麦资源,并加强对材料少、代表性较差的地区,如中国西北和西南地区的裸燕麦地方品种的收集,以丰富我国的裸燕麦基因源.     

AFLP analysis of genetic variability among Stylosanthes guianensis accessions resistant and susceptible to the stylo anthracnose

Stylosanthes guianensis, belonging to the genus Stylosanthes, is one of the most important tropical forage legumes and is native to South and Central America and Africa. Anthracnose, caused by the fungus Colletotrichum gloeosporioides (Penz.) Sacc., is a major constraint to the extensive use of Stylosanthes as tropical forage. Forty‐two accessions of S. guianensis were assessed with amplified fragment length polymorphism (AFLP) for genetic diversity and for resistance to anthracnose. In AFLP analysis, four selective primer combinations screened from 96 primer combinations were used to analyse these accessions, and a total of 225 clear bands were used for genetic similarity (GS) analysis, showing a 95.5% level of polymorphism on average. GS from 31.0% to 95.0% among the accessions was calculated with ntsys ‐pc software. The dendrogram was constructed with unweighted pair group method of averages (UPGMA) based on the AFLP data, and five clusters were defined at 48% GS. Two typical strains of C. gloeosporioides from Stylosanthes in China were used for anthracnose resistance screening. Most of the plant accessions showed variation in the reaction to two strains, and the correlation of resistance had a value of 0.904 (P < 0.01), suggesting common resistance to the two strains. The resistance accessions were randomly distributed in different groups of UPGMA clustering. These results demonstrate that AFLP analysis is an efficient method for evaluating the genetic diversity among S. guianensis accessions.     

大白菜亲缘关系研究

为更有效地利用现有资源、挖掘优异基因为育种和生产服务,对大白菜种质资源进行亲缘关系研究在理论和应用上都具有重要意义。本研究利用6对AFLP引物对89份来源和特性不同的大白菜（Brassica campestris L. ssp. pekinensis）种质进行系统评价和分类研究。6对AFLP引物共扩增出286条带,123条为多态性带,占总数的43.01%。不同引物组合扩增出的总带数和多态性带数差异较大,其中,引物组合E-ACT/M-CAC的扩增效果最好,多态性条带最多,占总带数的54.35%。聚类分析显示：中国大白菜种质资源遗传多样性丰富,参试种质间最大遗传距离为0.359,遗传背景广泛。以0.73的相似系数为阈值,所有材料被分为四大组群,其中浙江黄芽菜单独构成一个组群,明显地区分于其他大多数大白菜品种（“主体”组群）。在“主体”组群中,参试的台湾种质和大陆种质分为两个亚组,日本和韩国种质均被聚到了大陆种质亚组,表明供试的日本和韩国材料与中国大陆种质遗传亲缘关系密切。     

Amplified fragment length polymorphism for variety identification and genetic diversity assessment in oleander (Nerium oleander L.)

Oleander is a Mediterranean evergreen shrub found along watercourses, gravelly places and damp slopes. It is grown widely as an ornamental for its abundant and long-lasting flowering as well as its moderate hardiness. Genetic relatedness among 71 accessions, including commercial varieties, different sources of the same varieties, and selections from the wild were investigated using amplified fragment length polymorphism (AFLP). Nine primer combinations yielded a total of 603 bands of which 241 were polymorphic. Genetic similarities among accessions were calculated according to Jaccard's Similarity Index and used to construct a dendrogram based on the unweighted pair group method using arithmetic averages. Our results show that the AFLP technique, which can simultaneously and assay a large number of loci randomly distributed in the genome, is much more informative on the genetic relationship and origin of accessions than the limited number of morphological characters conventionally used for variety discrimination. Up to about 9% molecular genetic differentiation was detected among morphologically indistinguishable provenances of the same variety; this can be partly attributed to scoring error but mainly to somatic variation occurring during vegetative propagation. On the other hand lower genetic distance values were sometimes found among varieties which differ in morphological characters and are thus commercialised with different names. The possibility of considering the amount of genetic variation within a variety as the threshold value for discrimination of initial varieties and essentially derived varieties is discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

AFLP genetic diversity analysis in Russian wheat aphid resistant wheat accessions

Russian wheat aphid (RWA), Diuraphis noxia (Mordvilko), is an important insect pest which causes severe economic losses in wheat (Triticum spp.). Among the various U.S. RWA biotypes, biotype 1 (RWA1) and biotype 2 (RWA2) are the most prevalent and most virulent on cultivated genotypes. Although many sources of resistance to these biotypes are available among landraces, their relatedness should be characterized to permit their more efficient use in breeding programs. In this study, 38 hexaploid accessions resistant to biotype 1 and/or biotype 2 were evaluated for genetic diversity based on amplified fragment length polymorphisms (AFLP). Fifteen AFLP selective primer combinations were used to genotype these accessions, resulting in 893 amplicons. Of these, 274 (30.6%) informative polymorphic bands were used for genetic diversity analysis. Genetic similarity coefficients ranged from 0.47 to 0.87 among the resistant accessions, indicating high genetic diversity among them. Cluster analysis grouped the 38 accessions into two major clusters, I and II, including resistant lines for RWA1 and RWA2. The study indicated that accessions in the National Small Grains Collection conferring RWA1 or RWA2 resistance comprise a diversified population which should support introgression efforts and provide genetic diversity for future breeding for RWA resistance.