Allozyme Variability in the Tepary Bean, Phaseolus acutifolius A. Gray

The tepary bean (Phaseolus acutifolius) is the object of renewed interest because it possesses some interesting agronomic attributes such as tolerance to drought and salinity. In order to gain a better understanding of the organisation of its genetic diversity, we have examined patterns of diversity for nine polymorphic enzyme systems representing 12 loci, in a sample of 55 wild and 8 cultivated accessions. Several geographic patterns were identified, including an East-West differentiation, (across the Sierra Madre Occidental mountains), a North-South differentiation, and a localized differentiation. Wild teparies displayed a higher number of polymorphic loci, as well as a higher number of alleles per polymorphic locus compared to cultivars, consistent with previous observations indicating a reduction of phaseolin diversity on domestication. No clearcut separation with respect to isozyme differentiation was observed between P. acutifolius var. tenuifolius and var. acutifolius, further questioning the validity of this taxonomic separation based on leaflet shape.     

Genetic diversity of natural populations of Atriplex halimus L. in Morocco: An isoenzyme-based overview

Based on polyacrylamide gel electrophoresis, nine natural populations of Atriplex halimus L., a perennial shrub, collected in different regions of Morocco, were studied for their genetic variation using isoenzyme polymorphism of the highly active enzyme systems: esterases (EST), acid phosphatases (ACP) and glutamate oxaloacetate transaminase (GOT). Different allozyme frequencies from 7 different loci were obtained for all populations of this halophyte species. High levels of genetic diversity were revealed. The mean number of alleles per locus (A = 1.9–2.0), the percentage of polymorphic loci (p = 71.4–85.7) and the mean expected heterozygosity (H_e = 0.339–0.385) showed an important variability in all populations. Gene diversity was essentially explained by the within population component. The between populations differentiation accounted for 8% of the whole diversity (F_ST, averaged over all loci, is 0.08). The relationships among the 9 populations were inferred from the Nei’s genetic distances. Four major groups were formed. The northern population ‘Tanger’, forming a unique group, was highly divergent from the other groups. It appeared that the genetic distance between all groups was related to the geographic distance that separates them. This revised version was published online in August 2006 with corrections to the Cover Date.     

用SSR标记分析1949—2005年华南地区常规籼稻主栽品种遗传多样性及变化趋势

利用均匀分布于水稻基因组的300个SSR标记对95个华南地区不同年代常规籼稻主栽品种进行分析,研究该地区常规稻品种的遗传多样性及其变化趋势。检测到236个SSR标记有多态性,共获得776个等位基因,每个位点等位基因2~12个,平均3.29个,共有206个位点的等位基因数介于2~4个,占全部多态性位点的87.3%。多态性标记位点的PIC值平均为0.42,变化范围为0.041~0.790。不同染色体的位点多态性差异显著,其中第10染色体的位点平均等位基因数最多,PIC值最高,而第5染色体的位点平均等位基因数最少,PIC值最低;6个年代中,50~70年代育成品种包含等位基因数呈显著的上升趋势,70年代达最高值2.83,随后逐渐下降。分子方差分析(AMOVA)结果显示不同年代间遗传变异仅占总体变异的3.77%,但仍达极显著水平(P0.001)。不同年代育成品种的遗传距离(GD)呈下降趋势。聚类分析结果显示,在遗传相似系数(GS)为0.685处可将品种区分为5大类,表明华南地区各时期的常规稻品种遗传改良都是围绕少数骨干亲本进行的。试验结果显示,华南地区籼稻品种的遗传多样性狭窄且随年代而变化,70年代以后呈下降趋势,在今后的育种中应扩大亲本选材范围、拓宽育种亲本的遗传基础以提高育成品种的遗传多样性。     

Genetic relationships in cultivars of hop, Humulus lupulus L., determined by RAPD analysis

Random amplified polymorphic DNA (RAPD) was used to evaluate the genetic variability and relationship of 65 hop cultivars from all the major hop-growing regions in the world. Twenty-eight selected random primers used in the RAPD reaction generated an average of 38.6%) polymorphic fragments, which was sufficient to produce 47 different RAPD profiles among the cultivars examined. The level of genetic variability was much higher than previously reported. Genetic similarity was estimated and UPGMA cluster analysis was performed using the RAPD data. Cluster analysis separated the cultivars into genetically related RAPD groups which were compared with pedigree data and grouping of the hop cultivars by essential oil type. The RAPD groups, strongly supported by pedigree data, gave more precise information on the level and distribution of genetic variability within hop cultivars than characterization by essential oils. Cultivars were divided into American and European groups, supporting the distinction between two geo-graphically distinct hop germplasms. Five genetically distinct groups revealed differences within the European germplasm, reflecting past hop breeding practices which have been adopted in different regions. The use of RAPD markers for hop germplasm characterization and genetic diversity study is discussed.     

梨属植物等位酶遗传多样性研究

利用超薄平板微型聚丙烯酰胺凝胶的等电聚焦电泳技术,对286份梨材料进行了等位酶遗传变异分析。在8个酶系中共检测到19个清晰位点和82个等位基因,19个位点均为多态位点,位点最大等位基因数为6,体现出梨丰富的遗传种质多样性;不同的居群具有特有等位基因;通过82个等位基因可以将286份材料完全区分开,表明等位酶基因型指纹可用作梨品种区分与鉴定的依据。     

Genetic variation within and between two cultivars of red clover (Trifolium pratense L.): Comparisons of morphological,isozyme, and RAPD markers

Summary Morphological, isozyme and random amplified polymorphic DNA (RAPD) markers were used to estimate genetic variation within and between cultivars of red clover (Trifolium pratense L.), an important temperate forage legume. Two cultivars of red clover, Essi from Europe and Ottawa from Canada, were evaluated. Six monogenic morphological characters were observed for 80 plants from each of these two cultivars. All six morphological loci were polymorphic in the cultivar Essi whereas only four loci were polymorphic in the cultivar Ottawa. Forty plants from each cultivar were assayed for isozyme markers. A total of 21 enzyme-coding loci with 43 alleles was detected using twelve enzyme systems. Thirteen and nine of these loci were polymorphic in Essi and Ottawa, respectively. The mean number of alleles per locus was 1.81 in Essi and 1.67 in Ottawa. Seventeen random 10-mer primers were screened for RAPD markers. Nine primers which gave clear and consistent amplified products were used to assay 20 individuals from each cultivar. Each primer gave from 7 to 20 amplified bands with an average of 14.8 bands per primer. One hundred and eight of 116 putative loci were polymorphic in Essi and 90 of 98 loci were polymorphic in Ottawa. High within-cultivar variation was observed in both cultivars using both isozyme and RAPD markers. This high polymorphism makes these markers useful for germplasm characterization and genetic studies in red clover.     

河北省和中棉所育成陆地棉品种的遗传多样性分析

分别选取河北省历年育成的 1 9个陆地棉品种 ,中国农业科学院棉花研究所自 2 0世纪 70年代末到 90年代中期选育的品种中的 1 6个陆地棉品种 ,利用 RAPD分子标记研究各自育成品种的遗传多样性。 41个带型清晰、重复性好的多态性引物用于 RAPD分析 ,河北省育成的 1 9个品种得到 6 6个多态性位点 ,而中棉所育成的 1 6个品种扩增到 6 4个多态性位点。分析采用 Jaccard' s相似系数 ,使用 NTSYS- pc1 .80数据分析软件 ,非加权组平均法 (UPGMA)聚类。成对相似系数比较表明 ,中棉所在 2 0世纪 70年代末到 90年代中期育成的品种的遗传多样性水平高于河北省育成的品种。河北省育成的 1 9个品种的遗传基础很狭窄 ,斯字棉和岱字棉是河北省棉花两个最重要的基础种质 ,其中的 1 5个品种在聚类图上可以被分为两类 ,分别属于岱字棉和斯字棉系统。中棉所育成的 1 6个品种在聚类图上可被划分为三个类群。     

新疆彩色棉23个品种指纹图谱的构建及遗传多样性分析

以新疆截止2012年审定的23份新彩棉品种为材料,利用SSR标记进行DNA指纹图谱的构建和遗传多样性分析。从5000对SSR引物中,挑选出多态性高、稳定性好、均匀分布在棉花26条染色体上的52对引物,在23份新彩棉品种中筛选出核心引物47对,SSR扩增检测到多态性基因型位点数共计162个,每个标记检测到的基因型位点数在2~7之间,平均为3.45个;引物多态信息量(PIC)值介于0.4537~0.8686之间,平均值为0.7096。结果显示:在23份新彩棉品种中,14份品种采用特异或特征引物可以一次性区分开,其余9份品种需要采用引物组合来实现区别该品种与其他品种。最少选用18对特异引物及组合引物就可以完全区分开新彩棉1~23号品种。利用18对SSR标记构建了新彩棉1号至23号品种的指纹图谱。利用NTSYS-pcV2.10软件聚类分析表明:23个新彩棉品种遗传相似系数变化范围是0.3781~0.9298,平均为0.5511,表明新彩棉品种之间存在着丰富的遗传多样性。     

Genetic diversity among elite Bulgarian barley varieties evaluated by RFLP and RAPD markers

Genetic variation among five elite winter barley cultivars (H. vulgare L.) currently grown in Bulgaria was assessed at the molecular level using restriction fragment length polymorphism (RFLP) and randomly amplified polymorphic DNA (RAPD) markers. The present study sampled RFLPs in four well characterized multigene families in barley: the seed storage protein loci; the 18S, 5.8S and 26S ribosomal DNA loci; the loci coding for 5S ribosomal RNA and the loci coding subunit α of ATP-A complex in the mitochondrial genome. RFLPs were detected in three out of five investigated chromosomal loci in the barley cultivars studied. RAPD assay using arbitrary 10-base primers was applied to generate amplified length polymorphic markers in barley. Overall a total of 15 polymorphic phenotypes were found among the studied barley cultivars by using 11 out of 25 tested primers. All RAPDs were considered as dominant genetic markers except for two, where PCR and Southern blot analysis indicated the presence of codominant amplification products. Five RAPD polymorphisms in F₁ and F₂ progenies of the cross between Alpha and Obzor were inherited in Mendelian fashion. The determined values for the genetic variation proved a high genetic similarity among the tested cultivars. Genetic similarity (GS) calculated from RFLP and RAPD data ranged from 0.888 to 0.997 with a mean GS – 0.933. This revised version was published online in July 2006 with corrections to the Cover Date.     

野生稻与亚洲栽培稻的遗传多样性

为评价野生稻与亚洲栽培稻的遗传多样性及其变异关系,56对SSR引物被用于研究广泛地理分布的55份普通野生稻(其中32份O. rufipogon和23份O. nivara)和25份亚洲栽培稻(14份indica和11份japonica)样本。298个多态性位点被检出,占总扩增等位点的98.68%。野生稻多态性位点的百分比(平均达91%)及Nei’s遗传多样性值(h)明显高于亚洲栽培稻,表明普通野生稻比亚洲栽培稻具更丰富的遗传多样性。UPGMA聚类分析显示野生稻的两个类群(O. rufipogon和O. nivara)关系密切,但在遗传上存在明显的分化,支持其作为两个独立物种的分类观点。许多普通野生稻中籼粳分化尽管不很明显,然而亚洲栽培稻的籼粳亚种分化是明显的。亚洲栽培稻与多年生普通野生稻(O. rufipogon)关系更为密切,符合异源起源的遗传分化模式。     

Appraisal of RAPD and ISSR markers for genetic diversity analysis among cowpea (Vigna unguiculata L.) genotypes

The genetic variability and relationships among 11 cowpea genotypes representing two cultivars and nine elite genotypes were analyzed using 22 random amplified polymorphic DNA (RAPD) and nine inter-simple sequence repeat (ISSR) markers. ISSR markers were more efficient than RAPD assay with regards to polymorphism detection. But the average numbers of polymorphic loci per primer and resolution power were found to be higher for RAPD than for ISSR. Also, the total number of genotype specific marker loci, Nei’s genetic diversity, Shannon’s information index, total heterozygosity, and average heterozygosity were prominent in RAPD as compared to ISSR markers. The regression test between the two Nei’s genetic diversity indices showed low regression (0.3733) between ISSR and RAPD + ISSR-based similarities but maximum (0.9823) for RAPD and RAPD + ISSR-based similarities. The RAPD- and ISSR-generated cultivar- or genotype-specific unique DNA fingerprints able to identify the most diverse genotypes. A dendrogram constructed based on RAPD and ISSR combined data indicated a very clear pattern of clustering according to the groups (cultivars and elite genotypes). The results of principal coordinate analysis were comparable to the cluster analysis. Cluster analysis showed that most diverse genotypes (GP-125 — small size with good seed quality; GP-129, GP-90L — big size with poor seed quality) were separated from moderately diverse cultivars and genotypes. The genetic closeness among GP-129 and GP-90L, JCPL-42, and JCPL-107 could be explained by the high degree of commonness in these genotypes.     

Genetic Relationships in Malus Evaluated by RAPD 'Fingerprinting' of Cultivars and Wild Species

The potential use of RAPD markers for taxonomic studies in Malus was investigated using 18 accessions of wild species and 27 apple cultivars. 29 preselected random decamer primers were applied to three sets of Malus genotypes. Random amplified polymorphic DNA (RAPD) ‘fingerprints’ were analysed for polymorphic amplification fragments, and coefficients estimating genetic similarity were calculated on the basis of about 50 polymorphic RAPD loci in each set of genotypes. Cluster analysis by an unweighted pair-group method with arithmetic averages (UPGMA) revealed that, in the cultivars, the molecular classification was in good agreement with the known lineage. A dendrogram generated for the wild species gave relationships that were, in principle, in accordance with the known phylogenetic information. Closely related species from section I were clearly distinguishable from those of sections III and IV. On the molecular level, a high degree of genetic diversity was found among both different apple cultivars and wild species of the genus Malus. The results gave additional evidence for the hypothesis that M. pumila and M. sylvestris were involved in the origin of the cultivated apples.     

Molecular characterization of novel resynthesized rapeseed (Brassica napus) lines and analysis of their genetic diversity in comparison with spring rapeseed cultivars*

Resynthesized (RS) rapeseed generated from interspecific hybridization between suitable forms of Brassica rapa L. (syn. campestris; genome AA, 2n = 20) and B. oleracea L. (CC, 2n = 18) represents a potentially important resource to expand genetic diversity in the narrow gene pool of oilseed rape (B. napus L., AACC, 2n = 38). In this study 165 RS rapeseed lines originating from crosses between an Indian Yellow Sarson (B. rapa ssp. trilocularis) and five different cauliflower (B. oleracea convar. botrytis) cultivars were studied using amplified fragment length polymorphism (AFLP) markers and their genetic diversity was compared in relationship to an assortment of 40 diverse spring oilseed and fodder rape varieties. Using three AFLP primer combinations, a total of 467 polymorphic bands were scored. Cluster analysis allowed differentiation among the different RS lines, which, as expected, were genetically highly divergent from the cultivars. The genetic diversity of the material is discussed in relation to its morphological variability with a view to the implementation of RS lines in oilseed rape breeding.     

RAPD和AFLP标记分析中国马铃薯主要品种的遗传多样性

采用RAPD 和AFLP两种方法分析71份中国各地马铃薯主要品种,均可将其完全区分,并可对其进行分子鉴定;证明中国马铃薯主要品种遗传组成上差异小,遗传多样性差。由于标记方法的原理差异和栽培马铃薯遗传组成复杂性,用2种方法分类的结果有所差异。AFLP标记检测获得的Shannon-weaver指数和Simpson指数均高于RAPD标记检测的结果,AFLP标记检测多态性的能力远高于RAPD标记。AFLP标记平均每个引物组合检测到100.1个位点,其中54.9条为多态性位点,而RAPD标记的相应数据分别为12.5和9.8个。不同的标记方法在马铃薯遗传多样性研究中存在差异,聚类结果从分子水平反映了中国现有主要马铃薯品种遗传基础的狭窄。     

Genetic diversity among tea cultivars from China, Japan and Kenya revealed by ISSR markers and its implication for parental selection in tea breeding programmes

Tea plant [Camellia sinensis (L.) O. Kuntze] is an important beverage crop in the world. In recent years many clonal tea cultivars have been released, and they play major roles in improving the production and quality of tea. It is important to understand the genetic diversity and relatedness of these cultivars to avoid inbreeding and narrow genetic basis in future tea breeding. In the present study, genetic diversity and relationship of 48 tea cultivars from China, Japan and Kenya were evaluated by inter‐simple sequence repeat (ISSR) markers. A total of 382 ISSR bands were scored, of which 381 (99.7%) were polymorphic. The ISSR primers showed high ability to distinguish between tea cultivars according to their high Resolving Power (R_P) with an average of 7.4. The mean of Nei’s gene diversity (H) and Shannon’s information index (I) were 0.22 and 0.35, respectively. More abundant diversity was revealed among cultivars in China than those in Japan and Kenya. Within Chinese populations, the level of diversity in east China was higher than that in other regions. The coefficient of genetic differentiation (G_ST) was 0.202, which indicates a high degree of genetic variation within populations. This result was further confirmed by analysis of molecular variance, which revealed the variance component within the populations (92.07%) was obviously larger than that among populations (7.93%). The level of gene flow (N_m) was estimated to be 2.0. This could be explained by frequent natural cross‐pollination and seed dispersal among tea populations. The pairwise similarity coefficient between the cultivars varied from 0.162 to 0.538. A dendrogram of 48 tea cultivars was constructed where all the tested cultivars were divided into two groups. Our data show that the genetic relationship among tea cultivars can be determined by the ISSR markers. This will provide valuable information to assist parental selection in current and future tea breeding programmes.     

黑龙江省主栽大豆品种遗传多样性和群体结构分析

利用187对SSR标记对近25年（1992-2017）在黑龙江省栽培的202个大豆品种进行遗传多样性和群体结构分析。结果表明,从试验材料的基因组DNA中扩增出多态性位点808个,平均每对引物扩增出多态性位点4.42个;多态性位点最多的引物是satt703和satt311,均为10个;等位变异频率最高的引物是satt417和satt575,等位变异频率均为99.5%。供试品种间的遗传相似系数为0.283~0.930,平均值为0.519。同一个育种单位育成的部分品种具有较高的遗传相似性。群体结构、主坐标分析和NJ聚类将202个品种划分的结果是一致的,均为3个类群。类群中的部分材料血缘不是独立的,而是相互渗透的。     

红花品种遗传多样性的SRAP分析

利用相关序列扩增多态性SRAP(Sequence-related amplified polymorphism)技术对8份国内不同来源以及8份国外不同国家的,共计16份红花栽培种进行遗传多样性分析。选用20对多态性较高的引物组合对材料进行PCR扩增,而后进行聚丙烯酰胺凝胶电泳,获得多态性条带。共获得354条清晰的条带,其中,135条为多态性条带,多态性比率为38.14%。遗传相似系数在0.3~0.9之间,平均相似系数为0.65,表明红花种内不同栽培种之间具有较高的遗传多样性和相似性。     

Assessment of genetic diversity in cabbage cultivars using RAPD and SSR markers

Genetic relationship and diversity among seven cabbage cultivars were analyzed using RAPD and SSR markers. These cultivars are of great commercial value in India and are confirmed for their reaction to black rot caused by Xanthomonas campestris pv. campestris. However, so far the extent of genetic diversity and relatedness has not been studied in these cultivars. A total of 17 selected RAPD primers generated 90 bands, 76 of which were polymorphic (84.44%). In addition, 27 selected SSR primers generated 67 amplified bands with 59 of which were polymorphic (87.6%). Though both the marker techniques were able to discriminate the cultivars effectively, analysis of combined data of markers (RAPD and SSR) resulted in better distinction of cultivars. By combining both the markers, a total of 157 bands were detected of which 135 bands (85.98%) were polymorphic, i.e. an average of 5.95 bands per primer. High level of polymorphism (> 85%) recorded with two different marker systems indicated a high level of genetic variation existing among the cultivars. Genetic relationship estimated using similarity co-efficient (Jaccard’s) values between different pairs of cultivars varied from 0.21 to 0.77 in RAPD, 0.42 to 0.82 in SSR, and 0.43 to 0.89 with combined markers. A high correspondence had been recorded between the values of genetic variations generated by UPGMA, clustering, and scatter plot diagrams. The cultivars ‘January King Sel. Improved’ and ‘Golden Acre’ are highly divergent cultivars as demonstrated by both the marker systems.     

栽培罗汉果遗传多样性的RAPD分析

罗汉果(Siraitiagrosvenorii)是单性、雌雄异株的葫芦科球根多年生植物,是特产于我国南部的重要经济植物。其果实是传统中药,果实中还含有一种低热量的天然甜味剂。我们从罗汉果的主产地广西永福县、临桂县和融安县采集了13份雄株和8个品种的62份雌株栽培罗汉果样品,用21个随机引物对这75份样品进行了RAPD分析。共得到130个位点,其中92个(70.77%)是多态性的。用POPGENE软件统计了主要品种及雄株的多态位点百分率和Shannon信息多样性指数;根据样品间的Dice相似性系数,分别对雌株和雄株做了主成分分析(PCA);用UPGMA法进行了单株聚类分析。结果表明主要栽培品种青皮果、红毛果和爆棚果的遗传多样性很低,它们的品种内样品间相似性系数平均值分别为0.961、0.945和0.966,Shannon信息多样性指数分别为0.0997、0.1014和0.0571,这种遗传多样性很低的现状是由于栽培罗汉果主要采用营养繁殖方法进行繁殖的结果。而茶山果和冬瓜果具有相对较高的遗传多样性,样品间相似性系数平均值分别为0.868和0.857,Shannon信息多样性指数分别为0.1876和0.1855。雄株遗传多样性较高,样品间相似性系数平均值为0.873,Shannon信息多样性指数为0.2135。有必要扩大栽培罗汉果的遗传基础。     

新疆冬小麦地方品种与育成品种基于SNP芯片的遗传多样性分析

新疆是我国西北重要的小麦优势产区和消费区。解析新疆冬小麦地方品种与育成品种之间的遗传多样性和亲缘关系,对新疆冬小麦育种中杂交组合的合理选配以及后代选择具有重要的指导意义。本研究利用小麦55K SNP(single nucleotide polymorphism)芯片对134份新疆冬小麦地方品种及54份育成品种进行全基因组扫描,估算其品种间的遗传距离,揭示其遗传多样性。结果表明,所有SNP位点的多态性比率达到95.62%(50,743/53,063)。每条染色体分布1068～2616个多态性位点,多态性标记在基因组间分布呈现ABD。188个品种间的两两遗传距离在0.002～0.723之间,平均为0.378。其中134个地方品种两两之间的遗传距离在0.002～0.400之间,平均为0.070; 54个育成品种两两之间的遗传距离在0.004～0.337之间,平均为0.114; 134个地方品种与54个育成品种之间的遗传距离在0.605～0.723之间,平均为0.699。聚类结果显示可将所有材料分为10个不同类群。综合SNP和系谱分析,育成品种与地方品种之间的遗传差异最大,其次是育成品种之间,而地方品种之间遗传差异最小。鉴于育成与地方品种之间较大的遗传差异,新疆冬小麦品种可以利用地方种来丰富其育种的种质基础,拓宽遗传背景,进而提高当地小麦育种水平。本研究为新疆冬小麦品种选育和改良提供了重要的理论指导。