Genetic diversity of hexaploid wheat cultivars estimated by RAPD markers, morphological traits and coefficients of parentage

Estimates of genetic diversity can be based on different types of data. The aim of this research were to study genetic diversity among Croatian wheat cultivars by random amplified polymorphic DNA (RAPD) markers, morphological traits and pedigree records; to analyse differences between wheat cultivars from two breeding centres; and to evaluate usability of RAPD markers for estimation of genetic diversity among wheat cultivars in comparison with morphological traits and pedigree record data. Studies were conducted on 14 wheat cultivars and breeding lines from two breeding centres in Croatia. For the RAPD analysis 36 primers were screened and the 14 most polymorphic ones yielded 341 polymorphic bands. Twelve morphological traits were used for morphological analysis. Pedigrees were composed of seven generations of ancestors. RAPD markers showed a high level of polymorphism among the cultivars examined and the breeding lines. No significant correlations were observed among the methods tested.     

我国陆地棉品种的遗传多样性研究初报

选用１８个随机引物，对２１个陆地棉品种作ＲＡＰＤ分析，并对各品种的指纹图谱进行了聚类和相似性分析。结果表明大部分品种与其系谱吻合。研究结果充分展示了ＲＡＰＤ技术可以作为棉花品种分类和遗传多样性研究的可行方法。     

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.     

Genetic diversity in soybean as determined by RAPD and microsatellite analysis

The random amplified polymorphic DNA (RAPD) and microsatellite techniques were used to evaluate the genetic diversity among 18 soybean genotypes selected for a breeding programme to increase the protein content of varieties adapted for central European growing conditions. Out of 33 random primers used in RAPD reactions, only 12 showed polymorphism useful for characterization of these genotypes. In contrast, all 12 microsatellite primer pairs used in this study detected polymorphism with 2–6 alleles per locus. Similarity measures and cluster analysis were made using RAPD and simple sequence repeat (SSR) data, separately and together. The resulting dendrograms were compared with each other and with the available pedigree information as a control. The dendrogram derived from RAPD data showed some divergence from the pedigree information available for the lines. The dendrograms based on SSR data and SSR data combined with RAPD gave very good agreement with pedigree information. It can be concluded that the combined use of a limited number of RAPD and SSR markers is a useful and reliable means of evaluating genetic relationships of genotypes in the absence of pedigree data.     

A comparative analysis of the genetic relationships between rye cultivars using RFLP and RAPD markers

Summary The genetic similarities of eight closely related rye cultivars were estimated using two molecular marking techniques: restriction fragment length polymorphism (RFLP) and random amplified polymorphic DNA (RAPD). Cultivars were evaluated for variation by 11 random cDNA and genomic clones used in combination with four restriction enzymes and 40 decamer primers. A total of 53 polymorphic RFLP fragments and 94 polymorphic RAPD fragments were observed. Based on the presence/absence of fragments, two genetic similarity matrices were calculated which were then used in cluster analysis. Differences between pair of cultivars were observed in RFLP and RAPD dendrograms. RFLP analysis produced estimates of genetic relationships more in accordance with the partially known pedigree of the cultivars than did RAPD analysis. The use of bulk samples of DNA in these analyses affected the sensitivity of RAPD assays more strongly. Dendrograms which took into account all fragments produced, either by RFLP or RAPD, reflected better the relationships between cultivars than did dendrograms based on only one type of marker. This reflects the importance of the number of markers used in determining the genetic relationships between genotypes.     

Study of genetic diversity in safflower genotypes using agro-morphological traits and RAPD markers

This research was conducted to study the genetic diversity in safflower (Carthamus tinctorius L.) using agro-morphological traits and RAPD markers. Sixteen selected lines derived from landraces growing in various agro-climatic regions of Iran along with four exotic genotypes were evaluated in a randomized complete block design with three replications under field conditions. Days to emergence, days to initial flowering, days to flowering, days to maturity, plant height, branches per plant, capitula per plant, seeds per capitulum, 1,000-seed weight, seed yield per plant, seed yield, and reaction to powdery mildew (Leveillula taurica Arnaud) were evaluated in this study. Genetic diversity of the genotypes was assessed by RAPD markers. The results indicated significant differences among genotypes for the agro-morphological traits and clustering based on these traits classified the genotypes into five groups. Analysis of the RAPD markers revealed 15 polymorphic primers out of 50 used primers. Based on RAPD data, the highest genetic similarity was observed between the cultivars of “AC Sunset,” “AC Sterling” from Canada and the lowest relatedness observed between a local breeding line “E₂₄₂₈” and genotype “GE₆₂₉₂₃” from Germany. Cluster analysis based on RAPD markers and 54% coefficient of similarity divided the genotypes into five distinct groups. Comparing the clusters based on agro-morphological traits with those from molecular markers showed slight similarities. The finding of high genetic variation for agro-morphological traits and polymorphism at DNA level reveal that agronomic traits can be improved by selection programs.     

山东核桃良种SSR指纹图谱及分子身份证的构建——基于毛细管电泳分析

通过分析山东省内核桃良种的遗传背景,并构建其分子身份证,以期为核桃品种的选育、良种的质量追溯提供技术支持。本研究以17个核桃品种为试材,利用荧光SSR分子标记方法构建其指纹图谱,在分子层面进行数据分析。17个核桃品种间Nei’s遗传距离0.181~0.720,遗传相似系数0.267~0.867;筛选出5对SSR核心引物构建品种的指纹图谱,核心引物可分成HF9+HF15,HF3+HF9+HF14两组,每组均可完全区分17个品种;以指纹图谱为基础,与商品信息结合,数字化编码生成核桃品种分子身份证。17个核桃品种遗传基础较广,其聚类结果与地理位置相关性不大;5对核心引物多态性较好,可用于构建核桃指纹图谱;编码生成的分子二维码为核桃良种鉴定提供基础。     

Random amplified polymorphic DNA variation among German and Dutch asparagus cultivars

DNA polymorphism among nine cultivars of Asparagus officinalis L. was measured using random amplified polymorphic DNA (RAPD). Of 69 reproducible amplification products from 12 arbitrary decamer primers, 49 RAPD markers were polymorphic and could be used to distinguish six German and three Dutch asparagus cultivars. Even with very small sample sizes, genetic similarity measurements based on the RAPD data allowed accurate grouping of the nine cultivars into distinct clusters, with the exception of two individuals which clustered to closely related varieties. Two German cultivars showed high genetic similarity and were distinct from the remaining German varieties. The German and Dutch cultivars were clearly separated by a relatively large genetic distance.     

Prediction of breeding values for average fruit weight in mango using a multivariate individual mixed model

Mango is an important horticultural fruit crop and breeding is a key strategy to improve ongoing sustainability. Knowledge of breeding values of potential parents is important for maximising progress from breeding. This study successfully employed a mixed linear model methods incorporating a pedigree to predict breeding values for average fruit weight from highly unbalanced data for genotypes planted over three field trials and assessed over several harvest seasons. Average fruit weight was found to be under strong additive genetic control. There was high correlation between hybrids propagated as seedlings and hybrids propagated as scions grafted onto rootstocks. Estimates of additive genetic correlation among trials ranged from 0.69 to 0.88 with correlations among harvest seasons within trials greater than 0.96. These results suggest that progress from selection for broad adaptation can be achieved, particularly as no repeatable environmental factor that could be used to predict G × E could be identified. Predicted breeding values for 35 known cultivars are presented for use in ongoing breeding programs.     

我国短季棉品种的RAPD指纹图谱分析

本文选用了１８个随机引物，对２５个我国主要的短季棉品作物了ＲＡＰＤ多态分析。 各品种的指纹图谱进行了矛类和相似性分析。结果表明，大部分短季棉品种与其系谱吻合。主要从来自美国的金字棉口选育而成，它再次反映了我国现在推广的短季棉品种遗传基因比较狭窄，亟等发掘新的早熟棉基因资源。研究结果可对杂种优势利用的亲本选配提供理论依据。     

部分烟草种质遗传多样性与亲缘关系的ISSR标记分析

烟草遗传资源多样性与亲缘关系研究，是烟草遗传育种与起源演化研究的重要基础，本文首次应用ISSR标记，对烟草属（Nicotiana）4个种30份材料的遗传多样性进行分析。从70个ISSR引物中共筛选出16个多态性明显、条带清晰、反应稳定的引物，对30个样品DNA共扩增出309条谱带，平均每个引物扩增出19.31条带，多态性条带比率（PPB）达93.20%。种间遗传相似系数在0.26~0.96之间，表现出丰富的遗传多态性。系统聚类结果显示，N. glutinosa、N. suaveolens、N. gossei 3个野生种间存在较大的遗传差异，遗传相似系数在0.29~0.52之间；27份栽培品种种内遗传相似性相对较高，在0.54~0.96之间，显示出栽培种内的遗传基础相对比较狭窄，但其中白肋21、台烟7号与其他供试材料有较大的遗传差异。ISSR聚类分析表明，当L1取值为D = 0.475时，可将3个野生种与27份烟草栽培品种明显区分开，反映出种间的遗传差异；当L2取值为D = 0.776时，可将30份材料分为2个大类、3个小类和6个独立的个类，较好地揭示了烟草属种间或栽培种品种类型间的遗传多样性与亲缘关系，可为烟草遗传育种和遗传连锁图谱构建的杂交亲本选择提供科学依据。本研究还表明ISSR标记比RAPD标记具有更高的稳定性，在植物遗传多样性的分子标记或克隆研究中，可优先使用ISSR标记。     

Assessment of genetic diversity among wheat (Triticum aestivum L.) cultivars from a range of localities across Pakistan using random amplified polymorphic DNA (RAPD) analysis

Genetic diversity among 20 wheat genotypes/cultivars from diverse locations of Pakistan was studied using random amplified polymorphic DNA (RAPD) analysis. A total of 445 DNA fragments were amplified with50 random decamer primers 64.38% of which were polymorphic. Genetic similarity matrix based on Nei & Li's (1979) index detected coefficients ranging from 75.60% to 92.74%. These coefficients were used to construct a dendrogram using unweighted pair group of arithmetic means (UPGMA). The wheat genotypes were clustered into one major group (A) and two small groups (B and C). The most distant genotype in the dendrogram was PARC-1 that was 75.60% to 84.94% genetically similar with the other genotypes and clustered with PARC-3 which formed a group distantly related with the other clusters. Moreover, most of the wheat genotypes developed from the same breeding centre clustered in one group. It has been clearly shown that most of the cultivars except PARC-1possessed narrow genetic background. The information would be helpful for future genome mapping programs as well as for the application of intellectual breeder rights in the country. The study will also work as indicator for wheat breeders to evolve varieties with diverse genetic background to achieve sustainability in wheat production in the country. This revised version was published online in August 2006 with corrections to the Cover Date.     

Analyses of genetic diversity in Cuban rice varieties using isozyme, RAPD and AFLP markers

A survey of the genetic diversity among the major cuban rice cultivars was conducted using isozyme, RAPD and AFLP markers. Polymorphisms were detected for esterases, peroxidases, alcohol dehydrogenases and polyphenoloxidases systems; 21 RAPD primers and four AFLP primer combinations. Heterozygosity arithmetic mean value (H_av(p)), the effective multiplex ratio (EMR) and the marker index (MI), were calculated for isozyme, RAPD and AFLP markers. The mean value of genetic similarity among the different varieties was 0.92 for isozyme, 0.73 for RAPD and 0.58 for AFLP analyses. Thus, AFLP were able to detect polymorphisms with higher efficiency than RAPD (+15%) and isozyme (+34%). Data from the isozyme, RAPD and AFLP analyses were used to compute matrices of genetic similarities. The efficiency of the UPGMA for the estimation of genetic relatedness among varieties was supported by cophenetic correlation coefficients. The resulting values indicated that the distortion level for the estimated similarities was minimal. The correlation coefficients obtained by the Mantel matrix correspondence test, which was used to compare the cophenetic matrices for the different markers, showed that estimated values of genetic relationship given for isozyme and RAPD markers (r = 0.89), as well as for AFLP and RAPD markers (r = 0.82) were properly related. However, AFLP and isozyme data showed only moderate correlation (r = 0.63). Although the genetic variability found among the different cultivars was low, both RAPD and AFLP markers proved to be efficient tools in assessing the genetic diversity of rice genotypes. This revised version was published online in July 2006 with corrections to the Cover Date.     

Genetic variability among elite red clover ( Trifolium pratense L.) parents used in Chile as revealed by RAPD markers

Red clover (Trifolium pratense L.) is one of the main forage species of temperate regions. Cultivars of red clover are heterogeneous which makes their genetic analysis difficult. We applied RAPDs (Random Amplifed Polymorphic DNA) in order to assess the genetic relationship and levels of genetic variability existing among a group of 16 elite red clover parents organised in four subsets of 4 parents each. Out of 55 primers 21 provided reproducible results. A total of 135 reliable and polymorphic RAPD bands were detected which were used to estimate genetic distances among pair-wise combinations of elite parents. Nei and Li's similarity values ranged from 0.60 to 0.77, with a mean of 0.66, which reflects a rather high genetic variability among the genotypes evaluated. Lower levels of genetic variability, as detected by polymorphic loci and mean heterogeneity values, were detected in a subset of parents selected for resistance to the stem nematode. Cluster analyses resolved the different sets of parents in a manner consistent with what is known from their breeding origins. An Analysis of Molecular Variance detected substantial levels of variation within subsets of parents. RAPDs represent a valuable source of genetic information for red clover breeding programmes. This revised version was published online in August 2006 with corrections to the Cover Date.     

Identification and classification of aromatic rices based on DNA fingerprinting

Aromatic rices are preferred by the consumers all over the world due to its flavour and palatability. Although a large number of these collections are available, little systematic analysis of genetic diversity has been carried out. With the objective of identification and classification of aromatic rice genotypes, RAPD profiling was employed using 58 random decamer primers. Most of these primers (96.5%) detected polymorphism among the genotypes. Of the 465 amplified bands, 314 were polymorphic. Cluster analysis based on Jaccard's similarity coefficient using UPGMA grouped all the traditional tall, photosensitive, low yielding, long grained ‘basmati’ aromatics together. The short grained aromatic cultivars, formed a different cluster with high level of average similarity among themselves. The dendrogram based on 58 primers was highly similar to that based on 10 and 15 primers with matrix correlation (r) of 0.88 and 0.91, respectively. This suggested that a set of 10 primers can be employed for an initial assessment of genetic diversity in a large number of collections. All the rice genotypes included in the study could be distinguished from each other at the level of 19 to 186 polymorphic bands between individuals in pair wise comparison over all the 58 primers. Probability of identical profiles by chance suggested that about 10⁴¹ genotypes can be unambiguously differentiated by RAPD fingerprints obtained by 58 primers. A diagrammatic mode of presentation of DNA fingerprints of the aromatic rices based on 10 of the informative primers was developed. This revised version was published online in July 2006 with corrections to the Cover Date.     

用SSR标记分析‘赞皇大枣’遗传变异

研究旨在从分子水平揭示‘赞皇大枣’的遗传变异,以期为‘赞皇大枣’新品种选育提供有效的理论基础。利用SSR标记对395份‘赞皇大枣’样品进行PCR扩增,扩增产物经毛细管电泳技术检测,通过分析样品的指纹图谱数据,来评价样品间的遗传关系。16对SSR标记共扩增出51个等位基因,每个位点的等位基因数从2~9不等,平均每位点3.2个。产物片段大小范围为104~304 bp。稀有等位基因为16个,多态位点百分率为75%。395份样品中共得到28种两两相异的‘赞皇大枣’指纹图谱数据,即28种不同的基因型。对28种基因型进行聚类分析的结果表明,它们的遗传相似系数在0.7500~0.9804之间,平均值为0.9047。在遗传相似系数0.90处这些基因型被划分为5个类群。结果表明,‘赞皇大枣’群体内存在较丰富的的遗传变异,还有较大的选育出新品种的潜力。     

Criteria for the definition of similarity thresholds for identifying essentially derived varieties

The concept of essential derivation refers to cases where a new variety is obtained from an initial variety (IV), of which it retains the expression of the essential characteristics resulting from the genotype or combination of genotypes of the IV. Protecting plant breeder's rights can be facilitated by identifying a species‐specific threshold level of similarity above which the new variety could be suspected as essentially derived (EDV). Amplified fragment length polymorphisms and simple sequence repeats proved suitable for measuring genetic similarity. Three approaches have been proposed for the definition of thresholds. According to the ‘calibration principle’, the critical level to discriminate EDVs is defined based on similarities among closely related genotypes. Adopting the ‘tail principle’, the threshold is placed at a given percentile of the distribution of the similarities within a reference population of independent genotypes. Finally, with the ‘pedigree principle’, the threshold is set starting from the a priori definition of breeding techniques that lead to EDVs. The tools and the approaches described in this review represent the basis for developing protocols useful in handling disputes of essential derivation.     

Evaluation of genetic diversity among commercial cultivars, hybrids and local mango (Mangifera indica L.) genotypes of India using cumulative RAPD and ISSR markers

An assessment of genetic diversity studies was undertaken to understand the level and pattern of diversity in 65 mango (Mangifera indica L.) genotypes of India including 20 commercial cultivars, 18 hybrids, 25 local genotypes and two exotic cultivars based on qualitative and quantitative fruit characters as well as RAPD and ISSR profiles. A considerable variation was observed in respect of three important qualitative characters namely table quality, fruit attractiveness and storage life of ripe fruits and potentially superior genotypes for the above traits were identified. A wide variation was noticeable regarding metabolite composition of pulp of ripe mango fruit with respect to total soluble solids, total sugar, reducing sugar, acidity, sugar:acid ratio, ascorbic acid and phenolic content. Fifteen RAPD primers yielded 27 monomorphic and 129 polymorphic bands with percent polymorphism averaging 82.7%. Of a total 70 ISSR bands generated from eight ISSR primers, 60 bands (85.71%) were found to be polymorphic. Cumulative band data from these two methods precisely arranged accessions into eight clusters which correspond well with their pedigree relationship. UPGMA dendrograms drawn using RAPD, ISSR and cumulative data showed highly similar grouping of genotypes on the basis of their parental origin. No clear-cut geographical separation was revealed among East, West, North and South Indian mango cultivars by neither of these molecular markers nor their combinations. This supports the common gene pool origin of mango as well as operation of similar selection pressure as the cultivar preferences in these areas are largely similar.     

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。     

Relationship between heterosis and genetic divergence in 'Tongil'-type rice

Improving grain yield and quality of ‘Tongil’‐type rice (indica/japonica) continues to be a major breeding objective in Korea. In this study, genetic divergence among 13‘Tongil’‐type rice cultivars was evaluated and the relationship between genetic distance and hybrid performance in all possible nonreciprocal crosses between them assessed. The 78 F₁ hybrids together with the 13 parents were evaluated for eight traits of agronomic importance, including yield, in a replicated field trial. The 13 parents were examined for DNA polymorphism using 71 micro‐satellite or simple sequence repeats and 46 random decamer oligonucleotide primers. A total of 319 polymorphic variants were generated and, based on the polymorphism data, genetic distances (GDs) ranged from 0.021 to 0.437. Cluster analysis based on GDs revealed associations among cultivars which was in agreement with the pedigree data. Heterosis was observed in hybrids for most of the traits, and yield exhibited the highest heterosis among the eight traits examined. The correlation values of GDs with F₁ performance were mostly nonsignificant, except for yield, culm length and spikelets per panicle. The correlations of GDs with midparent and better‐parent heterosis were not significant enough to be of predictive value. These results indicate that GDs based on the microsatellite and random amplified polymorphic DNA (RAPD) markers may not be useful for predicting heterotic combinations in ‘Tongil’‐type rice and support the idea that the level of correlation between hybrid performance and genetic divergence is dependent on the germplasm used.