Evaluation of genetic diversity among some Iranian wild asparagus populations using morphological characteristics and RAPD markers

An evaluation of genetic diversity in 39 wild asparagus populations was carried out using morphological and RAPD markers. A combination of morphological traits and random RAPD primers was used to examine the level of genetic variation and polymorphisms among the populations. A factor analysis using Ward's method on mean values of morphological characteristics indicated seven main factors resulting in four groups. Analysis of polymorphic bands using Jaccard's similarity coefficient indicated that genetic similarity ranged between 0.71 and 0.29. At a similarity level of 0.64, the populations were divided in three sub-clusters, containing 34, four and one populations, respectively. Significant regression associations were found between 21 morphological characteristics and 18 RAPD markers, revealing some informative markers associated with some traits. The highest R² was related to 18 RAPD markers associated with gender (53.5%) that among them BA-04₂₀₀₀ had a maximum R². The results showed that Iranian wild asparagus with its high levels of genetic variation could be considered as a valuable gene pool for future asparagus breeding programs. Furthermore, it could be inferred that morphological characteristics and RAPD markers are suitable tools to discriminate asparagus populations for the evaluation of genetic diversity.     

Genetic structure of Buxus sinica var. parvifolia, a rare and endangered plant

Buxus sinica var. parvifolia, a rare and endangered tree species in some semitropics alpine areas of China, plays an important role in the maintenance of the landscape and ecosystem. In this study, RAPD and ISSR markers were used to investigate the genetic diversity and structure of five natural populations and one tamed population of B. sinica var. parvifolia. 21 RAPD primers amplified 209 bands with 167 (79.90%) polymorphic and 21 ISSR primers amplified 518 bands with 467 (90.15%) polymorphic. The genetic diversity, estimated by Shannon’ index, was 0.4343 (by RAPDs) and 0.3661 (by ISSRs). Both RAPD and ISSR analyses revealed a high level of genetic diversity in natural populations of B. sinica var. parvifolia. Furthermore, analysis of molecular variance (AMOVA) was used to apportion the variation within and between populations. The proportion of variation attributable to within-population differences was very high (69.2% by RAPDs; 84.51% by ISSRs). Moderate differentiation was detected among populations using RAPDs (30.80%), while only a small amount of variation (15.49%) was detected among populations using ISSRs. We suggest that the present genetic structure is due to high levels of environmental variability and gene flow, which still need further study to confirm. Conservation measures are suggested, including in situ and ex situ strategies, based on the observed population genetic information.     

Phylogenetic relationships in Origanum spp. based on rDNA sequences and intra-genetic variation of Greek O. vulgare subsp. hirtum revealed by RAPD

Origanum species are among the most widely spread herbs in the Mediterranean basin. Eventhough they are used as a spice, evaluation of their genetic diversity and evolution has only recently drawn attention. In order to study phylogenetic relationships, 14 ITS1-5.8S-ITS2 clones belonging to the most common Origanum species were sequenced and a parsimony tree was constructed, using the approximate likelihood ratio test. All Origanum species were clearly separated from allied genera of the Mentheae tribe while a clear distinction between the Greek and the Spanish accessions was revealed. In addition the germplasm variability of the most common Greek oregano (O. vulgare subsp. hirtum) was investigated using the RAPD markers. The use of 10 random decamers resulted in 133 unambiguous and reproducible bands detected across 27 entries. Two main groups were identified by the UPGMA clustering using Jaccard's similarity coefficient, and major genetic dissimilarities among Greek O. vulgare subsp. hirtum populations and O. onites/O. virens species were detected. Analysis of molecular variance revealed that genetic variability is distributed mainly within populations; however, significant Φ_st values were detected between different geographical localities, supporting noteworthy genetic differentiation among O. vulgare subsp. hirtum populations.     

中国樱桃地方种质资源表型性状遗传多样性分析

以源自中国樱桃主产区7个群体,80份地方种质资源为试材,采用巢式方差及聚类分析法对其叶片、果实及果核26个表型性状进行多样性分析。结果表明:(1)质量性状之间存在较大的多样性差异,Shannon-Wiener多样性指数范围在0.61~1.64之间,其中果实形状、果皮颜色、叶片及果核剖面形状等表现出较高的遗传多样性。(2)数量性状无论是在群体间还是群体内均表现出极显著差异,14个数量性状的平均变异系数为9.93%,其中果柄长度的变异系数最高(24.80%),果核顶面观指数最低(6.61%)。在7个群体中,云南的种质材料变异系数最大,为12.92%;安徽的最小,为7.42%。(3)对14个数量性状的主成分分析表明,前5个主成分的累积贡献率达80.9434%,能反应数量性状的基本特征。(4)基于Nei’s遗传距离的聚类分析以及主坐标分析将80份材料分为2个大类,分布于山东、河南和安徽群体的种质资源聚为一类,为华北类群,分布于四川、云南、贵州和重庆群体的聚为另一类,为西南类群。研究结果表明不同地理分布群体的中国樱桃表型多样性存在差异,而这种遗传差异可能是由于来自不同的驯化位点而造成的。     

Genetic variability in wild vs. cultivated Eruca vesicaria populations as assessed by morphological,agronomical and molecular analyses

In this study, the genetic diversity of 50 individuals of rocket, Eruca vesicaria, from five accessions, four of them wild type collected from different parts of Spain and one commercial, were evaluated using morphological, agronomical and inter simple sequence repeat DNA (ISSR) data. Molecular analysis was carried out using the ISSR technique with 20 primers. Out of these 20 primers, nine were polymorphic, producing a total of 395 DNA bands, 247 of which were polymorphic among the accessions. A dendrogram drawn on the basis of a similarity matrix using the UPGMA algorithm revealed that the 50 samples of rocket plants could be classified into three major clusters at a Nei's genetic distance of 0.36. The experiment shows that molecular markers such as ISSR are a good instrument for distinguishing and selecting rocket accessions to group different wild populations. In general, a high variation was observed for most of the 16 morphological and 6 agronomical traits showing significant differences. Some morphological traits such as leaf length, petiole length and lamina width explained 69.1% of the whole variation observed in the populations, and some agronomical traits such as leaf area, nitrate and chlorophyll contents accounted for 65.7%, but the clusters generated by means of agronomical and morphological variables were less evident than when ISSR markers used. Some accessions showed good qualities, such as small leaves, high chlorophyll content, late-flowering or low nitrate content. All these parameters, together with the high degree of genetic homogeneity found, could make the local accessions good candidates for a future breeding programme.     

珙桐遗传多样性的AFLP 分析

 采用7对AFLP引物对中国珍稀濒危植物珙桐（Davidia involucrata Baill.）的6个天然居群和2个人工居群的229份材料进行扩增,共检测到100 ~ 500 bp的位点506个,其中多态性位点488个,多态性位点百分率96.44%;各居群多态位点百分率为52.77% ~ 79.25%,平均为67.00%。在物种水平上,珙桐的Nei’s基因多样性H_s = 0.3429,Shannon’s遗传多样性信息指数I = 0.5107。而居群水平上,各居群的Nei’s基因多样性介于0.2229 ~ 0.3389,各居群的平均Nei’s基因多样性H_s = 0.2748,各居群Shannon’s遗传多样性信息指数（I）介于0.3219 ~ 0.4877,平均Shannon’s遗传多样性信息指数I = 0.3995。居群间基因间分化度（G_st）为21.60%,基因流N_m为1.8。说明珙桐的遗传多样性较高,这8个居群间存在一定的遗传分化和基因流动,但遗传分化主要存在于居群内。在珙桐与其变种光叶珙桐之间未找到差异标记。     

观赏海棠‘王族’自然杂交后代的遗传多样性分析

 利用叶、花形态性状分析及AFLP标记技术,对50份‘王族’（Malus‘Royalty’）观赏海棠的自然杂交后代进行遗传多样性分析。结果表明：自然杂交后代叶和花18个形态指标的变异系数在3.44% ~ 42.68%之间,其中托叶长、托叶宽、叶面积、叶片鲜样质量、花瓣宽、花梗长的变异系数在20%以上,多样性特征显著。9对AFLP分子标记引物共扩增得到349条带,其中多态性条带303条,占总数的86.8%。Nei’s遗传多样性指数在0.24 ~ 0.36之间,Shannon信息指数在0.36 ~ 0.53之间。分析认为,‘王族’自然杂交后代不论是在形态水平上还是在分子遗传水平上都具有丰富的多样性,尤其是形态上的差异更为显著。这些变异为观赏海棠选择育种提供了良好材料。     

蜡梅天然群体遗传多样性的SRAP标记分析

 在蜡梅[Chim onanthus praecox (L.) Link ] 分布区内选取5个天然群体, 采用筛选出的7对SRAP (sequence-related amplified polymorphism) 引物进行遗传多样性分析。结果显示: 多态性条带百分率为23.4% , 多态位点比率( PPL ) 为83.41% , Nei's基因多样性指数(h) 为0121～0125 (均值为0.22) ,Shannonps信息指数( I) 为0.34, 群体分化系数(Gst) 为0.17, 群体间总的基因流的估算值Nm为2.40。AMOVA分析显示群体内方差分量的贡献率占79.89% , 而群体间方差分量的贡献率占20.11% , 区域间占32.39% , 区域内67.61%。Nei's (72) 群体遗传距离的UPGMA聚类分析显示群体间的遗传距离与群体的地理距离关系一致, Mantel分析相关系数为0.89。基于试验分析结果, 蜡梅种质资源保护和利用策略为在优先保护现有群体的基础上, 加强筛选和保存群体内的变异样本。     

Genetic diversity of bitter gourd (Momordica charantia L.) genotypes revealed by RAPD markers and agronomic traits

Bitter gourd or bitter melon (Momordica charantia L.) is considered as minor cucurbitaceous vegetable in spite of having considerable nutritional and medicinal properties. Although some reports on genetic diversity based on morphological characterization are available, no work has been conducted to estimate genetic diversity using molecular markers in this crop. In the present study, 38 genotypes of M. charantia including few commercially cultivars collected from different parts of India based on agro-ecological zones were analysed for diversity study both at morphological and molecular levels. Genomic DNA was extracted from young healthy leaves following the procedure of Doyle and Doyle [Doyle, J.J., Doyle, J.L., 1990. A rapid DNA isolation procedure from small quantity of fresh leaf material. Phytochem. Bull. 119, 11–15]. Pair-wise comparison of genotypes was calculated as per the procedure of Jaccard [Jaccard, P., 1908. Nouvelles recherches sur la distribution florale. Bull. Soc. Vaud. Sci. Nat. 44, 223–270]. Dendrogram was constructed using the unweighted pair group method with arithmetic averages (UPGMA) and the computation for multivariate analysis was done using the computer programme NTSYS-pc Version 2.0 [Rohlf, F.J., 1998. NTSYS-pc Numerical Taxonomy and Multivariate Analysis System, Version 2.01. Exeter Software, Setauket, NY, USA]. Diversity based on yield related traits and molecular analysis was not in consonance with ecological distribution. Among 116 random decamer primers screened 29 were polymorphic and informative enough to analyse these genotypes. A total of 208 markers generated of which 76 (36.50%) were polymorphic and the number of bands per primer was 7.17 out of them 2.62 were polymorphic. Pair-wise genetic distance (GD) based on molecular analysis ranged from 0.07 to 0.50 suggesting a wide genetic base for the genotypes. The clustering pattern based on yield related traits and molecular variation was different.     

紫丁香天然群体遗传多样性的AFLP分析

 抽取内蒙古大黑山、辽宁北票和山西五老峰3个紫丁香(Syringa oblata Lindl. ) 天然群体为拟似群体, 用筛选出的8对引物, 对群体家系组成的72个样品进行AFLP分析。Nei (1973) 基因多样性指数h = 0.2287, Shannon's信息指数I = 0.3464; 种级遗传多样性Ht = 0.3522, 群体内遗传多样性Hs =0.2287, 群体间遗传多样性Dst = 0.1235, 群体分化系数Gst = 0.3508, 群体间总的基因流的估算值Nm =0.9253, 群体遗传变异的AMOVA分析表明群体的遗传多样性主要分布在群体间, 群体间方差分量的贡献率占51.53%。基于Nei (1972) 遗传距离的UPGMA聚类分析显示群体间的遗传距离与群体的地理距离关系一致。结果表明: 紫丁香群体间的遗传分化较大, 其天然群体缺失造成的现有分布的间断性和地理隔离以及高自交率是导致其群体间的高度分化的主要因素。在目前分布残缺, 群体丢失严重的情况下, 紫丁香种质资源保护和利用策略应为采取优先保存尽可能多的群体。     

Characterization of pea accessions by SRAP's markers

Sequence-related amplified polymorphism (SRAP) and morphological markers to estimate the genetic relations among forty pea varieties (Pisum sativum L.) were studied. Data on 15 morphological traits were collected and analyzed. A total of 162 polymorphic SRAP's bands were scored using seven combinations of primers. Cluster analysis and both principal component and principal coordinate analysis were carried out. The varieties were grouped in four clusters through procrustes generalized analysis. Relationships among varieties revealed by molecular markers were significantly correlated with those based on the agronomic traits, suggesting that the two systems give similar estimates of genetic relations among the varieties. Parents selection depend on the specific objectives in further breeding programs.     

Genetic diversity and population genetic structure of pomegranate (Punica granatum L.) in Iran using AFLP markers

Pomegranate (Punica granatum L.) is one of the oldest known edible fruits. It is native to Iran and spread from Iran to other areas. In this study amplified fragment length polymorphism (AFLP) was used to detect intra- and inter-population genetic diversity of pomegranate. A group of 67 accessions belonged to 4 populations from Iran was studied using eight primer combinations. A total of 221 scorable bands were amplified, of which, 118 (54.13%) were polymorphic. Resolving power (Rp) ranged from 5.70 to 9.21, and the average of polymorphism information content (PIC) per primer pair was 0.40. According to Nei's gene diversity and allelic statistics, Isfahan population had a highest genetic diversity (H = 0.3646, I = 0.5327, N_e = 1.6467). Coefficient of gene differentiation between populations (GST) was 0.124, indicated that mainly proportion of genetic variation (87.6%), was within populations and the remaining (12.4%) of the variation was among populations that, also supported by analysis of molecular variance (AMOVA). The gene flow (Nm) varied from 0.969 to 10.404 between pair-wise populations and was 3.504 among all of the populations. The Jaccard similarity coefficient between individuals ranged from 0.26 to 0.88. The UPGMA dendrogram clustered all 67 accessions into 6 groups. In some cases accessions from same region were grouped together but in most cases, there was gene exchange. To study the genetic relationships among populations, a principal coordinate analysis (PCoA) based on Nei's genetic distances was performed. Results of this study showed that AFLP marker can be a useful tool for investigating the genetic diversity of pomegranate genotypes.     

广西野生濒危植物掌叶木遗传多样性的ISSR与SRAP分析

采用ISSR和SRAP技术对中国野生濒危植物掌叶木[Handeliodendron bodinieri（Lévl.）Rehd.]的5个野生居群的95份材料进行分析,10条ISSR引物共扩增得到114个条带,其中多态性条带68个,多态性条带百分率（PPB）59.65%,掌叶木在物种水平上的Nei’s基因多样性（H）为0.1817,Shannon’s遗传多样性信息指数（I）为0.2792;观测等位基因数（Na）为1.5965,有效等位基因数（Ne）为1.2984,居群间基因间分化度（Gst）为35.17%。15组SRAP引物共扩增出292个条带,其中多态性条带269个,多态性条带百分比为92.12%,物种水平上,掌叶木的Nei’s基因多样性（H）为0.3171,Shannon’s遗传多样性信息指数（I）为0.4758;观测等位基因数（Na）为1.9212,有效等位基因数（Ne）为1.5380,居群间基因间分化度（Gst）为23.17%。结果显示两种标记均能检测到掌叶木具有较高的遗传多样性,掌叶木不同居群间存在一定的遗传分化和基因流动,但遗传分化主要存在于居群内。     

Assessment of genetic diversity in cashew germplasm using RAPD and ISSR markers

Diversity and genetic relationship in 100 cashew germplasm accessions were analyzed by using RAPD and ISSR markers. Using 10 selected RAPD primers 60 bands were generated, of which 51 bands were polymorphic (85%), and with 10 selected ISSR primers 67 amplified bands were observed with 58 polymorphic bands (86.6%). Though both kinds of markers discriminated the accessions effectively, analysis of combined data of markers (RAPD + ISSR) resulted in better distinction of accessions. By combining markers, a total of 127 bands were detected, of which 109 bands (85.8%) were polymorphic and produced on an average of 5.45 polymorphic bands per primer. Primers with high polymorphic information content and marker index were identified for discriminating accessions. High percentage of polymorphism (>85%) observed with different markers indicated high level of genetic variation existing among the accessions. Genetic relationship estimated using similarity co-efficient (Jaccard’s) values between different pair of accessions varied from 0.43 to 0.94 in RAPD, 0.38 to 0.89 in ISSR and 0.43 to 0.87 with combined markers suggested a diversity (dissimilarity) ranging from 6 to 57%, 11 to 62% and 13 to 57% respectively and the diversity skewed around 50% indicated moderate diversity. The cluster analysis with UPGMA method separated the accessions broadly into 13 clusters and in that three into smaller clusters. Some correspondence between the molecular groupings and the morphological clusters were observed. Among the accessions, NRC-142 and NRC-12 were highly divergent and NRC-231 and NRC-232 were genetically similar.     

Morphological and molecular variability in some Iranian almond genotypes and related Prunus species and their potentials for rootstock breeding

In this study, in order to know the variability for a rootstock breeding program genetic diversity and relationships among 55 Iranian almond genotypes and seven related Prunus species were investigated. Morphological and molecular analyses were used. Principal component analysis showed that three components explained 67.6% of the total morphological variation for the first year and 68.06% for the second year of the study. Leaf traits were predominant in the first component and contributed most of the total variation. Leaf length and width, as well as, leaf area were highly correlated with each other and correlated to vigor. Also a negative correlation was found between leaf length/width ratio and vigor. Ward's method was used to construct cluster from morphological data which allocated individuals into their respective species. Out of 100 pre-screened RAPD primers, 16 with reproducible bands and maximum polymorphism were selected. Two-hundred and sixty bands were scored of which 250 of them were polymorphic. Average value of polymorphism per primer was 95.81% and maximum value for polymorphism (100%) was obtained from TIBMBA-14, TIBMBA-17, TIBMBB-05, TIBMBB-08, TIBMBD-09, and TIBMBD-10. On the other hand, the minimum value was obtained from TIBMBB-16 (86%). Primer TIBMBB-5 gave the maximum number of bands (25 fragments) and the minimum obtained from TIBMBE-18 (11 fragments). Genetic similarity based on Jaccard's coefficient ranged from 0.28 to 0.79 with an average of 0.53. Molecular analysis revealed a high degree of separation among samples regarding their geographical origin. Correlation between two approaches was low (R = −0.38). High molecular and morphological variability indicated that this collection includes rich and valuable plant materials for almond rootstock breeding.     

云南蔷薇属部分种质资源的SSR遗传多样性研究

利用简单重复序列SSR（Simple Sequence Repeat）标记技术对42份蔷薇属（Rosa L.）种质资源（包括13份野生种、变种、变型及29份栽培品种）的遗传多样性进行了研究。用筛选出的18对SSR引物对42份材料DNA进行PCR扩增,在18个位点共检测到148个等位基因,每一位点的等位基因变幅为6~14个,平均8.2个。材料间遗传相似系数变化范围为0.282~0.892,表明在分子水平上云南省蔷薇属植物具有丰富的遗传多样性。本研究发现,在相似系数为0.456时,基于SSR标记的聚类分析可以将 13个蔷薇野生种明显分为5个组,这与植物形态学分类结果大体一致。在遗传相似系数为0.43水平上,聚类分析将42份供试材料分为5大组群;同时初步探讨了野生种之间以及野生种与栽培品种之间的遗传亲缘关系。     

Genetic survey of 10 Indian coconut landraces by simple sequence repeats (SSRs)

A rich genetic diversity of coconut exists in farmer's fields, which represent valuable genetic resource for breeding. The study was conducted to assess the pattern of diversity in 102 coconut palms representing 10 landraces from 3 coconut-growing communities of India using 14 simple sequence repeat (SSR) markers. A total of 90 alleles were detected with an average of 6.42 alleles per locus and an average polymorphism information content of 0.61. Expected heterozygosity (H_e) was highest for the two tall landraces from Pallikkara community, while the least heterozygosity was observed for the dwarf coconut landraces from Vayalar community. Mean fixation index (F_ST) of 0.42 indicates a high level of population differentiation. A low gene flow (N_m) of 0.37 was observed. Based on molecular data, genetic similarities were calculated. The unweighted pair group method with arithmetic averages (UPGMA) cluster analysis grouped the landraces according to their geographical locations and breeding behaviour. The practical implications of this study in farmer participatory evaluation and conservation of coconut genetic resources are highlighted.