Molecular diversity and population structure of the Ethiopian lentil (<Emphasis Type="Italic">Lens Culinaris</Emphasis> Medikus) genotype assessment using SSR markers

Knowledge of genetic diversity in germplasm is essential for formulating effective germplasm collection, conservation, utilization strategies in and crop improvement programs. It also provides an opportunity to take corrective steps infusing new genes to avoid risks associated with a narrow genetic bases. Genetic diversity analysis of 119 lentil genotypes of including 83 germplasm and 36 exotic genotypes from International Center for Agricultural Research in the Dry Areas was studied using 27 primers of simple sequence repeat (SSR) marker. Molecular analysis of variance showed variations of 82% within and 18% of the among population variance was explained. Degree of polymorphism observed among the populations was 100%. A total 122 alleles were detected, with 2 to 7 alleles per locus, with a mean of 4.52 alleles per locus. The estimated gene diversity value for 27 loci was 0.64. The average Shannon’s information index value of 1.19 was obtained showed the existence of high genetic variation within the genotypes. The genetic similarity indices ranged from 0.21 to 1.00. The SSR markers showed an average polymorphic information content (PIC) value of 0.58. Cluster analysis grouped the genotypes into five major clusters as distinct genetic populations. Diversity analyses revealed the existence of a high level of genetic variation among genotypes. This molecular diversity information provides a basis for future germplasm collection, utilization, and conservation strategies in gene banks and introducing exotic germplasm to widen the genetic base of the current lentil breeding population.     

Genetic diversity and population structure of four Iranian alfalfa populations revealed by sequence-related amplified polymorphism (SRAP) markers

In the present study, genetic diversity of 48 individual plants from four Iranian cultivated populations of alfalfa (Medicago sativa L.) was evaluated using sequence-related amplified polymorphism (SRAP) markers. Fourteen SRAP primer combinations produced 193 fragments of which 95 were polymorphic. The number of polymorphic fragments detected per primer combination ranged from 3 to 10 bands with an average of 6.78 bands. Average polymorphic information content (PIC) was 0.343 for all primer combinations. Although the AMOVA (analysis of molecular variance) results showed a significant difference in the genetic diversity among the populations (P < 0.0001), the genetic variation mainly caused by the variation of intra population accounted for 93.17% of the total genetic variation. Unweighted pair-group method arithmetic average (UPGMA) analysis of the marker data clearly separated the populations of subtropical (Yazdi) and semi-cold (Hamadani and Nikshahri) as well as Kodi, an improved population. It can be concluded that SRAP markers are useful for studying diversity and relationships among and within alfalfa populations.     

Genetic Diversity of Farmers’Preferred Sorghum Accessions and Improved Lines from ICRISAT Reveal a Disconnect Between Innovation and Technology Transfer

The genetic diversity of 65 accessions of sorghum [Sorghum bicolor (L.) Moench] collected from various farmers and germplasm lines from ICRISAT-Kenya were analyzed. Simple sequence repeats (SSR) markers were used in order to determine the extent and distribution of its genetic diversity. Twenty-nine (29) SSRs markers were polymorphic and a total of 192 alleles were detected which showed diversity. The number of alleles per primer ranged from 2 to 17, with an average of 6.62. The range of polymorphism information content (PIC) ranged from 0.03 to 0.86, with total average of 0.82. According to the results analyzed, estimates of the mean allelic pattern across the two populations was generated; expected heterozygosity (He; 0.45, 0.54), average observed alleles (Na; 3.40, 6.20), number of private allele (0.23, 3.03), and Shannon information index (I; 0.85, 1.13) for farmer and ICRISAT-Kenya germplasm, respectively. The expected heterozygosity (He) varied from 0 to 0.26 with an average of 0.05. The Neighbor-joining phenogram based on Nei’s genetic distance grouped the 65 accessions into three main groups. The analysis of molecular variance (AMOVA) revealed that 99% of the total genetic variation was within accessions in a population whereas the genetic variation among populations in accessions accounted for 1% of the total genetic variation. Genetic diversity in ICRISAT sorghum material compared to the farmer’s collection suggested little infiltration of improved germplasm to the farmers.     

亚洲棉种质资源的SSR遗传多样性分析

 对我国棉花中期库保存的200份不同地理来源的亚洲棉代表性样本进行了SSR遗传多样性分析,结果表明：亚洲棉分子水平的遗传多样性较高。83个多态性位点共检测到368个等位基因变异,其中多态性的等位基因数为329个,平均每个SSR位点3.964个。位点多态性信息量（PIC）变幅为0.010～0.882,平均0.578,PIC值大于0.7的标记有33个（占39.8%）。基因多样性（H′）变幅为0.031～2.163,有效等位基因数（Ne）变幅为1.010～8.496。华南棉区基因遗传多样性最高,其次为长江流域棉区、黄河流域棉区,从理论上支持被广泛接受的亚洲棉在我国的传播路线是由南到北,华南棉区是中棉种系的遗传多样性富集中心。利用软件NYSTS-pc2.20,采用类平均法(UPGMA)进行聚类分析,种质间SSR相似系数变幅为0.58～0.997,平均0.745,在阈值0.73处200份亚洲棉聚为8个类群,贵池小子棉白子单独聚为一群,与其他种质遗传距离较远。遗传距离和地理距离没有必然联系,但种质间亲缘关系处于极端远或极端近时,则地理距离一般也趋于较远或较近。     

Genetic structure of putative heterotic populations of alfalfa

Semi‐hybrids between genetically distant alfalfa (Medicago sativa subsp. sativa) populations may display heterosis whose extent is affected by the structure of genetic diversity across populations. This study aimed to assess the genetic diversity across three putative heterotic populations, one Italian, one Egyptian and one of semi‐erect germplasm from Eastern Europe, Canada and Spanish Mielga (EECM population). Each population was bred from ten parents after various selection cycles. Fifteen genotypes per population were characterized by 20 polymorphic SSR markers. The among‐population variance was over eightfold smaller than the average within‐population variance (2.05 vs. 17.24) and accounted for 10.6% of the total variation. G’_ST = .090 across markers indicated modest population differentiation. Various diversity measures, multidimensional scaling, and cluster analysis of the genetic structure indicated that the Italian population was more distant from the EECM population than the Egyptian one. The EECM and Egyptian populations were the most distant geographically and genetically. EECM displayed widest intrapopulation variation, accordingly to its constitutive geographical diversity. In conclusion, this study indicates modest genetic differentiation between alfalfa populations even for geographically distant germplasm.     

Molecular characterization of genetic diversity in European germplasm of perennial ryegrass

Summary Perennial ryegrass (Lolium perenne L.) is the most important grass species for temperate grassland agriculture. The level and distribution of genetic variation in gene bank ecotype collections is still largely unknown but of great interest for the planning of breeding programs. The objectives of this study were to (i) assess the molecular diversity of Polish ecotypes of perennial ryegrass, and (ii) compare the relationship between this group and German ecotypes and European cultivars investigated previously. A total number of 166 polymorphic marker bands were detected among the 171 individual plants of the 9 Polish ecotypes. In a joint analysis with 9 Polish and 22 German ecotypes, and 22 European cultivars 172 polymorphic RAPD markers could be found. Genetic distance among the Polish ecotypes ranged from 0.31 to 0.51, while for all 53 populations a broader range was detected (0.25–0.67). An analysis of molecular variance (AMOVA) revealed a much larger variation within populations (71%) than among them (29%). The Polish ecotypes contained the highest within population variation (74%). The largest among group difference (15%) was found between the Polish ecotypes versus all other accessions. We conclude that the Polish ecotypes represent a valuable genetic resource for enlarging the genetic variation in the West European germplasm pool of perennial ryegrass.     

陕西省野生大豆种质资源的SSR遗传多样性研究

为了研究陕西地区野生大豆的遗传多样性特点,利用SSR分子标记分析了陕西省6个野生大豆(Glycine soja)天然种群和1个栽培大豆(Glycine max)种群的遗传结构与遗传多样性。结果显示：13个位点共检测出113个等位基因,平均每个位点的等位基因数(A)为8.69个,等位基因数目范围为4~13个,有效等位基因数(N_e)范围为2.135(Satt590)~9.385 (Satt487),平均有效等因基因数为5.623;观察杂合度(H_o)变化范围为0.033~0.121,平均为0.080;预期杂合度(H_e)的变化范围为0.312~0.658,平均为0.482;种群平均Shannon遗传多样性指数(I)为0.657;野生大豆种群基因多样度比率(F_ST)为0.465。该研究显示,陕西省野生大豆具有较高水平的遗传多样性,野生大豆的遗传多样性普遍高于栽培大豆;随着海拔的不断升高,野生大豆遗传多样性变低;陕西中部、南部的野生大豆种质资源丰富、种群具有较高的遗传多样性,推测该区域为陕西省野生大豆的遗传多样性中心。     

Genetic variation and gene flow estimation of <Emphasis Type="Italic">Nepenthes khasiana</Emphasis> Hook. F- A threatened insectivorous plant of India as revealed by RAPD markers

Random Amplified Polymorphic DNA (RAPD) markers were utilized for determination of diversity within and among the three populations of Nepenthes khasiana Hook f., a threatened insectivorous plant of Meghalaya (India). A total of 90 bands were generated from 10 random amplification polymorphic DNA (RAPD) primers of which 71 were found to be polymorphic (78.89%). Nei’s gene diversity (h) ranged between 0.124–0.201 with overall diversity of 0.228 while Shannon’s information index I) values recorded between 0.187–0.308 with an average value of 0.352. The values of gene flow (Nm = 1.284) and the diversity among populations (0.280) recorded demonstrates higher genetic variation within the population. AMOVA analysis revealed a low level of genetic variation (21.96%) among the populations. This study indicates that some variation still exists within and between the existing populations of N. khasiana, thus, these populations could provide materials for re-establishing of this important rare and threatened species.     

Use of RAPD and microsatellite (SSR) variation to assess genetic relationships among populations of tetraploid alfalfa, Medicago sativa

The level of random amplified polymorphic DNA (RAPD) and microsatellite variation present in four ecotypes and two varieties of alfalfa (lucerne) from Italian and Egyptian germplasm sources was evaluated. A sample of 100 plants from 10 populations was analysed by means of 41 RAPD markers and 37 simple sequence repeat (SSR) markers. Both molecular approaches revealed a high degree of genetic diversity within each of the cultivated populations and enabled each of the plants considered to be uniquely fingerprinted. The genetic relationships among plants and populations were analysed by computing AMOVA (analysis of molecular variance) and F_ST analyses. RAPDs were able to separate the Italian populations from the Egyptian variety. SSRs allowed strong separation of the four Italian alfalfa ecotypes. It was concluded that RAPD and microsatellites could be useful and powerful tools for assessing genetic variation and genetic relationships in tetraploid alfalfa.     

基于线粒体Cyt b基因的皖南山区温州光唇鱼种群遗传结构

为探讨皖南山区温州光唇鱼的种群遗传结构,采用线粒体细胞色素b基因(Cyt b)对该区5个野生种群(祁门、黟县、休宁、旌德和宁国)进行群体遗传变异分析。131个样本的Cyt b基因(1141 bp)中共检出38个变异位点(变异率3.33%)、14种单倍型。序列碱基的平均含量分别为A(28.2%)、C(29.7%)、T(27.0%)、G(15.1%),A+T含量(55.2%)明显大于G+C含量(44.8%)。5个地理种群的单倍型多样性(0.0000~0.6799)和核苷酸多样性(0.0000~0.00759)普遍较低。群体分化系数(FST:0.2916-0.9782)和AMOVA分析中高达52.74%的遗传变异来自地理种群间,说明温州光唇鱼地理种群间已产生显著遗传分化;但不同水系的种群间没有显著遗传差异。群体间系统进化树显示:5个群体聚为两大进化枝,祁门和旌德种群为一枝,其余种群为另一枝。温州光唇鱼的这种种群遗传结构与地理隔离及其生态习性相关。     

Analysis of the molecular diversity of Kenyan sorghum germplasm using microsatellites

The aim of this study was to estimate the extent of the genetic diversity present in sorghum germplasm grown in Kenya using simple sequence repeat markers. A total of 139 accessions were genotyped using 11 microsatellite markers or simple sequence repeats (SSRs) and the genetic diversity was estimated. The markers showed a wide range of differences in quality index from 0.005 to 0.39. The average Polymorphic Information Content value observed was 0.6241 indicating a high level of diversity. The gene diversity index ranged between 0.2419 and 0.9313 with a mean of 0.6627 per locus. A total of 105 alleles were observed with an average of 10.4 alleles per locus. The average heterozygosity level per locus was low at 0.1717. The variability within accessions among the populations was 74.85% and within individual accessions was 18.67%. The results showed that genetic diversity within Kenyan sorghum germplasm accessions is higher than that between the different populations. It is implied that such genetic diversity can be exploited as such or in hybridization programs to improve sorghum varieties currently grown by subsistence farmers in Kenya.     

不同地理来源旱稻种质资源的遗传多样性分析

用38对SSR引物对144份不同地理来源的旱稻种质资源进行遗传多样性分析。结果表明,共检测到137个等位变异,平均每对引物检测到3.6个,等位变异范围2~9。Nei基因多样性指数（He）在0.440（RM162）~0.854（RM335）之间,平均0.598。旱稻种质资源亚种间SSR多样性差异明显,籼稻的等位基因数目和Nei基因多样性指数（Na = 3.5,He = 0.558）明显高于粳稻（Na = 3.2,He = 0.415）。Nei基因多样性指数以亚洲最高,非洲最低,亚洲其他（He = 0.594）>中国（He = 0.593）>南美（He = 0.545）>非洲（He = 0.512）。AMOVA分析表明,旱稻种质资源的遗传变异主要来自亚种内（占总变异的76.3％）,亚种间遗传分化极显著。系统聚类能较好地区分籼粳亚种,但不能区分地理组。旱稻种质资源丰富的遗传多样性为水稻节水抗旱品种的选育提供了条件。     

Genetic diversity and population structure of Indian soybean [<Emphasis Type="Italic">Glycine max</Emphasis> (L.) Merr.] revealed by simple sequence repeat markers

Genetic diversity in 90 Indian soybean cultivars was assessed using 45 SSR markers distributed on 20 soybean chromosomes. Forty-five SSR markers generated 232 alleles with an average of five alleles/locus. The observed frequencies of the 232 alleles ranged from 0.01 to 0.94 with an average of 0.19. The polymorphic information content (PIC) value of the SSR markers varied from 0.10 to 0.83 with an average of 0.61 and about 71% markers have a PIC value of >0.5. In this study, 54 rare alleles including 19 genotype specific alleles were also identified. The observed hetrozygosity for SSR markers ranged from 0 to 0.11 with a mean of 0.10. Cluster analysis grouped the 90 soybean cultivars into three major clusters and principal coordinates analysis (PCoA) results were similar to those of the cluster analysis. A combination of eight SSR markers successfully differentiated all 90 soybean cultivars. The population structure analysis distributed the 90 soybean genotypes into two populations with mean alpha (α) value of 0.1873. In AMOVA analysis, proportion of variation within population was high (88%), whereas only 12% occurred among populations. In cluster and structure analyses, most of the genotypes with similar pedigree were grouped together. Soybean cultivars DS228, MACS-13, LSb-1, Hardee, Improved Pelican, and Pusa-24 were the six most genetically distinct cultivars identified. The study reported a moderate genetic diversity in Indian soybean cultivars and findings would be useful to the soybean breeders in selecting genetically distinct parents for a soybean improvement program.     

Molecular evaluation of genetic variability in wild populations of mulberry (Morus serrata Roxb.)

Sixteen populations of the wild mulberry, Morus serrata Roxb., were analysed for their genetic diversity with the aim of using them in introgressive breeding programmes with cultivated relatives. Five genets from each population were collected from different natural populations of M. serrata present in Uttaranchal and Himachal Pradesh in India, and diversity of morpho‐anatomical traits and inter‐simple sequence repeat (ISSR) markers were studied. Significant amounts of genetic diversity were observed among these populations for morpho‐anatomical as well as DNA markers. The 17 ISSR primers generated a total of 95 DNA markers, 51 of which were polymorphic, revealing 67% polymorphism among the populations. The pair‐wise genetic distance, estimated from these DNA markers varied from 0.091 between Urgam‐3 and Kathpuria to 0.258 between Dakrakao‐1 and Dunda with an average genetic distance of 0.165. Clustering analysis grouped these 16 populations into three broad groups. The grouping showed a moderate correlation with the geographical distances. Based on the morphological traits and molecular genetic variability, plants of Urgam‐1, Bhowali farm, Nainitikar, Dunda or Korwa‐2 can be selected for breeding and conservation programmes.     

Molecular genetic diversity within and among German ecotypes in comparison to European perennial ryegrass cultivars

Perennial ryegrass (Lolium perenne L.) is the most important grass species for temperate grassland agriculture. The level and distribution of genetic variation in gene bank ecotype collections is still largely unknown but of great interest for the planning of breeding programmes. The objectives of this study were to (i) assess the molecular variance and population structure of German ecotypes at the regional and population level, (ii) assign ecotypes to germplasm pools and (iii) compare the relationship between German ecotypes and previously‐investigated European cultivars of perennial ryegrass, A total of 22 ecotypes originating from three geographic areas in Germany. each with a sample size of 20 individual plants, were investigated with 156 polymorphic RAPD markers. Genetic distance among ecotypes ranged from 0,27 to 0.48, An analysis of molecular variance (AMOVA) revealed a much larger variation within populations (71%) than among them (29%). Ecotypes from North Germany were significantly different from those of South and Middle Germany. Thus, two distinct germplasm pools could be identified. The 22 ecotypes and 22 previously investigated cultivars shared 98% of the molecular variance.     

Evaluating genetic variation and relationships among two bromegrass species and their hybrid using RAPD and AFLP markers

The two most widely grown bromegrass species in North America are smooth bromegrass (Bromus inermis Leyss.), a hay type grass, and meadow bromegrass (Bromus riparius Rehmann), a pasture type grass. Recently a hybrid bromegrass population between the two species has been produced as a dual-purpose hay-pasture grass. Molecular markers have the potential to improve selection procedures to enhance bromegrass breeding. The objective of this study was to use RAPD and AFLP markers to determine genetic relationships and variations among bromegrass populations. Forty-three RAPD markers from 21 primers and 83 AFLP markers from seven primer combinations were used. Both marker types were able to group the individuals into their respective populations. The relationships among the individuals within each of the populations were not similar between the two marker types. Analysis of molecular variance (AMOVA) detected greater within-population variation than among-population variation for both marker types. The highest variation was observed in the hybrid population followed by meadow and then smooth bromegrass. The inter-population distance from both markers indicated that the highest genetic distance was between meadow and smooth bromegrass and lowest between smooth and hybrid bromegrass, which reflect the breeding history of the hybrid population. This study showed that both markers are capable of differentiating bromegrass genotypes into their respective populations, detecting genetic variation and relationships of the populations. Results of this study suggest that these two markers can be used in the future to enhance the current breeding practices in bromegrass, however, AFLP markers would be the marker of choice due to the high number of polymorphic markers. This revised version was published online in July 2006 with corrections to the Cover Date.     

Genetic diversity of feral alfalfa (<Emphasis Type="Italic">Medicago</Emphasis> <Emphasis Type="Italic">sativa</Emphasis> L.) populations occurring in Manitoba,Canada and comparison with alfalfa cultivars: an analysis using SSR markers and phenotypic traits

Feral populations of cultivated crops may act as reservoirs for novel traits and aid in trait movement across the landscape. Knowledge on the genetic diversity of feral populations may provide new insights into their origin and evolution and may help in the design of efficient novel trait confinement protocols. In this study, the genetic diversity of 12 feral alfalfa (Medicago sativa) populations originating from southern Manitoba (Canada) and 10 alfalfa cultivars and a M. falcata germplasm were investigated using eight SSR markers (i.e., microsatellites) and 14 phenotypic traits. We found that the genetic diversity observed in feral populations was similar to the diversity detected among the 10 cultivars. Analysis of molecular variance revealed that there was great genetic variation within (99.8%) rather than between different feral populations. Cluster analysis (unweighted pair-group method using arithmetic average) revealed no differentiation between feral populations and cultivars for neutral loci. High levels of population differentiation for phenotypic traits (and not for neutral markers) suggest the occurrence of heterogeneous selection for adaptive traits. The phenotypic traits we studied did not distinctly separate feral populations from cultivars but there was evidence of natural selection in feral populations for traits including winter survivability, rhizome production, and prostrate growth habit. Our results suggest that feral alfalfa populations need to be considered in the risk assessment of alfalfa containing novel genetically modified (GM) traits. Further, feral alfalfa populations may be regarded as a source of new germplasm for plant improvement.     

楸树EST-SSR标记开发及种质资源的遗传多样性分析

利用转录组数据开发SSR标记是一种经济高效的DNA分子标记开发策略。本研究利用高通量测序技术开发楸树(Catalpa bungei)EST-SSR标记,了解其在转录组序列中的分布类型,并利用41对多态性较好的引物对来自安徽(AH)、河南(HN)、湖北(HB)和山东(SD)的4个楸树居群的48个无性系的遗传多样性进行分析。结果表明:在大于1kb的14634条序列中,共鉴定出3999个SSR位点,580条序列包含2个及以上位点;单核苷酸重复(1957,48.94%)是最常见的SSR类型,其次是二核苷酸重复(1164,29.11%),三核苷酸重复(834,20.86%),四核苷酸重复(41,1.03%)和五核苷酸重复(3,0.08%);共扩增出243个等位基因,每个位点的等位基因数为3~13个,平均为4.85个,观测杂合度为0.14?1.00,平均为0.63,期望杂合度为0.42?0.84,平均为0.67,大部分位点偏离哈迪-温伯格平衡;AH和HN居群在有效等位基因数和期望杂合度的数值较高,表明其遗传多样性较丰富;分子方差分析(AMOVA)表明,遗传变异主要发生在居群内。本研究为楸树及同属其他树种种质资源鉴定、遗传多样性提供依据,且在指导楸树良种选育等方面具有重要的意义。     

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.