Collection and evaluation of grasspea (Lathyrus sativus L.) germplasm of central Italy

Characterization of 16 grasspea (Lathyrus sativus L.) accessions collected in central Italy (Marche region) was performed for morphological and agronomic traits and for 3‐(‐N‐oxalyl)‐L‐2,3 diaminoprop‐ionic acid (ODAP) content. Field trials, carried out in two locations, revealed high among‐population genetic variation. In particular, an evident differentiation between commercial populations, cultivated by market‐oriented farms, and household populations (not market‐oriented) was found. Genotype × environment interaction was negligible. Medium‐high values of ODAP content were observed following colorimetric and capillary electrophoresis analyses. A high positive correlation between the two methods was found (r = 0.83**), but the colorimetric values showed, on average, significant 14% lower ODAP values. This research represents a pre‐breeding activity aimed at developing a breeding programme for the preservation and utilization of Italian grasspea germplasm within sustainable or organic agricultural systems.     

Genetic similarity among European winter triticale elite germplasms assessed with AFLP and comparisons with SSR and pedigree data

Genetic similarities (GS) based on molecular markers are well suited for direct exploration of relationships within a germplasm pool. The objectives of this study were to: (i) assess the genetic diversity in the European winter triticale germplasm by using AFLP markers, and (ii) compare the GS estimates of AFLP markers, simple sequence repeat (SSR) markers and MALÉCOT's coancestry coefficient (f). A representative set of 127 European winter triticale varieties and breeding lines, previously investigated with SSR, was assessed with 10 PstI/TaqI primer combinations (PC). AFLP analysis identified 344 polymorphic fragments with an average polymorphic information content per PC of 0.25 and a marker index of 8.56. GS‐values between genotypes (calculated after DICE) averaged 0.61 for AFLP and 0.43 for SSR. The mean f‐value was 0.06. Dendrograms based on ‘unweighted pair‐group method and arithmetic average’ showed no clear groupings within the triticale germplasm pool, but smaller clusters were consistently found. Both molecular marker systems were superior to the coancestry coefficient for genetic diversity assessment within the elite triticale germplasm.     

Genetic diversity of garlic (Allium sativum L.) germplasm from China by fluorescent‐based AFLP,SSR and InDel markers

Garlic (Allium sativum L.), an asexually propagated crop, is an important vegetable and medicinal plant. China is the biggest garlic producer in the world; however, the genetic background of garlic from China is not well understood. In this study, population structure and clustering analysis of garlic germplasm was performed using amplified fragment length polymorphism (AFLP), simple sequence repeat (SSR) and insertion–deletion (InDel) markers. Among 212 accessions of garlic, genetic diversity analysis identified 546 alleles amplified by AFLP, SSR and InDel primers, and 492 of these were polymorphic. All accessions were divided into five groups by structure analysis and neighbor‐joining clustering. Most traits, including allicin content, were only slightly affected by population structure, which indicated that this germplasm can be used as populations for association mapping. The results provide a molecular basis for understanding the genetic diversity of the garlic germplasm preserved in China.     

Evaluation of an Italian germplasm collection of Festuca arundinacea Schreb. through a multivariate analysis

Summary Forty-eight accessions of tall fescue (Festuca arundinacea Schreb.), collected from Northern to Southern Italy, were analyzed using both univariate and multivariate analysis applied to a set of 12 quantitative traits. Four principal components were found to explain 77% of the total variation in the dependence structure. Productivity characters together with heading time and dimensions of flag leaf appeared to be the major sources of diversity among tall fescue populations. On the basis of the 4 principal components similar populations were clustered according to minimal distance analysis. Seven clusters were identified. The results of cluster analysis confirmed the presence of a remarkable diversity within the germplasm collection and explained why results of a univariate analysis of variance did not reveal significant differences among groups of accessions collected in Northern, Central and Southern Italy. The multivariate approach seemed to point out a problem of genetic erosion of the local germplasm in the Po Valley (Northern Italy) and, on the whole, appeared to be a valid system for tall fescue germplasm evaluation.Research supported by C.N.R., Italy, special grant I.P.R.A., Subject 1.5.1., paper n. 1146.     

DNA分子标记在银杏中的应用

DNA分子标记广泛用于生物研究的许多方面。本文简要介绍了RFLP、RAPD、AFLP、SSR及ISSR等几种目前常用分子标记的原理,归纳总结了分子标记在银杏中的应用研究进展。(1)获得了2个雄性特有的RAPD标记和2个雌性特有的AFLP标记,为银杏的早期性别鉴定及相关基因克隆奠定了基础;(2)采用RAPD标记和ISSR标记对我国部分栽培品种进行了分子鉴别和分类的研究,编制了一些品种的DNA指纹检索表;(3)利用RAPD和ISSR标记对一些群体、个体及栽培品种或变异类型进行了遗传分化和遗传多样性研究,结果发现银杏具有较高的遗传多样性,群体间、个体间、栽培品种及类型间都存在不同程度的遗传分化;(4)利用ISSR标记对一些个体的遗传杂合性进行了研究,结果显示个体的平均杂合率为43.53%;(5)构建了包含62个RAPD标记、19个连锁群的银杏分子遗传图谱;(6)探索了银杏优先保护种群的确定。分子标记在银杏其它方面的应用还很少。今后,除了继续对上述方面进行深入系统的研究外,还应充分运用DNA分子标记技术,开展银杏的分子标记辅助选择育种、种质评价与鉴别及保育生物学等方面的研究。     

Molecular marker diversity and bacterial wilt resistance in wild <Emphasis Type="Italic">Solanum commersonii</Emphasis> accessions from Uruguay

Solanum commersonii is a wild tuber-bearing species native to Uruguay with high potential for use in potato breeding programs. Little is known about the genetic diversity within this wild species and the relationship with the resistance to the bacterial pathogen Ralstonia solanacearum. We studied 30 S. commersonii clonal accessions, 20 of which were collected from geographically different areas across the country, while the other ten were grown from seeds from a single plant. Resistance against R. solanacearum was tested and different levels of resistance were found, ranging from delayed wilting to asymptomatic reactions. The genetic variation and the relationships among individuals in this germplasm collection were studied by different molecular markers: Random Amplified Polymorphic DNA (RAPD), Amplified Fragment Length Polymorphism (AFLP) and Microsatellites or Simple Sequence Repeats (SSR). AFLP markers generated the largest number of total and polymorphic fragments per assay unit while SSR revealed the highest frequency of polymorphic bands (100%), followed by AFLP (96.2%) and RAPD (89.4%). In contrast, when comparing the number of different genetic profiles generated, the SSR markers exhibited the lowest discriminatory power. The clustering pattern obtained with the three marker systems showed a similar distribution of the S. commersonii germplasm revealing a high correlation between the three methods employed. All three dendrograms grouped most of the accessions into two main clusters, containing the same accessions regardless of the marker type. Bacterial wilt resistant accessions were present in both clusters. Accessions originated from different seeds of the same plant were grouped within one of the major clusters, and differed in the response to R. solanacearum revealing segregation of resistance. Furthermore, the distribution in two main clusters showed high correspondence with the geographical origin of the accessions, from the north and south of the country, and with the subspecies malmeanum and commersonii morphologically identified.     

Genetic variability measures among Bromus catharticus Vahl. populations and cultivars with RAPD and AFLP markers

The objectives of this study were to optimize RAPD and AFLP techniques in B. catharticus, and to determine the genetic variability of populations and commercial prairie grass cultivars with the aforementioned molecular markers. Two populations with contrasting morphological characteristics were evaluated from individual and bulked DNA samples using RAPD markers. Both analyses showed a similar information about inter population variability. Each accession was sampled by a single leaf bulk of 10 plants. Accession similarities were established with 276 RAPD and 714 AFLP bands using Jaccard similarity coefficient. The dendrogram of the accessions using RAPD markers showed that they shared high similarity values (>94%). A similar result was obtained with AFLP markers (similarity values >98%), revealing a narrow genetic basis in the analyzed accessions. Consequently, molecular characterization of germplasm should be considered in addition to morphological criteria, to choose the parental genotypes for breeding programs of this forage crop. The AFLP technique was more efficient to detect DNA polymorphism in our experiments and unique fingerprints were detected for all the accessions. RAPD is a simple and non expensive technique, suitable to estimate genetic similarity. This revised version was published online in August 2006 with corrections to the Cover Date.     

Genetic variation among lentil (Lens culinaris Medik) landraces from Southeast Turkey

The molecular characterization of cultivated plant genepools is of foremost importance for germplasm utilization in plant breeding. However, no comprehensive genetic fingerprinting of Turkish lentil landraces existed so far. To overcome this gap, 38 lentil landraces from southeast Turkey, together with six commercial varieties, were molecularly characterized using inter simple sequence repeat (ISSR) and amplified fragment length polymorphism (AFLP) molecular markers. The ISSR analysis, performed with 14 primers, yielded 105 polymorphic bands and the AFLP analysis, carried out with six primer combinations, amplified 119 polymorphic fragments. Even though the AFLP produced more bands per primer combinations, the ISSR detected more polymorphisms. Unweighted pair-group method with arithmetic means dendrograms based on Jaccard similarities obtained from three data sets: (i) ISSR, (ii) AFLP and (iii) combined ISSR and AFLP data, were similar and separated the landraces into two main groups. Turkish lentil landraces exhibited considerable genetic diversity. One landrace from Karacadag/Diyarbakir region was significantly different from the rest of the germplasm analysed. Jaccard distances highlighted sharp differences among landraces over short geographic distances. The knowledge of regional differentiation has practical utility in the management of germplasm and in breeding programmes.     

用RAPD、ISSR和AFLP标记分析系谱关系明确的甘薯品种的亲缘关系

用RAPD、ISSR和AFLP标记对系谱关系明确的7个甘薯品种进行了亲缘关系分析。24个RAPD引物、14个ISSR引物和9对AFLP引物分别扩增出173、174和168条多态性带。3种分子标记在检测甘薯品种间遗传差异上相关程度高，其中RAPD与ISSR之间的相关系数最大为0.9328。用ISSR标记估计的品种间遗传距离为0.1286~1.0932，平均0.4883，大于其余2个标记的估计值。3种分子标记皆可揭示甘薯品种的亲缘关系，其中ISSR标记产生的聚类图与系谱图最吻合，认为ISSR标记更适于分析甘薯品种的亲缘关系。     

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.     

Targeted BSA mapping of Scmv1 and Scmv2 conferring resistance to SCMV using PstI/MseI compared with EcoRI/MseI AFLP markers

In a previous study, two chromosome regions (Scmv1 and Scmv2), conferring sugarcane mosaic virus (SCMV) resistance in maize, were enriched with EcoRI/MseI AFLP (Eco‐AFLP) markers (methylation insensitive) by targeted bulked segregant analysis (tBSA). The objective of the present study was to further saturate these two regions with PstI/MseI AFLP (Pst‐AFLP) markers (methylation sensitive) using the same tBSA approach, and to compare the genomic distribution of both Pst‐AFLP and Eco‐AFLP markers. Out of 470 PstI/MseI primer combinations screened, four Pst‐AFLP markers were identified in the Scmv1 region (chromosome 6), and none in the Scmv2 region (chromosome 3). These Pst‐AFLP markers were more closely linked to the Scmv1a gene than any of the Eco‐AFLP markers, and could be useful for marker‐assisted selection and even map‐based cloning. In addition, Pst‐AFLP and Eco‐AFLP markers were dissimilarly distributed in both target regions. Pst‐AFLP markers were equally distributed across both regions, while Eco‐AFLPs were clustered in the Scmv2 region.     

基于AFLP遗传多样性和叶片内含物的银杏特异种质资源分析

本研究以南京林业大学银杏种质资源库中新收集30份特异种质资源为材料,通过对AFLP遗传关系和叶片黄酮总量和3个主要单体(槲皮素,山奈酚,异鼠李素)及可溶性糖含量分析,探究各特异种质资源间遗传联系。研究表明各种质资源间存在不同程度的遗传差异,不同种质资源黄酮总量和单体组成比例均具有较大差异,筛选出具有较高特异性的种质资源C16。以黄酮单体和可溶性糖含量进行系统聚类,与UPGMA聚类结果不能完全吻合,说明UPGMA聚类分析结果和叶片内含物含量聚类具有一定的差别。通过结合AFLP分子标记技术和生理特征探究特异种质资源遗传多样性,发掘银杏种质遗传变异,可以为选育银杏特异种质资源提供思路,后续种质资源筛选与鉴定提供依据。     

Effectiveness of different classes of molecular marker for classifying and revealing variation in rice (Oryza sativa) germplasm

We have examined the effectiveness of similar numbers of markers from four molecular marker systems (AFLP, isozymes, ISSR and RAPD) for revealing genetic diversity and discriminating between infraspecific groups of Oryza sativa germplasm. Each marker system classifies the germplasm into three major groups (most effectively with isozymes and AFLPs), but with differences (primarily with ISSR) between the precise classifications generated. However, at the highest levels of genetic similarity there was only partial agreement as to relationships between individual accessions when different markers were used. When variance was partitioned among and within the three subspecific groups, although the differences were not significant, greater variation was found among than within groups using AFLP and isozymes, with the reverse for RAPD and ISSR. Measurement of polymorphism using average heterozygosity and effective number of alleles gave similar results for each marker system. These results are discussed in relation to various genetic resources conservation activities, and the advisability of extrapolating to other sets of germplasm particularly of other crop species. This revised version was published online in July 2006 with corrections to the Cover Date.     

宁夏枣树品种品系的AFLP分析

摘 要： 应用基因组DNA-AFLP分子标记技术,对原产宁夏回族自治区的8个枣树品种品系和原产甘肃靖远县的1个枣树品种的亲缘关系进行了研究。研究结果表明,5对引物（E1/M12、E1/M4、E4/M11、E2/M10和E5/M4）扩增出了466条带,其中多态性条带为130条,多态性比率达27.89%,表明遗传多样性不是很丰富。聚类结果证明灵武长枣2号与中卫大枣、同心圆枣聚为一类,彼此间近缘关系较近,而与灵武长枣品种的亲缘关系较远,是一个新种质;同心圆枣与中卫大枣的遗传距离仅为0.015,亲缘关系最近,是同物异名。     

罗布麻遗传多样性的AFLP分析

利用AFLP分子标记技术,对产于内蒙古通辽、包头与巴彦淖尔的罗布麻遗传多样性进行分析.用筛选出的4对适宜引物组合共扩增出5 359个AFLP位点,其中多态性位点4 889个,多态性位点比率高达91.22%;产于通辽地区的罗布麻遗传变异较大,与包头和巴彦淖尔两地的罗布麻亲缘关系较远,而包头与巴彦淖尔两地间的罗布麻亲缘关系相对较近.聚类结果与材料的来源地有一定相关性,较好地反映了不同地区间的遗传背景及亲缘关系.     

Identification of commercial chicory cultivars for hydroponic forcing and their phenetic relationships revealed by random amplified polymorphic DNAs and amplified fragment length polymorphisms

One‐hundred and twenty‐four amplified fragment length polymorphism (AFLP) and 49 random amplified polymorphic DNA (RAPD) markers have been used to distinguish between 20 and 23 commercial chicory cultivars, respectively. These were all Cichorium intybus var. foliosum F₁ hybrids, currently used in hydroponic forcing. Five‐hundred and twenty RAPD primers (OPERON) were tested, of which 156 resulted in reproducible patterns and 26 yielded polymorphisms. Two‐hundred and fifty‐six AFLP primer‐combinations were tested and six combinations were selected for identification purposes. Similarity indices were measured and clustering has been done using pairwise comparison. Both types of marker provide similar conclusions. Two major clusters are formed, representing late and early cultivars. All cultivars were identified using 10 informative RAPD primers or three AFLP primer combinations. A low degree of polymorphism was detected between some early cultivars, suggesting a narrow genetic base in their breeding strategy.     

Development of SCAR and CAPS markers linked to a recessive male sterility gene in lettuce (<Emphasis Type="Italic">Lactuca sativa</Emphasis> L.)

Genetic male sterility (GMS) has been a useful system for the production of hybrid varieties in self-pollinated plants. We obtained a GMS line developed from a spontaneous mutation in lettuce (Lactuca sativa L.). Genetic analysis in our previous study revealed that the sterility was controlled by a recessive gene which was named ms-S. For simple and quick screening of individuals showing male sterility, we attempted molecular mapping of the ms-S locus using an amplified fragment length polymorphism (AFLP) technique. From the examination of 4,096 AFLP primer combinations, 63 AFLP markers were found to be linked to the gene and nine of them were successfully converted into sequence characterized amplified region (SCAR) markers and cleaved amplified polymorphic sequence (CAPS) markers. Linkage analysis indicated that these nine markers were closely linked to the ms-S gene and all were located on the same side of the gene. The minimum genetic distance between the ms-S gene and a marker was 3.1 cM. These results provide additional information for map-based cloning of the ms-S gene and will be of great help for lettuce breeding using GMS to produce F₁ hybrids.     

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.     

Genetic diversity in mustard (Brassica juncea L.) germplasm from Pakistan as determined by RAPDs

The genetic diversity and the relationships among a collection of mustard (B. juncea) germplasm, including 41 accessions collected from Pakistan, 6 oilseed cultivars/ lines and 5 Japanese vegetable cultivars, were evaluated using random amplified polymorphic DNA (RAPD) markers. A total of 198 polymorphic amplified products were obtained from 30 decamer primers. Of these, 14 were unique to the accession PAK-85835 and 37 were specific to PAK-85839. Based on pair-wise comparisons of RAPD amplification products, genetic similarity was estimated using similarity coefficients of Nei & Li (1979) and a dendrogram was constructed using the unweighted pair-group method with arithmetic averages (UPGMA). Cluster analysis based on these genetic similarities placed most of the collected germplasm and oilseed cultivars/lines close to each other, showing a low level of polymorphism between the oilseed accessions collected in Pakistan. However, the clusters formed by the oilseed collections and cultivars were distinct from those formed by the vegetable cultivars. A low level of genetic variability of oilseed mustard in Pakistan was attributed to the selection for similar traits and horticultural uses. The farmers' preference for more remunerative crops and perhaps the close parentage of these accessions further contributed towards their little diversity. The study demonstrated that the RAPD is a simple and fast technique to compare the genetic relationships and the patterns of variation among accessions of this crop. This revised version was published online in August 2006 with corrections to the Cover Date.     

中国西双版纳水稻老品种在育性恢复基因Rf-1位点的遗传多样性

云南西双版纳素有"物种基因库"的美称,是稻种资源的多样性中心区和优异种质富集区之一.本研究利用8个位于Rf-1位点的PCR分子标记对53份西双版纳地方籼稻、粳稻和光壳稻种质资源的遗传多样性和遗传分化进行了分析,共检测到15种多态性扩增片段.Shannon's信息指数和Nei'S遗传多样性指数分析表明西双版纳水稻地方品种在Rf-1位点具有较高的遗传多样性,这些遗传变异主要来自于不同品种之间的遗传差异.分析表明西双版纳籼稻的遗传多样性要显著高于粳稻和光壳稻.结果还表明籼稻与粳稻和光壳稻之间的分化要显著大于粳稻与光壳稻之间的分化,光壳稻与粳稻的亲缘关系更近.这一结果可能表明水稻Rf-1位点的遗传分化与水稻的籼粳分化紧密相关,还暗示水稻Rf-1位点遗传分化可能是与籼粳分化同时进行的.