Using genetic diversity information to establish core collections of Stylosanthes capitata and Stylosanthes macrocephala

Stylosanthes species are important forage legumes in tropical and subtropical areas. S. macrocephala and S. capitata germplasm collections that consist of 134 and 192 accessions, respectively, are maintained at the Brazilian Agricultural Research Corporation Cerrados (Embrapa-Cerrados). Polymorphic microsatellite markers were used to assess genetic diversity and population structure with the aim to assemble a core collection. The mean values of H_O and H_E for S. macrocephala were 0.08 and 0.36, respectively, whereas the means for S. capitata were 0.48 and 0.50, respectively. Roger’s genetic distance varied from 0 to 0.83 for S. macrocephala and from 0 to 0.85 for S. capitata. Analysis with STRUCTURE software distinguished five groups among the S. macrocephala accessions and four groups among those of S. capitata. Nei’s genetic diversity was 27% in S. macrocephala and 11% in S. capitata. Core collections were assembled for both species. For S. macrocephala, all of the allelic diversity was represented by 23 accessions, whereas only 13 accessions were necessary to represent all allelic diversity for S. capitata. The data presented herein evidence the population structure present in the Embrapa-Cerrados germplasm collections of S. macrocephala and S. capitata, which may be useful for breeding programs and germplasm conservation.     

Genetic variability in wild genotypes of Passiflora cincinnata based on RAPD markers

The genetic diversity and characteristics of commercial interest of Passiflora species make it useful to characterize wild germplasm, because of their potential use for fruit, ornamental and medicinal purposes. We evaluated genetic diversity, using RAPD markers, of 32 genotypes of Passiflora cincinnata collected from the wild in the region of Vitória da Conquista, Bahia, Brazil. Thirteen primers generated 95 polymorphic markers and only one monomorphic marker. The mean genetic distance between the genotypes estimated by the complement of the Dice index was 0.51 (ranging from 0.20-0.85), and genotype grouping based on the UPGMA algorithm showed wide variability among the genotypes. This type of information contributes to identification and conservation of the biodiversity of this species and for the identification of pairs of divergent individuals for maximum exploitation of existing variability.     

Isolation and characterization of six novel microsatellite markers for mulberry (Morus indica)

Genetic characterization of germplasm resources is necessary for their effective management and efficient utilization, especially for species like mulberry in which the available germplasm exhibits rich phenotypic diversity with almost no information about its genetic base. Here we present the first report on the isolation of six novel microsatellite markers of mulberry, developed from an enriched genomic library of Morus indica. These markers revealed a high degree of polymorphism (14–26 alleles per locus; polymorphic information content, 0.85–0.90) and a broad cross‐species affinity when tested on a set of 43 elite genotypes including 13 related Morus species. The data thus demonstrate their utility as potentially efficient genetic markers for germplasm characterization, crop improvement and molecular systematics of mulberry.     

Molecular characterization of sawtooth oak (Quercus acutissima) germplasm based on randomly amplified polymorphic DNA

Sawtooth oak (Quercus acutissima) is a predominant tree species in the deciduous broad-leaved forest in China. It distributes in a large landscape area and can disperse in various ecology types. Molecular study on sawtooth oak can provide valuable information about the genetic diversity level and genetic relatedness on this important tree species. Insight into the genetic structure also provides resources of a species with its current feature and future evolutionary potential. The genetic structure of sawtooth oak was investigated by randomly amplified polymorphic DNA (RAPD). Twelve RAPD markers were used to assess genetic diversity of 408 individuals from 17 provenances enveloping most of the current distribution area of sawtooth oak. A total of 66 amplification products were detected, of which 49 bands (74.24 %) were polymorphic. Nei’s gene diversity, 0.2409, indicated a relatively high level of genetic variation in sawtooth oak germplasm. Analysis of molecular variance showed that most of the genetic diversity (87 %) was allocated within provenances. A combination of UPGMA dendrogram and STRUCTURE analysis was employed to estimate the genetic relationships of sawtooth oak germplasm; interestingly, the two methods presented similar grouping pattern with few discrepancies. Results revealed that 16 out of 17 provenances were clustered into one group, while the other 1 (LQ provenance) constituted a separate cluster. The data presented in this study suggested that the RAPD method was a valuable tool for estimation of genetic diversity and genetic relatedness of sawtooth oak germplasm. The present study also gave useful implications for germplasm conservation and new cultivar development for this promising energy tree species.     

Genetic fingerprinting of germplasm accessions as an aid for species conservation: a case study with Borderea chouardii (Dioscoreaceae), one of the most critically endangered Iberian plants

BACKGROUND AND AIMS: Molecular markers have changed previous expectations about germplasm collections of endangered plants, as new perspectives aim at holding a significant representation of all the genetic diversity in the studied species to accomplish further conservation initiatives successfully. Borderea chouardii is a critically endangered allotetraploid dioecious member of Dioscoreaceae, known from a single population in the Iberian pre-Pyrenees. This population was reported to be highly structured into two genetically distinct groups of individuals corresponding to their spatial separation along the vertical cliff where it grows. In 1999, the Spanish Government of Aragón launched the first conservation programme for the ex situ preservation of this species, and since then a seed collection has been conserved at the Germplasm Bank of the Universidad Politécnica de Madrid. However, as some seed samples had not been labelled clearly at the time of collection, their origin was uncertain. METHODS: Genetic variation in germplasm accessions of B. chouardii was investigated using microsatellite (simple sequence repeat; SSR) markers. KEY RESULTS: The 17 primer pairs used detected 62 SSR alleles in the 46 samples analysed from five different germplasm stocks. Eight alleles scored from the wild population were not detected in the germplasm samples analysed. The relatedness of the germplasm samples to the wild subpopulations through neighbour-joining clustering, principal coordinates analysis (PCO) and assignment tests revealed a biased higher representation of the genetic diversity of the lower cliff (43 samples) subpopulation than that of the upper cliff (three samples). CONCLUSIONS: The collection of additional samples from the upper cliff is recommended to achieve a better representation of the genetic diversity of this subpopulation. It is also recommended that these stocks should be managed separately according to their distinct microspatial origin in order to preserve the genetic substructuring of the wild population.     

Genetic variation and cultivar identification in Cymbidium ensifolium

As a popular flowering species with many cultivars, Cymbidium ensifolium (L.) is commercially important in horticulture. However, so far little has been known about genetic diversity and conservation genetics of this species. Understanding of the genetic variation and relationships in cultivars of C.?ensifolium is a prerequisite for development of future germplasm conservation and cultivar improvement. Here we report assessment of genetic variations in C.?ensifolium cultivars using the DNA fingerprinting technique of inter-simple sequence repeats (ISSR). A total of 239 ISSR loci were identified and used for evaluation of genetic variation with a selection of 19 ISSR primers. Among these ISSR loci, 99.16% were polymorphic with wide genetic variation as shown by Nei??s gene diversity (H?=?0.2431) among 85 tested cultivars. ISSR fingerprinting profiles showed that each cultivar had its characteristic DNA pattern, indicating unequivocal cultivar identification at molecular level. Eighteen cultivar-specific ISSR markers were identified in seven cultivars. The cultivar Sijiwenhan was confirmed as hybrid by four ISSR primers. Several cultivars with same name but different geographical origins were distinguished based on their ISSR profiles. A dendrogram generated with ISSR markers could group 73 of 85 cultivars into four major clusters. Further analysis of ISSR variation revealed that about 69% of total genetic variation in this species is due to genetic divergence inside geographical groups. Our results suggest that both germplasm collection and in?situ conservation are important for future planning of C.?ensifolium species conservation.     

云南茶树种质资源的研究进展及发展重点

云南是世界茶树的原产地和起源中心,茶树种质资源种类众多,遗传多样性丰富,在茶学研究中占有非常重要的地位。本文总结了60多年来云南茶树种质资源在考察征集、保存保护、鉴定评价与共享利用等方面的研究进展,阐述了云南茶树种质资源研究存在的问题及对今后发展方向的建议。提出今后应加强珍稀濒危茶树种质的收集保存、生态型及遗传多样性的研究和利用、生物技术在优良茶树种质创新的应用、优良基因的发掘和功能研究以及利用平台的构建、物种或种群保护的生物学基础等重点领域的研究。     

用SSR标记分析辣椒属种质资源的遗传多样性

利用21对引物在33个辣椒材料中共检测到54个等位基因,每对SSR引物检测到2~4个等位基因变异,平均为2.6个,说明辣椒种质资源的遗传多样性相对较少。通过MVSP3.13f软件对SSR数据进行聚类分析,把33个辣椒材料分为五类,同个种的辣椒种质资源基本聚在一起,说明用SSR标记来区分辣椒种质是可行的,是研究辣椒种质资源遗传多样性的有效手段。     

苦荞地方种质资源的遗传多样性分析

利用SSR分子标记技术对中国苦荞主产区陕西、云南、四川、西藏等地的82份苦荞地方种质的遗传多样性进行了分析,以揭示中国特有的作物种质--苦荞地方种质资源的遗传多样性,促进苦荞优良品种的选育.结果显示:(1)所用25个SSR引物中有13个引物在苦荞地方品种中具有多态性,且扩增条带的稳定性较好,共扩增出208条条带,其中多态性条带200条,占总数的96.2%;(2)聚类分析结果显示,82份苦荞材料的遗传相似系数(GS)分布于0.52～0.85之间,平均值为0.69,在GS值为0.722的水平上,82份材料被聚为10大类群.研究表明82份苦荞品种间遗传多样性明显,具有丰富的遗传基础.     

Siberian wild rye (Elymus sibiricus L.): Genetic diversity of germplasm determined using DNA fingerprinting and SCoT markers

Elymus sibiricus is a perennial, self-pollinating, allotetraploid grass native to northern Asia. It is widely used in cultivated pastures and natural grassland due to excellent cold and drought tolerance, good forage quality, and adaptability to a variety of habitats. Information on the genetic diversity and variation among worldwide E. sibiricus germplasm is limited but necessary for germplasm collection, conservation and effective commercial use. In this study we ana lyzed genetic diversity and variation of 69 E. sibiricus accessions from the species range and constructed DNA fingerprinting profiles of 24 accessions using SCoT markers. A total of 173 bands were generated from 16 SCoT primers, 154 of which were polymorphic with 89.0% of polymorphic bands (PPB) occurring at the species level. The PPB within 8 geographical regions ranged from 2.3 to 54.3 %. Genetic variation was greater within geographical regions (57.9%) than between regions (42.1%). The 24 accessions from Qinghai-Tibet Plateau, Mongolia Plateau, Kazakhstan, and Russia were distinguished by their unique fingerprinting. This is the first report using SCoT markers for identifying cultivars and accessions of E. sibiricus. The DNA fingerprinting profiles of E. sibiricus were useful in germplasm collection and identification. The genetic diversity of worldwide E. sibiricus germplasm has been substantially affected by ecogeographical factors. Our results suggest that collecting and evaluating E. sibiricus germplasm from major geographic regions and unique environments broadens the available genetic base and illustrates the range of variation.     

近缘物种的遗传多样性及其亲缘关系的取样策略研究

取样策略的问题目前大多停留于对单一物种的种质资源研究.然而,对野生近缘植物取样策略的研究,不仅有利于准确快捷地阐明物种间的系统发育关系,而且对于了解遗传多样性分布状况,制定野外考察、材料收集取样及保护策略均有重要的理论指导意义.以巴山松及其近缘种为例,利用cpSSR(叶绿体微卫星)和AFLP2种分子标记对其取样策略和统计方法进行分析,揭示居群取样个体数和基因位点数与遗传多样性的关系以及基因(引物)和系统树构建方法对亲缘关系确定的影响.研究表明:(1)居群取样个体数和基因位点数差异对遗传多样性影响并不显著,但当居群取样个体数为30个左右,单引物基因位点数为60个以上,总位点数为480个以上时,所得遗传多样性及亲缘关系较为准确可靠.(2)较多数目的基因(引物)对于得到较为准确可靠的亲缘关系分析是十分必要的.(3)系统树构建方法对近缘种亲缘关系的确定有一定的影响.     

Genetic diversity of Crotalaria germplasm assessed through phylogenetic analysis of EST-SSR markers.

The genetic diversity of the genus Crotalaria is unknown even though many species in this genus are economically valuable. We report the first study in which polymorphic expressed sequence tag-simple sequence repeat (EST-SSR) markers derived from Medicago and soybean were used to assess the genetic diversity of the Crotalaria germplasm collection. This collection consisted of 26 accessions representing 4 morphologically characterized species. Phylogenetic analysis partitioned accessions into 4 main groups generally along species lines and revealed that 2 accessions were incorrectly identified as Crotalaria juncea and Crotalaria spectabilis instead of Crotalaria retusa. Morphological re-examination confirmed that these 2 accessions were misclassified during curation or conservation and were indeed C. retusa. Some amplicons from Crotalaria were sequenced and their sequences showed a high similarity (89% sequence identity) to Medicago truncatula from which the EST-SSR primers were designed; however, the SSRs were completely deleted in Crotalaria. Highly distinguishing markers or more sequences are required to further classify accessions within C. juncea.     

Genetic relationships among cherry species with transferability of simple sequence repeat loci

Sweet and sour cherries are two economically important species in the world. The capability to distinguish among cherry genotypes in breeding, cultivation and germplasm collection is extremely important for scientific as well as economic reasons. In the present research, sixteen simple sequences repeat (SSR) loci were used to estimate the relationships among sweet, sour, duke and wild cherries. All of the SSR markers showed high transferability across the studied species that allowed us to study genetic diversity in them. Totally 96 alleles were generated with SSR loci, of which 93 were found polymorphic with 97.57 % polymorphism. Values of genetic similarity between genotypes varied from 0.16 to 0.97 which indicated high level of genetic diversity. On the basis of their genetic similarities, SSR analysis allowed to group the genotypes into three main clusters according to their species. These results have an important implication for cherry germplasm characterization, improvement, and conservation.     

Isolation and characterization of microsatellite loci in Paspalum notatum Flüggé (Poaceae)

Paspalum notatum is a forage grass recognized as one of the major constituents of the native grasslands in the New World. The knowledge of the genetic diversity and structure of P. notatum populations is fundamental for the conservation and germplasm management of this species. About 11 microsatellite markers were isolated from P. notatum and characterized in 25 accessions. The average number of alleles per locus was 7.9 and the PIC ranged from 0.36 to 0.89. The data demonstrated that the most of markers are suitable to detect polymorphism and to study the genetic diversity in the P. notatum species. Moreover, the transferability of these microsatellite were tested on other three congeneric species.     

A set of microsatellite markers for Arrabidaea chica (Bignoniaceae), a medicinal liana from the Neotropics

? Premise of the study: Microsatellite markers were developed, optimized, and characterized for Arrabidaea chica (Humb. & Bonpl.) Verl. (Bignoniaceae), a Neotropical liana extensively used in folk medicine. The aim of this study was to develop molecular tools to investigate the genetic structure and diversity of natural populations and germplasm collections of this species. ? Methods and Results: Eight highly polymorphic microsatellite markers revealed a multibanded pattern, suggesting that the species is polyploid. The total number of bands per locus ranged from 9 to 17, revealing high levels of polymorphism. ? Conclusions: The high level of polymorphism detected with these markers indicates their utility in devising conservation strategies and rational exploitation of A. chica.     

中国作物种质资源多样性

本文综述了中国作物种质资源的物种多样性和遗传多样性。按农艺学和用途可将中国作物分为8大类即粮食作物、经济作物、蔬菜作物、果树作物、饲用和绿肥作物、花卉作物、药用作物和林木作物。汇集了中国作物总计有840种（类）,涉及栽培物种1251个和野生近缘植物物种3308个,它们隶属176个科和619个属,这充分说明中国作物种质资源物种多样性相当丰富。依据中国作物的类型或变种多和性状变异幅度大,阐明了中国作物种质资源遗传多样性十分丰富。为中国作物种质资源的收集、保护、高效利用、创新、分类和遗传研究奠定了坚实基础,为生物多样性保护提供了科学依据。     

Analysis of genetic diversity of southern Spain fig tree (Ficus carica L.) and reference materials as a tool for breeding and conservation

The common fig tree (Ficus carica L.) is a Mediterranean crop with problematic cultivar identification. The recovery and conservation of possible local varieties for ecological production requires the previous genetic characterization of the available germplasm. In this context, 42 lines corresponding to 12 local varieties and two caprifigs, in addition to 15 reference samples have been fingerprinted using 21 SSR markers. A total of 77 alleles were revealed, detecting a useful level of genetic variability within the local germplasm pools. UPGMA clustering analysis has revealed the genetic structure and relationships among the local and reference germplasm. Eleven of the local varieties could be identified and defined as obtained clusters, showing that SSR analysis is an efficient method to evaluate the Andalusian fig tree diversity for on-farm conservation.     

Genetic diversity and genetic structure of black alder (<Emphasis Type="Italic">Alnus glutinosa</Emphasis> [L.] Gaertn) in the Belgium-Luxembourg-France cross-border area

Due to its beneficial effects on river ecosystems, black alder (Alnus glutinosa) is one of the tree species selected for planting on riverbanks in the cross-border area encompassing Wallonia in Belgium, Lorraine in France, and Luxembourg. The preservation of this species, however, is threatened by an invasive pathogen that particularly targets and kills young alder individuals. The objectives of this study were to characterize the genetic diversity and the genetic structure of A. glutinosa at this local level with the aim of assisting the conservation and replanting strategies and to determine if a germplasm collection comprising individuals from the same cross-border area captures the diversity present in the region. Nuclear simple sequence repeat (SSR) and chloroplastic DNA (cpDNA) markers were used to analyze four local wild populations and the germplasm collection which is representative of two river catchments and six legal provenance regions. Three populations distant from the studied area were also included. A panel of 14 nuclear SSR loci revealed high allelic diversity and very low differentiation among wild populations (mean F _ST?=?0.014). The germplasm collection displayed a range of alleles that were representative of the different populations, and no significant differentiation between the germplasm collection and the local wild populations was observed, making this collection, as far as allelic diversity is concerned, suitable for providing trees for riverbank replanting programs. Using SSR markers, various statistical approaches consistently indicated the lack of a significant geographical structure at the level of the river catchments or provenance regions. In contrast, two cpDNA haplotypes were detected and displayed a cross-border geographically structured distribution that could be taken into account in defining new cross-border provenance regions.     

Genetic diversity analysis in sorghum germplasm as estimated by AFLP, SSR and morpho-agronomical markers

A comparison of the different methods of the estimation of genetic diversity is important to evaluate their utility as a tool in germplasm conservation and plant breeding. Amplified fragment length polymorphism (AFLP), microsatellites or SSR and morphological traits markers were used to evaluate 45 sorghum germplasm for genetic diversity assessment and discrimination power. The mean polymorphism information content (PIC) values were 0.65 (AFLPs) and 0.46 (SSRs). The average pairwise genetic distance estimates were 0.57 (morphological traits), 0.62 (AFLPs) and 0.60 (SSRs) markers data sets. The Shannon diversity index was higher for morphological traits (0.678) than AFLP (0.487) and SSR (0.539). 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 AFLP and SSR markers, as well as for morphological and SSR markers were significantly related (p <0.001). However, morphological and AFLP data showed non-significant correlation (p >0.05). Both data sets from AFLP and SSR allowed all accessions to be uniquely identified; two accessions could not be distinguished by the morphological data. In summary, AFLP and SSR markers proved to be efficient tools in assessing the genetic variability among sorghum genotypes. The patterns of variation appeared to be consistent for the three marker systems, and they can be used for designing breeding programmes, conservation of germplasm and management of sorghum genetic resources.     

Geographical trends within a diverse spring barley collection as identified by agro/morphological and electrophoretic data

For ex-situ germplasm conservation purposes, the concept of genetic diversity being concentrated in certain geographic regions is useful for the conscious selection of diverse forms. Numerous studies of barley and other major corp species often confirm the concentration of simply-inherited, phenotypicallyobvious markers within the Vavilovian centres of diversity/origin. However, more recent studies of electrophoretic patterns and or more complexly-inherited traits do not always confirm the same geographic patterns. Unfortunately, few studies of world germplasm collections have screened a range of agro/morphological/electrophoretic patterns using the same germplasm collection as a consistent base for evaluation purposes, making precise genetic estimates of diverse geographic areas difficult. A diverse collection of 1 118 spring-sown barley cultivars was, therefore, evaluated for both agro/morphological and biochemical genetic markers in an effort to identify appropriate criteria for the construction of a comprehensive ex-situ germplasm collection. On the basis of both agro/morphological and biochemical data, countries whose cultivated barley germplasm was identified as diverse and genetically distinct were Algeria, Afghanistan, Argentina, Ethiopia, India, Peru and Turkey. However, within broad limits, separate cluster analyses of the agro-morphological and electrophoretic patterns identified dissimilar groups of countries, which demonstrated that a collection strategy based solely on country of origin is inappropriate.