Genetic diversity of a <Emphasis Type="Italic">Coffea</Emphasis> Germplasm Collection assessed by RAPD markers

Genetic diversity and relationships within and among nine species of Coffea, one species of Psilanthus and the Piatã hybrid from the Coffee Germplasm Collection of Instituto Agronômico de Campinas (IAC), Brazil were assessed using RAPD markers. Genetic diversity and relationships were evaluated by proportion of polymorphic loci (P), Shannon’s genetic index (H′ and G′_ST) and clustering analysis. The overall RAPD variation among all accessions was mostly partitioned between rather than within species. However, C. canephora and C. liberica showed a high genetic diversity within the species (\({\underline{\hbox{H}'}} \) _sp = 0.414 and \({\underline{\hbox{H}'}} \) _sp = 0.380, respectively) and this was highly structured (high \({\underline{\hbox{G}'}} \) _ST). Genetic diversity from C. congensis and C. arabica was also structured, but with lower levels of genetic diversity (\({\underline{\hbox{H}'}} \) _sp = 0.218 and \({\underline{\hbox{H}'}} \) _sp = 0.126, respectively). The results were consistent with agronomic and molecular studies and demonstrated that the IAC Coffea Collection is representative of the phylogenetic structure observed in the genera. This study devises sampling strategies for coffee germplasm collections and provides genetic diversity parameters for future comparisons among them.     

AFLP fingerprinting in <Emphasis Type="Italic">Capparis</Emphasis> subgenus <Emphasis Type="Italic">Capparis</Emphasis> related to the commercial sources of capers

A genetic fingerprinting technique (AFLP) was used to determine the relationships among Capparis spp. Genetic distances, based on AFLP data were estimated for 45 accessions of Capparis species, from Spain, Morocco and Syria. The results of this analysis support the differentiation of four of the five taxa involved. The group of plants recognised as C. spinosa on the basis of morphological characters, includes several cultivars and appears in an intermediate position between C. orientalis and C. sicula and overlaps with C. orientalis. The other two species C. aegyptia and C. ovata are separate from the rest. Capparis spinosa had a low number of unique bands in comparison with the other species. Although these results cannot confirm the hybrid origin of C. spinosa, the distribution of the bands supports this hypothesis, the most likely parental species being C. orientalis and C. sicula.     

Genetic Diversity of the Greater Yam (<Emphasis Type="Italic">Dioscorea alata</Emphasis> L.) and Relatedness to <Emphasis Type="Italic">D. nummularia</Emphasis> Lam. and <Emphasis Type="Italic">D. transversa</Emphasis> Br. as Revealed with AFLP Markers

Amplified fragment length polymorphism markers were used to assess the genetic relatedness between Dioscorea alata and nine other edible Dioscorea. These species include D. abyssinica Hoch., D. bulbifera L., D. cayenensis-rotundata Lamk. et Poir., D. esculenta Burk., D. nummularia Lam., D. pentaphylla L., D. persimilis Prain. et Burk., D. transversa Br. and D. trifida L. Four successive studies were conducted with emphasis on the genetic relationship within D. alata and among species of the Enantiophyllum section from Vanuatu. Study 1 was carried out to select a set of polymorphic primer pairs using 11 combinations and eight species belonging to five distinct sections. The four most polymorphic primer pairs were used in study 2 among six species of the Enantiophyllum section. Study 3 focussed mainly on the genetic relationship among 83 accessions of D. alata, mostly from Vanuatu (78 acc.) but also from Benin, Guadeloupe, New Caledonia and Vietnam. The ploidy level of 53 accessions was determined and results indicated the presence of tetraploid, hexaploid and octoploid cultivars. Study 4, included 35 accessions of D. alata, D. nummularia and D. transversa and was conducted using two primer pairs to verify the taxonomical identity of the cultivars `langlang', `maro' and `netsar' from Vanuatu. The overall results indicated that each accession can be fingerprinted uniquely with AFLP. D. alata is an heterogeneous species which shares a common genetic background with D. nummularia and `langlang', `maro' and `netsar'. UPGMA cluster analysis revealed the existence of three major groups of genotypes within D. alata, each assembling accessions from distant geographical origins and different ploidy levels. The analysis also revealed that `langlang', `maro' and `netsar' clustered together with the cultivar `wael' (D. transversa) from New Caledonia. Results are discussed in the paper.     

Analysis of genetic variability in <Emphasis Type="Italic">Couepia</Emphasis> accessions using AFLP markers

Couepia is a genus distributed in tropical regions of America. The nut of some Couepia species is used as fresh fruit and oil source for local communities. Despite its critical situation of conservation and its economic potential, there is a lack of information on the genetic variability of Couepia species. This study examines AFLP variation among 40 accessions of Couepia collected in the Colombian Amazonian region, representing two species: Couepia dolichopoda and Couepia subcordata. The individuals were examined for 96 markers generated from four EcoRI/MseI primer pairs, with 80% polymorphism across all accessions studied. According to cluster analysis, 40 accessions were grouped into two major clusters, corresponding to the two species analyzed, except one case whose situation is discussed. In C. dolichopoda accessions, significant correlation between the clustering pattern and the geographical origin was detected; the extent of variation within and among its collect sites was examined by AMOVA. The knowledge about the genetic variability of the accessions examined contributes to development of Couepia conservational efforts.     

RAPD-based Analysis of Genetic Diversity and Selection of Lingonberry (<Emphasis Type="Italic">Vaccinium vitis-idaea</Emphasis> L.) Material for <Emphasis Type="Italic">ex situ</Emphasis> Conservation

Random amplified polymorphic DNA markers were used to assess relatedness and genetic diversity for 15 lingonberry (Vaccinium vitis-idaea) populations. Seven primers yielding 59 polymorphic bands were used to analyse 13 populations, representing ssp. vitis-idaea from Sweden, Finland, Norway, Estonia and Russia, and two populations, representing ssp. minus from Japan and Canada. A cluster analysis and a multidimensional scaling analysis (MDS) showed similar phenetic patterns among populations, with a pronounced geographic grouping in most cases. Significant correlations were obtained between geographic and genetic distances for the entire set of populations as well as for the 13 ssp. vitis-idaea populations. Mean within-population diversity was 0.206 when estimated with Lynch and Milligan's index, and 0.431 when estimated with Shannon's index, which is in agreement with the mixed mating system reported for lingonberry. Within-population variability accounted for 68.6% of the total variance when all populations were included, and for 78.8% when only populations of ssp. vitis-idaea were analysed. Two different approaches were applied to the selection of plant material for a potential gene bank: (1) a hierarchical sampling strategy based on a cluster analysis and (2) the Maximum genetic diversity program, developed for the establishment of core collections. Random sampling was undertaken for comparisons with the selected data sets. The most diverse and representative set of lingonberry specimens was obtained when samples were selected with the Maximum diversity program.     

AFLP and RFLP linkage map in <Emphasis Type="Italic">Coix</Emphasis>

Coix is a genus in the grass family placed in the tribe Maydeae. It is closely related to maize and is also used as a crop plant. Since many valuable traits have been identified recently in Coix, it is considered to be a valuable genetic resource, particularly for maize improvement. In this study, a Coix genetic linkage map was constructed using an F2 population of 131 individuals. Eighty AFLP and 10 RFLP markers were mapped, covering a total length of 1339.5 cM with an average interval of 14.88 cM. The map consisted of 10 linkage groups, were consistent with the chromosome numbers observed cytogenetically. Both AFLP and RFLP markers were used first for genetic analysis in Coix. AFLP markers were generated by two restriction enzyme combinations, EcoRI/MseI and PstI/MseI. A total of 1349 bands were amplified, of which 140 were polymorphic. The polymorphism detection efficiency of the two enzyme combinations was compared, and utility of AFLP markers to construct the linkage map was discussed. Ten RFLP markers detected by three different probes were distributed on eight different linkage groups. The results provide a foundation to map and isolate important genes in Coix, and to investigate its genomic architecture, possible origins, and relationship with maize at the DNA level.     

Assessment of genetic diversity and genetic relationships among 29 populations of <Emphasis Type="Italic">Azadirachta indica</Emphasis> A. Juss. using RAPD markers

Randomly amplified polymorphic DNA (RAPD) analysis was employed to assess genetic divergence among 29 neem accessions collected from two agro-ecological regions of India (11 agro-climatic sub-zones), which cover three states, Punjab, Haryana and Rajasthan. Out of 24, 10-mer random primers used for studying genetic divergence, 14 were polymorphic, generating a total of 73 amplification products with an average of 5.21 products per polymorphic primer and estimated gene diversity of 0.49. Genetic relationships among accessions were evaluated by generating a similarity matrix based on Jaccard’s coefficient, ranging from 0.70 to 0.96. The phenetic dendrogram generated by UPGMA analysis grouped accessions into five clusters. RAPD performed within accessions (individual seedlings collected from the same mother plant) showed no variation indicating homogeneous population within accessions. Primers OPA-18, OPC-08 and OPI-03 were found most informative based on their resolving power. The degree of genetic variation detected among the 29 accessions with RAPD analysis suggests that RAPD can be used for studying genetic diversity in neem. The study also demonstrated that neem germplasm collected from northwestern plains of India shows no eco-geographical isolation based on sub-zones because accessions collected from different sub-regions are grouping together in the genetic tree.     

Genetic Analysis of Egyptian Date (<Emphasis Type="Italic">Phoenix dactylifera</Emphasis> L.) Accessions Using AFLP Markers

Forty-seven samples of date palm (Phoenix dactylifera L.) collected from eight locations in Egypt were studied using four sets of amplified fragment length polymorphism (AFLP) markers with near infrared fluorescence labeled primers. These samples belonged to 21 named accessions and 9 of unknown pedigrees. A total of 350 bands were scored and 233 (66.6%) were polymorphic. Twenty-seven Egyptian accessions and ‘Medjool’and ‘Deglet Noor’accessions from California could beclassified into the major cluster. This major cluster may represent a major group of date palm germplasm in North Africa. There were four other clusters, each containing one or two accessions. The variety ‘Halawy’and one accession of unknown provenance were most likely from hybridization between two clusters. Six groups of accessions of which had the same names, revealed similar but not identical AFLP profiles suggesting these accessions might derive from seedlings rather thanthrough clonal offshoot propagation.     

Genetic Characterization of Brazilian Annual <Emphasis Type="Italic">Arachis</Emphasis> Species from Sections <Emphasis Type="Italic">Arachis</Emphasis> and <Emphasis Type="Italic">Heteranthae</Emphasis> using RAPD Markers

The genus Arachis is divided into nine taxonomic sections. Section Arachis is composed of annual and perennial species, while section Heteranthae has only annual species. The objective of this study was to investigate the genetic relationships among 15 Brazilian annual accessions from Arachis and Heteranthae using RAPD markers. Twenty-seven primers were tested, of which nine produced unique fingerprintings for all the accessions studied. A total of 88 polymorphic fragments were scored and the number of fragments per primer varied from 6 to 17 with a mean of 9.8. Two specific markers were identified for species with 2n = 18 chromosomes. The phenogram derived from the RAPD data corroborated the morphological classification. The bootstrap analysis divided the genotypes into two significant clusters. The first cluster contained all the section Arachis species, and the accessions within it were grouped based upon the presence or absence of the ‘A’ pair and the number of chromosomes. The second cluster grouped all accessions belonging to section Heteranthae.     

Discrimination among subterranean clover (<Emphasis Type="Italic">Trifolium subterraneum</Emphasis> L. complex) genotypes using RAPD markers

RAPD markers were applied to subterranean clover aiming at: (i) assessing the genetic relationships among the subspecies subterraneum L., brachycalycinum Katzn. et Morley, yanninicum Katzn. et Morley, as their taxonomic status is still debated; (ii) verifying the adoption of RAPDs to supplement the common morphological markers used for cultivar identification and protection; (iii) assessing the possible genetic diversity in relation to the geographic origin. Eight primers were selected for genetic analysis of 18 genotypes: 10 subsp. yanninicum (five from Greece and five from Sardinia), six subsp. subterraneum (forming three pairs, each one difficult to distinguish by morphological markers), and two subsp. brachycalycinum. Cluster analysis, performed on the Jaccards coefficients of association computed across the eight primers, formed three groups of genotypes, corresponding to the three subspecies. The results supported at the DNA level previous inferences, made at cytological, karyological, and isoenzymatic levels, on the ongoing speciation process within the subterranean clover complex, although not warranting yet the full species rank to the three forms. The genotypes of subsp. yanninicum were genetically closer to those of subsp. subterraneum than either group was to the subsp. brachycalycinum genotypes. Within the subsp. yanninicum cluster, the Sardinian genotypes appeared fairly distinct from those from Greece, suggesting a possible, independent evolution going on in different centres of diversity of this subspecies. In two pairs of subsp. subterraneum genotypes, the members could be unequivocally distinguished, thus supporting the role of RAPD fingerprinting in cultivar identification. In the third pair, the two genotypes appeared to be the same, inadvertently duplicated within the germplasm collection.     

AFLP-based genetic diversity assessment among Chinese vegetable mustards (<Emphasis Type="Italic">Brassica juncea</Emphasis> (L.) Czern.)

Amplified fragment length polymorphism (AFLP) analysis was used to evaluate the genetic relationships and diversities of Chinese vegetable mustards. Fourteen pairs of primers generated a total of 366 scorable fragments among 16 accessions of Brassica juncea studied, of which 296 bands were polymorphic with an average of 21.1% polymorphic bands per primer combination. Genetic similarities were obtained using Nei and Li similarity coefficients, and a dendrogram of the 16 accessions was made by UPGMA clustering method. The Nei and Li Similarity coefficient value ranged from 0.63 to 0.88. This result indicated that the 16 accessions of B. juncea possessed high level genetic variations. The cluster analysis showed that the vegetable mustards could be grouped into two main groups and some minor rami, which was partially in accordance with the traditional classification that based on different edible organs of vegetable mustards. The incongruity between morphological and molecular classification might be attributed to the high selection pressure during domestication of Chinese vegetable mustards, producing some accessions with similar genetic backgrounds evolving into abundant morphological variations. The great diversification among Chinese vegetable mustards not only provides an excellent object for molecular evolution research of B. juncea but also is of great value for widening the genetic basis of breeding programs and breeding materials selection. Besides, our study also indicates that AFLP are informative and can provide significant insights for genetic diversity research in B. juncea.     

Determining genetic diversity between lines of <Emphasis Type="Italic">Vigna unguiculata</Emphasis> subspecies by AFLP and SSR markers

AFLP (Amplified Fragment Length Polymorphisms) and SSR (Simple Sequence Repeat) markers were utilized to assess genetic diversity and relatedness between Vigna unguiculata subspecies. Three AFLP primer combinations and 10 SSR primer sets successfully identified closely related accessions, and the presence of heterogeneity in some accessions. AFLP methodology was successful in separating different species of Vigna. However, the level of intra-subspecies variation was as great as was the interspecies variation with both marker methods. The number of markers employed was insufficient to successfully group the subspecies into distinct clades.     

Pheno-morphological,agronomic and genetic diversity among natural populations of sulla (<Emphasis Type="Italic">Hedysarum coronarium</Emphasis> L.) collected in Sicily,Italy

Sulla (Hedysarum coronarium L.) is a short-lived perennial forage legume that plays a key role in cereal-based systems in semi-arid Mediterranean regions, particularly in organic production and low-input oriented agriculture. In Sicily, the species is widespread both as a wild and cultivated plant. The present study assessed the phenotypic and genetic variation among natural populations of sulla collected from different environments throughout Sicily and analysed how the patterns of phenotypic diversity varied according to the environmental parameters of each collection site. Two commercial varieties and two Sicilian agro-ecotypes were also included in the study as controls. Principal components analysis (PCA) was performed on the sites using geographic, climatic, and pedological data to assess the differences in types of collection sites. PCA was also performed on the accessions (using pheno-morphological and agronomic data) to establish the importance of different traits in explaining multivariate polymorphisms. The results showed a large degree of genetic diversity (based on ISSR markers) and variability in pheno-morphological and agronomic traits. PCA did not clearly differentiate the accessions according to their habitats of origin, but in some cases accessions from the same habitat had a tendency to group together. The agronomic attributes of several populations were more pronounced than those of the controls. The observed variability may be valuable when selecting for H. coronarium varieties suitable for various uses (e.g., hay production, grazing, soil protection).     

Origin and ancestry of Egyptian clover (<Emphasis Type="Italic">Trifolium alexandrinum</Emphasis> L.) As revealed by AFLP markers

The origin and ancestry for Egyptian clover, Trifolium alexandrinum, was examined using AFLP data. The data support a close relationship of T. alexandrinum accessions from Syria and Egypt to T. apertum, T. berytheum, and T. salmoneum. However, crossability and geographic distributions suggest that T. apertum is an unlikely progenitor. In contrast, T. salmoneum appears to be the most probable progenitor for Syrian material of Egyptian clover, although a close relationship to T. berytheum was also revealed. The ability of these species to cross freely indicates that T. salmoneum and T. berytheum may be regarded as the primary ancestors from, which man domesticated Egyptian clover through artificial selection in Syria. Following domestication, the earlier forms of the crop species could have been taken into rain-fed cultivation in Palestine and irrigated cultivation in Egypt. In this regard, the domestication of Egyptian clover may be analogous to other crops, such as barley and wheat, which were also domesticated in the Fertile Crescent and taken into cultivation in the Nile Valley. It appears that genetic improvement of the crop occurred in Egypt after cultivation, and that the varieties that were developed in Egypt were later distributed worldwide.     

Genetic diversity of the D-genome in <Emphasis Type="Italic">T. aestivum</Emphasis> and <Emphasis Type="Italic">Aegilops</Emphasis> species using SSR markers

Simple sequence repeats (SSRs), highly dispersed nucleotide sequences in genomes, were used for germplasm analysis and estimation of the genetic relationship of the D-genome among 52 accessions of T. aestivum (AABBDD), Ae. tauschii (D^tD^t), Ae. cylindrica (CCD^cD^c) and Ae. crassa (MMD^cr1D^cr1), collected from 13 different sites in Iran. A set of 21 microsatellite primers, from various locations on the seven D-genome chromosomes, revealed a high level of polymorphism. A total of 273 alleles were detected across all four species and the number of alleles per each microsatellite marker varied from 3 to 27. The highest genetic diversity occurred in Ae. tauschii followed by Ae. crassa, and the genetic distance was the smallest between Ae. tauschii and Ae. cylindrica. Data obtained in this study supports the view that genetic variability in the D-genome of hexaploid wheat is less than in Ae. tauschii. The highest number of unique alleles was observed within Ae. crassa accessions, indicating this species as a great potential source of novel genes for bread wheat improvement. Knowledge of genetic diversity in Aegilops species provides different levels of information which is important in the management of germplasm resources.     

Diversity analysis of Central Asia and Caucasian lentil (<Emphasis Type="Italic">Lens</Emphasis><Emphasis Type="Italic">culinaris</Emphasis> Medik.) germplasm using SSR fingerprinting

Diversity analysis was performed among 39 cultivated lentil (Lens culinaris Medik.) accessions of Central Asia and Caucasian origin using five highly polymorphic microsatellite markers. A total of 33 alleles determined ranging from 3 to 8 per locus. Estimated gene diversity value for 33 loci was 0.66. Genetic similarity indices among 39 accessions ranged from 0.24 to 1.0. Cluster analysis using the unweighted pair group method with arithmetic mean method classified accessions into six major groups at 0.5 similarity coefficient. More than half accessions from Tajikistan formed large cluster. On the other hand, a few accessions from each country showed unique genotypes. Overall, most of the accessions, except ones with closely related origin, were distinguished by the present high quality DNA fingerprinting. This molecular diversity information gives important basis for conservation strategy in gene bank and exotic germplasm introduction in breeding programs in Central Asia and Caucasian countries.     

Genetic diversity among <Emphasis Type="Italic">Jatropha</Emphasis> species as revealed by RAPD markers

The genus Jatropha is native of tropical America with more than 200 species that are widely distributed in tropics with a promise for use as an oil crop for biodiesel. This investigation was carried out to assess the genetic diversity of 12 Jatropha species based on random amplified polymorphic DNA markers. From 26 random primers used, 18 primers gave reproducible amplification banding patterns of 112 polymorphic bands out of 134 bands scored accounting for 80.2% polymorphism across the genotypes. Three primers viz., OPA 4, OPF 11, and OPD 14 generated 100% polymorphic patterns. The polymorphic information content was highest for the primer OPD 14 (0.50) followed by the primers OPF 11 and OPAD 11 (0.48). Jaccard’s coefficient of similarity varied from 0.00 to 0.85, indicative of high level of genetic variation among the genotypes studied. UPGMA cluster analysis indicated three distinct clusters, one comprising all accessions of J. curcas L., while second included six species viz., J. ramanadensis Ramam., J. gossypiifolia L., J. podagrica Hook., J. tanjorensis J. L. Ellis et Saroja J. villosa Wight and J. integerrima Jacq. J. glandulifera Roxb. remained distinct and formed third cluster indicating its higher genetic distinctness from other species. The overall grouping pattern of clustering corresponds well with principal component analysis confirming patterns of genetic diversity observed among the species. The result provides valid guidelines for collection, conservation and characterization of Jatropha genetic resources.     

Comparative Study of Common Bean (<Emphasis Type="Italic"> Phaseolus vulgaris</Emphasis>L.) Landraces Conserved <Emphasis Type="Italic">ex situ</Emphasis> in Genebanks and <Emphasis Type="Italic">in situ</Emphasis> by Farmers

Genetic diversity of populations stored ex situ or in situ can be altered due to the management practices they are subjected to. In this paper, we compare populations of two common bean (Phaseolus vulgaris L.) landraces grown on farms with material collected from the same farms and now kept in two ex situ collections (CIAT and REGEN) with the purpose to monitor any changes that have occurred due to ex situ conservation. The diversity was measured using seven bean microsatellite markers. Further phenotypic and developmental traits were registered in a field experiment. Compared with the in situ populations, the ex situ ones had a lower level of gene diversity and we suggest that this is due to the regeneration process. Most of the phenotypic traits did not differ significantly between ex situ and in situ populations, although for yield and 100-seed weight, the CIAT material showed significant lower values. We assume that these populations have gone through an adaptational change. Overall, the conservation ex situ has been successful in maintaining the majority of the adaptations found in the landraces studied, however, the probable loss of genetic diversity that we have observed, suggest that protocols for the regeneration process must be carefully worked out if the majority of alleles are to be preserved for the future. This study also highlights the complementarity of ex situ and in situ conservation methods in order to preserve landrace adaptations and to capture new, useful diversity generated in in situ populations.     

Genetic Diversity and Relationships of Japanese Peach (<Emphasis Type="Italic">Prunus persica</Emphasis> L.) Cultivars Revealed by AFLP and Pedigree Tracing

To evaluate the genetic diversity and to clarify the genetic relationships of Japanese peach cultivars, we analyzed the amplified fragment length polymorphism (AFLP) and traced the pedigree of 17 Japanese commercial peach cultivars and six traditional accessions. Sixteen AFLP primer combinations produced a total of 837 fragments and 146 polymorphic bands with a polymorphism percentage of 17.5%. All of the peach accessions could be identified from differences in at least 10 polymorphic bands. A cluster analysis showed that all the Japanese commercial peach cultivars, except ‘Kiyomi’ and ‘Jichigetsuto’, formed a major group consisting of three sub-groups. Of the six traditional accessions, four were genetically distant from the Japanese commercial peach cultivars while two accessions from China were classified into the Japanese commercial peach cultivars group. Both the AFLP analysis and pedigree tracing suggested that Japanese commercial peach cultivars are mainly derived from ‘Shanhai Suimitsuto’, one of the traditional accessions from China. Although the genetic relationships revealed by AFLP were generally in agreement with those shown by the pedigree information, some contradictions were found. Combining the AFLP results and pedigree information can provide a better understanding of the genetic relationships of Japanese peach cultivars.     

Exploitation of <Emphasis Type="Italic">Malus</Emphasis> EST-SSRs and the utility in evaluation of genetic diversity in <Emphasis Type="Italic">Malus</Emphasis> and <Emphasis Type="Italic">Pyrus</Emphasis>

A total of 8117 suitable SSR-contaning ESTs were acquired by screening from a Malus EST database, among which dinudeotide SSRs were the most abundant repeat motif, within which, CT/TC followed by AG/GA were predominant. Based on the suitable sequences, we developed 147 SSR primer pairs, of which 94 pairs gave amplifications within the expected size range while 65 pairs were found to be polymorphic after a preliminary test. Eighteen primer pairs selected randomly were further used to assess genetic relationship among 20 Malus species or cultivars. As a result, these primers displayed high level of polymorphism with a mean of 6.94 alleles per locus and UPGMA cluster analysis grouped twenty Malus accessions into five groups at the similarity level of 0.6800 that were largely congruent to the traditional taxonomy. Subsequently, all of the 94 primer pairs were tested on four accessions of Pyrus to evaluate the transferability of the markers, and 40 of 72 functional SSRs produced polymorphic amplicons from which 8 SSR loci selected randomly were employed to analyze genetic diversity and relationship among a collection of Pyrus. The 8 primer pairs produced expected bands with the similar size in apples with an average of 7.375 alleles per locus. The observed heterozygosity of different loci ranged from 0.29 (MES₉₆) to 0.83 (MES₁₃₈), with a mean of 0.55 which is lower than 0.63 reported in genome-derived SSR marker analysis in Pyrus. The UPGMA dendrogram was similar to the previous results obtained by using RAPD and AFLP markers. Our results showed that these EST-SSR markers displayed reliable amplification and considerable polymorphism in both Malus and Pyrus, and will contribute to the knowledge of genetic study of Malus and genetically closed genera.