Molecular Characterization by AFLPs of Capsicum Germplasm from the Amazon Department in Colombia

Using AFLPs, 71 peppers (Capsicum) accessions from the species C. chinense Jacq., C. baccatum L., C. annuum L. and C. frutescens L. from indigenous communities of the Amazon Department (Colombia) were studied to assess the genetic diversity of the collections, and delineate species gene groups. Ten additional accessions were included as a reference species group. Three clusters were identified in the Amazonian accessions by Multiple Correspondence Analyses (MCA) and a dendrogram from the UPGMA analyses of Nei – Li genetic similarity. The clusters correspond to gene groups of the species Capsicum chinense (the majority of the accessions), C. baccatum and the complex annuum - frutescens. A fourth cluster corresponds to the reference accession C. pubescens. The MCA analyses accounted for 95% of the total variation. The total genetic variation was low (Ht 0.119) and a genetic diversity index (Gst) of 0.331 was obtained. This suggests a limited genetic diversity of the accessions and a close relatedness of the species. This study is the first molecular marker assessment of genetic diversity for peppers from the Colombian Amazon, and provides information of biodiversity that can be employed in the preservation and use of Capsicum germplasm.     

Lablab purpureus (L.) Sweet – diversity, potential use and determination of a core collection of this multi-purpose tropical legume

Two germplasm collections of Lablab purpureus L. Sweet totalling 249 accessions were grown in the field in Australia and Ethiopia and characterised using a common set of morphological and agronomic (M-A) attributes. Data from each site were analysed separately using multi-variate analysis and a classification constructed for each collection. There was considerable diversity within both collections, especially that maintained in Ethiopia. Time to flowering, seed weight, and plant height were the most important attributes in determining group allocation in both classifications. Both collections had a high proportion of L. purpureus subsp. purpureus, especially accessions similar to the Australian cultivars Highworth and Rongai. They also had good representation of the less common L. purpureus subsp. uncinatus Verdc. less common landraces and wild collections from either Africa or India. When combined, the two classifications provided an overview of diversity and highlighted the similarities and dissimilarities between the two collections. The vast range in plant types supported the view that lablab has the capacity to be a multi-purpose legume for both commercial and smallholder agriculture. Some important and less common germplasm identified were Ethiopian domesticated subsp. uncinatus, Ethiopian subsp. purpureus landraces collected from regional markets, semi-domesticated and wild accessions from southern Africa, and wild accessions from India. Using this overview, a core collection of germplasm was selected, which provides researchers with a sound basis for future plant breeding and agronomic studies with this important tropical legume.     

Morphological differentiation in four species of the genus Lolium

Three hundred and sixteen accessions belonging to four species of the genus Lolium were analysed for morphological differentiation following field trials. Principal component analysis (PCA) was shown to explain over 74% of the variation with the date of ear emergence, spike height and flag leaf length and width being responsible for the major differentiation. L. perenne separated into two groups following PCA, cluster analysis and k-means clustering, as a result of floral initiation in the sowing year. Those accessions that did not attain floral initiation until the second year were found to be from northern Europe. The inbreeder L. temulentum was separated from the three outbreeding species, L. perenne, L. multiflorum and L. rigidum, which are themselves distinct from each other although there is some introgression between them, especially between L. multiflorum and L. rigidum. The results are discussed in relation to selection of a core collection of Lolium accessions.     

Collection of Lupinus angustifolius L. Germplasm and Characterisation of Morphological and Molecular Diversity

Lupinus angustifolius L. is a Mediterranean species, domesticated in the 20th century, representing an important grain legume crop in Australia and other countries. This work is focused on the collection of wild germplasm and on the characterisation of morphological and molecular diversity of germplasm accessions. It reports the collection of 81 wild L. angustifolius accessions from the South and Centre of Portugal, available at the ‘Instituto Superior de Agronomia Gene Bank’, with subsequent morphological and molecular characterisation of a selection of these and other accessions. A multivariate analysis of morphological traits on 88 L. angustifolius accessions (including 59 wild Portuguese accessions, 15 cultivars and 14 breeding lines) showed a cline of variation on wild germplasm, with plants from Southern Portugal characterised by earlier flowering, higher vegetative development and larger seeds. AFLP and ISSR molecular markers grouped modern cultivars as sub-clusters within the wider diversity of wild germplasm, revealing the narrow pool of genetic diversity on which domesticated accessions are based. The importance of preserving, characterising and using wild genetic resources for L. angustifolius crop improvement is outlined by the results obtained.     

Chinese Cornus officinalis: genetic resources, genetic diversity and core collection

Cornus officinalis is plant species that is widely used in traditional Chinese medicine. To help the efficient utilization and conservation of this species, the genetic diversity of 73 germplasm resources collected from Zhejiang, Henan and Shaanxi provinces in China were used in a primary core collection. The germplasm resources were classified nine types and the analysis of morphological traits and chemical components showed wide variations among the germplasm types. The primary core collection of C. officinalis was evaluated by using 18 inter-simple sequence repeat (ISSR) markers and a mini core collection was developed based on three strategies on construction of a core germplasm resources. The ISSR results showed that a total of 233 alleles were identified in these 73 accessions, the genetic diversity (P?=?91.02%, H?=?0.293, I?=?0.446) revealed with an average of 2 alleles per locus. The genetic diversity index analysis suggested that there is a great richness and uniqueness of genetic variation in this primary core collection. Cluster analysis by UPGMA and STRUCTURE placed the 73-germplasm resources into four linkage clusters. Almost all samples from one sampling location were clustered together. Compared to the RS and SW strategy, the LDSS strategy proved to be more representative in the core collection of C. officinalis. Finally, a mini core collections by LDSS, composed of 18 accessions (24.66% of the primary core collection), let more than 94.85% of polymorphic loci remained, 97.55% of the observed number of alleles and 99.28% of the effective number of alleles of the mini core collection, respectively. The Nei’s gene diversity and Shannon’s Information index of core collection were 0.294 and 0.443, respectively. Taken all together, the results indicated that mini core collections constructed by the LDSS strategy represented their primary core collections and provided a rational framework for intensive surveys of natural variation in C. officinalis genetic resources.     

Development of an algorithm identifying maximally diverse core collections

The development of a core collection, one which represents the genetic diversity of a crop with minimal redundancy and increases utility of the collection as a whole, is especially important as the funding for germplasm collections decreases. With limited resources, it is difficult to manage large germplasm collections and disperse genetically diverse germplasm to plant breeders. An algorithm was developed to assist in selection of core collections based on estimates of genetic distance. The criteria for selection of the maximum genetically diverse set were based on rankings of genetic distance between an accession with respect to all other accessions. Depending on the size core which a user wished, a zone around each selected accession was determined and no other accession within these limits was selected. The premise for the algorithm was that the genetic variability represented in the core must be representative of the distribution of genetic distances within the population of interest. In the present study, the algorithm was used with RAPD-marker-based estimates of genetic distance for 270 Theobroma cacao L. accessions and 134 Capsicum accessions that chose a set representing 18.5% of the population and representing the breadth of RAPD-based variation.     

Assessments of genetic diversity within a world collection of cultivated hexaploid oat (<Emphasis Type="Italic">Avena sativa</Emphasis> L.) based on qualitative morphological characters

A characterization of 10,105 accessions of hexaploid cultivated oat (Avena sativa L. sensu lato) from 85 countries of the Plant Gene Resources of Canada (PGRC) collection was conducted at Saskatoon, Saskatchewan, Canada. Eight environmentally stable morphological characters (panicle type, panicle erectness, panicle density, lemma colour, dorsal awn of lemma, ligula, major infraspecific group), differentiated into 18 character states, were used to define genetically distinct morphological groups. Comparisons of oat diversity from different countries, among the major infraspecific groups and among Canadian oat cultivars registered between 1886 and 2002 were possible. The 10,105 accessions represented 118 different morphological groups. The number of accessions in each morphological group was unevenly distributed with the 13 most frequent morphological groups including 90% of the accessions. The most frequent morphological groups in the PGRC collection were identical with the most frequent types of Canadian oat cultivars. The greatest richness of diversity was found in oat from countries with temperate climates and intensive oat breeding programmes. The oat accessions comprised 8,754 accessions of common hulled oat, 183 accessions of hull-less oat and 1,168 accessions of red oat. For red oat (A. byzantina C. Koch), West Asia was richest in diversity. The USA could be considered a secondary centre of diversity for red oat and Canada a secondary centre of diversity for hull-less oat. Morphological diversity of oat cultivars released in Canada increased during the twentieth century. The morphological groups were related to formal taxonomical infraspecific classifications of A. sativa. Applications of the concept of defining morphological groups for phenotyping a large germplasm collection are demonstrated discussed.     

Genetic diversity in Ethiopian hexaploid and tetraploid wheat germplasm assessed by microsatellite markers

The genetic diversity of a subset of the Ethiopian genebank collection maintained at the IPK Gatersleben was investigated applying 22 wheat microsatellites (WMS). The material consisted of 135 accessions belonging to the species T. aestivum L. (69 accessions), T. aethiopicum Jacubz. (54 accessions) and T. durum Desf. (12 accessions), obtained from different collection missions. In total 286 alleles were detected, ranging from 4 to 26 per WMS. For the three species T. aestivum, T. aethiopicum and T. durum on average 9.9, 7.9 and 7.9 alleles per locus, respectively, were observed. The average PIC values per locus were highly comparable for the three species analysed. Considering the genomes it was shown that the largest numbers of alleles per locus occurred in the B genome (18.4 alleles per locus) compared to A (10.1 alleles per locus) and D (8.2 alleles per locus) genomes. Genetic dissimilarity values between accessions were used to produce a dendrogram. All accessions could be distinguished, clustering in two large groups. Whereas T. aestivum formed a separate cluster, no clear discrimination between the two tetraploid species T. durum and T. aethiopicum was observed.     

Genetic diversity of the African hexaploid species Solanum scabrum Mill. and Solanum nigrum L. (Solanaceae)

Two hexaploid species of Solanum sect. Solanum are present in Africa: Solanum scabrum and S. nigrum. Solanum scabrum is a widely cultivated species and is used as a leafy vegetable, as a source of medicine and as a source of ink dye. In previous studies a wide range of morphological diversity has been reported in this species and in some studies subspecies have been proposed. Subspecies are also recognized in S. nigrum. However, it has not been established whether or not the morphological differences are reflected at the genomic level. The present study applies AFLPs to study the genetic diversity in S. scabrum and its relationship to geographical provenance, morphological differences and the possible existence of subspecies within S. scabrum and S. nigrum. The data obtained were analyzed with cluster analysis (using UPGMA and NJ). The results indicate that the genetic variation within S. scabrum was higher within accessions than between accessions. Accessions did not cluster according to their geographical provenance, indicating that accessions from different geographical areas were not significantly different genetically. The clustering reflected neither morphological differences nor domestication status (cultivated or wild). The morphological differences exhibited by S. scabrum could be due to selection by farmers for different plant types. The AFLP derived clustering pattern did not segregate the subspecies recognized in S. scabrum and S. nigrum into separate subclusters.     

Extent and Pattern of Genetic Diversity for Morpho-agronomic Traits in Ethiopian Highland Pulse Landraces: I. Field Pea (<Emphasis Type="Italic">Pisum sativum</Emphasis> L.)

An experiment was conducted in 2001 at Holetta and Kulumsa, Ethiopia, to study the extent and pattern of genetic diversity in Ethiopian field pea (Pisum sativum L.) landraces. One hundred forty-eight germplasm accessions were grown in an alpha lattice design with 2 replications. Data on 12 traits were collected and analyzed. Differences among the accessions were significant for most of the traits (except number of seeds/ pod) at each location even though differences pooled over location were mostly non-significant. The accessions were grouped into five clusters of different sizes. Accessions from the southern part of the country (Arsi) distributed overall clusters while those from the northern half (North and South Wello, North Gonder and North Shewa) fell into clusters C₁ to C₃. Cumulative effects of a number of characters dictated differentiation of the accessions into clusters. There was no definite relationship between geographic diversity and genetic diversity as overlapping was encountered in clustering pattern among accessions from different parts of the country. Accessions from different regions might have similar genetic background and those from the same origin might also have different genetic background. Therefore, geographic diversity should not necessarily be used as an index of genetic diversity and parental selection should be based on a systematic study of genetic diversity in a specific population. Genetic distances among most of the clusters were significant that crosses between parents selected out of them are expected to generate desirable genetic recombination. Selection should also consider the special advantages of each cluster and each accession within a cluster. Future germplasm collection, conservation and breeding efforts should focus not only on inter-regional diversity but also on intra-regional diversity.     

Germplasm characterization of Sesbania accessions based on multivariate analyses

Nineteen Sesbania accessions were investigated, in a total of six species, to characterize the accessions based on morphological and agronomic data using multivariate methods, to identify a core set of morphological and agronomic attributes to be used in germplasm characterization of Sesbania spp., and to evaluate the forage potential of these accessions. Plots, in randomized block design, consisted of single 16 m-long rows of eight plants, spaced at 2 m between plants and 3 m between rows. Morphological data were obtained from the first four plants in each plot. The four remaining plants, submitted to four cuts at a height of 1 m, provided agronomic data. Principal component analysis indicated that variance accumulated by the first two components for morphological and agronomic data was 74.4% and 77.0%, respectively. Two large groups of accessions were formed in the cluster analysis for morphological data, one consisting of S. virgata, subgenus Daubentonia and the second group comprising the other five species. Cluster analysis using agronomic data classified higher yielding perennial species in three groups, and the annual species, which stood out principally in relation to seed production, in three other groups. Accession n° 8 of S. sesbanwas classified in an isolated group due to its forage superiority. A character discard resulted in the selection of eight characters: floral initiation, pod and leaf length, number of leaflet pairs, plant regrowth height, leaf dry matter yield, seed production and vigour, indicated as a core set of attributes for Sesbania germplasm characterization.     

Taxonomic status and geographic provenance of germplasm accessions in the Solanum nigrum L. complex: AFLP data

Amplified Fragment Length Polymorphisms (AFLPs) were employed within the taxonomically difficult Solanum nigrum L. complex in order to characterize the genetic diversity present in a collection of the Gatersleben Genebank, to classify taxonomically unknown material, and to correlate the clustering of the examined accessions with their geographic origin. The results from AFLP analyses using two primer combinations on 44 entries from five species led to the detection of four major clusters, simultaneously uncovering significant differences in the levels of genetic diversity within or between species. S. americanumexhibited the highest infraspecific variation despite close geographic origins, simultaneously being placed in a clearly separated cluster in comparison to the other examined species of the complex. In addition, these other species showed even less interspecific variation than was found at the infraspecific level in S. americanum. In terms of taxonomy, the application of AFLPs helped in the classification of 13 black nightshade accessions formerly only listed as Solanum sp. This also was confirmed by morphological determination. Furthermore, one accession formerly classified as S. physalifolium var. nitidibaccatum i) clustered with AFLPs and ii) was identified morphologically as S. villosum. Contradictory classifications remain for two further entries from the same species, found within the S. nigrum cluster after AFLP analyses, while belonging to S. physalifolium var. nitidibaccatum according to herbarium specimen. Finally, as indicated by the information on provenance in geographically separated subclusters in S. americanum and partially in S. villosum, clues on the currently unknown origin of accessions from the genebank seem feasible by AFLP data.     

RAPD Polymorphism of the White-Flowered Gourd (Lagenaria siceraria (Molina) Standl. Landraces and its Wild Relatives in Kenya

The present study was performed to investigate genetic diversity of Kenyan landraces of the white-flowered gourd (Lagenaria siceraria), which exhibits tremendous morphological variation. RAPD analyses were performed on 53 landraces of the cultivated species L. siceraria and 42 accessions of three wild species (40 L. sphaerica, 1 L. abyssinica, and 1 L. breviflora). A total of 432 polymorphic bands were detected using 54 primers. The four species were clearly differentiated from one another. Intra-specific variations were investigated with L. siceraria and its wild relative L. sphaerica. Landraces of the cultivated species collected from different ethnic communities or regions were differentiated. Morphological variations were not associated with RAPD variations. Bitter landraces collected in Maasai communities showed two specific RAPD bands. In the wild species, accessions collected from the eastern and western sides of the Great Rift Valley were genetically differentiated from each other. In both species, genetic and geographical distance matrices computed among all pairs of accessions were significantly correlated, implying that the observed geographical variation can be explained by the 'Isolation by distance model'. Progeny plants derived from a common mother in L. siceraria showed a low level of segregation in RAPD pattern, suggesting that collected landraces are cultivated, maintaining their inherent traits although they are monoecious and insect-pollinated, whereas the wild relative L. sphaerica showed a higher level of segregation. The morphological diversity observed among landraces of L. siceraria is the result of human selection and their genetic identities are maintained by inbreeding probably resulting from frequent self-pollination.