Morphological and isozyme variation in genebank accessionsof Aegilops umbellulata Zhuk., a wild relative of wheat

Morphological variation and genetic variation at 15 enzyme lociwere studied in genebank accessions of Aegilopsumbellulata Zhuk., the diploid genome donor to all thepolyploid species of the section Aegilops ofAegilops. Accessions from the Greek Islands hadshorter spikes with smaller number of spikelets and smaller number ofawns on the empty glume. The number of alleles per locus (A= 2.01) and the proportion of polymorphic loci (P= 0.627) were similar to those of the other species ofAegilops so far reported. Genetic distancescalculated from isozyme variations among five regions revealed thataccessions from the Greek Islands are more distantly related to theother four continental regions, Iran and Iraq, Southeast Turkey,Central Turkey and South and West Turkey. The present resultsindicated the importance of collection covering the whole range ofgeographical distribution to capture the genetic variation present inAe. umbellulata.     

Genetic identity and relationships of Iranian apple (Malus?×?domestica Borkh.) cultivars and landraces, wild Malus species and representative old apple cultivars based on simple sequence repeat (SSR) marker analysis

In order to shed light on the role of Iran in apple evolution and domestication, we chose to investigate the relationships of a collection of 159 accessions of wild and domesticated apples including Iranian indigenous apple cultivars and landraces, selected wild species, and old apple scion and rootstock cultivars from different parts of the world. The majority of the wild species belonged to M. sieversii, which is widely believed to be the main maternal wild ancestor of domestic apples, from Kazakhstan and M. orientalis, which is one of the probable minor ancestors of domestic apples, from Turkey and Russia located on the east and west of Iran, respectively. The accessions were assigned into six arbitrary populations for the purpose of generating information on genetic parameters. Nine simple sequence repeat (SSR) loci selected from previous studies in apple were screened over DNA extracted from all the accessions. Results showed that all SSR loci displayed a very high degree of polymorphism with 11–25 alleles per locus. In total, there were 153 alleles across all loci with an average of 17 alleles per locus. The SSR allelic data were then used for estimation of population genetic parameters, including genetic variation statistics, F-statistics, gene flow, genetic identity, genetic distance and then cluster analysis using POPGENE 1.32 software. The F-statistics and gene flow in particular, showed that there was more intra-population than between population variation. The genetic identity and genetic distance estimates, and the dendrogram generated from the un-weighted pair group arithmetic average (UPGMA) method of cluster analysis showed that the Iranian cultivars and landraces were more closely related to M. sieversii from Central Asia (east of Iran) and M. orientalis native to Turkey and Russia than to other accessions of Malus species. Also, the old apple cultivars from different parts of the world have a closer genetic relationship to M. sieversii, M. orientalis and the Iranian apples, than to other wild species. Based on these results, we suggest that the Iranian apples may occupy an intermediate position between the domesticated varieties and wild species. We propose that Iran could be one of the major players in apples’ domestication and transfer from Central Asia to the western countries.     

Genetic relationships among perennial and annual Cicer species growing in Turkey as revealed by allozymes

Allozyme polymorphisms were used to assess genetic variation and relationships among ten Cicer species (annuals and perennials) growing in Turkey. Using seven enzyme systems, 12 putative scorable loci were detected and surveyed for polymorphism in an accession collection including wild and cultivated forms. Variation was generally low within accessions and species, but common between species. Cluster analysis based on the pairwise genetic distance coefficients among accessions and species using UPGMA revealed two species clusters; one includes three perennials (C. montbretii, C. isauricum and C. anatolicum) and the other contains six annuals (C. pinnatifidum, C. bijugum, C. judaicum, C. echinospermum, C. reticulatum and C. arietinum) and one perennial species (C.incisum). Grouping obtained in allozyme analysis appears to be consistent with the classification these species into three sections. However, contrary to relationships obtained in previous studies, three perennial species from section Polycicer were relatively distant to the group containing annuals. One perennial species, C. incisum from section Chamaecicer, clustered with annuals showing a close similarity. The grouping of six annual species was consistent with the previous reports of relationships. The relationships deduced between perennials and annuals appear to shed light on the evolution of annual habit from perennial habit.     

Estimation of Variability of Five Enzyme Systems Among Wild and Cultivated Species of <Emphasis Type="Italic">Trifolium</Emphasis>

The genus Trifolium comprises of 290 annual and perennial species of which the species such as T. repens, T. hybridum, T. pratense, T. ambiguum, T. resupinatum, T. alexandrinum are economically important. Boundaries between species in many cases have been difficult to define because of wide range of diversity caused by primary polymorphism. Hence, inter- and intraspecies variation in Trifolium, for zymogram pattern of five enzyme system was made to work out estimate of variability for isozymic banding pattern and get an insight into the species relationship. A total of 25 species represented by 134 accessions were compared for 5 enzymes viz. peroxidase, esterase, superoxide dismutase, acid phosphatase, and glutamate oxalo acetate transaminase using starch gel electrophoresis. Forty-six types of zymograms for Est isozyme pattern were observed amounting to 4.38 estimate of variability. The estimates of variability revealed maximum variation in T. resupinatum (4.24) followed with 3.02 in T. nigrescens. Estimate of variability for superoxide dismutase ranged from 0.46 to 2.67 among species amounting to 1.08 total variability across species. A total of 28 types of glutamate oxalo acetate transaminase zymograms were observed accounting for 2.48 estimate of variability. All but one band attributed to an estimate of variability of 1.43 in the genus for ACP and 16 different types of ACP zymograms were noticed. Highest variation for ACP was observed in T. resupinatum (4.53). Estimate of variation for peroxidase was 4.83 and 51 types of zymogram were observed. The species differed markedly for zymograms. The species distributed both in temperate and tropical parts like T. resupinatum had more variability as compared to cultivated species like T. alexandrinum and T. pratense. The rich variability present among these species can provide good source of gene transfer from wild to cultivated species which otherwise have no specific zymogram and exhibit low variability. The species sharing zymogram pattern for one or more enzymes with cultivated species were considered to have affinity and can further be utilized in attempting interspecific hybridization.     

Analysis of Microsatellite Diversity in Ethiopian Tetraploid Wheat Landraces

The extent and patterns of microsatellite diversity in 141 Ethiopian tetraploid wheat landraces consisting of three species Triticum durum Desf., T. dicoccon Schrank and T. turgidum L. were analyzed using 29 microsatellite markers. A high level of polymorphism and a large number of alleles unique for each species were detected. Compared to emmer (T. dicoccon) and poulard (T. turgidum) wheats, a higher genetic diversity was observed in T. durum. The A-genome was more polymorphic than the B-genome in all the three species. Microsatellites with (GA) _n -repeats had a higher number of alleles than (GT) _n -repeats. A species pairwise comparison was made to determine the percentage of shared alleles and a large number of common alleles among species were observed. Average gene diversity, across the 29 microsatellite loci, was 0.684 for T. durum, 0.616 for T. dicoccon and 0.688 for T. turgidum. Genetic distances were lower between T. durum and T. turgidum (0.26) than between T. durum and T. dicoccon (0.34) or between T. turgidum and T. dicoccon (0.38). A significant correlation (p < 0.01) was found between the number of alleles per locus and the gene diversity in all the three species. Allelic frequency variation was highest between T. turgidum and T. dicoccon (10.62%) and lowest between T. durum and T. turgidum (4.86%). A genetic similarity coefficient of 0.34, 0.46 and 0.37 was found in T. durum, T. dicoccon, and T. turgidum, respectively. The dendogram, which was constructed on the basis of a similarity matrix using the UPGMA algorithm, distinguished all accessions represented in the study.     

Use of GIS for Optimizing a Collecting Mission for a Rare Wild Pepper (<Emphasis Type="Italic">Capsicum flexuosum</Emphasis> Sendtn.) in Paraguay

This paper presents an effective method for prioritizing areas within a country for acquisition of germplasm of a crop gene pool for ex situ conservation. The method was applied to the rare wild pepper species, Capsicum flexuosum Sendtn., in south-east Paraguay. A model to prioritize areas for collecting germplasm was constructed by combining (1) a prediction of the species' geographic distribution based on the climate at previous collection points, (2) the distribution of forest margins (the species' natural habitat) and (3) areas accessible by road. The model was then tested in the field by visiting 20 sites having both high and low predicted probability of occurrence of C. flexuosum. Six new populations were found, representing a significant improvement over two previous collecting missions for the species in the same region, undertaken without the use of GIS targeting. Using the most optimistic analysis of model performance, C. flexuosum was found at five out of seven points predicted to harbour the species and not found at four of five points predicted not to harbour the species. The model was then improved by the use of higher resolution climate surfaces. It is recommended that future explorers use more recent and higher resolution satellite images to locate suitable habitats. The method is replicable for different species in different geographic regions and is offered as a means of optimizing efficiency in financially constrained, national plant genetic resources programs.     

Genetic variation within and among species of five sections of the genus Arachis L. (Leguminosae) using RAPDs

Some Arachis species are widely used as commercial plants, e.g. the groundnut A. hypogaea, an important source of good quality protein and oil, and A. pintoi and A. glabrata, that are utilized as forage species. Germplasm of most Arachis species is available in germplasm banks. However, little it is known about the genetic attributes of this germplasm, and mainly about its genetic variability, which is very important for its maintenance. In the present study RAPDs were used to assay the genetic variation within and among 48 accessions of five sections of the genus Arachis and to establish the genetic relationships among these accessions. Ten of 34 primers tested were selected for DNA amplification reactions since they yielded the largest numbers of polymorphic loci. A dendrogram was constructed based on data from the 10 primers selected. Eighty RAPD polymorphic bands were analyzed among the accessions studied. The relationships among species based on RAPDs were similar to those previously reported based on morphological, cytological and crossability data; demonstrating that RAPDs can be used to determine the genetic relationships among species of the different sections of the genus Arachis. In general, wide variation was found among accessions and low variation was found within the accessions that had two or more plants analyzed. However, higher polymorphism was found in the section Trierectoides and in one accession of A. major, indicating that generalizations should be avoided and each species should be analyzed in order to establish collection and maintenance strategies.     

Preliminary study of genetic diversity in Swedish flax (Linum usitatissimum)

To investigate the genetic diversity of Linum usitatissimumL. in Sweden, 18 accessions, including 13 cultivars and five landraces, were analysed. This study was based on genetic variation in three enzyme systems (i.e., PGD, GPI and MDH) by using horizontal starch gel electrophoresis. The total genetic diversity of the studied flax material was very high (H _T= 0.62). Even though the highest genetic diversity lies within the accessions (G _ST= 0.07), a clear differentiation between fibre and oil flax was found with respect to three polymorphic loci (Pgd-1, Gpi-2 and Mdh-1). A phenogram, based on Nei's genetic distances between the accessions studied, showed five clearly defined groups but with low variation within the groups. The unexpected high genetic diversity found within accessions in the studied flax material may indicate that flax is more outbreeding than earlier believed.     

RAPD and AFLP assessment of genetic variation in a landrace of pepper (Capsicum annuum L.), grown in North-West Italy

In several regions of Italy as well as other parts of southern Europe, the heterogeneity of the land, the climate and the soil favour the survival in cultivation of a large number of landraces specifically adapted to local conditions. Knowledge on the level and distribution of their genetic variation can help to develop appropriate strategies, in order to suistainably manage in situ these germplasm resources at risk of genetic erosion. C. annuum is an herbaceous diploid species and is considered to be self-pollinating, although different rates of out-crossing have been recorded. We used random amplified polymorphic DNA (RAPD) and amplified fragment length polymorphism (AFLP) markers to assess genetic diversity within and between five populations of a landrace of Capsicum annuum L., grown in a limited area in north-west Italy and locally known as Cuneo pepper. Partitioning the genetic variation with Shannon's diversity index revealed that 41.6% occurred between and 58.4% within populations. Analogous results were obtained when the analysis was based only on RAPD or AFLP markers. However, AFLP was more reliable, since a lower range of variation was observed among primer combinations in detecting the two components of genetic variation. Notwithstanding the rather high level of within genetic variation detected, the five populations were clearly differentiated and differed in the frequency of alleles exclusive and/or present at very low frequencies. Our results show the need for accurate estimation of allele frequencies, in order to identify populations to which priority should be given for dynamic conservation of landraces.     

Mapping the potential distribution of five species of Passiflora in Andean countries

Geographic location data for 383 accessions of five Passiflora species from five Andean countries (P. cumbalensis, P. mixta, P. tripartita var. mollissima, P. natistipula, and P. manicata) were linked to interpolated continental surfaces of monthly mean rainfall, monthly mean temperature, and monthly mean diurnal temperature range. This permitted us to identify the potential distribution of each species and document its climatic adaptation. Maps are presented showing regions where the climate is similar to that in areas where each species has been found, but from which no collections have yet been made, thus representing possible geographical gaps in collections. Some species showed evidence of intra-specific variation in their climatic adaptation. The study of Passiflora diversity in Andean countries continues and these maps based on passport data are expected to be useful tools for the planning of both ex situ and in situ conservation activities.     

Inter-simple sequence repeat (ISSR) and RAPD variation among wild barley (Hordeum. vulgare subsp. spontaneum) populations from west Turkey

Inter-Simple Sequence Repeat (ISSR) and Randomly Amplified Polymorphic DNA (RAPD) markers were used to analyze genetic distance among H. vulgare subsp. spontaneum populations from west Turkey. Fifty-five RAPD and 10 ISSR primers were used to detect variation among sample. A total of 55 polymorphic loci were found using 65 primers. Two distinct cluster groups were clearly established among populations. The minimum variation was detected between Pinarbasi and Bornova (GD = 0.192) populations and the maximum was found between Icmeler and Aydin populations (GD = 0.926). As two dominant markers, RAPD and ISSRs are effective and promising marker systems for detecting genetic variation.     

Genetic structure among earthworms (Lumbricus terrestris L.) from different sampling sites in western Germany based on random amplified polymorphic DNA

Earthworms are being used as bio-indicators to assess terrestrial pollution. However, it is often not known whether their populations possess a uniform genetic structure, which would allow comparison of residues or biological properties of earthworms from different sampling locations. In order to investigate this point, random amplified polymorphic DNA (RAPD) variation was surveyed in earthworms (Lumbricus terrestris) from five different sampling sites in Germany. Forty oligonucleotide RAPD primers (10 base pairs in length) were screened, three of which produced high polymorphic band patterns. A total of 61 DNA fragments were detected in 90 individuals of L. terrestris from five sampling sites with 49 (80.3%) RAPD markers being polymorphic. The genetic similarities within (band sharing rates between 0.756 and 0.795) and among the L. terrestris populations (0.635) were similar even at widely separate locations. Inter-population variation in the RAPD pattern for all five earthworm populations accounted for 37.9% of the total variation, while intra-population variation for three adjacent Saarland populations accounted for only 18.0% of the total variation. Principal component analysis (PCA) and the genetic distances of the populations confirm these results. Twenty-four percent of the genetic distance is caused by geographical isolation as shown by a test for isolation by distance. These results show that L. terrestris fulfils the genetic qualifications for a bio-indicator particularly at closely located sampling sites. However, the results also suggest that earthworm studies of widely separated locations should include genetic characterisation of the earthworm samples.     

Argania spinosa (L.) Skeels flowering phenology

Argan (Argania spinosa (L.)Skeels) flowering was observed at Ait Melloul, Ait Baha andArgana, three sites in the south-west of Morocco during twoseasons, 1994–95 and 1995–96. Glomerule count wasmonitored on first-year and on last-year growth shootevery twenty days. Season was the primary source of variation ofglomerule count on argan first-year and last-yeargrowth shoots. The first season was very dry when flowering beganlate April, while the second was very humid and flowering startedearly December. In both cases flowering precocity was associated toprecipitation precocity, i.e. flowering would commence whenapproximately 100 mm rain was accumulated. High positivecorrelation between glomerule count and precipitation were observed.But, considering the two seasons, glomerule count always attained itspopulation maximum in spring even if differences were observedbetween sites. However, intensity and duration of the peak were alsodistinct depending on shoot type: on first-year growth shoot,glomerules were abundant but would last for a short period, whereason last–year growth shoot, they were less frequent but wouldlast for a longer period. Not all trees flowered during a season,although some trees flowered every season, showing the extremes ofthe large variability observed for glomerule count abundance andduration within argan populations.     

Genetic variation analysis in the genus Lathyrus using RAPD markers

RAPD analysis was applied to five species belonging to the genusLathyrus (Fabaceae): L.sativus, L.cicera, L.ochrus, L.sylvestris and L.latifolius. All the species under study belong to thesection Lathyrus except L.ochrus which is in section Clymenum.Nine populations representing these species were used and ten random10-mer primers were sampled. A total of 129 amplification products,ranging in size from 0.3 to 3 Kb, were generated. Partitioning ofvariation was studied using the AMOVA technique. The genetic variation proved tobe nearly equally distributed among species and among populations withinspecies. A similar approach was carried out to distinguish populations, whichproved to be efficient. The between population dissimilarities were calculatedand a dendrogram of genetic relationships was drawn. The RAPDs obtained weresufficient to distinguish between the two accessions of a species and toseparate these accessions by clustering them according to species. A highgenetic similarity between the populations of L.sylvestris and L.latifolius was established. A similar result is also shownfor the populations of L. sativus andL. cicera.     

Population genetic structure of Milicia speciescharacterised by using RAPD and nucleotide sequencing L.

Milicia species(M. excelsa andM. regia, Moraceae, theirokos) are among the most useful indigenous rain forest treespecies in Africa. They are not grown in plantations but extractedfrom the natural forest at an unsustainable rate. The majorconstraint on cultivation and afforestation is thegall-forming insect Phytolyma lata, whichdestroys seedlings through gall formation, causing dieback of theshoot. Genetic diversity studies should aid the design ofconservation and resistance-breeding strategies. This study,therefore, aimed to investigate the genetic variability inMilicia from three West African countries andthe power of bulk-family DNA in partitioning of geneticdiversity. Genetic variation within and between 5 natural populationsof Milicia from Ghana, Côte d'Ivoireand Sierra Leone was investigated using random amplified polymorphicDNA (RAPD) and non-coding chloroplast DNAsequencing. RAPD analysis of 41 DNA samples with 8 primers produced atotal of 74 bands with 40 bands (54.1%) beingpolymorphic. Dendrogram analysis produced 2 major clusters separatingdry and moist/wet ecotypes. Sub-clustering furtherseparated accessions by forest type and/or geographic region. Agreater proportion of the total genetic variability was attributed tovariation between populations (62.2%) than withinpopulations (37.8%). Sequencing of chloroplasttRNA^Leu intron (cptrnL) and intergenic spacer(cpIGS) between cptrnL and tRNA^Phe(trnF) did not show any variation. The RAPD analysissuggests that trees from wet/moist sites are mostlyM. regia, while those fromdry sites are mostly M.excelsa. Sequencing results, however, suggestthat the two species are closely related. These results are discussedwith reference to conservation strategies.     

A-genome cotton as a source of genetic variability for Upland cotton (<Emphasis Type="Italic">Gossypium hirsutum</Emphasis>)

Since Upland cotton (Gossypium hirsutum) is known to have relatively low levels of genetic diversity, a better understanding of variation and relationships among possible sources of novel genes would be valuable. Therefore, analysis of genetic variation of the genus Gossypium, especially the diploids, which are the putative donors of the A and D genomes for the commercially important allotetraploid cottons (AADD), G. hirsutum and G. barbadense, could provide important information about the feasibility of using these genetic resources for cotton improvement. The primary objective of this study was to analyze the genetic diversity in A-genome diploid cotton species, G. herbaceum (A1) and G.␣arboreum (A2) by using microsatellite markers. Forty-one A-genome germplasm accessions were evaluated with 32 microsatellite loci. Genetic similarities between A1 and A2 ranged from 0.62 to 0.86 with a mean of 0.70. Within each A-genome species similarities ranged from 0.80 to 0.97 with a mean of 0.89 for A1 and from 0.82 to 0.98 with a mean of 0.89 for A2. A UPGMA tree and principal coordinate analysis based on genetic similarity matrices showed distinct clusters consistent with the genomic groups.     

Comparison of the genetic structure of populations of wild emmer wheat, <Emphasis Type="Italic">Triticum turgidum</Emphasis> ssp. <Emphasis Type="Italic">dicoccoides</Emphasis>, from Israel and Turkey revealed by AFLP analysis

The aim of the present study was to assess the genetic variation in several Israeli and Turkish populations of wild emmer wheat, Triticum turgidum ssp. dicoccoides, the progenitor of most domesticated wheat. Single spikes were collected in 2002 from 60 plants that grew in six different habitats in Ammiad, northeastern Israel (8–12 plants from each habitat), and in 1998 from 56 plants that grew in seven different habitats in Diyarbakir, southeastern Turkey (8 plants from each habitat). Seeds were planted in a nursery and DNA was extracted from every plant and analyzed by the fluorescent-based amplified fragment length polymorphism (AFLP) method. Seven primer combinations produced 788 discernible loci of which 48.6% were polymorphic in Israel and 40.5% in Turkey. The genetic diversity estimates P (frequency of polymorphic loci) and He (gene diversity) were higher in Ammiad than in Diyarbakir (means of P = 0.34 and He = 0.13 in Ammiad vs. P = 0.20 and He = 0.08 in Diyarbakir). Ammiad populations contained more unique alleles than Diyarbakir populations. The relative genetic diversity estimates (θ) values were 0.188 in Ammiad and 0.407 in Diyarbakir, suggesting better differentiation of the populations in Turkey. Genetic distance was larger between Israeli and Turkish populations than between populations of each country. The data indicate that the Israeli and Turkish populations are considerably diverged and that the Israeli populations are more polymorphic than the Turkish ones, having a larger within-populations genetic variation than among-populations one. The significance of the results in relation to the differentiation pattern of wild emmer in the Near East is discussed.     

Analysis of Genetic Variation Among Accessions of <Emphasis Type="Italic">Pueraria montana</Emphasis> (Lour.) Merr. var. <Emphasis Type="Italic">lobata</Emphasis> and <Emphasis Type="Italic">Pueraria phaseoloides</Emphasis> (Roxb.) Benth. based on RAPD Markers

Pueraria montana var. lobata and P. phaseoloides originating from tropical Asia and parts of Oceania are ecologically and economically important legumes that are used as green manure, cover crop or forage plants. Conservation and use of plant genetic resources require an understanding of the extent and distribution of genetic diversity in any given region. In this study, genetic variation of five P. montana var. lobata and 16 P. phaseoloides accessions was analysed developing a random amplified polymorphic DNA (RAPD) marker methodology for Pueraria species and thereby creating basic data for follow-up research and the development of conservation strategies. Seeds were collected from native populations in Bac Kan Province, a mountainous region in Northeast Vietnam. P. montana var. lobata presented a high level of variation with 54.3% of the detected markers being polymorphic, whereas P. phaseoloides exhibited an intermediate to high level of variation (45.5%). The P. montana var. lobata accessions clustered in congruence with their eco-geographical origin. For P. phaseoloides no correspondence between sampling sites and genetic differentiation was found. Inter-population differentiation was measured as Jaccard's similarity coefficient (JSC). Mean JSC amounted to 0.35 in P. montana var. lobata and 0.52 in P. phaseoloides. Results are compared to other genetic studies of herbaceous legumes and conservation strategies are suggested.     

Population diversity of the Vicia sativa agg. (Fabaceae) in the flora of the former USSR deduced from RAPD and seed protein analyses

The genetic diversity of 58 wild and weedy populations representing taxa within the V. sativa aggregate from the former USSR, 4 cultivars of V. sativa, 2 accessions of V. cordata and 3 accessions of V. macrocarpa from Mediterranean countries were analysed using randomly amplified DNA fragments (RAPDs) and seed protein electrophoresis (SDS-PAGE). Interspecific variation between taxa in the V. sativa aggregate could readily be detected using both techniques. RAPDs and seed protein patterns were found to be an effective means of identifying accessions that cannot be identified clearly by morphological criteria alone. RAPD and seed protein analysis revealed a clear relationship between observed genetic variation of populations and their geographical distribution. Populations from each region had their own gene pools. Geographical variation was detected in V. segetalis. The degree of genetic divergence between local populations was usually related to proximity. In several locations where wild and weedy populations of different V. sativa agg. taxa grow sympatrically, intermediate forms could be detected at the DNA and protein levels. Both RAPD and seed protein analysis support the view that the V. sativa aggregate consists of 8 taxa warranting recognition at the species level. Several species in this aggregate are evolving intra-specific groups which can readily be detected at the molecular level.