Study of allozyme diversity in guar [Cyamopsis tetragonoloba (L.) Taub.] germplasm

Guar [Cyamopsis tetragonoloba (L.) Taub.] is an important commercial crop of India, mainly cultivated in the northwestern part and neighboring areas of Pakistan. High morphological diversity is reported in guar. The present study attempts to analyze the genetic diversity in cultivated guar using allozyme polymorphism and compare it with reported morphological diversity. Accessions for the study were selected from a set of guar germplasm conserved at the National Bureau of Plant Genetic Resources, New Delhi. The morphological data of 108 selected accessions was subjected to UPGMA clustering. Fifty-five accessions were then picked randomly from the clusters generated. Tris-borate system (pH 8.3) was found to be most effective for detection of allozyme polymorphism in guar. Ten enzymes were selected for the study out of 18 systems tried, namely, malate dehydrogenase, malic enzyme, phosphogluco isomerase, glutamate oxaloacetate transaminase, esterase, peroxidase, alcohol dehydrogenase, glutamate dehydrogenase, glucose 6-phosphate dehydrogenase and shikimate dehydrogenase. A total of 20 loci were detected with 34 alleles. The number of polymorphic loci in different accessions was observed to be highly variable (0–9). An average of 1.27 allozyme per polymorphic locus was observed. The allele frequency data of the 55 accessions was used to determine genetic diversity parameters. The dendrogram generated on genetic distance showed three clusters in the distance range of 0–0.28. Most of the accessions were in the first clusters in a narrow distance range of 0–0.08. The second cluster consisted of improved varieties and accession from Pakistan. The third cluster of accessions was genetically more distant but there was no definite segregation of accessions according to the place of collection. The accessions from south west Rajasthan were scattered in all the three clusters showing higher variability in this region, also supported by higher morphological variation. The genetic distance was low among accessions of guar, indicating low diversity in India.     

Nature of taro (Colocasia esculenta (L.) Schott) genetic diversity prevalent in a Pacific Ocean island, Vanua Lava, Vanuatu

Taro (Colocasia esculenta (L.) Schott), cultivated in Vêtuboso, a village of northern Vanuatu, Melanesia, was surveyed to: (1) assess the extent of morphological and molecular variation being maintained by growers at the village level and, (2) compare this diversity with the diversity found in the crops in Vanuatu. Ethnobotanical data were combined with AFLP analysis to elucidate possible sources of variation. Folk assessment of variation is based on: (a) morphological characteristics (11 characters), (b) names and (c) classification according to habitat, uses, origin and agronomic adaptation. This 3-fold approach allowed growers to differentiate 96 morphotypes, all of which are given distinct vernacular names. AFLP fingerprints successfully differentiated all these 96 morphotypes which do not present a significant intra-clonal variation. But genetic results showed no clear groupings according to geographic origin or habitat of morphotypes and stated that the diversity found within the village was comparable with the overall diversity found in Vanuatu. Local nomenclature and stories associated with each cultivar suggested three sources of diversity: introductions (38%), somatic mutations (15%) and sexual recombinations (48%). AFLP results confirm folk beliefs about origin at least for three pairs of mutants. The 11 so-called wild forms analysed by AFLP were suggested to be feral, escapes from domestication. A dynamic in situ conservation strategy (DISC), favouring a broadening of the national genetic base, was discussed for taro.     

Extensive introgression of Middle American germplasm into Chilean common bean cultivars

Summary The genetic diversity of 95. representative Chilean common bean (Phaseolus vulgaris L.) landraces was analyzed using phaseolin seed protein and eight isozyme systems as genetic markers. Four types of phaseolin were found, “C”, “T”, “S” and “H”, in decreasing order of frequency. Each type had a different distribution between the Northern and Southern regions of the country. Nei’s genetic distance based on isozyme diversity indicated that a high percentage of the total variation found in this sample occurred between landraces and only a small percentage of the variation was detected within populations. Cluster analysis based on Nei’s genetic distance and a principal component analysis of isozyme frequencies did not detect a clear association between the geographic distribution of the landraces and their isozyme constitution. However, Nei’s genetic distance analysis clustered the bean landraces into two major groups which had a specific isozyme pattern, seed color, and seed size. The genetic analysis also detected a rare polymorphism for theMdh-2 locus, a null allele at theDiap-2 locus, and polymorphism for theAco-2 locus. The principal component analysis of isozyme frequencies showed that only 30% of the genotypes analyzed were similar to the Andean check and 5% of the samples were similar to Middle American check. This finding suggests a high frequency of hybridization between the Middle America and Andean gene pools in cultivated common bean from Chile.     

Geographic differentiation of morphological traits and isozymes in the Mediterranean island endemic Anchusa crispa: implications for the conservation of a protected species

Understanding population differentiation and genetic diversity within population is critical to the development of conservation programmes for threatened species. The protected species, Anchusa crispa Viv. occurs on coastal sand dunes in three disjunct geographical groups of populations: two populations on the east coast of Corsica, several populations around a single estuary on the west coast of Corsica and at a number of sites in north-west Sardinia, where two different subspecies occur. In this study we quantify differentiation of reproductive traits and isozymes among regions and populations on Corsica and between subspecies on Sardinia. No isozyme variation was detected within any of the studied populations nor among populations from a given geographic region. Two loci showed geographic differentiation between regions on Corsica. On Sardinia, the two subspecies showed no isozyme differentiation, and, depending on the loci examined, are similar to one or other of the two regions on Corsica. Reproductive traits show significant differentiation between geographic regions on Corsica, but little variation among populations within a region. In populations on the west coast of Corsica, flowers are small and have a stigma situated at the same level as the apex of the anthers whereas on the east coast, flowers are larger and have the stigma below the anthers. On Sardinia, A. crispa subsp. crispa has a floral morphology similar to the Corsican plants, whereas subspecies maritima has larger flowers with the stigma positioned above the anthers (approach-herkogamy). Quantitative variation in floral morphology thus agrees with the separation of two taxonomic entities on Sardinia and geographic variation in reproductive traits and isozymes has important ramifications for the sampling of populations for conservation programmes.     

Isozyme analysis of indigenous cassava germplasm for identification of duplicates

Isozyme analysis was utilized for identification of duplicates in indigenous collection of cassava germplasm. 786 indigenous accessions of cassava were screened for 11 key morphological characters and the morphological duplicates were identified. Two hundred and eighteen accessions of cassava consisting of ninety one sets of morphological duplicates were analysed for esterase isozyme polymorphism. Thirty seven polymorphic bands were obtained, showing high polymorphism for this enzyme. Thirty seven double sets and 17 multiple sets were found to be similar at isozyme level. These isozyme duplicates will be further analysed for DNA similarity before eliminating them from the field germplasm. The dissimilar sets showed variation in minor morphological characters, thus confirming the relation between isozyme polymorphism and phenotypic variation.     

Isozyme diversity and analysis of the mating system of the wild rice Oryza latifolia Desv. in Costa Rica

Oryza latifolia is the most abundant and widespread wild relative of rice in the lowlands of Costa Rica. It also has the most diverse morphology. However, little is known about the genetic diversity and mating system of this allotetraploid species. Genetic analyses of nine populations from different life zones from the Pacific and Atlantic slopes were performed using six isozyme loci. Differences in genetic variability were observed among populations, of which Palo Verde and Santa Rosa were the most diverse. Most Atlantic populations clustered together, and a similar result was obtained with the Pacific populations. High levels of interpopulation diversity were observed while most populations were monomorphic for at least one genotype, suggesting little genetic flow within populations. Even in polymorphic populations no variation was observed within progeny groups, combined with Hardy-Weinberg disequilibria in most populations, commonly observed in autogamous species or in species with clonal reproduction. However, the high frequency of heterozygous-like patterns may suggest that the reproductive system of O. latifolia might be more complex. Future research may explain the genetic patterns as well as the reproductive biology of this species. The knowledge of the genetic diversity and mating system of O. latifolia could contribute to the implementation of genetic resource conservation strategies and gene flow analyses, as well as of breeding programs for rice improvement.     

High genetic diversity revealed in barley (Hordeum vulgare) collected from small-scale farmer’s fields in Eritrea

Remarkable morphological variation has been found within small Eritrean barley fields. Barley was collected from fields approximately 50 m² in size. Spike shape, type, and colour were observed to vary both between and within fields. A set of 39 Simple Sequence Repeats (SSR) markers were used to explore the genetic diversity of the Eritrean barley collected from small-scale farmer’s fields. Significant genetic diversity was found within the barley fields. Out of 240 spikes collected from 24 fields (10 spikes per field), only two spikes from geographically distant fields were genetically similar. Based on the SSR data, individual farmers’ fields were found to possess 97.3% of the genetic variation present in the Eritrean barley. We discuss a strategy to improve the barley yield in Eritrea, and to facilitate the in situ conservation of barley genetic diversity. Gunter Backes and Jihad Orabi contributed equally to this work.     

Capturing genetic diversity of wild populations for <Emphasis Type="Italic">ex situ</Emphasis> conservation: Texas wild rice (<Emphasis Type="Italic">Zizania texana</Emphasis>) as a model

Genebanks complement other conservation programs because they preserve genetic diversity needed for future breeding and restoration. We evaluated efficiency of capturing genetic diversity, using endangered Zizania texana (Texas wild rice) as a model for plants with recalcitrant seeds. This perennial aquatic grass is restricted to 4 km of the San Marcos River in Texas. An early conservation collection included plants from stands throughout the river, based on the assumption stands would be unique genotypes. Using microsatellite markers, we found that genetic diversity was concentrated in five of 15 large, demographically stable stands; 96 stands smaller than 2 m² contributed no unique alleles. High heterozygosity and few duplicate genotypes suggested that sexual reproduction occurs more often than presumed. Simulations of stratified sampling of large stands captured all alleles in only 45 individuals, while random sampling along the river captured much less diversity. The early conservation collection captured as much diversity as expected from random sampling. Texas wild rice stands resemble a mainland-island metapopulation; our analyses suggest that stratified sampling maximizes genetic diversity for this population dynamic. Demographic and genetic information is important for validating the design of efficient ex situ collections and guiding in situ conservation.     

Allozyme Diversity and Population Structure of Caragana korshinskyi Kom.in China

Caragana korshinskyi Kom. is a long-lived shrub species indigenous to northwestern China, and important in vegetation rehabilitation of widely degraded and degrading semiarid and arid regions because of its high ecological and economic values. Information at molecular level on its genetic diversity, however, is not available. Accordingly, the extent and distribution for genetic diversity and population structure in 11 populations of C. korshinskyi were assessed using polyacrylamide gel electrophoresis for seven enzymes including aminopeptidase, aspartate aminotransferase, glucose-6-phosphate dehydrogenase, malate dehydrogenase, phosphoglucoisomerase, phosphogluconate dehydrogenase, and peroxidase. The seven-enzyme systems produced 11 loci encompassing 19 alleles demonstrating high genetic variation at both species and population levels. A considerable excess of heterozygotes relative to Hardy–Weinberg expectations was detected at the both levels as well. G_ST ranged from 0.0074 for AMP-1 to 0.4646 for PGD with a mean of 0.1517, indicating that approximately 84.8% of the total allozyme variation occurred within populations. An indirect estimate of the number of migrants per generation indicated that gene flow was high among populations of the species.     

Genetic Diversity Among Traditional Ethiopian Highland Maize Accessions Assessed by Simple Sequence Repeat (SSR) Markers

Over the past three centuries, maize has become adapted to complex environmental conditions in the highlands of Ethiopia. We analyzed 62 traditional Ethiopian highland maize accessions, using 20 simple sequence repeat (SSR) markers and 15 morphological traits, to assess genetic diversity and relationships among these accessions and to assess the level of correlation between phenotypic and genetic distances. The accessions varied significantly for all of the measured morphological traits. The average number of alleles per locus was 4.9. Pair-wise genetic dissimilarity coefficients ranged from 0.27 to 0.63 with a mean of 0.49. Ward minimum variance cluster analysis showed that accessions collected from the Northern agroecology were distinct from the Western and Southern agroecologies. However, there was no differentiation between the Western and Southern accessions. This suggested gene flow between these regions. The relationship between morphological and SSR-based distances was significant and positive (r = 0.43, p = 0.001). The high genetic diversity observed among these set of accessions, suggests ample opportunity for the development of improved varieties for different agroecologies of Ethiopia. From conservation perspective, sampling many accessions from all agroecologies would be an effective way of capturing genetic variation for future collections and conservation.     

Population evaluation in crop wild relatives for <Emphasis Type="Italic">in situ</Emphasis> conservation: a case study for raspberry <Emphasis Type="Italic">Rubus idaeus</Emphasis> L. in the Leningrad region,Russia

Many crop wild relatives are usually widely distributed species, and in this relation a question arises about selecting populations of special importance for conservation. Using Rubus idaeus as an example, we propose a compound selection of wild populations for in situ conservation. Twelve raspberry populations in the Leningrad region have been evaluated from the point of view of their significance for in situ conservation. To exclude the run-wild forms from the evaluation process, we tried to identify them applying a mathematical model developed by statusmetry method. The constructed model is based on morphological features of leaves of primocanes and helps to distinguish the wild forms from the cultivated and run-wild ones. The wild raspberry populations were screened for plants with features useful for cultivated raspberry improvement such as waxy cover presence, yellow fruiting plants, spinelessness, fruiting primocanes and others. The diversity of wild raspberry populations was assessed by variation of morphological features, presence of botanical varieties and polymorphism of seed storage proteins. The results of viability status assessment for the chosen populations reflect the perspectives of their development and permit to determine the strategy and measures for optimal conservation.     

Variation and phylogeny of the triploid cultivated potatoSolanum chaucha Juz. et Buk. based on RAPD and isozyme markers

Ninety four accessions of the cultivated triploid potatoS. chaucha were analyzed and classified in genotypic groups using 9 isozyme loci and RAPD markers disclosed by 20 arbitrary 10-mer primers. Eight isozyme loci out of nine were polymorphic. A total of 22 allozymes were analyzed but none of them were specific for any genotypic group. About half (52%) of the 102 RAPD markers scored, were polymorphic, all of them showing polymorphism among groups and rarely within groups. Eighteen RAPD markers were specific for certain genotypes. The isozyme markers showed a certain amount of intra group variation which made classification less reliable than with RAPD markers. A total of 10 triploid genetic groups were discriminated using both techniques together. A single primer was found to be sufficient to distinguish all 10 groups. All varieties of a single group are considered to have been derived from the same cross and then clonally propagated, even though there is a high amount of morphological variation within a single genotypic group due probably to somatic mutations. RAPD markers have been shown to be more reliable in the classification of triploid potato varieties than other genetic markers like isozymes, proteins and morphological traits.     

Isozyme analysis of three species ofRoegneria C. Koch from China

To collect and exploitRoegneria genetic resources, isozyme variation on 7 different enzymes encoded by 26 presumptive loci were analyzed in leaf extracts ofR. kamoji, R. ciliaris, R. nakaii andTriticum aestivum cv. Chinese Spring (control species) using polyacrylamide gel electrophoresis. No isozyme polymorphism was detected within accession of the threeRoegneria species, all having self-pollinating reproduction. However, extensive variations at isozyme loci were observed among accessions ofR. kamoji — even among accessions distributed at different altitudes within the same collection area. These results suggested that: (1) a select few individuals collected at a collection site may be enough to represent the genetic variability of that population; and (2) collection and maintenance ofR. kamoji materials from different altitudes may be required to increase genetic diversity. Furthermore, the banded phenotypes at EST 1 and EST-2 loci may be used as a biochemical marker associated with a morphological character, pubescences covered at leaf margin inR. kamoji. The banded characters at EST- 1, SOD-1, SOD-2, and SOD-3a loci may be used as biochemical markers to identify theR. kamoji chromosomes carrying these loci in aT. aestivum ×R. kamoji hybridization program.     

Intra-specific DNA polymorphism in pineapple (<Emphasis Type="Italic">Ananas comosus</Emphasis> (L.) Merr.) assessed by AFLP markers

Pineapple (Ananas comosus (L.) Merr.) cultivars, often derived from somatic mutations, are propagated vegetatively. It has been suggested by isozyme data that there is little genetic variation among Smooth Cayenne cultivars. A thorough investigation of the genetic variation within the cultivated speciesAnanas comosus, particularly among commercial cultivars, will provide critical information needed for crop improvement and cultivar protection. One-hundred and forty-eight accessions ofA. comosus and 14 accessions of related species were evaluated with AFLP markers. The average genetic similarity ofA. comosus was 0.735 ranging from 0.549 to 0.972, suggesting a high degree of genetic variation within this species. With AFLP markers, discrete DNA fingerprints were detected for each commercial cultivar, breeding line, and intra-specific hybrid. Self-incompatibility, high levels of somatic mutation, and intraspecific hybridization may account for this high degree of variation. However, major cultivar groups of pineapple, such as Cayenne, Spanish, and Queen, could not be distinctively separated. These cultivar groups are based on morphological similarity, and the similar appearance can be caused by a few mutations that occurred on different genetic background. Our results suggest that there is abundant genetic variation within existing pineapple germplasm for selection, and discrete DNA fingerprinting patterns for commercial cultivars can be detected for cultivar protection. The genetic diversity and relationships of fourAnanas species are also discussed.     

Genetic variation of five species of Trifolium L. from south-west Turkey

Following a legume collection mission to south-west Turkey in 1996, five species of Trifolium were analysed for genetic variation within and between species in eleven morphological and flowering characters. The five species included two outcrossing species, T. michelianum and T. resupinatum, and three inbreeding species, T. clypeatum, T. glomeratum and T. tomentosum. The genetic diversity found was related to climate and edaphic factors. All five species showed significant amounts of genetic differentiation between sites and the species could be separated morphologically by principal components analysis and cluster analysis. The most significant source of genetic variation was found to be related to geographical distribution with those species which were widely distributed across south-west Turkey exhibiting much greater amounts of genetic variation between sites, than those which had a narrow distribution. The breeding system was found to be less important, but only the morphology of the outbreeding species showed any environmental clines in relation to climate. A multiple regression analysis was computed to estimate the effect of growing season on the days to flowering of each of the species.     

Diversity of <Emphasis Type="Italic">Ullucus tuberosus</Emphasis> (Basellaceae) in the Colombian Andes and notes on ulluco domestication based on morphological and molecular data

Ullucus tuberosus (common name ‘ulluco’) is a popular tuber crop of Andean highlands. Until now attention has been focused on ulluco from Bolivia, Peru and Ecuador, but little is known about its diversity in Colombia. Thirty-six accessions of cultivated ulluco preserved in the Gene Bank of the Universidad Nacional de Colombia were studied to assess the genetic diversity and spatial genetic structure of Colombian ulluco. We used morphological characters, molecular markers (total proteins, isozymes and RAPDs) and ploidy level. High morphological variability, especially for tuber shape and colour characters, was found. Eight accessions from north-east Colombian Andes showed some typical character-states of wild ulluco, suggesting that they could be partially domesticated forms. Genetic analysis using RAPD markers indicated that these accessions were very similar to other cultivated ullucos belonging to the same region. The regional structure observed in isozyme and RAPD dendrograms was confirmed by AMOVA results (52.6% among-region variation) and the spatial correlogram, showing the presence of two gene pools of ulluco in Colombia. Our results and the accumulated data suggest that ulluco was introduced to Colombia at least two times. Initially, semi-domesticated forms would have come from the central Andes to north-east Colombian Andes, where the native inhabitants would have completed the domestication process. Again, fully domesticated ullucos might have been introduced to south-west Colombian Andes.     

Using SSR markers to determine the population genetic structure of wild apricot (<Emphasis Type="Italic">Prunus armeniaca</Emphasis> L.) in the Ily Valley of West China

Genetic structure of three wild populations (Xinyuan, Gongliu and Daxigou) of apricot in the Ily Valley, Xinjiang Uygur Autonomous Region of China, was investigated with microsatellite (simple sequence repeat, SSR) markers. The higher polymorphism and greater transportability of these markers between Prunus species proved SSR markers were much efficient for conducting genetic diversity studies in wild apricot. Nei's gene diversity (He) and Shannon's index of diversity (I) were 0.287 and 0.458, respectively. This indicated that the wild apricot in the Ily Valley still maintained a relatively high level of diversity. The Gst of 0.137 and Fst of 0.164 revealed that genetic variation mainly resided among individuals within populations (83.6–86.3%). Population differentiation could also be found according to the distribution of SSR alleles between the populations. Mantel test showed the genetic distance between populations was significantly correlated to the geographical distance. The modest amount of gene flow (2.684) would reduce the disjunction between wild apricots. The long-distance dispersal of pollen by insects was probably the main way of gene flow between populations. Based on the study of population genetic structure, an effective conservation strategy of the species was discussed.     

Diversity in Phenotypic Profiles in Landrace Populations of Vietnamese Rice: A Case Study of Agronomic Characters for Conserving Crop Genetic Diversity on Farm

Phenotypic variation of agronomic characters in aromatic rice (Oryza sativa L.) was analyzed in order to elucidate the genetic diversity of the populations of rice landraces. Thirteen populations in Namdinh province, in the Red River Delta of Vietnam, were studied in both of farmers fields and an experimental field. Only small differences were found in agronomic characters among the populations, including populations containing different varieties. Variations among the populations in the frequency distributions of several characters suggested different degrees of diversity among the populations even among populations with the same variety name. The phenotypic profiles of farmers fields were different from those in an experimental field suggesting the environmental differences between them. One variety cultivated in the Myloc district had different phenotypic characters from the other varieties in the Haihau district more clearly in the experimental field. Since the small phenotypic variation in each of farm might be due to the genetic drift and selection by the farmers, on farm conservation of the landraces of rice is considered to be under a force to decrease phenotypic diversity. The present study suggests that the targeting several farms with different phenotypic profiles contribute to the conservation of regional genetic diversity of the landraces of rice.     

Towards a methodology for on-farm conservation of plant genetic resources

The signing of the Convention on Biological Diversity at the ``Earth Summit' in 1992, its ratification and its subsequent entering into force have highlighted the need for an approach to biodiversity conservation that employs both ex situ and in situ techniques in a complementary manner. Though much research has focused on ex situ techniques, less progress has been made in developing methodologies for the conservation of genetic diversity in situ. The definition of in situ conservation used in the Convention on Biological Diversity encompasses two distinct processes: conservation of wild species in genetic reserves and of crops on-farm. Of these two, the latter, where the genetic diversity of crop land races is conserved within traditional farming systems, has been the less studied and remains less well understood. While there are still relatively few practical examples of on-farm genetic resources conservation, most genetic diversity of immediate and potential use to plant breeders is found among land races, and there is evidence that it is being rapidly eroded. This paper attempts to set on-farm conservation within the context of plant genetic resource conservation as a whole, to introduces a possible generalised model for the conservation of genetic diversity on-farm and to promote debate around the science of on-farm conservation.