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.     

Genetic diversity in grasspea (Lathyrus sativus L.)

Genetic diversity was investigated in 348 accessions and subaccessions of grasspea (Lathyrus sativus L.) from 10 geographical regions. Polymorphism for 20 isozymes of 13 enzyme systems was studied to estimate the genetic diversity. The Near East and North Africa regions included the most variability for these isozyme systems, suggesting that the center of diversity (center of origin) for grasspea is in this general area. The lowest variability was found in accessions and subaccessions from South America, followed by those from Sudan–Ethiopia. Diversity was measured for individual loci over regions and EST-1 and SKDH had the highest genetic diversity. The closest genetic diversity was observed for LAP-2, followed by AAT-1 and PGM. The closest genetic distance existed between populations from the Near East and North Africa. Populations from South Asia and Sudan–Ethiopia, though geographically widely separated, exhibited a closer genetic distance from each other than from other regions.     

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.     

Germplasm characterization of Sesbania accessions based on isozyme analyses

Twenty-two accessions belonging to seven species of Sesbania Scop. (S. emerus, S. exasperata, S. grandiflora, S. rostrata, S. sesban, S. tetrapteraand S. virgata), were evaluated in order to characterize the isozyme electrophoretic patterns and to estimate their genetic variability and outcrossing rates. Eight isozyme systems were used: acid phosphatase (ACP), isocitrate dehydrogenase (IDH), phosphoglucomutase (PGM), malate dehydrogenase (MDH), phosphoglucoisomerase (PGI), glutamate oxaloacetate transaminase (GOT), peroxidase (PRX) and catalase (CAT), with two gel/electrode buffers and distilled water as the extraction buffer. Cotyledons were chosen for enzyme extraction. The accessions were compared considering the presence/absence of bands for each system. A dendrogram was obtained using the Jaccard similarity index and the UPGMA clustering method, classifying the accessions into eight groups, evidencing both inter- and intraspecific variability. Annual species were classified in one major group separate from the perennials (S. grandiflora, S. sesban, S. virgata), which were allocated into two other main groups. S. virgata accessions (subgenus Daubentonia) were clearly separated from the other species. Isozyme banding patterns for the accessions are presented for each system. A high incidence of intraspecific monomorphism was observed. Interpretation in terms of loci and alleles was made for S. sesbanand S. virgataaccessions, which presented polymorphism for the systems PGI, GOT (S. sesban) and MDH (S. virgata). The average expected heterozygosity estimates varied from 0.0 to 0.231 for S. sesban and 0.0 to 0.20 for S. virgata. A mean number of 2.0 alleles per locus was observed for both species. Population multilocus outcrossing rate (t _m) of 0.62 was estimated for S. sesban, indicating a partial allogamy mode of reproduction. Individual families presented multilocus rates varying from 0.00 to 1.00, indicating both self-pollination and cross-pollination.     

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.     

Genetic diversity of 38 spinach (<Emphasis Type="Italic">Spinacia oleracea</Emphasis> L.) germplasm accessions and 10 commercial hybrids assessed by TRAP markers

Target region amplification polymorphism (TRAP) markers were used to assess genetic variability among 38 germplasm accessions and 10 commercial hybrids of spinach (Spinacia oleracea L.), an economically important leafy vegetable crop in many countries. Germplasm accessions with different geographic origins and 10 commercial hybrids were examined. For assessing genetic diversity within accessions, DNA was extracted from 12 individual seedlings from six germplasm accessions and two hybrids. A relatively high level of polymorphism was found within accessions based on 59 polymorphic TRAP markers generated from one fixed primer derived from the Arabidopsis-like telomere repeat sequence and two arbitrary primers. For evaluating interaccession variability, DNA was extracted from a bulk of six to 13 seedlings of each accession. Of the 492 fragments amplified by 12 primer combinations, 96 (19.5%) were polymorphic and discriminated the 48 accessions from each other. The average pair-wise genetic similarity coefficient (Dice) was 57.5% with a range from 23.2 to 85.3%. A dendrogram indicated that the genetic relationships among the accessions were not highly associated with the geographic locations in which the germplasms were collected. The seven commercial hybrids were grouped in three separate clusters, suggesting that the phenotype-based breeding activities tended to reduce the genetic variability. This preliminary study demonstrated that TRAP markers are effective for fingerprinting and evaluating genetic variability among spinach germplasms. Mention of trade names or commercial products in this article is solely for the purpose of providing specific information and does not imply recommendation or endorsement by the US Department of Agriculture. The U.S. Government's right to retain a non-exclusive, royalty-free license in and to any copyright is acknowledged.     

Genetic diversity analysis in Boerhavia diffusa L. of different geographic locations in India using RAPD markers

Boerhavia diffusa is extensively used in herbal medicines as well as in the Ayurvedic system, because it contains a set of clinically important compounds. In the present study, the genetic variability in Boerhavia diffusa between accessions of different geographical origin within the Indian territory is assessed through random amplified polymorphic DNA (RAPD) markers. Twenty-eight accessions of Boerhavia were screened with eighteen primers of which nine were found to be the most informative. The degree of polymorphism was found to be high in accessions collected from different places of Uttar Pradesh (Set II) in comparison to other states of India (Set I). A relatively lower level of polymorphism was recorded in accessions collected from diverse locations around Lucknow (Set III). Accessions from neighbouring geographical regions exhibited more similarity than those from distant regions (as revealed by the set I analysis). Certain diagnostic markers may be correlated with morphological character(s) such as plant type. BDL appeared most distinct and divergent from the rest of the accessions and the BDJ plant in set II also showed least similarity estimate. Fragments of 5.62 Kb and 4.47 Kb with primer GN59 was found to be unique for BDP and BD2 having ovate leaf character, whereas ovoid leaf genotype exhibited 0.79 Kb (GN34 primer) fragment. Similarly a unique band type (0.35 Kb) with primer GN83 was present in BDL and BD1 that share light pink flower. Jaccard's, and Nei and Li similarity coefficient values amongst the accessions were in the range of 0.22 to 0.89 and 0.33 to 0.93, respectively. Association of RAPD markers with the leaf characteristics, flower colour as well as with geographical locations has been made. This shows that RAPD markers are also useful for the study of genetic structure of Boerhavia populations.     

Use of the bulked genomic DNA-based RAPD methodology to assess the genetic diversity among abandoned Korean tea plantations

The variability of 20 abandoned Korean tea plantations was investigated using the RAPD methodology and thebulked DNA procedure. Out of 50 primers screened, 13 revealed diversity among the populations and generated26 polymorphic markers in all. Two and two populations were indistinguishable from each other and in general,the diversity detected among the populations was relatively low. The genetic distance calculated from theJaccard's variability index varied from 0 to 0.79 with an average of 0.44. The low genetic diversity found in thisstudy could be explained by the narrow genetic base of the Korean tea populations introduced from MainlandChina, by the short history and relatively homogeneous environment of the populations, by mass destruction oflocal tea plantations in the 14th century and by the reproductive mode of Camellia sinensis. Indeed, Camellia sinensis being a highly outcrossing species maintains high intrapopulationnal variability as compared to itsinterpopulation variability. In the light of the results obtained, a strategy for local tea conservation is discussed.     

Molecular characterization of mulberry genetic resources indigenous to India

Mulberry (Morus L.) is essential for the sericulture industry as the primary feed for the silkworm Bombyx mori L. in India, with its long tradition of practicing sericulture, has a large number of indigenous cultivars. Since knowledge on the genetic divergence of these cultivars/varieties is essential for proper conservation and utilization, Inter Simple Sequence Repeat (ISSR) profiling was employed to assess genetic relationships among 34 mulberry accessions, collected from different regions of India. By using 12 ISSR primers, which produced 72 markers displaying a high degree of polymorphism (94.4%), genetic dissimilarity coefficients were calculated for each pair of the accessions and clustering of the accessions with Unweighted Pair Group Method using Arithmetic average (UPGMA) analysis was done to unravel the genetic diversity among the accessions. The dissimilarity coefficients varied from 0.111 to 0.692. UPGMA analysis generated a dendrogram with six groups and five isolates. Clustering of the accessions did not correspond with the information on the geographic origin of many of the accessions. Two-dimensional representations of the relative positions of the accessions with regards to divergence using the first two canonical variates as co-ordinate axes revealed considerable variability among the cases in each group. Further, Discriminant Function Analysis (DFA) showed changes in the geographic origin of 11 accessions and species status of 20 accessions.     

Morphological and molecular diversity analysis among the Indian clones of <Emphasis Type="Italic">Sesuvium portulacastrum</Emphasis> L.

Sesuvium portulacastrum L. (seapurslane) is a halophyte used as pioneer species in sand dune fixation and stabilization of saline soil. Studies on the morphological and molecular diversity were carried out for the 14 clones of Sesuvium collected from the different coastal regions of India. Significant differences were observed for morphological traits viz., length, width, diameter and area of leaf, internodal distance and stem diameter for different clones when compared with the clone from Gujarat state (GJ1). A UPGMA dendrogram for morphological traits based on the Pearson’s similarity coefficient clustered the clones into three groups considering 80% polymorphism as criteria. Molecular diversity among the clones was studied using Randomly Amplified Polymorphic DNA (RAPD), Internal Transcribed Spacer (ITS) and markers specific to Ac homologous region. Of the total 749 RAPD loci amplified with 70 random primers, 294 were polymorphic with 39.25% diversity. A phylogenetic tree constructed with UPGMA and SHAN, grouped the clones into three major clades based on RAPD data. The molecular diversity studied with ITS and markers specific to Ac homologous region revealed 37.50% and 66.66% polymorphism and clustered the clones into three and four clades, respectively. The genetic diversity analysis revealed wide variations among the S. portulacastrum clones, reflecting a high level of diversity within the species which might be due to anthropogenic impact and geographic environmental conditions. Further, the various clones from the different eco-geographic coastal localities might have originated from native places of wild abundance. To the best of our knowledge, this is the first attempt to evaluate both morphological and genetic diversity among the Sesuvium clones collected from the distant habitats of the coastal regions of the India.     

Study of clonally propagated cassumunar ginger (<Emphasis Type="Italic">Zingiber montanum</Emphasis> (Koenig) Link ex Dietr.) and its relation of wild <Emphasis Type="Italic">Zingiber</Emphasis> species from Thailand revealed by RAPD markers

The genetic relatedness among 51 accessions, 14 species of the genus Zingiber and genetic variability of a clonally propagated species, Zingiber montanum (Koenig) Link ex Dietr., from Thailand were studied using random amplified polymorphic DNA (RAPD) profiles. Twenty-nine random primers gave reproducible amplification banding patterns of 607 polymorphic bands out of 611 scored bands accounting for 99.40% polymorphism across the genotypes. Jaccard’s coefficient of similarity varied from 0.119 to 0.970, indicative of distant genetic relatedness among the genotype studied. UPGMA clustering indicated eight distinct clusters of Zingiber, with a high cophenetic correlation (r = 1.00) value. Genetic variability in Z. montanum was exhibited by the collections from six regions of Thailand. High molecular variance (87%) within collection regions of Z. montanum accessions was displayed by AMOVA and also explained the significant divergence among the sample from six collection regions. Our results indicate that RAPD technique is useful for detecting the genetic relatedness within and among species of Zingiber and that high diversity exists in the clonally propagated species, Z. montanum.     

Genetic Diversity in Accessions of Wild Rice Oryza granulata from South and Southeast Asia

Oryza granulata, an upland wild rice species, represents an unique germplasm for possessing abilities of tolerance to shade and drought, immune to bacterial blight and resistance to brown planthopper. Although low degree of genetic variability has been revealed within its populations, little genetic information at the species level is available in determining rational conservation strategies. Here we used dominant DNA marker random amplified polymorphism DNA (RAPD) to assess the genetic variability among 23 accessions of O. granulata that collected from main distribution areas worldwide. Twenty decamer primers generated a total of 243 bands, with 83.5% of them (203 bands) being polymorphic. Calculation of Shannon index of diversity revealed an average value of 0.42 ± 0.25, indicating that O. granulata maintains a relatively high degree of genetic diversity on the species level. Analysis of genetic dissimilarity (GD) showed that genetic differentiation occurred among studied accessions, which supports the feasibility of current ex situ conservation strategies. We also suggested that information based on population studies, which could be achieved by international co-operation, is needed to conserve this widespread germplasm more effectively.     

Genetic Diversity of Glutenin Protein Subunits Composition in Durum Wheat Landraces [Triticum turgidum ssp. turgidum Convar. durum (Desf.) MacKey] from the Mediterranean Basin

The genetic diversity of high and low molecular weight glutenin subunits of 63 durum wheat landraces from different geographical regions in the Mediterranean Basin was studied using SDS-PAGE. Great variability in glutenin composition was found, with 42 high and low molecular weight glutenin haplotypes, 20 allele combinations at the HMW-GS loci, and 18 at the LMW-GS. All five possible LMW models were detected in all Mediterranean regions. Rare alleles were found at Glu-B1 locus in high frequencies and a priori related alleles to grain quality were also observed. Global genetic diversity index was relatively high (0.67); it ranged from 0.33 to 0.66. Cluster analysis on the frequency patterns of origins grouped genotypes following a geographical structure. Rogers’ distance coefficient on frequency pattern for each region of origin showed two germplasm pools with distinct quality profiles, where South West Asian landraces were very different from the landraces of other Mediterranean areas. The relationship between different regions of origin is discussed and two possible ways of introduction of wheat in the Iberian Peninsula (N Africa and SE Europe) are hypothesized. The use of Mediterranean durum wheat landraces as source of genetic variability for grain quality improvement is highly recommended.     

Genetic variability of Coffea arabica L. accessions from Ethiopia evaluated with RAPDs

The genetic diversity of 50 wild and semi-wild accessions of the Coffea arabica L. germplasm collection, gathered by the FAO and ORSTOM missions to Ethiopia, and maintained in Colombia by CENICAFE, was evaluated with RAPD markers. The evaluation was carried out in two phases: In phase one, the polymorphism of 8 Ethiopian accessions of different geographic origin, plus the cultivated variety 'Caturra' was assessed with the RAPD technique with forty-two 10-mer oligonucleotides. In phase two, 51 accessions were assessed with a set of 5 polymorphic primers that reproduced, with a correlation of 95%, the groups generated by the 24 polymorphic primers found in phase one. Principal Coordinate Analysis of molecular data revealed that a closely related group consisting of 86% of the Ethiopian C. arabica accessions evaluated are significantly different from the Caturra variety and could be used in a genetic breeding initiative to increase the variability of cultivated varieties. The results also indicate that a larger polymorphism is present in the Colombian replica of FAO Ethiopian coffee germplasm collection than previously reported.     

Genetic relationships and diversity among Brazilian cultivars and landraces of common beans (Phaseolus vulgaris L.) revealed by AFLP markers

The genetic variation and relationships among 31 accessions of Phaseolus vulgaris L., and two representatives of Vigna unguiculata L., were evaluated by AFLP analysis. A total of 263 DNA fragments across all materials were scored using nine primer combinations, averaging 32 per primer. More than 95% of the amplification products showed polymorphism, indicating high variation at the DNA level among these accessions. Pair-wise genetic similarity (Jaccard's coefficient) ranged from 0.553 to 0.840, with a mean of 0.765. Twenty-three accessions (70%) clustered into three groups. A majority of the commercial cultivars (91%) clustered within a single group, whereas the landraces were distributed along all the variation. An apparent correlation with phaseolin types was detected. Results of this study suggest that Brazilian landraces truly represent the overall genetic variability of Phaseolus vulgaris, confirming the multiple origins of these materials, and their potential as a source of variation for breeding programs.     

Phenotypic diversity of durum wheat (<Emphasis Type="Italic">Triticum durum</Emphasis> Desf.) from Jordan

Durum wheat landrace genotypes are disappearing from the main wheat areas in Jordan, because of spreading of new uniform cultivars and the serious reduction in wheat cultivation. This study was conducted to evaluate genetic diversity in durum wheat landraces from Jordan and to identify desirable agronomic traits. Landraces were collected from two target areas: Ajloun and Karak. The collected material was grown under rainfed conditions using an augmented design with five blocks and four repeated check cultivars. Data were collected for 14 morphological and agronomic traits. Phenotypic diversity index (H′) was estimated, and the relationships among accessions were measured using cluster analysis and dendrogram similarity matrix. The results revealed the presence of a wide range of variability among landraces., which possess high levels of variability for biological yield, fertile tillers, number of seeds per spike, seed weight per spike and weight of 1000 seeds. These landraces must be considered as a reservoir of genes that plant breeders need in their wheat improvement programs and should be conserved both ex situ and in situ.     

Genetic diversity of cowpea [<Emphasis Type="Italic">Vigna unguiculata</Emphasis> (L.) Walp.] in four West African and USA breeding programs as determined by AFLP analysis

Cowpea is an important grain legume and hay crop of many tropical and subtropical regions, especially in the dry savanna region of West Africa. The cowpea gene pool may be narrow because of a genetic bottleneck during domestication. Genetic variation within specific breeding programs may be further restricted due to breeding methods, ‘founder effects’ and limited exchange of germplasm between breeding programs. Genetic relationships among 60 advanced breeding lines from six breeding programs in West Africa and USA, and 27 landrace accessions from Africa, Asia, and South America were examined using amplified fragment length polymorphism (AFLP) markers with six near infrared fluorescence labeled EcoRI + 3/1bases/MseI + 3/1bases primer sets. A total of 382 bands were scored among the accessions with 207 polymorphic bands (54.2%). Despite a diverse origin, the 87 cowpea accessions shared a minimum 86% genetic similarity. Principal coordinates analysis showed clustering of breeding lines by program origin, indicating lack of genetic diversity compared to potential diversity. Accessions from Asia and the Americas overlapped and were distinct from West African breeding lines, indicating that germplasm from Asia and the Americas have common origins outside West Africa. US and Asian breeding programs could increase genetic variability in their programs substantially by incorporating germplasm from West Africa, while national programs in West Africa should consider introgression of Asian germplasm and germplasm from other parts of Africa into their programs to ensure long-term gains from selection.     

Genetic variation of Ethiopian and Eritrean sorghum (Sorghum bicolor (L.) Moench) germplasm assessed by random amplified polymorphic DNA (RAPD)

The extent and patterns of distribution of genetic variation among 80 sorghum (Sorghum bicolor (L.) Moench) germplasm accessions from Ethiopia and Eritrea were investigated using RAPD with 20 oligonucleotide primers. The primers generated a total of 147 polymorphic bands across the 80 accessions with a mean of 7.35 bands per primer. Estimation of the extent of variation by the Shannon-Weaver diversity index revealed an intermediate level of overall variation (H = 53), although the levels varied among regions of origin of the accessions. Partitioning of the total variation revealed considerable variation (77%) within the region of origin of the accessions and the remainder (23%) among regions of origin. Similarly, a large portion (94%) of the total variation was found within the adaptation zones compared to among the adaptation zones (6%). The results suggest a weak differentiation of the sorghum material both on regional and agro-ecological bases, which could be ascribed to the high rate of outcrossing in cultivated sorghum and its free natural hybridization with its wild and weedy relatives, as well as to seed movement by humans. The average genetic dissimilarity was found to be 36% among the 80 accessions and 13% among the 15 regions of origin. Cluster analysis failed to group accessions of the same region or the same adaptation zone, which further confirmed the weak differentiation of the material studied. The clustering pattern of the regions of origin was broadly concordant with previous clustering patterns obtained using morphological characters, in which regions with broad agro-climatic conditions were grouped together.     

Inter-Simple Sequence Repeat fingerprints to assess genetic diversity in Tunisian fig (Ficus carica L.) germplasm

The genetic diversity of 18 Tunisian fig cultivars was investigated at the DNA level using the Inter Simple Sequence Repeat (ISSR) associated with the Polymerase Chain Reaction (PCR). Using a set of primers, the most informative ones were selected that were characterized by an important Resolving power value of 29.6. A total of 47 discernible fragments were scored from samples, with a mean of 11.7 fragments per primer. The 90.4% of sample that were polymorphic were scored as molecular markers to examine the Tunisian fig germplasm polymorphism at DNA level. A large genetic diversity as related to ISSR patterns was found within the local Tunisian fig germplasm. An UPGMA dendrogram exhibits the unstructured variability in this crop. Moreover, the principal component analysis shows that the observed diversity was typically continuous. Our data provide a large number of ISSR markers that are useful in the fingerprinting of Ficus carica L. cultivars, and in the understanding of the genetic relationships among these accessions.