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.     

Genetic relationships among Arachis hypogaea L. (AABB) and diploid Arachis species with AA and BB genomes

The cultivated peanut (Arachis hypogaea L.) is an allotetraploid, with two types of genomes, classified as AA and BB, according to cytogenetic characters. Similar genomes to those of A. hypogaea are found in the wild diploid species of section Arachis, which is one of the nine Arachis sections. The wild species have resistances to pests and diseases that affect the cultivated peanut and are a potential source of genes to increase the resistance levels in peanut. The aim of this study was to analyze the genetic variability within AA and BB genome species and to evaluate how they are related to each other and to A. hypogaea, using RAPD markers. Eighty-seven polymorphic bands amplified by ten 10-mer primers were analyzed. The species were divided into two major groups, and the AA and the BB genome species were, in general, separated from each other. The results showed that high variation is available within species that have genomes similar to the AA and the BB genomes of A. hypogaea.     

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.     

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

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

Characterization of genetic variation and relationships among Choix germplasm accessions using RAPD markers

Choix, a plant in the tribe Maydeae of the grass family, has been cultivated in Asia for several thousand years. It is a potential gene resource for improvement of other cereal crops because of its nutritional value and tolerance to stress. Genetic variation and relationships among 21 Choix lachryma-jobi L. accessions were characterized by random amplified polymorphic DNA (RAPD) markers. A total of 205 DNA fragments across all materials were amplified with 31 random primers, averaging 6.61 per primer. Among amplified fragments, 115 showed polymorphism averaging 3.71 per primer. Of amplified markers, 56.1% were polymorphic, indicating considerable variation at the DNA level among these accessions. Some fragments were accession-specific. Pair-wise genetic similarity (GS) among 21 accessions ranged from 0.809 to 0.301. The 21 accessions clustered into two major groups. Three exotic Choix accessions clustered together. Three other Choix accessions, collected from Guangxi, China, clustered into a cohesive subgroup. Four wild types of Choix clustered into the same subgroup. These results indicated that the classification by RAPD data reflected the differences in geographic origins and evolution in Choix.     

Interspecific relationship and genetic diversity in wild beets in section Corollinae genus Beta: Isozyme and RAPD analyses

Interspecific relationship among wild beets in section Corollinae was analyzed. Isozyme allele composition supports the concept of three basic species consisting of two diploid species, Beta lomatogona and Beta macrorhiza and the tetraploid species, Beta corolliflora. The polyploid and apomictic species referred to as Beta trigyna and Beta intermedia were found as hybrids. Based on isozyme and RAPD results, B. corolliflora was also found more related to B. macrorhiza than to B. lomatogona. From the phylogenetic trees constructed from isozyme data, B. corolliflora and B. macrorhiza seem to be the end members of the evolutionary line. Analysis of genetic diversity showed that B. corolliflora was more polymorphic and heterozygous than B. lomatogona and B. macrorhiza. Differences between B. macrorhiza accessions from Turkey and Dagestan were found.     

RAPD and seed coat morphology variation in annual and perennial species of the genus Cicer L.

Molecular and morphological variation of six perennial and five annual species including domesticated chickpea, C. arietinum, were inferred on the basis of RAPD and S.E.M. seed coat features using three outgroup taxa (Lens ervoides, Lathyrus japonica and Pisum sativum). Of the 66 decamer arbitrary primers tested, eight primers revealed 87 informative fragments. Neighbor-joining cluster analysis using Jaccard's coefficient of similarity on the basis of polymorphic fragments indicated a narrow variation in C. arietinum and recognized two major clusters in the genus Cicer. The first one included four species of sect. Monocicer: C. echinospermum, C. arietinum, C. reticulatum and Iranian material of C. bijugum. The second cluster contained annual and perennial species belonging to sections Chamaecicer, Polycicer and Acanthocicer. The character state of morphological and ecological traits on the RAPD phenogram indicated no monophyletic incision. Our results suggested that the high genetic difference between annual and perennial species might be regarded as a rapid speciation of section Monocicer. Reconsideration of traditional classification of sections Polycicer and Acanthocicer is necessary. The Desi and Kabuli types of C. arietinum could not be separately grouped at the DNA level, and the low level of genetic variation of C. arietinum may result from a bottleneck during the domestication process.     

Isozyme variation in species of the section Comopyrum of Aegilops

Genetic variation at 21 enzyme loci was studied in Aegilops comosa and Ae. uniaristata, the two species belonging to section Comopyrum of Aegilops. In Ae. comosa, the mean number of alleles per locus was 2.00 and the proportion of polymorphic loci was 0.667; in Ae. uniaristata they were 1.19 and 0.143, respectively. The two species were genetically distant from each other (I=0.561) supporting the previously assigned different genome symbols, M and N.     

Assessment of genetic relationships among Pyrus species and cultivars using AFLP and RAPD markers

Twenty-five Pyrus communis L. cultivars including eight traditional Portuguese pears, and four commercial Pyrus pyrifolia (Burm.) Nak. (Japanese pear or `nashi') cultivars were analysed by RAPD and AFLP techniques focusing on their molecular discrimination and the assessment of their genetic relatedness. Twenty-five primers generated 324 RAPD markers, among which 271 (84%) were polymorphic. The AFLP technique, using seven primer combinations, revealed a similar level of molecular polymorphisms (87%), representing 418 polymorphic bands among a total of 478 scored in autoradiographs. The high reproducibility of RAPD and AFLP techniques was confirmed comparing DNA samples from different extractions and different digestions of DNA from the same plant. Three genetic similarity matrices and respective dendrograms were elaborated on using RAPD, AFLP or joint RAPD and AFLP data. Both molecular marker techniques proved their reliability to assess genetic relationships among pear cultivars. P. pyrifolia cultivars exhibit a closer genetic relatedness, clustering apart from P. communis cultivars. Within P. communis, `William's', as well as `Doyenne du Comice', cluster close to their hybrids. Most of the Portuguese cultivars tend to cluster together, indicating to constitute a relatively independent genetic pool, which can be of interest in pear breeding programs.     

Genetic Diversity within and Between Populations of Lathyrus genus (Fabaceae) Revealed by ISSR Markers

In order to evaluate the genetic diversity in Lathyrus genus, the Inter Simple Sequence Repeats method (ISSR) was exploited in five populations. These consisted of two cultivated species belonging to section Lathyrus (L. sativus L. and L. cicera L.) and a wild one belonging to the section Clymenum (L. ochrus DC.). Two 3′anchored ISSR primers and two unanchored ones, generated a total of 60 useful polymorphic DNA bands. Our data provide evidence of high molecular polymorphism at the intra- and the inter-specific levels showing that both wild and cultivated forms constitute an important pool of diversity. Moreover, among the generated DNA bands, a 500 bp band, totally absent in the banding patterns of the section Clymenum, appears to be a molecular marker of section Lathyrus. Results provided for lineage and suggest recent origin of these species that might have evolved from a common ancestor producing both L. ochrus species and the two other species L. sativus and L. cicera. These relationships support previous studies based on morphological variation and molecular analysis.     

Potential of wild species for genetic enhancement of some semi-arid food crops

Discovery and incorporation of genes from wild species provide means to sustain crop improvement, particularly when levels of resistance in the cultigens are low and virulent strains of pests and pathogens overcome the host plant resistance. The extent of utilization and the potential of the wild genepool for genetic enhancement were reviewed in five important food crops viz. sorghum, pearl millet, chickpea, pigeonpea and groundnut grown in the semi-arid tropics. Introgression from compatible wild germplasm in the primary gene pool resulted in transfer of new cytoplasmic male sterility systems in pearl millet and pigeonpea, development of high protein, cleistogamous flower and dwarf pigeonpea lines and foliar disease resistant groundnut cultivars. Utilization of wild species in secondary and tertiary gene pools has been generally limited due to sterility, restricted recombination or cross incompatibility. Nevertheless, these species are extremely important as they contain high levels of resistance to several important biotic and abiotic stresses. Several of them, like those belonging to the Parasorghum section and the rhizomatous Arachis species are sources of multiple resistances and hold great promise to sustain crop productivity.     

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.     

Competition among strains of Bradyrhizobium and vesicular-arbuscular mycorrhizae for groundnut (Arachis hypogaea L.) root infection and their effect on plant growth and yield

Summary Strains of Bradyrhizobium influenced root colonization by a species of vesicular-arbuscular mycorrhizae (VAM), and species of VAM influenced root nodulation by strains of Bradyrhizobium in pot experiments. In a field experiment, the effects of VAM on competition amongst inoculated bradyrhizobia were less evident, but inoculation with Bradyrhizobium strains increased root colonization by VAM. Certain VAM/Bradyrhizobium inoculum strain combinations produced higher nodule numbers. Plants grown without Bradyrhizobium and VAM, but supplied with ammonium nitrate (300 g ml^–1) and potassium phosphate (16 g ml^–1), produced higher dry-matter yields than those inoculated with both symbionts in the pot experiment. Inoculation with either symbiont in the field did not result in higher pod and haulm yields at harvest.ICRISAT Journal Article No. 886     

Close Genetic Proximity Between Cultivated and Wild Bactris gasipaes Kunth Revealed by Microsatellite Markers in Western Ecuador

Bactris gasipaes Kunth (peach palm or Pejibaye) is the only domesticated palm of the Neotropics. The genetic relationships between the crop and its wild relatives are still unclear. We undertook field and laboratory work in order to describe differentiation and relationships between the wild and cultivated populations of the species in Western Ecuador, and their possible interactions. A volumetric study was undertaken on the fruits of both populations, as well as a population genetic analysis in order to clarify these relationships. Fruits from cultivated plants collected in the region of sympatry of wild and cultivated plants in North-West Ecuador showed intermediate volumes between those of reference samples for the wild and cultivated plants in allopatry. Using 8 microsatellite loci, we assessed 83 wild and cultivated individuals from Western Ecuador and cultivated plants from Amazonia and Central America as a reference for the cultivated form. We detected high polymorphism in the wild and cultivated samples and low, but significant level of genetic differentiation between wild and cultivated populations. The cultivated population in North-Western Ecuador showed close genetic proximity with the sympatric wild population, consistent with the volumetric study. These results have implications for hypotheses on evolution of this crop and for strategies of genetic conservation of the wild forms.     

Genetic diversity of introduced and local Spanish persimmon cultivars revealed by RAPD markers

Persimmon culture has become an alternative for fruit growers in Italy, Israel and Spain. However, there is some confusion concerning cultivar identification in non-Asian countries were the species was introduced. Authenticating the identity of germplasm resources of persimmon would be a great value for breeding. Due to the floral biology of the persimmon species the expected genetic variability is high. RAPD markers were chosen for this study of persimmon germplasm resources in Spain. Nineteen markers were obtained from 8 primers. The average of 2.4 markers/primer was higher than that obtained in other woody fruit species. The availability of data referenced from isozymes and pomological traits revealed complete agreement among groups identified by RAPDs, isozymes and pomological traits. Results suggest that RAPD technology is adequate for fingerprinting persimmon and the data is consistent with persimmon's hypothetical origins, adaptation history and previous classification by isozymes or pomological traits. Many synonyms have been identified, mostly in cultivars from local surveys, suggesting that a few new genotypes arose in Spain since the species was introduced to Europe. By means of RAPD markers, a better identification of the European persimmon germplasm is available.     

Genetic Diversity Among Brazilian Cultivars and Landraces of Tomato Lycopersicon Esculentum Mill. Revealed by RAPD Markers

Random amplified polymorphic DNA markers (RAPD) were used to estimate the variability of 35 tomato accessions (Lycopersicon esculentum Mill.). A total of 257 reproducibly scorable bands were obtained from 20 primers, 78.6% of which were polymorphic. The percentage distribution of RAPD markers shows a bimodal distribution, and the frequency of rare alleles is similar in commercial and landrace accessions. Genetic distances among accessions were calculated and a dendrogram showing the genetic relationships among them was constructed allowing for the separation of four groups. Twenty out of 23 Brazilian landraces fell within one group, whereas commercial cultivars were distributed in the four groups. AMOVA analysis of RAPD data showed that, despite the high within Brazilian landraces and commercial cultivars variation, these two groups are significantly different, indicating that landraces can be a source of variation for breeding programs.     

An AFLP diversity study of the genus Passiflora focusing on subgenus Tacsonia

DNA variation was studied at the intra- andinterspecific levels within the genus Passiflora, with particular emphasis on the subgenusTacsonia and its relationship with the subgenusPassiflora. AFLP fingerprints were generated for35 accessions from 21 species, 14 of which belong to subgenusTacsonia, and four to subgenusPassiflora. The morphologically intermediateP. manicata (subgenusManicata) was also included, as wereP. trinervia (subgenusPsylanthus) and P.alnifolia (subgenus Decaloba). The two latter were used as outgroup species. Twoprimer combinations generated 34–80 scorable fragments pergenotype for a total of 260 fragments. Polymorphism analyses spannedthe levels from individual genotype to section. SubgenusTacsonia presented wide genetic variability atboth intra- and interspecific levels, particularly within wildspecies. The classification based on these markers generally agreeswell with current taxonomic definitions of common species, but lesswith definitions of sections. A geographical component was evident inthe intraspecific variation of P.mixta, P.tripartita var. mollissimaand P. tarminiana, withwider variation in Ecuador and Peru than in Colombia and Venezuela.These same three species, plus P.cumbalensis, formed adjacent clusters, showingcloser affinity with each other than with the other species ofsubgenus Tacsonia. As expected, P. manicata clustered at anintermediate position between subgenera Tacsoniaand Passiflora.     

Phyletic relationships within the genus Hordeum using PCR-based markers

Fifteen Hordeum species and subspeciesbelonging to groups with different genomes were studied usingPCR-based markers to establish phylogenetic relationshipswithin the genus. Two hundred and seventeen RAPD and STS markers wereused to calculate genetic distances and construct phylogenetic trees.The phenetic analysis clearly separated the primary gene pool,represented by H. vulgaressp. vulgare, H.vulgare ssp. vulgare convar. vulgare f. agriochriton andH. vulgare ssp.spontaneum, from the secondary gene pool,represented by H. bulbosumand the tertiary gene pool, represented by American wild barleys andH. bogdanii. Data obtainedfrom PCO analysis are in complete agreement with taxonomicclassifications proposed previously, which were comparisons ofnumerous morphological, cytological and reproductivecharacters.     

Sources of resistance to cyst nematode in cultivated and wild Cicer species

Summary Among the nematodes infesting chickpea (Cicer arietinum L.) plants in Syria, cyst nematode (Heterodera ciceri Vovlas, Greco et Di Vito) is the most important. It is uneconomical to grow chickpea in fields infested with cyst nematode and to control this nematode with nematicide. Therefore, investigations were conducted at ICARDA, Syria from 1987 to 1991 to identify sources of resistance to cyst nematode in 7258 lines of C. arietinum and 102 lines of eight annual Cicer species including C. bijugum K.R. Rech. (13 lines), C. chorassanicum (Bge) M. Pop. (3 lines), C. cuneatum Hochst. ex Rich. (3 lines), C. echinospermum P.H. Davis (8 lines), C. judaicum Boiss. (18 lines), C. pinnatifidum Jaub. & Sp. (18 lines), C. reticulatum Ladiz. (36 lines), and C. yamashitae Kitamura (3 lines). All lines were grown in a greenhouse at 15–25°C in pots containing soil infested with 20 eggs of the nematode g^-1 soil. Nematode infestation was evaluated on a 0 to 5 scale based on number of females and cysts on roots. Resistance was found in one line of C. bijugum, six lines of C. pinnatifidum, and one line of C. reticulatum. No lines of C. arietinum, C. chorassanicum, C. cuneatum, C. echinospermum, C. judaicum, or C. yamashitae was resistant to cyst nematode. Plants with resistance have been recovered in the F₃ generation from crosses between the cultigen and C. reticulatum, indicating the possibility of transfer of gene(s) for resistance to cyst nematode from wild to cultivated Cicer species.Joint contribution from Istituto di Nematologia Agraria, ICARDA and ICRISAT (International Crops Research Institute for the Semi-Arid Tropics), Patancheru P.O., A.P. 502 324, India.