Wild Arachis Species Research Articles

A linkage map for the B-genome of Arachis (Fabaceae) and its synteny to the A-genome

BackgroundArachis hypogaea (peanut) is an important crop worldwide, being mostly used for edible oil production, direct consumption and animal feed. Cultivated peanut is an allotetraploid species with two different genome components, A and B. Genetic linkage maps can greatly assist molecular breeding and genomic studies. However, the development of linkage maps for A. hypogaea is difficult because it has very low levels of polymorphism. This can be overcome by the utilization of wild species of Arachis, which present the A- and B-genomes in the diploid state, and show high levels of genetic variability.ResultsIn this work, we constructed a B-genome linkage map, which will complement the previously published map for the A-genome of Arachis, and produced an entire framework for the tetraploid genome. This map is based on an F2 population of 93 individuals obtained from the cross between the diploid A. ipaënsis (K30076) and the closely related A. magna (K30097), the former species being the most probable B genome donor to cultivated peanut. In spite of being classified as different species, the parents showed high crossability and relatively low polymorphism (22.3%), compared to other interspecific crosses. The map has 10 linkage groups, with 149 loci spanning a total map distance of 1,294 cM. The microsatellite markers utilized, developed for other Arachis species, showed high transferability (81.7%). Segregation distortion was 21.5%. This B-genome map was compared to the A-genome map using 51 common markers, revealing a high degree of synteny between both genomes.ConclusionThe development of genetic maps for Arachis diploid wild species with A- and B-genomes effectively provides a genetic map for the tetraploid cultivated peanut in two separate diploid components and is a significant advance towards the construction of a transferable reference map for Arachis. Additionally, we were able to identify affinities of some Arachis linkage groups with Medicago truncatula, which will allow the transfer of information from the nearly-complete genome sequences of this model legume to the peanut crop.

Genomics: An Evolving Science in Peanut

Abstract Genomic spcience offers new research tools to explore the function of genes and their effects on plants and animals. Arachis hypogaea is a polyploid species of relatively recent origin and molecular analyses with technologies available in the 1980s and 1990s resulted in little progress in the cultivated species because of apparent lack of molecular variation. Large numbers of polymorphisms existing in wild Arachis species led to evolutionary and gene introgression studies. High throughput genomic sequencing technologies have greatly expanded the possibilities for investigating gene function, but techniques are sufficiently expensive that most federal funding has been directed toward model species and ‘major’ crops. Peanut has lagged behind many other crops, but the number of researchers working on the species in the U.S. and internationally has greatly increased during recent years. In an effort to bring researchers who work with a number of legume crops together to discuss common goals, a national strategic planning workshop was held in 2001 which led to the U.S. Legume Crops Genomics Initiative. A second workshop was held in 2004 to develop a plan with specific objectives for cross-legume genomics research and to outline milestones for accomplishments. Specifically for peanut, a genomics strategic planning workshop was organized at Atlanta in 2004 by the American Peanut Council. A broad view of genomic science was adopted and goals were set by participants to include (a) improving the utility of genetic tools for peanut genomics research, (b) improving the efficacy of technology for gene manipulation in genomics, (c) developing a framework for assembling the peanut genetic blueprint, (d) improving knowledge of gene identification and regulation, and (e) providing bioinformatic management of peanut biological information. Teams of researchers, including molecular biologists, plant breeders, pathologists, and many other disciplines need to be developed to fully utilize the potential of genomics for peanut improvement.

Post‐infection development and histopathology of Meloidogyne arenaria race 1 on Arachis spp.

The reproductive behaviour of the root‐knot nematode Meloidogyne arenaria race 1 was compared on two wild species of Arachis (A. duranensis and A. stenosperma) and cultivated peanut (A. hypogaea cv. IAC‐Tatu‐ST). The three species were considered moderately susceptible, resistant, and susceptible, respectively. Penetration and development of the root‐knot nematode in the resistant species was reduced in comparison with that occurring in susceptible plants. Several cell features, including dark blue cytoplasm and altered organelle structure were observed in the central cylinder of A. stenosperma, indicating a hypersensitive‐like response (HR) of infested host cells. Neither giant cells, nor nematodes developed beyond the second stage, were found on A. stenosperma. Arachis duranensis showed a delay in the development of nematodes in the roots compared to A. hypogaea. The two wild peanut species were chosen to be the contrasting parents of a segregating population for mapping and further investigation of resistance genes.

Flavor Profiles of Wild Species-Derived Peanut Breeding Lines

Abstract Several diploid wild species of the genus Arachis L. have been used as sources of resistance to common diseases of cultivated peanut (Arachis hypogaea L.). Because flavor is among the most important quality attributes for commercial acceptance of roasted peanuts, sensory attributes of interspecific hybrid derived breeding lines were evaluated to determine if transfer of disease resistance from wild species is associated with concomitant changes in flavor. Sixteen interspecific hybrid derivatives with five diploid species in their ancestries and the commercial flavor standard, NC 7 were evaluated for sensory quality. Significant variation among entries was found for the roasted peanut, sweet, and bitter sensory attributes, but not for the overall contrast between NC 7 and the wild species-derived breeding lines. The variation was either between two groups of wild species-derived breeding lines or within one or both groups. Introduction of disease and pest resistance traits from Arachis species did not result in degradation or improvement of the flavor profile. This suggests that flavor of wild species-derived germplasm will not prevent its use either as parents in peanut breeding programs or as cultivars.

Development and use of single nucleotide polymorphism markers for candidate resistance genes in wild peanuts (Arachis spp)

The cultivated peanut (Arachis hypogaea L.) is an allotetraploid of recent origin, with an AABB genome and low genetic diversity. Perhaps because of its limited genetic diversity, this species lacks resistance to a number of important pests and diseases. In contrast, wild species of Arachis are genetically diverse and are rich sources of disease resistance genes. Consequently, a study of wild peanut relatives is attractive from two points of view: to help understand peanut genetics and to characterize wild alleles that could confer disease resistance. With this in mind, a diploid population from a cross between two wild peanut relatives was developed, in order to make a dense genetic map that could serve as a reference for peanut genetics and in order to characterize the regions of the Arachis genome that code for disease resistance. We tested two methods for developing and genotyping single nucleotide polymorphisms in candidate genes for disease resistance; one is based on single-base primer extension methods and the other is based on amplification refractory mutation system-polymerase chain reaction. We found single-base pair extension to be an efficient method, suitable for high-throughput, single-nucleotide polymorphism mapping; it allowed us to locate five candidate genes for resistance on our genetic map.

BAC libraries construction from the ancestral diploid genomes of the allotetraploid cultivated peanut

BackgroundCultivated peanut, Arachis hypogaea is an allotetraploid of recent origin, with an AABB genome. In common with many other polyploids, it seems that a severe genetic bottle-neck was imposed at the species origin, via hybridisation of two wild species and spontaneous chromosome duplication. Therefore, the study of the genome of peanut is hampered both by the crop's low genetic diversity and its polyploidy. In contrast to cultivated peanut, most wild Arachis species are diploid with high genetic diversity. The study of diploid Arachis genomes is therefore attractive, both to simplify the construction of genetic and physical maps, and for the isolation and characterization of wild alleles. The most probable wild ancestors of cultivated peanut are A. duranensis and A. ipaënsis with genome types AA and BB respectively.ResultsWe constructed and characterized two large-insert libraries in Bacterial Artificial Chromosome (BAC) vector, one for each of the diploid ancestral species. The libraries (AA and BB) are respectively c. 7.4 and c. 5.3 genome equivalents with low organelle contamination and average insert sizes of 110 and 100 kb. Both libraries were used for the isolation of clones containing genetically mapped legume anchor markers (single copy genes), and resistance gene analogues.ConclusionThese diploid BAC libraries are important tools for the isolation of wild alleles conferring resistances to biotic stresses, comparisons of orthologous regions of the AA and BB genomes with each other and with other legume species, and will facilitate the construction of a physical map.

Genetic and epigenetic analyses of in vitro-grown plants of Arachis villosulicarpa Hoehne (Leguminosae) obtained from seed explants through different regeneration pathways

SummaryIn vitro conservation techniques are considered suitable strategies for ex situ conservation of wild species of Arachis. However, there is a potential risk of genetic and epigenetic alterations induced by tissue culture conditions. The goal of this work was to determine the influence of explant type and regeneration pathway on sequence modifications and changes in the DNA methylation status of in vitro-grown plants of A. villosulicarpa using Amplified Fragment Length Polymorphism (AFLP) and Methylation-Sensitive Amplified Polymorphism (MSAP) analyses. Regeneration from embryo axes occurred through multiplication of pre-existing meristems. Cotyledons and embryonic leaflets displayed direct and indirect organogenesis, respectively. No polymorphic AFLP amplification fragments were detected among the regenerants. Conversely, MSAP analysis showed a decrease in the levels of DNA methylation in cotyledon- and embryonic leaflet-derived plants.

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.

Evaluation of wild Arachis species for abiotic stress tolerance: I. Thermal stress and leaf water relations

The Indian groundnut cultivars have a narrow genetic base. Hence, it was of interest to investigate the genetic variability among wild Arachis species and their accessions for tolerance to thermal stress. A wide variation was observed in leaf morphological characters such as colour, shape, hairiness, length and width and thickness (SLA). The temperature and time required for 50% leaf injury was worked out with limited number of genotypes and was found to be 54°C for 50 min. Among 36 genotypes (having SLA in the range of 66 and 161 cm2 g−1) screened, the inherent potential for cold as well as heat tolerance in terms of relative leaf injury (RI) was observed. Thus, based on RI-values, A. glabrata 11824 and A. paraguariensis 12042 were identified as heat-tolerant and cold-tolerant genotypes, respectively while A. appresipila 11786 was found to be susceptible to both heat and cold. Correlation between SLA and RI values for heat (r = 0.38, P < 0.05) and cold (r = 0.52, P < 0.05) tolerance was positive, indicating that thicker the leaf the lower the injury or higher the tolerance. Among six species and 13 accessions, comprising both heat-tolerant and heat- susceptible genotypes, the concentrations of various leaf chemical constituents such as total protein, phenols, sugars, reducing sugar, amino acids, proline, epicuticular wax load and chlorophyll varied significantly. The epicuticular wax load ranged between 1.1 and 2.5 mg dm2 among 13 A.glabrata accessions. These accessions were categorized into two groups, i.e. high-wax (range: 2.0–2.5 mg dm2) and low-wax types (range: 1.1–1.6 mg dm2). The high-wax type showed a higher diffusion resistance (dr) as compared to low-wax type; though the transpiration rate (tr) in high-wax type was moderate (between 9.5 and 11.6 μg cm−2 s−1). Genetic variability in parameters such as canopy temperature, dr and tr was also distinct. The fully turgid leaves with relative water content ≥91%, showed leaf water potential (ψleaf) between −0.7 and −1.2 MPa. Results indicated that the plants with thicker leaves are better protected from heat injuries. Further, epicuticular wax load seems to help in maintaining stomatal regulation and leaf water relations, thus affording adaptation to wild Arachis species to thrive under water-limited environments. The sources of tolerance, as identified in this study, could be utilized to improve thermal tolerance of the groundnut cultivars by intra-specific hybridization, following either conventional breeding using embryo rescue techniques, if required or utilizing biotechnological tools.

Assessment of Genetic Stability Among In Vitro Plants of Arachis retusa Using RAPD and AFLP Markers for Germplasm Preservation

Abstract Arachis retusa Krapov. et W. C. Gregory et Valls is endemic in the West‐central region of Brazil, occurring in areas endangered by human actions. The establishment of in vitro preservation methods for wild species of Arachis is an alternative to seed banks for germplasm storage, multiplication and distribution. The risk of genetic changes induced by tissue culture and the monitoring of the genetic stability of the biological material before, during and after storage must be considered in the context of conservation. Random amplified polymorphic DNA (RAPD) and amplified fragment length polymorphism (AFLP) fingerprinting were used to evaluate the genetic stability of in vitro plants originated from cotyledons and embryo axes of A. retusa. Cotyledons originated shoots through direct organogenesis and embryo axes displayed multishoot formation induced by 110 mmol/L and 8.8 mmol/L BAP, respectively. Ninety genomic regions (loci) generated from RAPD and 372 from AFLP analyses were evaluated. All amplified fragments detected by both techniques in plants derived from the two explant types were monomorphic. The results indicate that the recovered shoots are genetically stable at the assessed genomic regions.

ESTs from a wild Arachis species for gene discovery and marker development.

BackgroundDue to its origin, peanut has a very narrow genetic background. Wild relatives can be a source of genetic variability for cultivated peanut. In this study, the transcriptome of the wild species Arachis stenosperma accession V10309 was analyzed.ResultsESTs were produced from four cDNA libraries of RNAs extracted from leaves and roots of A. stenosperma. Randomly selected cDNA clones were sequenced to generate 8,785 ESTs, of which 6,264 (71.3%) had high quality, with 3,500 clusters: 963 contigs and 2537 singlets. Only 55.9% matched homologous sequences of known genes. ESTs were classified into 23 different categories according to putative protein functions. Numerous sequences related to disease resistance, drought tolerance and human health were identified. Two hundred and six microsatellites were found and markers have been developed for 188 of these. The microsatellite profile was analyzed and compared to other transcribed and genomic sequence data.ConclusionThis is, to date, the first report on the analysis of transcriptome of a wild relative of peanut. The ESTs produced in this study are a valuable resource for gene discovery, the characterization of new wild alleles, and for marker development. The ESTs were released in the [GenBank:EH041934 to EH048197].

In vitro regeneration and conservation of wild species of Arachis

SummaryWild species of Arachis are restricted to South America and generally occur in regions under intensive environmental disturbance. Both in situ and ex situ conservation strategies are required in order to maintain the availability of these genotypes. This work developed in vitro regeneration systems from seed explants of 17 wild species of Arachis from six Sections (Heteranthae, Caulorrhizae, Triseminatae, Erectoides, Procumbentes and Arachis). After seed disinfection, embryonic axes, leaflets and cotyledons were excised aseptically and cultured on Murashige and Skoog (MS) medium supplemented with 8.8 µM, 22 µM or 110 µM 6-benzylaminopurine (BAP). Cultures were maintained in a growth chamber at 28° ± 2°C with a 16 h photoperiod. Regeneration patterns from seed explants were similar among species from all Sections. Embryonic axes produced plants through meristematic amplification or multiple shoot formation, while cotyledons and embryonic leaflets produced shoots at significantly lower frequencies through direct and indirect organogenesis, respectively. Shoots obtained from all explants were transferred to MS medium without growth regulators to induce root formation.

Characterization and transferability of microsatellite markers of the cultivated peanut (Arachis hypogaea)

BackgroundThe genus Arachis includes Arachis hypogaea (cultivated peanut) and wild species that are used in peanut breeding or as forage. Molecular markers have been employed in several studies of this genus, but microsatellite markers have only been used in few investigations. Microsatellites are very informative and are useful to assess genetic variability, analyze mating systems and in genetic mapping. The objectives of this study were to develop A. hypogaea microsatellite loci and to evaluate the transferability of these markers to other Arachis species.ResultsThirteen loci were isolated and characterized using 16 accessions of A. hypogaea. The level of variation found in A. hypogaea using microsatellites was higher than with other markers. Cross-transferability of the markers was also high. Sequencing of the fragments amplified using the primer pair Ah11 from 17 wild Arachis species showed that almost all wild species had similar repeated sequence to the one observed in A. hypogaea. Sequence data suggested that there is no correlation between taxonomic relationship of a wild species to A. hypogaea and the number of repeats found in its microsatellite loci.ConclusionThese results show that microsatellite primer pairs from A. hypogaea have multiple uses. A higher level of variation among A. hypogaea accessions can be detected using microsatellite markers in comparison to other markers, such as RFLP, RAPD and AFLP. The microsatellite primers of A. hypogaea showed a very high rate of transferability to other species of the genus. These primer pairs provide important tools to evaluate the genetic variability and to assess the mating system in Arachis species.

Molecular analysis of Arachis interspecific hybrids

Incorporation of genetic resistance against several biotic stresses that plague cultivated peanut, Arachis hypogaea (2n = 4x = 40), is an ideal option to develop disease resistant and ecologically safe peanut varieties. The primary gene pool of peanut contains many diploid wild species (2n = 2x = 20) of Arachis, which have high levels of disease and insect resistances. However, transfer of resistant genes from these species into A. hypogaea is difficult due to ploidy level differences and genomic incompatibilities. This study was conducted to monitor alien germplasm transmission, using Random Amplified Polymorphic DNA (RAPD) markers, from two diploid wild species, A. cardenasii and A. batizocoi, into A. hypogaea. Triploid interspecific hybrids were produced by crossing two A. hypogaea cultivars (NC 6 and Argentine) with the two species and by colchicine-treating vegetative meristems, fertility was restored at the hexaploid (C(o)) level in the four hybrids. Hexaploids were allowed to self-pollinate for four generations, each referred to as a cycle (C1, C2, C3, and C4). At each cycle, a backcross was made with the respective A. hypogaea cultivar as the maternal parent and only lineages tracing back to a single hexaploid hybrid were used for RAPD analysis. Analysis of mapped, species-specific RAPD markers in BC1F1 to BC1F3 hybrids indicated that alien germplasm retention decreased every generation of inbreeding, especially in Argentine and in A. batizocoi crosses. A similar trend was also observed for every cycle in BC1F2 and BC1F3 families, possibly, due to the loss of alien chromosomes following selfing of hexaploids. RAPD marker analysis of 40-chromosome interspecific hybrid derivatives from the four crosses supported previous reports that reciprocal recombination and/or translocations are the predominant mechanisms for exchange of chromosomal segments. No evidence was found for preferential transfer of alien chromosomal regions to specific linkage groups. The implications for developing disease resistant peanut breeding lines are discussed in light of these findings.

Biogeography of wild Arachis (Leguminosae):distribution and environmental characterisation

Geographic Information System (GIS) tools are applied to a comprehensive database of 3514 records of wild Arachis species to assist in the conservation and utilisation of the species by: (a) determining the distributional range of species and their abundance; (b) characterising species environments; (c) determining the geographical distribution of species richness; and (d) determining the extent to which species are associated with river basins. Distributional ranges, climatic variables and indices of endemism for each species are tabulated. A. duranensis Krapov. & W.C. Gregory, the most probable donor of the A genome to the cultivated peanut, is distributed in close proximity to both the proposed donor of the B genome, A. ipaënsis, and the closest wild relative of the cultigen, A. monticola Krapov. & Rigoni. This region in the eastern foothills of the Andes and the adjoining chaco regions of Argentina, Bolivia and Paraguay, is a key area for further exploration for wild Arachis. An area of particularly high species richness occurs in the State of Mato Grosso, close to the Gran Pantanal in southwest Brazil. Seventy-one percent of the species were found to have some degree of association with water catchment areas, although in most cases it was difficult to determine whether this was due to climatic adaptation reasons, restricted dispersal due to geocarpic habit, or the role of watercourses as a principal dispersal agent. In only two cases could climatic adaptation be eliminated as the reason for species distribution.

Rooting performance from leaf petioles of accessions and hybrids of wild Arachis species

The genus Arachis has 69 species, the majority native to Brazil. Arachis spp. seedling root tips have been used to obtain dividing cells for cytological preparations. Several wild species produce small amounts of seeds and consequently few root meristems, making the cytogenetic characterization very difficult. The purpose of this study was to evaluate the rooting capacity of several wild species of Arachis using the detached leaves technique for obtaining roots from petioles. One hundred and thirty accessions and 27 interspecific hybrids were studied. Three evaluations were performed at 15, 31, and 49 days after planting. Leaf rooting observed in the wild species confirmed the great genetic variability typical of the Arachis genus, indicating that the number of days interferes with the rooting percentage. The Arachis Section presented the highest rate of rooting leaves. The Caulorrhizae Section showed the same pattern observed in the Erectoides Section. Accessions from Heteranthae, Trierectoides, Extranervosae, and Rhizomatosae Sections did not differ among each other. Differences were not observed in Erectoides Section when compared to Rhizomatosae and Extranervosae Sections. The number and morphology of roots obtained through this technique were promising to be used on cytological preparations.

Microscopical studies of Arachis pintoi root nodule with special reference to bacteroids and oleosomes at different stages of plant growth and nodule development

Nitrogen fixation in legume root nodules are believed to be supported by the supply of carbon compounds derived from host cells. The presence of storage organelles such as oleosomes in the infected cells may have an advantage as a supplementary source of carbon and energy as has been suggested for peanut ( Arachis hypogaea) root nodules. The present investigation explores the development of symbiosis in a wild diploid species of Arachis in relation to the bacteroids and oleosomes. Microscopical observations at different developmental stages of nodule and host plant have revealed that oleosomes were present only during early stages of infection process and nodule development before the establishment of symbiosis, and during senescence. Oleosomes were absent in the infected cells of effective nodules of A. pintoi whereas similar cells in the nodules of other wild diploid species of Arachis ( A. batizocoi, A. duranensis) and tetraploid A. hypogaea show presence of oleosomes. Another interesting feature in this species was the reversion of spherical bacteroids into rod-forms within the confines of the senescent nodule tissue.

Genetic diversity of peanut (Arachis hypogaea L.) and its wild relatives based on the analysis of hypervariable regions of the genome

BackgroundThe genus Arachis is native to a region that includes Central Brazil and neighboring countries. Little is known about the genetic variability of the Brazilian cultivated peanut (Arachis hypogaea, genome AABB) germplasm collection at the DNA level. The understanding of the genetic diversity of cultivated and wild species of peanut (Arachis spp.) is essential to develop strategies of collection, conservation and use of the germplasm in variety development. The identity of the ancestor progenitor species of cultivated peanut has also been of great interest. Several species have been suggested as putative AA and BB genome donors to allotetraploid A. hypogaea. Microsatellite or SSR (Simple Sequence Repeat) markers are co-dominant, multiallelic, and highly polymorphic genetic markers, appropriate for genetic diversity studies. Microsatellite markers may also, to some extent, support phylogenetic inferences. Here we report the use of a set of microsatellite markers, including newly developed ones, for phylogenetic inferences and the analysis of genetic variation of accessions of A. hypogea and its wild relatives.ResultsA total of 67 new microsatellite markers (mainly TTG motif) were developed for Arachis. Only three of these markers, however, were polymorphic in cultivated peanut. These three new markers plus five other markers characterized previously were evaluated for number of alleles per locus and gene diversity using 60 accessions of A. hypogaea. Genetic relationships among these 60 accessions and a sample of 36 wild accessions representative of section Arachis were estimated using allelic variation observed in a selected set of 12 SSR markers. Results showed that the Brazilian peanut germplasm collection has considerable levels of genetic diversity detected by SSR markers. Similarity groups for A. hypogaea accessions were established, which is a useful criteria for selecting parental plants for crop improvement. Microsatellite marker transferability was up to 76% for species of the section Arachis, but only 45% for species from the other eight Arachis sections tested. A new marker (Ah-041) presented a 100% transferability and could be used to classify the peanut accessions in AA and non-AA genome carriers.ConclusionThe level of polymorphism observed among accessions of A. hypogaea analyzed with newly developed microsatellite markers was low, corroborating the accumulated data which show that cultivated peanut presents a relatively reduced variation at the DNA level. A selected panel of SSR markers allowed the classification of A. hypogaea accessions into two major groups. The identification of similarity groups will be useful for the selection of parental plants to be used in breeding programs. Marker transferability is relatively high between accessions of section Arachis. The possibility of using microsatellite markers developed for one species in genetic evaluation of other species greatly reduces the cost of the analysis, since the development of microsatellite markers is still expensive and time consuming. The SSR markers developed in this study could be very useful for genetic analysis of wild species of Arachis, including comparative genome mapping, population genetic structure and phylogenetic inferences among species.

A large scale analysis of resistance gene homologues in Arachis.

Arachis hypogaea L., commonly known as the peanut or groundnut, is an important and widespread food legume. Because the crop has a narrow genetic base, genetic diversity in A. hypogaea is low and it lacks sources of resistance to many pests and diseases. In contrast, wild diploid Arachis species are genetically diverse and are rich sources of disease resistance genes. The majority of known plant disease resistance genes encode proteins with a nucleotide binding site domain (NBS). In this study, degenerate PCR primers designed to bind to DNA regions encoding conserved motifs within this domain were used to amplify NBS-encoding regions from Arachis spp. The Arachis spp. used were A. hypogaea var. Tatu and wild species that are known to be sources of disease resistance: A. cardenasii, A. duranensis, A. stenosperma and A. simpsonii. A total of 78 complete NBS-encoding regions were isolated, of which 63 had uninterrupted ORFs. Phylogenetic analysis of the Arachis NBS sequences derived in this study and other NBS sequences from Arabidopsis thaliana, Medicago trunculata, Glycine max, Lotus japonicus and Phaseolus vulgaris that are available in public databases This analysis indicates that most Arachis NBS sequences fall within legume-specific clades, some of which appear to have undergone extensive copy number expansions in the legumes. In addition, NBS motifs from A. thaliana and legumes were characterized. Differences in the TIR and non-TIR motifs were identified. The likely effect of these differences on the amplification of NBS-encoding sequences by PCR is discussed.

Registration of Rust and Late Leaf Spot Resistant Peanut Germplasm Lines

Four peanut (Arachis hypogaea L.) germplasm lines, ICGV 99001 (Reg. no. GP-118, PI 631072), ICGV 99003 (Reg. no. GP-119, PI 631073), ICGV 99004 (Reg. no. GP-120, PI 631074), and ICGV 99005 (Reg. no. GP-121, PI 631075), were released in 2001 by the Plant Materials Identification Committee of International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Patancheru, AP, India, for their resistance to foliar fungal diseases. These lines were derived from A. hypogaea and wild Arachis species crosses. ICGV#99001 and 99004 belong to subsp. fastigiata var. vulgaris while ICGV#99003 and 99005 belong to subsp. hypogaea var. hypogaea. ICGV# 99001 and 99004 possess resistance to late leaf spot (LLS) [(caused by Phaeoisariopsis personata (Berk. & Curt.) v. Arx = Cercosporidium personatum (Berk. & Curt.) Deighton], and ICGV# 99003 and 99005 have resistance to rust (caused by Puccinia arachidis Speg.).