RFLP analysis of genetic variation in species of section Arachis, genus Arachis (Leguminosae)

Four A-genome species of the genus Arachis (A. cardenasii, A. correntina, A. duranensis, A. kempff-mercadoi), three B genomes species (A. batizocoi, A. ipaënsis and A. magna),the AABB allotetraploid A. hypogaea (cultivated peanut) and introgression lines resulting from a cross between A. hypogaea and A. cardenasii were analyzed by RFLP. The A genome species (cytologically characterized by the presence of a small chromosome pair ‘A’) were closely similar to each other and shared a large number of restriction fragments. In contrast, the B genome species differed more from one another and shared few fragments. The results of this study indicate that the absence of the small chromosome pair is not a good criterion for grouping species of section Arachis as B genome species, since their genome might be quite distinct from the B genome of A. hypogaea.The lowest genetic variation was detected within accessions of A. duranensis (17 accessions), followed by A. batizocoi (4 accessions) and A. cardenasii (9 plants of accession GKP 10017).The high level of genetic variation found in A. cardenasii might indicate that not all accessions of wild species of Arachis are autogamous, as reported for A. hypogaea.     

Phylogenetic relationships in the genus Arachis based on seed protein profiles

Summary Seed protein profiles of 19 accessions representing seven sections of the genus Arachis were studied using sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The general profile showed appreciable homology between these taxa, supporting their classification based on morphology and cross-compatibility relationships. The accessions of section Arachis expressed a high variation confirming inferences from earlier studies. Variation between accessions of a species is limited. Accessions of the section Ambinervosae and Caulorhizae formed one cluster and accessions of sect. Erectoides and Procumbensae formed another. Whereas the representative accessions of sect. Triseminalae and Extranervosae formed two independent clusters. Using the percentage of dissimilarity in electrophoretic bands as a statistical genetic distance between accessions, sect. Arachis (containing the cultivated groundnut, A. hypogaea) is phylogenetically closest to sect. Erectoides followed by Procumbensae, Ambinervosae, Caulorhizae, Triseminalae and Extranervosae, respectively.Submitted as Journal Article No. 1485 by the International Crops Research Institute for the Semi-Arid Tropics (ICRISAT) Patancheru, A.P. 502 324, India.     

The genomes of Arachis hypogaea. 1. Cytogenetic studies of putative genome donors

Summary Cytological studies of wild diploid Arachis species in the same section of the genus (sect. Arachis) as the cultivated peanut A. hypogaea L. show, with one exception, a karyotype characterized by the presence of 9 pairs of larger chromosomes and one pair of small (A) chromosomes. The exceptional species A. batozocoi Krap. et Greg. has a more uniform karyotype. Interspecific hybrids between diploid species of similar karyotype have moderate to high pollen stainability, those involving A. batizocoi have zero pollen stainability and a very irregular PMC meiosis. Such infertile hybrids are the most likely to produce fertile, stable amphidiploids on doubling the chromosome complement. It is suggested that the cultivated peanut could have originated from such a sterile interspecific hybrid and on morphological and phytogeographic grounds the most likely genome donors are A. cardenasii (nomen nudum) and A. batizocoi of the species within section Arachis, which have been collected up to the present time.Paper number 5560 of the Journal Series of the North Carolina Agricultural Experiment Station, Raleigh, NC 27650     

Genetic variation and phylogenetic relationships based on RAPD analysis in section Caulorrhizae, genus Arachis (Leguminosae)

Wild Arachis germplasm includes potential forage species, such as the rhizomatous Arachis glabrata and the stoloniferous A. pinto and A. repens. Commercial cultivars of A. pintoi have already been released in Australia and in several Latin American countries, and most of these cultivars were derived from a single accession of A. pintoi (GK 12787). Arachis repens is less productive as a forage plant than is A. pintoi. However, it can be crossed with A. pintoi, and thus has good potential as germplasm for the improvement of A. pintoi. Arachis repens is also used as an ornamental plant and ground cover. Many new accessions of these two stoloniferous species are now available, and they harbor significant genetic variability beyond that available in the few older accessions, previously available. Therefore, these new accessions need to be conserved, documented and considered in terms of their potential for crop improvement and direct commercial use. Sixty-four accessions of this new germplasm were analyzed using RAPD analysis. Most of the accessions of A. repens grouped together into a clearly distinct group. In general, the accessions from the distinct valleys of the Jequitinhonha, São Francisco and Paranã rivers did not group together, suggesting there is not a tight relation between dispersion by rivers and the geographic distribution of genetic variation in these species.     

Introgression of disease resistance genes from Arachis kempff-mercadoi into cultivated groundnut

Arachis kempff‐mercadoi is a wild species from the section Arachis. All kempff‐mercadoi accessions originate from the Santa Cruz province of Bolivia and they represent Arachis species with the A genome. From molecular analysis it was found that although cultivated A. hypogaea is made up of A and B genomes, A. kempff‐mercadoi may not be as closely related to it as are some of the other A genome species. Arachis kempff‐mercadoi is of interest because it has multiple disease resistance. It was crossed with a Spanish A. hypogaea cultivar which is susceptible to foliar diseases and to the insect pest Spodoptera litura. The success rate of the cross A. hypogaea (2n = 40) ×A. kempff‐mercadoi (2n = 20) was very low, but it could be increased by culturing immature seeds in vitro. Although the hybrids were triploids, a few fertile pollen grains were obtained due to the formation of restitution nuclei in the F₁ plants. Interspecific derivatives at the BC₂F₂ generation were scored for early leaf spot, late leaf spot and to Spodoptera damage. Screening results showed that 29% of the derivatives had both early and late leaf spot resistance and that less than 5% of the derivatives had resistance to both the foliar diseases and to Spodoptera.     

Determination of genetic variation and taxonomy in lentil (Lens Miller) species by chloroplast DNA polymorphism

Summary Chloroplast DNA restriction fragment lenght polymorphisms (RFLP) were used to examine the taxonomic relationships of cultivated and wild lentil (Lens Miller) species and identify the extent of genetic variation in this genus. Twelve accessions representing all Lens subspecies were digested with four hexanucleotide-recognizing restriction endonucleases. These digests randomly surveyed 540 base pairs, or 0.4% of the approximately 125 kilobase lentil chloroplast genome. A high degree of gragment length conservation was seen among members of crossability group I, i.e., L. c. ssp. culinaris, L. c. ssp. orientalis and L. c. ssp. odemensis. Accessions of the two subspecies comprising crossability group II, i.e., L. n. ssp. nigricans and L. n. ssp. ervoides, showed the greatest amount of variation when compared to the cultivated lentil, L. c. ssp. culinaris. Limited variation was observed within subspecies except for L. n. ssp. nigricans, where accessions of the normal cytotype were highly polymorphic to those of the differentiated cytotype. Chloroplast DNA RFLPs reaffirm hypotheses that propose L. c. ssp. orientalis as the progenitor to the cultivated lentil. The implications of this study on taxonomy and genetic resources is also discussed.     

Morphological, cytological and disease resistance studies of the intersectional hybrid between Arachis hypogaea L. and A. glabrata Benth.

Arachis glabrata Benth, variety glabrata collector GK 10596 (PI276233; ICG 8176) belonging to section Rhizomatosae has resistance to rust, late leaf spot and viral diseases. A. hypogaea L. cv MK 374 (section Arachis) is susceptible to rust, late leaf spot and to the viral diseases peanut stripe, peanut mottle and peanut bud necrosis. Hybrids between A. hypogaea cv MK 374 and A. glabrata were produced after inter specific pollinations and embryo culture. The hybrids produced had morphological characters of both parents plus floral abnormalities not seen in either parent. It was possible to identify the hybrids by esterase isozyme analysis when still in culture. Cytological research showed variable chromosome association and also homeology between the genomes of A. hypogaea and A. glabrata. The hybrids inherited resistance to rust, late leaf spot, peanut bud necrosis and peanut stripe diseases from the pollen parent A. glabrata. This revised version was published online in August 2006 with corrections to the Cover Date.     

Aseptic culture of gynophores to obtain peanut intersectional hybrids

Summary Cross-incompatibility between cultivated peanuts and their wild relatives outside the section Arachis has impeded the utilization of many species possessing high resistances or good qualities. Despite the great efforts made to culture immature ovules or embryos, few hybrid offspring have been obtained. In this study, gynophores from Arachis hypogaea L. pollinated with A. glabrata Benth. were cultured and F₁ hybrids seeds were harvested, and F₂ and F₃ generations produced. The characters of F₂ generation exhibited a wide range of segregation. Leaf peroxidase isozyme PAGE analysis revealed that the hybrids were quite different from their parents in relation to band number, width and isozyme activity. The zymograms of the hybrids and their parents were partially alike. This verified the authenticity of the hybrids.     

野生花生脂肪酸组成的遗传变异及远缘杂交创造高油酸低棕榈酸花生新种质

以花生属19个近缘野生物种87份种质和113份栽野远缘杂交后代为材料, 系统分析野生花生脂肪酸组成的遗传变异及其在栽培种花生脂肪酸改良中的潜力。结果表明, 野生花生的棕榈酸含量与栽培种花生相似, 硬脂酸和油酸含量略低于栽培种花生, 亚油酸含量略高于栽培种。不同物种间以及同一物种内不同资源间的脂肪酸组成存在较大差异。A. rigonii棕榈酸含量较低, A. pusilla和A. duranensis油酸含量较高, A. batizocoi亚油酸含量较高, A. rigonii和A. duranensis油酸和亚油酸含量变幅较大。发掘出油酸含量达60%以上的野生资源2份(19-6, A. duranensis和23-1, A. sp.), 亚油酸含量达40%以上的资源7份, 其中A. rigonii(编号为11-4)亚油酸含量高达48%, 是目前所发现的花生资源中亚油酸含量最高的种质。远缘杂交后代脂肪酸的变异远远超过亲本间的差异, 而且不同组合间的棕榈酸、硬脂酸、油酸和亚油酸含量差异达显著或极显著水平。通过远缘杂交获得了6份油酸含量达64.0%以上且棕榈酸含量在8.5%以下的新种质, 其中yz8913-8油酸含量达67.85%, 比其栽培种亲本提高近30个百分点, 且棕榈酸含量仅7.60%。SRAP检测表明, 这6份远缘杂交后代除整合了亲本的DNA片段外, 还产生了新的DNA片段, 有的还丢掉了亲本的某些片段。农艺性状分析表明, 其中4份种质的综合农艺性状较好, 具有重要育种利用价值。     

Phenotypic diversity and identification of wild Arachis accessions with useful agronomic and nutritional traits

Wild relatives harbor novel sources of variation, which can be used to enhance the genetic base of a cultivar gene pool. A total of 269 accessions from 20 wild Arachis species belonging to six sections were evaluated for 41 morpho-agronomic traits and 89 selected accessions for oil, protein and total sugar content. Six plants from each accession were grown in an open Arachis house in large-cylindrical concrete structures during the 2004?C2005 season at Patancheru, India. REML analysis showed significant differences between species and accessions for most of the traits studied. Hierarchical cluster analysis, based on the first five principal component scores accounted for 82.5% variation, resulting in four clusters. Variation in genome relationships and ploidy levels had no bearing on the clustering pattern which was predominated by life forms: clusters 1 and 2, contained mostly annuals and clusters 3 and 4 perennials. A large range of variations were noticed among species for some of the agronomic traits: days to flowering, pod and seed characteristics, specific leaf area (SLA) and for SPAD chlorophyll meter reading (SCMR). Arachis duranensis showed the maximum intraspecific variation as revealed by a high diversity index for 23 of the 41 traits which included: days to flowering, primary branches, plant width, pod length, pod width, SCMR and SLA. The other species with desirable traits were A. pusilla (earliest flowering) and A. villosa (high SCMR at 60 and 80 days after sowing). The latter species is cross compatible with cultivated groundnut, thus, is a good source to enhance the trait value in the cultigen??s gene pool. The best 20 accessions with superior agronomic, nutritional quality and drought related trait combinations have been identified for their use in introgression of diverse and unique alleles from wild Arachis species into A. hypogaea.     

The classification of varieties of groundnut (Arachis hypogaea L.)

Summary An expanded scheme of groundnut variety classification is presented which is revised in the light of recently collected West African material and is designed in concordance with the most recent taxonomic treatment of the species Arachis hypogaea L. This species consists of two subspecies ssp. hypogaea and ssp. fastigiata Waldron. Each subspecies contains two botanical varieties. Those of ssp. hypogaea are var. hypogaea and var. hirsuta Kohler and those of ssp. fastigiata Waldron are var. fastigiata and var. vulgaris Harz. Each infraspecific taxon contains a morphologically distinct group of cultivated varieties.Grain Legume Research Laboratory, Agricultural Research Council of Malawi, POB215, Lilongwe, Malawi.Department of Agricultural Botany, University of Reading, Reading, England.     

Chloroplast DNA variation in the wild potato species, Solanum acaule and S. albicans

The chloroplast DNA of Solanum acaule (109 accessions) and S. albicans (9 accessions) was investigated by restriction endonuclease analysis. Unexpectedly, all the accessions analyzed had C type chloroplast DNA in common. This suggested that S. acaule originated from a species with C type chloroplast DNA. DraI restriction digestion revealed further differentiation of C type chloroplast DNA into 8 types. The DraI polymorphism indicated the province of Salta in Argentina and the nearby regions to be a center of diversity for S. acaule. Surprisingly, S. albicans as well as S. acaule both ssp. acaule and ssp. punae, from Peru were virtually indistinguishable, although by morphology and/or cytology all three taxa are easily distinguished. This revised version was published online in July 2006 with corrections to the Cover Date.     

Cytology and leafspot resistance in Arachis hypogaea x wild species hybrids

Summary Introgression of germplasm from diploid wild Arachis species to A. hypogaea has great potential for improving pest resistance in cultivated peanuts. This investigation evaluated methods for incorporating exotic germplasm into cultivated peanuts, especially for Cercospora arachidicola Hori resistance. Interspecific hybrids between A. hypogaea (cvs. NC 2 and NC 5) and the wild species A. cardenasii Krap. et Greg. nom. nud. and A. chacoense Krap. et Greg. nom. nud. were analyzed cytologically and for leafspot resistance. All F₁ hybrids were sterile, had irregular meiosis, and very few multivalents. They were highly resistant to C. arachidicola in field tests and had a 10-fold reduction of conidia per lesion in the greenhouse as compared to A. hypogaea cultivars. After colchicine treatments of F₁ hybrids, hexaploids (2n=60) and aneuploids (2n=54, 56, 63) were observed. The hexaploids had up to 18 univalents per pollen mother cell and very few multivalents, indicating a low frequency of intergenomic chromosome pairing. For C. arachidicola resistance, significant differences were not found among wild species parents, F₁ hybrids and two generations of hexaploids. Most hexaploids were stable at 2n=60 and embryos aborted when backcrosses with the respective wild species were attempted. However, when hexaploids were backcrossed to A. hypogaea, several fertile pentaploid (2n=50) offspring were obtained. Use of self-pollinating pentaploids is believed to be the quickest method to recover 40-chromosome hybrid derivatives in these hybrids.     

Interspecific hybridization in the genus Arachis through embryo culture

Summary The embryos of a cultivated tetraploid peanut (Arachis hypogaea), a wild diploid species (A. villosa), and their hybrid embryos, which generally abort in nature, were cultured in vitro and the plants have been successfully transferred to the soil. The hybrids showed triploid chromosome number (3x=30). The significance of wide hybridization in peanut-improvement programs is discussed.     

Variation in the ribosomal RNA units of the generaLens andCicer

Summary Investigation of the organization of the tandemly repeated 5S and 18–25S ribosomal RNA (rRNA) genes was carried out on members of the generaLens andCicer using restriction endonuclease digestion and Southern hybridization. The 5S rRNA unit (gene+spacer) inLens (0.38 to 0.50 kb) andCicer 0.50 to 0.44 kb) is the largest among the legumes. InL. nigricans andL. culinaris ssp.odemensis a second repeat size was detected. Restriction maps for the 18–25S units ofLens andCicer species were developed. The 18–25S rRNA unit varied from 8.3 to 9.8 kb inLens and 10.5 to 11.4 kb inCicer. The only detectable variability in theLens andCicer 18–25S rRNA unit was in the length of the intergenic spacer (IGS) region, except for a HindIII site in the IGS region ofL. nigricans ssp.nigricans and in allCicer species.Similarities in the size of both 5S and 18–25S rRNA units were noted betweenL. culinaris ssp.culinaris andL. culinaris ssp.orientalis, which supports previous evidence thatL. culinaris ssp.orientalis is the progenitor of the cultivated lentil. Differences in the arrangement of both 5S and 18–25S rRNA units were noted betweenL. nigricans ssp.nigricans andL. nigricans ssp.ervoides, indicating either divergent evolution of these subspecies or alternatively incorrect taxonomy.On the basis of the size of the 5S rRNA unit,C. bijugum, C. chorassanicum andC. echinospermum formed a group with a slightly smaller unit than the otherCicer species. Similarities in the length of both 5S and 18–25S rRNA units ofC. arietinum andC. reticulatum possibly confirmC. reticulatum as the progenitor ofC. arietinum. Cicer cuneatum has a smaller 18–25S rRNA unit than the otherCicer species as a result of a smaller intergenic spacer.     

The role of mutations in intraspecific differentiation of groundnut (Arachis hypogaea L.)

Summary Based on morphological diversity, cultivated groundnut (Arachis hypogaea L.) is classified into two subspecies (fastigiata and hypogaea) and further into four botanical types (Spanish bunch, Valencia, Virginia bunch and Virginia runner). In a cross between two Spanish cultivars belonging to ssp. fastigiata, a true breeding variant (Dharwad early runner) sharing some characters of both the subspecies was isolated. The variant, on mutagenesis with ethyl methane sulphonate (EMS) yielded a very high frequency of mutants resembling all four botanical types. Some of the mutants produced germinal reversions to Dharwad early runner in later generations indicating genetic instability. While most of the revertants bred true, some of the mutants continued to segregate, wherein each botanical group of mutants produced all other botanical types. A detailed analysis of the breeding behaviour of mutants revealed several unusual features (such as homozygous mutations, mutation outbursts, segregation distortions, somatic mutations and multiple character mutations) that could not be explained through conventional mutation theory. In the light of these findings, the role of mutations in evolutionary differentiation of the crop and the probable mode of their origin have been discussed.     

Reaction of wild species of the geneus Lens to drought

Summary Susceptibility to drought stress is a key factor in dry land lentil (Lens culinaris Medikus) production in the Mediterranean region of West Asia and North Africa. This study examined the response to drought stress of 121 accessions representing all subspecies of the genus Lens; cultivated, and the wild L. culinaris ssp. orientalis (Boiss.) Ponert, L. culinaris ssp. odemensis (Ladiz.), L. nigricans M.B. Godr. ssp. nigricans Godr. and L. nigricans ssp. ervoides (Brign.) Ladiz. for their potential use in breeding for dry land conditions. Accessions were grown under two moisture regimes (dry land and dry land plus supplemental irrigation) at Breda, Syria during the 1990–91 and 1991–92 seasons. The cultivated lentil had markedly superior seed and straw production than did the wild Lens species. Time to flowering accounted for less than 10% of the variation in yield of wild accessions under rainfed conditions in the two seasons, showing that, in contrast to the cultivated germplasm, drought escape was relatively unimportant in wild lentil. Performance under drought in wild lentil, measured in terms of dry land seed yield or drought susceptibility index (S), was randomly distributed among collection locations with little relation to collection site aridity. Direct selection of wild lentil germplasm for biomass yield under dry conditions is of little value and an evaluation of wild accessions in hybrid combination is needed.     

The probable genome donors to Arachis hypogaea L. based on arachin seed storage protein

Summary Interrelationships among six diploid species (A. chacoense, A. villosulicarpa, A. batizocoi, A. correntina, A. duranensis and A. cardenasii), tetraploid wild groundnut A. monticola and four accessions of A. hypogaea were studied by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) of total seed proteins and arachin immunoprecipitates. Arachin in A. monticola and A. hypogaea cv. Trombay Groundnut 9 (TG-9) was composed of four acidic subunits (47.5 kd, 45.1 kd, 42.6 kd and 41.2 kd) and one major basic subunit (21.4 kd). The arachin subunit pattern in cv. Spanish Improved (SP) and TG-1 was almost similar to the pattern observed in A. monticola with the exception that the SP lacked the 41.2 kd and TG-1 lacked the 42.6 kd acidic subunits of A. monticola. Seed extracts of all diploid species studied reacted with the anti-arachin antibodies raised against SP arachin. The electrophoretic analysis of immunoprecipitates of diploid species showed a range of acidic subunits from 46.2 kd to 41.2 kd and one or two basic subunits of 21.4 kd and 20.2 kd. None of the diploid species showed the 47.5 kd subunit found in A. monticola or A. hypogaea. Of the diploid species, A. duranensis and A. cardenasii demonstrated two acidic subunits of 45.1 kd and 41.2 kd and a basic subunit of 21.4 kd as found in A. monticola. Likewise A. batizocoi showed two acidic subunits of 45.1 kd and 42.6 kd and the basic subunit of 21.4 kd as was observed in A. monticola. Based on electrophoretic data, our research supports the earlier conclusion that the probable genome donors to A. monticola are A. batizocoi and A. duranensis or A. cardenasii.     

Use of the gene pools of Triticum turgidum ssp. dicoccoides and Aegilops squarrosa for the breeding of common wheat (T. aestivum), through chromosome-doubled hybrids

Summary Triticum turgidum ssp. dicoccoides (wild emmer wheat, AABB, 2n=28) and Aegilops squarrosa (goat grass, DD, 2n=14) comprise a rich reservoir of valuable genetic material, which could be useful for the breeding of common wheat (T. aestivum, AABBDD, 2n=42). Many accessions of both wild species, most of them selected for resistance to stripe rust, were used to make amphiploids. Two strategies were applied: (1) the production of autopolyploid cytotypes of the wild species, followed by hybridisation, and (2) the production of allotriploid interspecific hybrids, followed by doubling of the number of chromosomes. The first route was unsuccessful because of failure of the crosses between the autopolyploid cytotypes, possibly due to incongruity between the two species and to reduced fertility in the autopolyploid cytotypes. The second route yielded the desired synthetic hexaploids. However, the rate of success of the crosses was low and there were great differences between years, and within years between crosses. Embryo rescue was applied to obtain the primary hybrids (2n=21), which were highly sterile and had on average 0.3 bivalents and 20.4 univalents per pollen mother cell. Various abnormalities were recorded. Doubling of the number of chromosomes sometimes occurred spontaneously or was brought about by colchicine treatment. The large scale of the interspecific hybridisation programme ensured that one-third of the female and one-sixth of the male accessions were represented in the synthetic hexaploids.     

Reactions of Hordeum spontaneum to infection with two cultures of Puccinia hordei from Israel and United States

Summary The reactions to infection with two cultures of Puccinia hordei were determined for 292 Hordeum spontaneum (syn. H. vulgare ssp. spontaneum) accessions, collected at 16 sites which encompassed the ecological range of H. spontaneum in Israel. Culture Tel-Aviv was from Israel and culture 57.19 was from the United States. Fifty-two percent of the accessions were resistant to culture Tel-Aviv and 67% were resistant to culture 57.19. Forty-three percent of the accessions were resistant to both cultures. The average infection type (IT) of accessions within sites ranged from 2.7 to 7.5 on a 0–9 rating scale. The results showed that the presence of Ornithogalum species, the alternate hosts of P. hordei, may increase the percentage of H. spontaneoum accessions resistant to P. hordei. More accessions were resistant at sites where humidity at 1400 was higher, the annual evaporation was lower, and where the glumes were shorter. Kernel weight and annual rainfall was not correlated with resistance. A lower percentage of H. spontaneum accessions were resistant to P. hordei culture Tel-Aviv from Israel than to culture 57.19 from the United States. In a previous study a lower percentage of H. spontaneum accessions also was found to be resistant to a culture of Erysiphe graminis hordei from Israel than to cultures from other countries. Previous studies also have shown that cultures of P. hordei and E. graminis hordei from Israel have many genes for virulence on barley, and that H. spontaneum accessions from Israel have many genes for resistance to these two pathogens. Previous results and the results reported in this paper support the hypothesis of coevolution of resistant host genes and virulent pathogen genes where hosts and pathogens have coexisted for many thousand years.