<Emphasis Type="Italic">Rhizobium</Emphasis>,<Emphasis Type="Italic"> Bradyrhizobium</Emphasis> and<Emphasis Type="Italic"> Agrobacterium</Emphasis> strains isolated from cultivated legumes

The present study was conducted to isolate and characterize rhizobial strains from root nodules of cultivated legumes, i.e. chickpea, mungbean, pea and siratro. Preliminary characterization of these isolates was done on the basis of plant infectivity test, acetylene reduction assay, C-source utilization, phosphate solubilization, phytohormones and polysaccharide production. The plant infectivity test and acetylene reduction assay showed effective root nodule formation by all the isolates on their respective hosts, except for chickpea isolate Ca-18 that failed to infect its original host. All strains showed homology to a typical Rhizobium strain on the basis of growth pattern, C-source utilization and polysaccharide production. The strain Ca-18 was characterized by its phosphate solubilization and indole acetic acid (IAA) production. The genetic relationship of the six rhizobial strains was carried out by random amplified polymorphic DNA (RAPD) including a reference strain of Bradyrhizobium japonicum TAL-102. Analysis conducted with 60 primers discriminated between the strains of Rhizobium and Bradyrhizobium in two different clusters. One of the primers, OPB-5, yielded a unique RAPD pattern for the six strains and well discriminated the non-nodulating chickpea isolate Ca-18 from all the other nodulating rhizobial strains. Isolate Ca-18 showed the least homology of 15% and 18% with Rhizobium and Bradyrhizobium, respectively, and was probably not a (Brady)rhizobium strain. Partial 16S rRNA gene sequence analysis for MN-S, TAL-102 and Ca-18 strains showed 97% homology between MN-S and TAL-102 strains, supporting the view that they were strains of B. japonicum species. The non-infective isolate Ca-18 was 67% different from the other two strains and probably was an Agrobacterium strain.     

<Emphasis Type="Italic">Calamintha nepeta</Emphasis> (L.) Savi and <Emphasis Type="Italic">Micromeria thymifolia</Emphasis> (Scop.) Fritsch cultivated in Italy

Calamintha nepeta and Micromeria thymifolia have been traditionally used in the Mediterranean area as condiments and medicinal plants for a long time. Whereas in parts of Italy C. nepeta (special recipes have been developed in Lazio and Tuscany) is also an established garden plant showing different evolutionary products and their interaction among each other and the wild progenitor, M. thymifolia is being developed into a new crop plant. Both plants and their uses are described with regard to Italy. There is a marked tendency to broaden the use of condiments and spices which results in new crop plants which have to be documented and elaborated in further studies. Many species of Labiatae are predisposed to use by man and new items can be found even in areas which have to be considered as well studied.     

Salt tolerance in <Emphasis Type="Italic">Zea mays</Emphasis> (L). following inoculation with <Emphasis Type="Italic">Rhizobium</Emphasis> and <Emphasis Type="Italic">Pseudomonas</Emphasis>

This study aimed to investigate the effect of inoculation with plant growth-promoting Rhizobium and Pseudomonas species on NaCl-affected maize. Two cultivars of maize (cv. Agaiti 2002 and cv. Av 4001) selected on the basis of their yield potential were grown in pots outdoors under natural conditions during July. Microorganisms were applied at seedling stage and salt stress was induced 21 days after sowing and maintained up to 50% flowering after 120 days of stress. The salt treatment caused a detrimental effect on growth and development of plants. Co-inoculation resulted in some positive adaptative responses of maize plants under salinity. The salt tolerance from inoculation was generally mediated by decreases in electrolyte leakage and in osmotic potential, an increase in osmoregulant (proline) production, maintenance of relative water content of leaves, and selective uptake of K ions. Generally, the microbial strain acted synergistically. However, under unstressed conditions, Rhizobium was more effective than Pseudomonas but under salt stress the favorable effect was observed even if some exceptions were also observed. The maize cv. Agaiti 2002 appeared to be more responsive to inoculation and was relatively less tolerant to salt compared to that of cv. Av 4001.     

<Emphasis Type="Italic">Ziziphus </Emphasis><Emphasis Type="Italic">spina-christi</Emphasis> (L.) Willd.: a multipurpose fruit tree

Neglected and underutilized species often play a vital role in securing food and livestock feed, income generation and energy needs of rural populations. In spite of their great potential little attention has been given to these species. This increases the possibility of genetic erosion which would further restrict the survival strategies of people in rural areas. Ziziphus spina-christi is a plant species that has edible fruits and a number of other beneficial applications that include the use of leaves as fodder, branches for fencing, wood as fuel, for construction and furniture making, and the utilization of different parts e.g. Fruits, leaves, roots and bark in folk medicine. Moreover, the plant is adapted to dry and hot climates which make it suitable for cultivation in an environment characterized by increasing degradation of land and water resources. Lack of research in Z. spina-christi hinders its successful improvement and promotion. Therefore, studies are needed to fully exploit this species. This article aims at summarizing information on different aspects of Z. spina-christi to stimulate interest in this crop which is of importance in Sudan and other countries of the semi-arid tropics.

Diversity analysis of Central Asia and Caucasian lentil (<Emphasis Type="Italic">Lens</Emphasis><Emphasis Type="Italic">culinaris</Emphasis> Medik.) germplasm using SSR fingerprinting

Diversity analysis was performed among 39 cultivated lentil (Lens culinaris Medik.) accessions of Central Asia and Caucasian origin using five highly polymorphic microsatellite markers. A total of 33 alleles determined ranging from 3 to 8 per locus. Estimated gene diversity value for 33 loci was 0.66. Genetic similarity indices among 39 accessions ranged from 0.24 to 1.0. Cluster analysis using the unweighted pair group method with arithmetic mean method classified accessions into six major groups at 0.5 similarity coefficient. More than half accessions from Tajikistan formed large cluster. On the other hand, a few accessions from each country showed unique genotypes. Overall, most of the accessions, except ones with closely related origin, were distinguished by the present high quality DNA fingerprinting. This molecular diversity information gives important basis for conservation strategy in gene bank and exotic germplasm introduction in breeding programs in Central Asia and Caucasian countries.     

Identification of <Emphasis Type="Italic">Aegilops</Emphasis> L. species and <Emphasis Type="Italic">Triticum aestivum</Emphasis> L. based on chloroplast DNA

The genus Aegilops L. is a very important genetic resource for the breeding of bread wheat Triticum aestivum. Therefore, an accurate and easy identification of Aegilops species is required. Traditionally, identification of Aegilops species has relied heavily on morphological characters. These characters, however, are either not variable enough among Aegilops species or too plastic to be used for identification at the species level. Molecular markers that are more stable within species, therefore, could be the alternative strategy towards an accurate identification. Since the chloroplast DNA has a lower level of evolution compared to the nuclear genome, an attempt was made in this study to investigate polymorphism in the chloroplast DNA among 21 Aegilops species (including Ae. mutica that is now known as Amblyopyrum muticum) and between the latter and T. aestivum to generate markers for the diagnosis of all targeted species. Cleaved amplified polymorphic sequence (CAPS) applied on 22 coding and non-coding chloroplast regions using 80 endonucleases and sequencing of two of those regions revealed little polymorphism between T. aestivum and the various Aegilops species examined and to a less extent was the variation among Aegilops species. Polymorphism observed among species analysed allowed the discrimination of T. aestivum and 12 Aegilops species.     

Relationships among some cornelian cherry genotypes (<Emphasis Type="Italic">Cornus mas</Emphasis> L.) based on RAPD analysis

Turkey is an important producer of cornelian cherries (Cornus mas L.), especially in northern Anatolia. Seed propagation and long-term human selection has given rise to a great diversity of trees. Twenty-six cornelian cherry genotypes (CC1–CC26) from the Coruh Valley in northern Anatolia were evaluated for genetic relationships by using Randomly Amplified Polymorphic DNA (RAPD) markers, based on 56 decamer random primers, seven of which showed reliable polymorphisms. These seven primers generated 80 markers, with 77 (96.25%) displaying polymorphisms. Cluster analysis of the cornelian cherry genotypes was performed based on data from polymorphic RAPD bands, by using Jaccard’s similarity coefficient and the Unweighted pair-group method with arithmetic average (UPGMA) clustering method. A similarity matrix showed that the highest genetic similarity (0.913) was between CC15 and CC16 and the least (0.129) was between CC4 and CC16. The cophenetic correlation coefficient between the similarity matrix and the cophenetic matrix of the dendrogram was relatively high (r = 0.87), supporting the validity of the dendrogram. Based on these results, RAPD analysis can be used for the characterization and grouping of cornelian cherry genotypes. Genetically divergent genotypes identified in this study may be useful for future breeding programs. This is the first study demonstrating that RAPD analyses can be used to differentiate and classify cornelian cherry genotypes.     

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.     

The effect of rhizobiophages on<Emphasis Type="Italic"> Sinorhizobium meliloti</Emphasis> -<Emphasis Type="Italic"> Medicago sativa</Emphasis> symbiosis

Using Tn5 mutagenesis of lysogenic Sinorhizobium meliloti strain L5–30 (L54), three neomycin-resistant transconjugants differing in phage-resistance profiles were isolated. Two of them increased the dry weight of plants and were capable of establishing root nodules, whereas the third one was ineffective. The bacterium-phage interaction did not have observable consequences in the Medicago sativa - S. meliloti symbiosis because it did not affect the number of nodules on M. sativa or plant dry weight.     

A new mycorrhizal helper bacterium, <Emphasis Type="Italic">Ralstonia</Emphasis> species,in the ectomycorrhizal symbiosis between <Emphasis Type="Italic">Pinus thunbergii</Emphasis> and <Emphasis Type="Italic">Suillus granulatus</Emphasis>

In a Robinia-pseudoacacia-dominated coastal forest in Tottori prefecture Japan, the growth and survival of Pinus thunbergii seedlings and the natural regeneration of P. thunbergii was disturbed by R. pseudoacacia. In order to improve the growth of P. thunbergii seedling in the Tottori sand dune, we tried to find a mycorrhiza helper bacteria (MHB) from P. thunbergii mycorrhizosphere in a Tottori sand dune. Two MHB, Ralstonia sp. and Bacillus subtilis, were selected from the nine bacterial species isolated from the mycorrhizosphere of P. thunbergii. The bacterial effect on the ectomycorrhizal fungus Suillus granulatus was investigated by confrontation assay and a microcosm experiment. The confrontation assay showed that Ralstonia sp. promoted the hyphal growth of S. granulatus. Moreover, the S. granulatus–P. thunbergii symbiosis was significantly stimulated by Ralstonia sp. and B. subtilis. Ralstonia sp. and B. subtilis were regarded as MHB associated with P. thunbergii. This is the first report of Ralstonia sp. as an MHB.     

Development and utilization of diagnostic DAMD-PCR markers for <Emphasis Type="Italic">Capsicum</Emphasis> accessions

A polymerase chain reaction (PCR) based approach involving the directed amplification of minisatellite DNA region (DAMD-PCR) was used to identify accession specific DNA markers and study genetic relationships between and within 15 accessions corresponding to 11 species in genus Capsicum. A touch down PCR profile and unique chemical concentration of ingredients resulted in reproducible and reliable DNA amplifications. The number of amplified products varied from 1 to 12 fragments depending on the template DNA and the primers. The DAMD-PCR technique provided a total of 38 accession specific DNA markers (diagnostic DAMD-PCR) which can be utilized in accession identification, preservation and genetic studies of Capsicum germplasm. Based on 1,292 polymorphic and monomorphic DNA markers directed with 22 minisatellite specific primers, accessions were divided into four major groups, three of which corresponded to the three distinct Capsicum complexes. Capsicum chacoense was found to be the most distinct species.     

Transfer of leaf rust and stripe rust resistance from <Emphasis Type="Italic">Aegilops umbellulata</Emphasis> Zhuk. to bread wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.)

Aegilops umbellulata acc. 3732, an excellent source of resistance to major wheat diseases, was used for transferring leaf rust and stripe rust resistance to cultivated wheat. An amphiploid between Ae. umbellulata acc. 3732 and Triticum durum cv. WH890 was crossed with cv. Chinese Spring Ph ^I to induce homoeologous pairing between Ae. umbellulata and wheat chromosomes. The F₁ was crossed to the susceptible Triticum aestivum cv. ‘WL711’ and leaf rust and stripe rust resistant plants were selected among the backcross progenies. Homozygous lines were selected and screened against six Puccinia triticina and four Puccinia striiformis f. sp. tritici pathotypes at the seedling stage and a mixture of prevalent pathotypes of both rust pathogens at the adult plant stage. Genomic in situ hybridization in some of the selected introgression lines detected two lines with complete Ae. umbellulata chromosomes. Depending on the rust reactions and allelism tests, the introgression lines could be classified into two groups, comprising of lines with seedling leaf rust resistance gene Lr9 and with new seedling leaf rust and stripe rust resistance genes. Inheritance studies detected an additional adult plant leaf rust resistance gene in one of the introgression lines. A minimum of three putatively new genes—two for leaf rust resistance (LrU1 and LrU2) and one for stripe rust resistance (YrU1) have been introgressed into wheat from Ae. umbellulata. Two lines with no apparent linkage drag have been identified. These lines could serve as sources of resistance to leaf rust and stripe rust in breeding programs.     

Effect of mineral fertilizer,pig manure,and <Emphasis Type="Italic">Azospirillum rugosum</Emphasis> on growth and nutrient contents of <Emphasis Type="Italic">Lactuca sativa</Emphasis> L.

Benefits from the application of plant growth-promoting bacteria in agriculture largely depend on the complex interactions between several factors including the nature of fertilizers selected. This study was designed to determine the fine tuning between the inoculated bacteria and different fertilizers and their effect on the growth of lettuce plants (Lactuca sativa L.). Plant growth promotion by a novel species of the genus Azospirillum, namely A. rugosum IMMIB AFH-6, was tested by biochemical, bioassay, and greenhouse studies. The treatments used in the greenhouse study were; unfertilized control (Blank), half recommended dose of chemical fertilizer (1/2CF), full recommended dose of chemical fertilizer (1CF), pig manure fertilizer (PMF), pig manure fertilizer + half recommended dose of chemical fertilizer (PMF + 1/2CF), and pig manure fertilizer + full recommended dose of chemical fertilizer (PMF + 1CF). All these treatments when inoculated with A. rugosum IMMIB AFH-6 inoculation were, respectively, In-Blank, In-1/2CF, In-1CF, In-PMF, In-PMF + 1/2CF, and In-PMF + 1CF. Significant increase in plant biomass and shoot N, P, Ca, and Fe was shown in the In-Blank treatment. Plant growth in soil amended with PMF and A. rugosum IMMIB AFH-6 was significantly lower than in soil treated with the chemical fertilizer, but inoculation combined with chemical fertilizer significantly elevated the plant biomass. The In-PMF + 1/2CF treatment showed the highest yield. A. rugosum IMMIB AFH-6 facilitated the accumulation of trace minerals in higher concentrations when PMF was combined with 1CF. To examine the benefits of inoculation by A. rugosum IMMIB AFH-6, we have proposed a new type of data analysis which considers both biomass and nutrient content of plants. This new type of analysis has shown the importance of the mineral content of plant.     

Clustering of Chickpea (<Emphasis Type="Italic">Cicer arietinum</Emphasis> L.) Accessions

The variation in 20 morphological and agronomical traits has been evaluated in a set of chickpea genetic resources from four countries. Data indicated differences between accessions in leaf, flower, pod, and seed traits and characteristics, as well as in vegetation period. Multivariate analyses of these data segregated chickpeas into groups. Polymorphism in seed glutelines was absent, while variation in seed prolamines was very limited. DNA amplification patterns have been analyzed by semi-arbitrary primers and by specific microsatellite primers. Only some of semi-arbitrary primers generated usable DNA banding patterns, moreover interpretation of these patterns can be more or less difficult. On the contrary specific primers amplifying microsatellites at the specific loci generated unambiguous and reproducible differences between chickpeas.     

Mitigation of salt stress in wheat seedlings by a <Emphasis Type="Italic">gfp</Emphasis>-tagged <Emphasis Type="Italic">Azospirillum lipoferum</Emphasis>

Root colonization and mitigation of NaCl stress on wheat seedlings were studied by inoculating seeds with Azospirillum lipoferum JA4ngfp15 tagged with the green fluorescent protein gene (gfp). Colonization of wheat roots under 80 and 160 mM NaCl stress was similar to root colonization with this bacterial species under non-saline conditions, that is, single cells and small aggregates were mainly located in the root hair zone. These salt concentrations had significant inhibitory effects on development of seedlings, but not on growth in culture of gfp-A. lipoferum JA4ngfp15. Reduced plant growth (height and dry weight of leaves and roots) under continuous irrigation with 160 mM NaCl was ameliorated by bacterial inoculation with gfp-A. lipoferum JA4ngfp15. Inoculation of plants subjected to continuous irrigation with 80 mM NaCl or to a single application of either NaCl concentration (80 or 160 mM NaCl) did not mitigate salt stress. This study indicates that, under high NaCl concentration, inoculation with modified A. lipoferum reduced the deleterious effects of NaCl; colonization patterns on roots were unaffected and the genetic marker did not induce undesirable effects on the interaction between the bacterium and the plants.     

Characterization of fertile amphidiploid between <Emphasis Type="Italic">Raphanus sativus</Emphasis> and <Emphasis Type="Italic">Brassica alboglabra</Emphasis> and the crossability with <Emphasis Type="Italic">Brassica</Emphasis> species

A fertile amphidiploid × Brassicoraphanus (RRCC, 2n = 36) between Raphanus sativus cv. HQ-04 (2n = 18, RR) and Brassica alboglabra Bailey (2n = 18, CC) was synthesized and successive selections for seed fertility were made from F₄ to F₁₀. F₁₀ plants exhibited good fertility with 14.9 seeds per siliqua and 32.3 g seeds per plant. Cytological observation revealed that frequent secondary pairing occurred among 3 chromosome pairs in pollen mother cells of plants (F₄) with lower fertility, but not of plants with high fertility (F₁₀). GISH analysis indicated that these F₁₀ plants included the expected 18 chromosomes from R. sativus and B. alboglabra, respectively, but they lost approximately 27.6% R. sativus and 35.6% B. alboglabra AFLP (amplified fragment length polymorphism) bands. The crossability of the Raphanobrassica with R. sativus and 5 Brassica species (13 cultivars) were investigated. Seeds or F₁ seedlings were easy to be produced from crosses × Brassicoraphanus × R. sativus, and B. napus, B. juncea and B. carinata × Brassicoraphanus. Fewer seeds or seedlings were obtained from crosses × Brassicoraphanus × B. napus, B. juncea and B. carinata. However, few seeds were harvested in the reciprocals of × Brassicoraphanus with B. rapa and B. oleracea. The possible cause of fertility improvements and the potential of the present × Brassicoraphanus for breeding were discussed.     

Genetic diversity of the D-genome in <Emphasis Type="Italic">T. aestivum</Emphasis> and <Emphasis Type="Italic">Aegilops</Emphasis> species using SSR markers

Simple sequence repeats (SSRs), highly dispersed nucleotide sequences in genomes, were used for germplasm analysis and estimation of the genetic relationship of the D-genome among 52 accessions of T. aestivum (AABBDD), Ae. tauschii (D^tD^t), Ae. cylindrica (CCD^cD^c) and Ae. crassa (MMD^cr1D^cr1), collected from 13 different sites in Iran. A set of 21 microsatellite primers, from various locations on the seven D-genome chromosomes, revealed a high level of polymorphism. A total of 273 alleles were detected across all four species and the number of alleles per each microsatellite marker varied from 3 to 27. The highest genetic diversity occurred in Ae. tauschii followed by Ae. crassa, and the genetic distance was the smallest between Ae. tauschii and Ae. cylindrica. Data obtained in this study supports the view that genetic variability in the D-genome of hexaploid wheat is less than in Ae. tauschii. The highest number of unique alleles was observed within Ae. crassa accessions, indicating this species as a great potential source of novel genes for bread wheat improvement. Knowledge of genetic diversity in Aegilops species provides different levels of information which is important in the management of germplasm resources.     

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.     

Use and variation of <Emphasis Type="Italic">Pandanus tectorius</Emphasis> Parkinson (<Emphasis Type="Italic">P. fascicularis</Emphasis> Lam.) along the coastline of Orissa,India

Pandanus tectorius Parkinson ( P. fascicularis Lam.) of the family Pandanaceae constitutes one of the major bioresources of Ganjam coast, Orissa; used mainly in small scale perfume industry for aromatic compound extracted from the male inflorescences. In order to establish genetic diversity, if any related to perfume yield, samples of P. tectorius representing male populations from seven locations representing populations I–VII along the coastline of Orissa, India, were analysed for somatic chromosome number, 4C genomic DNA content, randomly amplified polymorphic DNA (RAPD) as well as phytochemicals. The somatic chromosome number in all the populations I–VII was 2n = 60. The chromosomes were of minute size without showing any remarkable structural variation. Like wise the average 4C DNA content was 5.09 pg (≅4,912 Mbp) that showed no intra- or inter-population differences. Out of 54 decamer primers tested, a total of 1,260 amplicons were obtained from 34 primers accounting 43.49% polymorphism. Molecular phylogenetic analysis of the seven populations revealed two distinct branches, with populations II and III in one and the rest populations in the other branch of the phylogenetic tree. It was important to note that the unique populations II and III confined to the Ganjam coast of Orissa having RAPD markers: OPA 09–940 bp, OPA 09–705 bp, OPC 14–1,500 bp, OPC 14–700 bp, OPC 20–1,475 bp, OPC 20–1,350 bp, OPC 20–920 bp and OPC 20–700 bp, were distinguished form the rest of the populations. The aforesaid populations (II and III) are well known to produce aroma of high quality and yield, composed of primarily phenyl ethyl methyl ether (66.8–83%) and terpinen-4-ol (5–12%) along with a number of other phyto-chemical compounds that support the flourishing perfume industry and livelihood of the local people in the region. The findings underscored the possible role of local eco-geography in contributing to the micro-evolution of unique high perfume yielding genotypes of P. tectorius that represented populations II and III at Ganjam coast, which were genetically distinct from the rest of the populations revealed by RAPD analysis.     

Classification and phylogenetic relationships in <Emphasis Type="Italic">Solanum</Emphasis> section <Emphasis Type="Italic">Lycopersicon</Emphasis> based on AFLP and two nuclear gene sequences

The classification and phylogeny of the species belonging to Solanum section Lycopersicon is a complex issue that has not yet reached a widely accepted consensus. These species diverged recently, are still closely related and, in some cases, are still even capable of interspecific hybridization, thereby blurring the difference between intra- and interspecific variation. To help resolve these issues, in the present study, several accessions covering the natural range for each species were used. In addition, to avoid biases due to the molecular method employed, both AFLP markers and two nuclear-gene sequences, CT179 and CT66, were used to characterize the plant materials. The data obtained suggest a classification similar to those previously proposed by other authors, although with some significant changes. Twelve species were recognized as distinct based on this dataset. According to the data presented, the recently proposed species, S. corneliomulleri, is indistinguishable from S. peruvianum s.str. In addition, both the sequence and the AFLP trees suggest that S. arcanum could represent a complex of populations composed of two cryptic species. With regard to phylogenetic relationships among these species, some clear groups were found: the Lycopersicon group formed by S. pimpinellifolium, S. lycopersicum, S. cheesmaniae and S. galapagense; the Arcanum group constituted by S. chmielewskii, S. neorickii, S. arcanum and S. huaylasense; and the Eriopersicon group made up of S. peruvianum and S. chilense. Solanum pennellii and S. habrochaites are not included in any group, but are the closest to the S. lycopersicoides outgroup. Jose Blanca and Fernando Nuez have contributed equally to this work and should be regarded as co-second authors.