Genetic relationships among Stylosanthes species revealed by RFLP and STS analyses

Genetic relationships between 6 unclassified taxa and 24 known species of the genus Stylosanthes were investigated by RFLP and STS analyses. This allowed the diploid taxa used in this study to be classified into nine basal (genome) groups. Representative species in these groups included ’S. seabrana’/S. hamata (Group A), S. viscosa (Group B), S. humilis (Group C), S. macrocephala/S. bracteata (Group D), S. pilosa (Group E), S. leiocarpa (Group F), S. guianensis (Group G), S. tomentosa (Group H) and S. calcicola (Group I). Polyploid taxa used were grouped into five classes based on their putative genomic structures. These are AABB for S. scabra, S. aff. scabra, S. sericeiceps, S. aff. hamata and S. tuberculata; AACC for S. mexicana, S. subsericea, S. sundaica and S. sp.A; DDEE for S. capitata; AAFF for S. sympodialis; and AABBXX for S. erecta, with XX representing an unknown genome. Of the 6 unclassified taxa, three were diploids and 3 tetraploids. Of the 3 diploids, the genome of S. sp. was markedly distinct from those of all other diploids analysed in this study, with that of S. leiocarpa being the closest. The genome of S. sp.B was similar to that of S. humilis, with an average dissimilarity value of 15% between them. The genome of S. aff. viscosa was very similar to that of S. viscosa. Genetic variation between these 2 taxa was not larger than that within each of the 2 taxa. Of the 3 tetraploids, the genomic structure of S. sp.A was similar to those of S. mexicana, S. sundaica and S. subsericea, and the genomic structures of S. aff. scabra and S. aff. hamata were similar to those of S. scabra and S. sericeiceps. Received: 18 September 1998 / Accepted: 23 March 1999     

Maternal origin,genome constitution and evolutionary relationships of polyploid <Emphasis Type="Italic">Elymus</Emphasis> species and <Emphasis Type="Italic">Hordelymus europaeus</Emphasis>

The trnS/psbC region of chloroplast DNA (cpDNA) was sequenced for 18 Elymus polyploid species, Hordelymus europaeus and their putative diploid ancestors. The objective was to determine the maternal origin and evolutionary relationships of these polyploid taxa. Phylogenetic analysis showed that Elymus and Pseudoroegneria species formed a highly supported monophyletic group (100 % bootstrap values), suggesting that Pseudoroegneria is the maternal genome donor to polyploid Elymus species studied here. The phylogenetic tree based on cpDNA sequence data indicates that E. submuticus contains a St-genome. Taking into consideration of our previously published RPB2 data, we can conclude that hexaploid E. submuticus contains at least one copy of St and Y genomes. Our Neighor-joining analysis of cpDNA data put Psathyrostachys juncea, Hordeum bogdanii and Hordelymus europaeus into one group, suggesting a close relationship among them.     

Stylosanthes sp. aff.S. scabra: a putative diploid progenitor ofStylosanthes scabra (Fabaceae)

Stylosanthes sp. aff.S. scabra is an undescribed taxon showing affinities with the allotetraploid speciesS. scabra, but distinct in a number of attibutes. Several collections show potential as forage for clay soils in northern Australia. Twelve accessions have been analysed using STS (sequence-tagged-sites) as genetic markers, and they all displayed STS phenotypes of typical diploid species. Taking into account their morphological similarities, the STS analysis provides strong evidence thatStylosanthes sp. aff.S. scabra might be a diploid progenitor of the allotetraploidS. scabra. This speculation was supported by cytological examinations. Somatic chromosome numbers of two of these accessions were counted and both were found to be diploid (2n = 20). The level of polymorphism among the 12Stylosanthes sp. aff.S. scabra accessions, estimated using randomly amplified polymorphic DNA (RAPD) as markers, was 7.8%, and the dissimilarity value betweenStylosanthes sp. aff.S. scabra andS. viscosa (the other putative progenitor ofS. scabra) was 89%.     

Phylogenetic relationships in the genusStylosanthes (Leguminosae) based upon chloroplast DNA variation

A detailed analysis of chloroplast DNA restriction fragment length variation was undertaken to reconstruct the maternal phylogeny of 18 taxa from both sections of the papilionoid tropical forage legume genusStylosanthes. Data were analysed by means of the computer program PAUP, using an heuristic search with Wagner parsimony. The resulting cladogram dividedStylosanthes into four separate clades, which comprised: (i) theS. guianensis complex and related species (i.e.S. gracilis, S. grandifolia andS. montevidensis); (ii)S. hispida, tetraploidS. hamata s. l.,S. sympodialis, S. humilis, S. leiocarpa, S. angustifolia and certain accesions ofS. scabra; (iii)S. calcicola, S. viscosa, diploidS. hamata s. str., andS. fruticosa, plus accessions ofS. scabra, S. capitata and one accession ofS. grandifolia; and (iv)S. macrocephala and other accessions ofS. capitata not included within clade 3. Results are generally congruent with previously established interspecific relationships and, moreover, enabled identification of putative maternal progenitors for four tetraploid taxa:S. humilis was identified as a likely maternal parent of bothS. sympodialis andS. hamata s. l.,S. viscosa as a maternal parent ofS. scabra, andS. macrocephala as a maternal parent ofS. capitata.     

Evidence for a low level of genomic specificity of sequence-tagged-sites in Stylosanthes

Genome-specific DNA markers are of great value in many applications. Recent work on different plants and animal species indicated that PCR- (polymerase chain reaction) based genetic marker systems using specific primers are highly genome-specific. To test the genome specificity of sequence-tagged-sites (STSs) as genetic markers in Stylosanthes, 20 pairs of primers were generated. Fifteen were from randomly selected single-copy Pstl genomic clones, and the other five were from two known gene sequences. These primer pairs were analysed against a set of 24 genotypes representing 12 different Stylosanthes species. Thirteen of these primer pairs amplified successfully. Overall, there was a low level of genome specificity, suggesting a low degree of genomic divergence within this group of Stylosanthes species. Of the 312 entries (24 genotypes by 13 primer pairs), PCR amplifications were unsuccessful (little or no products) in only 16 cases. The number of banding patterns detected by each of these primer pairs varied from 2 to 12 with an average pair-wise polymorphism of 44.3%. The level of intraspecific variation detected on normal agarose gels was only 3.8%. Further evidence that diploid S. hamata and diploid S. humilis are progenitors of tetraploid S. hamata and that S. viscosa is a progenitor of S. scabra, was obtained.     

Identification of Diploid <Emphasis Type="Italic">Stylosanthes seabrana</Emphasis> Accessions from Existing Germplasm of <Emphasis Type="Italic">S. scabra</Emphasis> Utilizing Genome-Specific STS Markers and Flow Cytometry,and Their Molecular Characterization

Stylosanthes seabrana (Maass and ‘t Mannetje) (2n = 2x = 20), commonly known as Caatinga stylo, is an important tropical perennial forage legume. In nature, it largely co-exist with S. scabra, an allotetraploid (2n = 4x = 40) species, sharing a very high similarity for morphological traits like growth habit, perenniality, fruit shape and presence of small appendage at the base of the pod or loment. This makes the two species difficult to distinguish morphologically, leading to chances of contamination in respective germplasm collections. In present study, 10 S. seabrana accessions were discovered from the existing global germplasm stock of S. scabra represented by 48 diverse collections, utilizing sequence-tagged-sites (STS) genome-specific markers. All the newly identified S. seabrana accessions displayed STS phenotypes of typical diploid species. Earlier reports have conclusively indicated S. seabrana and S. viscosa as two diploid progenitors of allotetraploid S. scabra. With primer pairs SHST3F3/R3, all putative S. seabrana yielded single band of ~550 bp and S. viscosa of ~870 bp whereas both of these bands were observed in allotetraploid S. scabra. Since SHST3F3/R3 primer pairs are known to amplify single or no band with diploid and two bands with tetraploid species, the amplification patterns corroborated that all newly identified S. seabrana lines were diploid in nature. Flow cytometric measurement of DNA content of the species, along with distinguishing morphological traits such as flowering time and seedling vigour, which significantly differ from S. scabra, confirmed all identified lines as S. seabrana. These newly identified lines exhibited high level of similarity among themselves as revealed by RAPD and STS markers (>92% and 80% respectively). Along with the enrichment in genetic resources of Stylosanthes, these newly identified and characterized accessions of S. seabrana can be better exploited in breeding programs targeted to quality.     

Phylogenetic analysis of Stylosanthes (Fabaceae) based on the internal transcribed spacer region (ITS) of nuclear ribosomal DNA

 Phylogenetic relationships in Stylosanthes are inferred by DNA sequence analysis of the ITS region (ITS1–5.8S–ITS2) of the nuclear ribosomal DNA in 119 specimens, representing 36 species of Stylosanthes and 7 species of the outgroup genera Arachis and Chapmannia. In all examined specimens of any particular diploid and (allo)polyploid species, only a single ITS sequence type was observed. This allowed us to identify a parental genome donor for some of the polyploids. In several diploid and polyploid species, different specimens contained a different ITS sequence. Some of these sequence types were present in more than one species. Parsimony analysis yielded several well-supported clades that agree largely with analyses of the chloroplast trnL intron and partially with the current sectional classification. Discordances between the nuclear and cpDNA analyses are explained by a process of allopolyploidization with inheritance of the cpDNA of one parent and fixation of the ITS sequences of the other. S. viscosa has been an important genome donor in this process of speciation by allopolyploidy. Received August 14, 2001; accepted March 4, 2002 Published online: November 14, 2002 Addresses of the authors: Jacqueline Vander Stappen, Steven Van Campenhout and Guido Volckaert (E-mail: guido.volckaert@agr.kuleuven.ac.be), Katholieke Universiteit Leuven, Laboratory of Gene Technology, Kasteelpark Arenberg 21, B-3001 Leuven, Belgium. Jan De Laet, American Museum of Natural History, Division of Invertebrate Zoology, Central Park West at 79th Street, New York 10024–5192, USA. Susana Gama-López, Universidad Nacional Autónoma de México, Unidad de Biología, Tecnología y Protipos (UBIPRO), FES-Iztacala, Laboratorio de Recursos Naturales, Av. de Los Barrios S/N, Colonia Los Reyes Iztacala, Municipio Tlalnepantla, Estado de México, C.P. 54090, México. Present address: Apartado Postal 154, Cto. Parque No. 3, C.P. 53102, México.     

Chloroplast DNA variation in diploid and polyploid species of Bromus (Poaceae) subgenera Festucaria and Ceratochloa

Summary Chloroplast DNA (cpDNA) restriction endonuclease patterns are used to examine phylogenetic relationships between Bromus subgenera Festucaria and Ceratochloa. Festucaria is considered monophyletic based on the L genome, while Ceratochloa encompasses two species complexes: the B. catharticus complex, which evolved by combining three different genomes, and the B. carinatus complex, which is thought to have originated from hybridization between polyploid species of B. catharticus and diploid members of Festucaria. All species of subgenus Ceratochloa (hexaploids and octoploids) were identical in chloroplast DNA sequences. Similarly, polyploid species of subgenus Festucaria, except for B. auleticus, were identical in cpDNA sequences. In contrast, diploid species of subgenus Festucaria showed various degrees of nucleotide sequence divergence. Species of subgenus Ceratochloa appeared monophyletic and phylogenetically closely related to the diploid B. anomalus and B. auleticus of subgenus Festucaria. The remaining diploid and polyploid species of subgenus Festucaria appeared in a distinct grouping. The study suggests that the B. catharticus complex must have been the maternal parent in the proposed hybrid origin of B. carinatus complex. Although there is no direct evidence for the paternal parent of the latter complex, the cpDNA study shows the complex to be phylogenetically very related to the diploid B. anomalus of subgenus Festucaria.     

Independent origins of tetraploid cryptic species in the fern <Emphasis Type="Italic">Ceratopteris thalictroides</Emphasis>

Ceratopteris thalictroides (L.) Brongn is a tetraploid fern species that contains at least three cryptic species, the south, the north and the third type. In this study we combined data from both chloroplast DNA (cpDNA) and nuclear DNA sequences of three diploid species and three cryptic species of C. thalictroides to unravel the origin of the cryptic species, particularly of the reticulate relationships among the diploid and tetraploid taxa in the genus Ceratopteris. Of the three diploid species examined, C. cornuta had cpDNA identical to that of the tetraploid third type plants, and this diploid species is a possible maternal ancestor of the tetraploid third type. Analysis of the homologue of the Arabidopsis thaliana LEAFY gene (CLFY1) identified ten alleles in the genus Ceratopteris, with six alleles found in C. thalictroides. The unrooted tree of the CLFY1 gene revealed four clusters. Each cryptic species showed fixed heterozygosity at the CLFY1 locus and had two alleles from different clusters of the CLFY1 tree. Consideration of the cpDNA sequences, CLFY1 genotypes of the cryptic species and CLFY1 gene tree in concert suggested that the cryptic species of C. thalictroides had originated through independent allopolyploidization events involving C. cornuta and two unknown hypothetical diploid species.     

DNA evidence for multiple origins of intergeneric allopolyploids in annualMicroseris (Asteraceae)

Seventy populations of North American annualMicroseris, Stebbinsoseris, andUropappus species were examined for chloroplast and nuclear ribosomal DNA restriction site variability to determine the origin of the allotetraploid speciesS. heterocarpa andS. decipiens. Previously identified chloroplast DNA restriction site variants were used in concert with restriction site variation forNco I in the nuclear-encoded ribosomal DNA repeat. The presence of two, mutually exclusive restriction site gains were observed in diploid populations ofM. douglasii; these same variants were also found in populations of allotetraploidS. heterocarpa, indicating mutiple origins of this species from different maternal diploid populations ofM. douglasii. Variation in the rDNA repeat between the diploid annual species and the putative paternal genome ofU. lindleyi was found to be additive inS. heterocarpa. A similar relationship was observed for the origin ofS. decipiens; cpDNA restriction site variants found inM. bigelovii andM. douglasii were present inS. decipiens. The rDNANco I variants also were additive in this purported allotetraploid. These results confirm the reticulate evolutionary pattern inStebbinsoseris and provide another example of multiple origins of intergeneric allopolyploids.     

Chloroplast-DNA variation in the genus Lotus (Fabaceae) and further evidence regarding the maternal parentage of Lotus corniculatus L.

 To resolve the maternal parentage of the tetraploid Lotus corniculatus, restriction-site variation of chloroplast DNA (cpDNA) was studied in several accessions of that species, in the four putative parental diploid species, L. tenuis, L. alpinus, L. japonicus and L. uliginosus, and in four phylogenetically more distant diploid species, L. hispidus, L. edulis, L. ornithopodoides and Tetragonolobus maritimus var. siliquosus. Evidence of cpDNA maternal inheritance was obtained by using reciprocal controlled crosses between plants of L. corniculatus and natural tetraploid individuals of L. alpinus showing very distinct restriction patterns. Interspecific cpDNA variation in the eight Lotus species and T. siliquosus was analysed by comparing cpDNA fragment patterns produced by five restriction endonucleases and totalling 304 distinct fragments. Genetic differentiation in cpDNA was very high between the L. corniculatus group and L. hispidus on the one hand, and the three other species on the other hand. Sixteen restriction-site mutations and eight length polymorphisms were identified among the five species of the L. corniculatus group and L. hispidus, Lotus uliginosus, L. alpinus and L. japonicus showed at least six DNA changes with regard to the molecule of L. corniculatus. Accordingly, these species should be excluded as maternal progenitors of L. corniculatus. Conversely, the cpDNA of L. tenuis differed from that of L. corniculatus by only two small-length mutations. As also suggested previously from an analysis of several nuclear markers, the results reported here show decisively that L. tenuis may be considered as the most probable maternal ancestor of L. corniculatus. Received: 23 February 1997/Accepted: 28 February 1997     

Carbon isotope discrimination function analysis and drought tolerance of stylo species grown under rain-fed environment

Limited germplasm and narrow genetic base in the past has hinders wider applicability of Stylosanthes in India. However, build up in Stylosanthes germplasm in recent years provided an opportunity to evaluate and use them under different agro-ecological situations preferably targeting drought tolerance. Rate reducing resistance (RRR) allo-tetraploid lines of S. scabra and diploid S. seabrana, a newly introduced species, observed well suited in hard and cracking soils under complete rainfed condition. Genotypes of S. scabra were more tolerant to drought over lines of other species as evidenced by high leaf thickness, more proline accumulation, contents of malondialdehyde (MDA), sugars, starch and chlorophyll and low carbon isotope discrimination (CID) values. Both in control and stress conditions, a positive relationship (r = 0.465 and 0.328) was observed between specific leaf area (SLA) and CID. Earlier reports have emphasized measurement of CID as an indirect way of measuring the transpiration efficiency (TE) in Stylosanthes (Thumma et al. Aust J Agr Res 49:1039–1045, 1998). The negative relationship observed between TE and CID suggested low CID bearing lines would have high TE, useful traits to select lines growing better in dry environments. Thus, selected lines having low CID provided better scope for the introduction of this important range legume for semi- arid regions of India.     

Genetic variation in agronomically important species of Stylosanthes determined using random amplified polymorphic DNA markers

Summary Random amplified polymorphic DNA (RAPD) markers were generated from 20 cultivars and accessions representing four agronomically important species of Stylosanthes, S. scabra, S. hamata, S. guianensis, and S. humilis. Approximately 200 fragments generated by 22 primers of arbitrary sequence were used to assess the level of DNA variation. Relatively low levels of polymorphism (0–16% of total bands in pairwise comparisons) were found within each species, while polymorphisms between the species were much higher (up to 46%). Very few polymorphisms (0–2%) were detected between the individuals of the same cultivar or accession. A phenogram of relationships among the species was constructed based on band sharing. Four main clusters corresponding to each species were readily distinguished on this phenogram. The allotetraploid species S. hamata and its putative diploid progenitor, S. humilis, were more similar to each other than to S. scabra and S. guianensis. No variation in RAPD markers was found between the two commercial S. hamata cvs Verano and Amiga. Cultivar Oxley in S. guianensis was considerably different from the other cultivars and accessions of this species. The phylogenetic distinctions obtained with RAPDs were in agreement with other studies from morphology, cytology, and enzyme electrophoresis. The low level of polymorphisms observed within each species suggested that interspecific crosses may be a better vehicle for the construction of RAPD linkage maps in Stylosanthes.     

Multiple independent formations of Tragopogon tetraploids (Asteraceae): evidence from RAPD markers

Polyploidy is widely recognized as a significant force leading to the formation of new plant species. Estimates of the number of angiosperm species with polyploid origins are as high as ≈ 50%; however, in spite of this prevalence, many aspects of polyploid evolution remain poorly understood. Recent studies have suggested that recurrent origins of polyploid species are the rule rather than the exception. The present study is one of only a few designed to quantify the number of independent origins of a polyploid species. The two tetraploid species Tragopogon mirus and T. miscellus (Asteraceae) arose within the past 50 years in the Palouse region of eastern Washington and adjacent northern Idaho. Previous work using morphology, cpDNA and rDNA restriction site analyses, allozymes, cytology, and flavonoid chemistry established that T. mirus had arisen at least five times, and T. miscellus at least twice, on the Palouse. To assess the frequency of multiple origins of these species more rigorously, seven populations of T. mirus and three populations of T. miscellus that were indistinguishable based on previous markers were surveyed using random amplified polymorphic DNA (RAPD) markers; populations of the diploid progenitor species from the same sites were also analysed. Each tetraploid population had a unique RAPD marker profile, suggesting that each population surveyed originated independently of the other populations in the region. Only two of the tetraploid populations combined the RAPD marker profiles of the diploid progenitors occurring at the same site. Both polyploid species, whose ranges and numbers have greatly increased since their formation in the early part of the twentieth century, have formed repeatedly on a local geographical scale and during a short time frame. Furthermore, each tetraploid species is spreading not primarily by dispersal of propagules from a single population of origin, but through repeated, independent polyploidization events that recreate the polyploid taxa.     

Chloroplast DNA variation in the cultivated and wild olive taxa of the genus Olea L.

Polymorphism in the lengths of restriction fragments of the whole cpDNA molecule were studied in 15 taxa (species or subspecies) of the genus Olea. From restriction analysis using nine endonucleases, 28 site mutations and five length polymorphisms were identified, corresponding to 12 distinct chlorotypes. From a phenetic analysis based on a Nei’s dissimilarity matrix and a Dollo parsimony cladistic analysis using, as an outgroup, a species of the genus Phillyrea close to Olea, the ten taxa of section Olea were distinguished clearly from the five taxa of section Ligustroides which appear to posses more ancestral cpDNA variants. Within the section Ligustroides, the tropical species from central-western Africa, Olea hochtetteri, showed a chlorotype which differed substantially from those of the other four Olea taxa growing in southern Africa, supporting a previous assessment according to which O. hochtetteri may have been subjected to a long period of geographical isolation from the other Olea taxa. Within the Olea section, three phyla were identified corresponding to South and East Africa taxa, Asiatic taxa, and a group including Saharan, Macaronesian and Mediteranean taxa, respectively. On the basis of cpDNA variation, the closest Olea taxa to the single Mediterranean species, Olea europaea, represented by its very predominant chlorotype, observed in both wild and cultivated olive, were found to be Olea laperrinei (from the Sahara), Olea maroccana (from Maroccan High Atlas) and Olea cerasiformis (from Macaronesia). These three taxa, which all share the same chlorotype, may have a common maternal origin. Received: 5 December 1999 / Accepted: 30 December 1999     

New chromosome counts and evidence of polyploidy in Haageocereus and related genera in tribe Trichocereeae and other tribes of Cactaceae

Chromosome numbers for a total of 54 individuals representing 13 genera and 40 species of Cactaceae, mostly in tribe Trichocereeae, are reported. Five additional taxa examined belong to subfamily Opuntioideae and other tribes of Cactoideae (Browningieae, Pachycereeae, Notocacteae, and Cereeae). Among Trichocereeae, counts for 35 taxa in eight genera are reported, with half of these (17 species) for the genus Haageocereus. These are the first chromosome numbers reported for 36 of the 40 taxa examined, as well as the first counts for the genus Haageocereus. Both diploid and polyploid counts were obtained. Twenty nine species were diploid with 2n=2x=22. Polyploid counts were obtained from the genera Espostoa, Cleistocactus, Haageocereus, and Weberbauerocereus; we detected one triploid (2n=3x=33), nine tetraploids (2n=4x=44), one hexaploid (2n=6x=66), and three octoploids (2n=8x=88). In two cases, different counts were recorded for different individuals of the same species (Espostoa lanata, with 2n=22, 44, and 66; and Weberbauerocereus rauhii, with 2n=44 and 88). These are the first reported polyploid counts for Haageocereus, Cleistocactus, and Espostoa. Our counts support the hypothesis that polyploidy and hybridization have played prominent roles in the evolution of Haageocereus, Weberbauerocereus, and other Trichocereeae.     

Comparison of four molecular markers in measuring relationships among the wild potato relatives Solanum section Etuberosum (subgenus Potatoe)

We evaluated chloroplast DNA (cpDNA), isozymes, single to low-copy nuclear DNA (RFLPs), and random amplified polymorphic DNAs (RAPDs) in terms of concordance for genetic distance of 15 accessions each of Solanum etuberosum and S. palustre, and 4 accessions of S. fernandezianum. These self-compatible, diploid (2n=24), and morphologically very similar taxa constitute all species in Solanum sect. Etuberosum, a group of non-tuber-bearing species closely related to Solanum sect. Petota (the potato and its wild relatives). Genetic distance and multidimentional scaling results show general concordance of isozymes, RFLPs and RAPDs between all three taxa; cpDNA shows S. etuberosum and S. palustre to be more similar to each other than to S. fernandezianum. Interspecific sampling variance shows a gradation of resolution from allozyme (low) to RAPD to RFLP (high); while intraspecific comparisons graded from RFLPs (low) to RAPDs (high; lack of sufficient allozyme variability within species precluded comparisons for allozymes). Experimental error was low in RFLPs and RAPDs.Names are necessary to report factually and available data; however, the USDA neither guarantees nor warrants the standard of the product, and the use of the name by USDA implies no approval of the product to the exclusion of others that may also be suitable     

Molecular systematics and biogeography of theCardamine pratensis complex (Brassicaceae)

Representatives of theC. pratensis complex were analysed for allozymes, ITS, non-coding cpDNA, and RAPDs to elucidate phylogenetic relationships and the historical biogeography of this species group. Our concepts differ in some important aspects from current ideas. Two diploid species from southeastern Europe form the Basal Group of the complex. A diploid from the Iberian Peninsula represents another old lineage. The phylogenetically younger Derived Group comprises diploid taxa and all known polyploid taxa. The two old lineages represent pleistocene relicts which were not involved in the formation of the Derived Group. All polyploids evolved in postglacial time from diploids of the Derived Group which may have survived the glaciations in refugia centered around and within the Alps. The arctic-circumpolarC. nymanii is of young age and migrated to Scandinavia in postglacial times from south to north.     

Phytogeny of Brassica and allied genera based on variation in chloroplast and mitochondrial DNA patterns: molecular and taxonomic classifications are incongruous

Summary Chloroplast DNA (cpDNA) variability of 60 taxa of the genus Brassica and allied genera comprising 50 species was studied. RFLPs for seven enzymes were generated and F values were estimated from five frequently cutting enzymes. Phenetic clusterings indicated a clear division of Brassica coenospecies into two distinct lineages referred to as the Brassica and Sinapis lineages. Two unexplored genera, Diplotaxis and Erucastrum, also exhibited two lineages in addition to the genera Brassica and Sinapis. This finding is inconsistent with the existing taxonomic classification based on morphology. Mitochondrial DNA (mtDNA) variability studied from EcoRI RFLP patterns, by hybridizing total DNA with four cosmid clones containing non-overlapping mtDNA fragments, did not show any congruence with cpDNA variation patterns. However, at the cytodeme level, the patterns of genetic divergence suggested by the cpDNA data could be correlated with mtDNA variation. In the Brassica lineage, Diplotaxis viminea was identified as the female parent of the allotetraploid D. muralis. The chloroplast DNAs of Erucastrum strigosum and Er. abyssinicum were found to be very closely related. In the Sinapis lineage, Brassica maurorum was found to be the diploid progenitor of autotetraploid B. cossoneana. B. amplexicaulis showed a very different cpDNA pattern from other members of the subtribe. Brassica adpressa was closest to Erucastrum laevigatum and could be the diploid progenitor of autotetraploid Er. laevigatum. Based on the close similarity of the cpDNA pattern of Diplotaxis siifolia with that of D. assurgens, we have proposed the retention of this species in the genus Diplotaxis. The taxonomic positions of some other species have also been discussed.     

Variation in Drought Tolerance of Different Stylosanthes Accessions

Twenty genotypes of Stylosanthes consisting four species were evaluated under rain fed condition employing biochemical and physiological attributes to select drought tolerant lines. Relative water content measured at 50 % flowering stage of the plants showed significant variations among the lines which ranged from 32.11 in S. scabra RRR94-86 to 83.33 % in S. seabrana 2539. The results indicated that S. scabra genotypes were more tolerant to drought over other lines as evidenced by high leaf thickness, proline accumulation, content of sugars and chlorophyll, and nitrate reductase activity.