Molecular phylogeny of genus <Emphasis Type="Italic">Guizotia</Emphasis> (Asteraceae) using DNA sequences derived from ITS

Complete sequences for the internal transcribed spacers of the 18s–26s nuclear ribosomal DNA were generated to establish phylogenetic relationships among five species of the genus Guizotia. Parsimony analysis and pairwise distance data produced a single tree with four clearly distinguished clades that accord with previously reported chromosomal data. The clades produced here have been discussed with reference to existing taxonomic treatments. It appears that Guizotia scabra ssp. scabra, G. scabra ssp. schimperi and Guizotia villosa have contributed to the origin of Guizotia abyssinica, the cultivated species of the genus. The present composition of the species of genus Guizotia and the subtribe the genus presently placed in are suggested to be redefined.     

Phylogenetic relationships within <Emphasis Type="Italic">Citrus</Emphasis> and its related genera as inferred from AFLP markers

We investigated the phylogenetic relationships among Citrus and its relatives, including 29 genotypes belonging to Citrus, Poncirus, Fortunella, Microcitrus, Eremocitrus, Atalantia and Severinia using AFLP analysis. Cluster analysis was conducted using neighbour joining (NJ) method. The results demonstrated that Poncirus, Microcitrus and Eremocitrus are distant from Citrus. A strong affiliation exists between C. halimii and Fortunella and the results did not support C. halimii as the fourth basic species. Further evidence was provided that P. polyandra should not be the hybrid between trifoliate orange and Citrus genotypes and it deserves species status. Neither the separation of subgenus Papeda and Citrus nor the separation of subgenus Archicitrus and Metacitrus were clearly resolved in the dendrogram. C. ichangesis is a distinct species very different from other Citrus genotypes, and the results showed it is improper to classify it into subgenus Metacitrus. C. reticulata, C. maxima and C. medica were separated into three distinct clusters. Therefore if only cultivated Citrus species are considered, the three basic species concept is acceptable. However, it is not applicable for whole genus Citrus.     

Genetic diversity of <Emphasis Type="Italic">rbc</Emphasis>L gene in <Emphasis Type="Italic">Elymus trachycaulus</Emphasis> complex and their phylogenetic relationships to several Triticeae species

Elymus trachycaulus complex species are known for their morphological variability, but little is known about their genetic basis. The phylogenetic relationships among the E. trachycaulus complex, and their systematic relation to other species in Triticeae remain unknown. Nucleotide diversity of ribulose-1,5 bisphosphate-carboxylase (rbcL) gene in E. trachycaulus complex species and several other Triticeae was first characterized and compared. A primary conclusion of the present study is that nucleotide diversity for rbcL gene in E. trachycaulus species was detected with the estimates of nucleotide diversity θ = 0.00039 and π = 0.00043. The estimate of nucleotide diversity in rbcL gene for species with different genome constitution here ranged from 0.00099 (π) and 0.00099 (θ) for the species with Ns genome to 0.00226 (π) and 0.00291 (θ) for the species with St genome. The phylogenetic relationships of these species were assessed using these rbcL sequences. A total of 47 variable positions including 19 parsimony-informative sites were detected among 24 accessions of 18 species/subspecies. The species with St, H/I and Ns genomes well separated from each other, and formed a three distinct clades with higher bootstrap values support for both Parsimony and NJ analyses. The St genome containing species is sister group of H/I genome containing species. Our result confirms that Pseudoroegneria is the maternal genome donor to these Elymus species studied here, regardless of their distribution. Elymus trachycaulus complex are more related to each other than to E. glaucescens, E. patagonicus, and E. solandri. This study suggested that Elymus species with StH genomes may form from multiple closely related sets of donors.     

Molecular phylogeny of Chinese vegetable mustard (<Emphasis Type="Italic">Brassica juncea</Emphasis>) based on the internal transcribed spacers (ITS) of nuclear ribosomal DNA

Sequence variation of nuclear internal transcribed spacer regions of ribosomal DNA (ITS1, 5.8S rRNA and ITS2) from Chinese vegetable mustards (AB-genome) and its putative parents Brassica rapa (the A-genome) and Brassica nigra (the B-genome) were used to investigate the molecular phylogeny and the probable evolutional pattern of this amphidiploid species that uniquely formed in China. Totally, 16 accessions of Chinese vegetable mustard those covering nearly all the diverse variations were included in this study, and together with three accessions of B. rapa and one accession of B. nigra. The results disclosed two strongly supported clades, one containing four accessions of vegetable mustard which have closer relationship with B-genome species “B.nigra” lineage and the other containing 12 accessions of B. juncea and three A-genome accessions. This classification was in disagreement with the evidence from chloroplast DNA, mitochondrial DNA, nuclear DNA restriction fragment length polymorphism (RFLP), which suggested that B. juncea was closely related to the A-genome type. For the incongruence, we speculated that the B. juncea crops derived from Chinese have evolved through different recombined events of the diploid morphutypes and evolved unidirectional concerted evolution. The traditional phenotypic classification of B. juncea was not wholly supported by ITS results, and hence the phylogenetic relationships among these subspecies need to be reconsidered on molecular level.     

Diversity among landraces of Indian snapmelon (<Emphasis Type="Italic">Cucumis melo</Emphasis> var. <Emphasis Type="Italic">momordica</Emphasis>)

Diversity among 36 snapmelon landraces, collected from 2 agro-ecological regions of India (9 agro-climatic sub-regions), was assayed using RAPD primers, morphological traits of plant habit and fruit, 2 yield-associated traits, pest and disease resistance and biochemical composition (TSS, ascorbic acid, titrable acidity). Typical differences among accessions were observed in plant and fruit characteristics and snapmelon germplasm with high titrable acidity and possessing resistance to downy mildew, Cucumber mosaic virus, Zucchini yellow mosaic virus, Papaya ringspot virus, Aphis gossypii and Meloidogyne incognita was noticed in the collection. RAPD based grouping analysis revealed that Indian snapmelon was rich in genetic variation and region and sub-region approach should be followed across India for acquisition of additional melon landraces. Accessions of var. agrestis and var. momordica clustered together and there was a separate cluster of the accessions of var. reticulatus. Comparative analysis of the genetic variability among Indian snapmelons and an array of previously characterized reference accessions of melon from Spain, Israel, Korea, Japan, Maldives, Iraq, Pakistan and India using SSRs showed that Indian snapmelon germplasm contained a high degree of unique genetic variability which was needed to be preserved to broaden the genetic base of melon germplasm available with the scientific community. N. P. S. Dhillon and Ranjana contributed equally to this work and are considered the first authors.     

Characterization of intergeneric hybrids of <Emphasis Type="Italic">Erianthus rockii</Emphasis> and <Emphasis Type="Italic">Saccharum</Emphasis> using molecular markers

Erianthus rockii, a wild relative of sugarcane, is drought and cold tolerant, and both are potentially important agronomic traits to the sugarcane industry worldwide. As such it is of interest as a source of parental germplasm to sugarcane breeders and is currently being used in sugarcane introgression programs in both China and Australia. To date morphological characters have been used to verify the putative hybrids produced. Two crosses were generated between two different Saccharum species and two E. rockii accessions. Over 400 AFLP markers were used to identify the intergeneric hybrid progeny as well as determine hybrid diversity. Both crosses generated hybrids but efficiency levels were very different and are probably related to the different Saccharum parent used in each cross. Cross 1 was between a Saccharum officinarum and E. rockii and generated 100% hybrid progeny. Cross 2, however, was between a sugarcane hybrid (S. officinarum × Saccharum spontaneum) and E. rockii and only 10% of the progeny were intergeneric hybrids. Inheritance of markers in the progeny was analysed and for both crosses there were equal numbers of markers from both parents indicating n + n transmission of chromosomes. This is the first verification of E. rockii hybrids with molecular markers. It may now be possible to exploit genes of value from E. rockii in sugarcane breeding programs.     

Isozyme Variation in <Emphasis Type="Italic">Passiflora</Emphasis> Subgenus <Emphasis Type="Italic">Tacsonia</Emphasis>: Geographic and Interspecific Differentiation Among the Three Most Common Species

In a general study of banana passion fruit genetic resources, diversity was analyzed in the two main cultigens, P. tripartita var. mollissima and P. tarminiana, and their closest wild relative, P. mixta, scoring isozyme bands (IDH,PGM,ACP,PGD,DIA,andPRX) on288 plants from 31 accessions. Polymorphismandallelic richness, Nei diversity indices, and neighbor joining clustering showed that variation was poor in the cultigens in northern and central Colombia, while P. mixta appeared much more polymorphic. The populations of P. tripartita var. mollissima and P. mixta from southern Colombia and Ecuador show higher diversity values and are clearly differentiated from those of central and northern Colombia. This geographic component of variation is even stronger than the interspecific one, which suggests a close relation and a regular gene flow between these two species. In contrast, all the accessions of P. tarminiana constitute a clearly differentiated group, even if some introgression with P. tripartita var. mollissima is also suspected. The high variation observed in the southern region indicates the proximity of a center of diversity for banana passion fruit and collecting in Ecuador, Peru and Bolivia is recommended. The high diversity of P. mixta and the evidence of gene flow with P. tripartita var. mollissima constitute a favorable context for the implementation of in situ conservation strategies.     

Production of Intergeneric Hybrids Between <Emphasis Type="Italic">Sinapis alba</Emphasis> and <Emphasis Type="Italic">Brassica carinata</Emphasis>

Reciprocal crosses were made between Brassica carinata and its related species Sinapis alba. Pollen germination studies indicated the presence post-fertilization barriers in both ways. Sequential ovary–ovule culture helped to realize the intergeneric hybrids from the cross S. alba × B. carinata. The F₁ obtained was confirmed as a hybrid based on morphology, cytology and isozyme studies. The hybrid was backcrossed to its male parent and obtained BC₁ seeds, which were used to raise BC₁ generation. The BC₁ generation plants were further backcrossed to B. carinata in order to develop alloplasmic lines.     

Resistance reaction to powdery mildew (<Emphasis Type="Italic">Erysiphe pisi</Emphasis>) in a germplasm collection of <Emphasis Type="Italic">Lathyrus cicera</Emphasis> from Iberian origin

Several Lathyrus species have a considerable potential as alternative pulses in sustainable dryland farming systems mainly due to their high tolerance to drought and disease resistance. Powdery mildew is a serious disease affecting several Lathyrus species. Little is known on the availability of resistance and the underlying resistance mechanisms against powdery mildew in the Lathyrus genus. The present study assessed and characterized the resistance reaction to powdery mildew, Erysiphe pisi, in a collection of Iberian Lathyrus cicera accessions. In general, a compatible reaction with no macroscopically visible necrosis was observed but accessions with reduced disease severity despite of a high infection type have also been identified. This Partial Resistance was in some accessions only expressed in the adult plant stage. The controlling genes of the Lathyrus resistance mechanisms can be of great interest not only for the Lathyrus improvement per se but also for related legume species, like field pea.     

Agromorphological and molecular variability in the genus <Emphasis Type="Italic">Sesbania</Emphasis>

Green manuring especially in rice wheat cropping systems of many tropical and subtropical countries is regaining importance due to increased fertilizer costs and environmental considerations. Many species of Sesbania genus have been reported as potential green manure crops. The genus is not well characterized and there is a lot of ambiguity over the nomenclature and classification of the species of this genus. A pilot study based on agro-morphological and molecular data was conducted to examine the interspecific and intraspecific variability in the selected species of Sesbania genus that were collected from diverse geographical locations and are recommended or traditionally used as green manures. The study revealed a lot interspecific variability at both morphological and molecular level. It was also shown that species-specific genetic markers could be generated. The potential use of such markers in Sesbania breeding program is discussed. The study also emphasizes the utilization of molecular profiling for proper identification and classification of the species of this genus. In addition, various limitations in breeding and utilization of Sesbania as a green manure crop and the potential of a radiation induced late flowering mutant (Trombay Sesbania rostrata-1; TSR-1) for crop improvement is also discussed.     

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.     

Analysis of Genetic Variation Among Accessions of <Emphasis Type="Italic">Pueraria montana</Emphasis> (Lour.) Merr. var. <Emphasis Type="Italic">lobata</Emphasis> and <Emphasis Type="Italic">Pueraria phaseoloides</Emphasis> (Roxb.) Benth. based on RAPD Markers

Pueraria montana var. lobata and P. phaseoloides originating from tropical Asia and parts of Oceania are ecologically and economically important legumes that are used as green manure, cover crop or forage plants. Conservation and use of plant genetic resources require an understanding of the extent and distribution of genetic diversity in any given region. In this study, genetic variation of five P. montana var. lobata and 16 P. phaseoloides accessions was analysed developing a random amplified polymorphic DNA (RAPD) marker methodology for Pueraria species and thereby creating basic data for follow-up research and the development of conservation strategies. Seeds were collected from native populations in Bac Kan Province, a mountainous region in Northeast Vietnam. P. montana var. lobata presented a high level of variation with 54.3% of the detected markers being polymorphic, whereas P. phaseoloides exhibited an intermediate to high level of variation (45.5%). The P. montana var. lobata accessions clustered in congruence with their eco-geographical origin. For P. phaseoloides no correspondence between sampling sites and genetic differentiation was found. Inter-population differentiation was measured as Jaccard's similarity coefficient (JSC). Mean JSC amounted to 0.35 in P. montana var. lobata and 0.52 in P. phaseoloides. Results are compared to other genetic studies of herbaceous legumes and conservation strategies are suggested.     

AFLP similarities among historic Danish cultivars of fodder beet (<Emphasis Type="Italic">Beta vulgaris</Emphasis> L. subsp. <Emphasis Type="Italic">vulgaris</Emphasis>var. <Emphasis Type="Italic">rapacea</Emphasis> Koch)

Dead seeds of a fodder beet cultivar ‘Elvetham’ stored under ambient conditions since 1880 were compared to a homonymous sample preserved in an on-farm situation in Denmark. DNA was isolated from single seeds and successfully applied to Amplified Fragment Length Polymorphism (AFLP) analysis of the accessions. Six primer pairs were used to determine the similarity between the two accessions based on 112 polymorphic bands. Furthermore, similarity among seven cultivars of fodder beets representing the main types used in Scandinavia at the end of the 19th century was determined. This analysis was based on 152 polymorphic bands. Differentiation among the seven cultivars was determined to a mean G _ST value of 0.438, while G _ST between the two ‘Elvetham’ accessions was 0.266. A principal coordinate analysis based on jaccards similarity index illustrates that the two ‘Elvetham’ accessions are different from each other. The differentiation is higher than the value found between two separate ‘Eckerndorfer’ accessions. The results indicate that the cultivated accession has changed. Additionally, the value of applying old dead seed material for documentation in gene banks is demonstrated. During the analysis it was found that DNA isolated from seeds and leaves behaved differently in the AFLP process, however, the two fractions assigned to their common accession.     

Seed inoculation with <Emphasis Type="Italic">Bacillus</Emphasis> spp. improves seedling vigour in oil-seed plant <Emphasis Type="Italic">Jatropha curcas</Emphasis> L.

Jatropha (Jatropha curcas L.) is a non-edible oil-seed plant with adaptability to marginal semi-arid lands and wastelands. The Indian Government is promoting jatropha to reduce dependence on the crude oil and to achieve energy independence by the year 2012, under the National Biodiesel Mission. Selected strains of Bacillus spp., either supplemented with or without chitin, were tested for their ability to promote growth of jatropha seedlings in pot culture studies. The strains supported growth of jatropha seedlings up to 42 days after sowing. Among all strains, Bacillus pumilus (IM-3) supplemented with chitin showed over all plant growth promotion effect resulting in enhanced shoot length (113%), dry shoot mass (360%), dry root mass (467%), dry total plant mass (346%), leaf area (256%), and chlorophyll content (74%) over control. Treating seeds with strain IM-3 without chitin resulted in enhanced dry shoot mass (473%), dry total plant mass (407%), and chlorophyll content (82%). However, Bacillus polymyxa (KRU-22) with chitin supported maximum root length (143%). Either strain IM-3 alone or in combination with other promising strains could be promoted further for enhanced initial seedling growth of jatropha.     

Characterization of Portuguese populations of <Emphasis Type="Italic">Vitis vinifera</Emphasis> L. ssp. <Emphasis Type="Italic">sylvestris</Emphasis> (Gmelin) Hegi

Wild populations of Vitis vinifera L.␣have been located in Portugal. Morphological characterization was carried out in three populations located in Alcácer do Sal, Castelo Branco, and Montemor-o-Novo, and then compared using multivariate discriminant analysis. These populations were from three different hydrological basins, therefore cross-pollination was not possible. It was verified that in each population all plants were different. The data suggest that the frequency of female and male plants is rather variable in wild populations. The morphology of the adult leaf, from the Alcácer do Sal population, had particular features when compared with Castelo Branco and Montemor-o-Novo populations, which were more homogeneous. The length of teeth compared with width at the end of the base, and the density of prostrate hairs between and on main veins (lower side) were the variables which allowed the best discrimination among populations.     

Production and analysis of reciprocal hybrids between <Emphasis Type="Italic">Asparagus officinalis</Emphasis> L. and <Emphasis Type="Italic">A. schoberioides</Emphasis> Kunth

The genus Asparagus (Asparagaceae) encompasses 100–300 species, including the important vegetable crop, A. officinalis (garden asparagus). Previous attempts to hybridize garden asparagus with A. densiflorus (Kunth) Jessop, with the aim of introducing disease resistance, were unsuccessful because of the failure of endosperm development. In the present study, reciprocal interspecific hybrids between garden asparagus and A. schoberioides Kunth were generated by hand pollination. The F1 hybrids were analyzed by using both morphological and molecular techniques. This is the first report describing the production of an interspecific hybrid between garden asparagus at the diploid level (2n = 2x = 20) and its diploid (2n = 2x = 20) wild relative, A. schoberioides. Morphological characteristics of candidate hybrids were a mixture of those found in the parents, and cytological and RFLP analyses confirmed that morphologically intermediate plants were indeed diploid hybrids of those two species. In other words, post-zygotic isolation is not complete between phylogenetically distinct these two species. Our results suggest that other Asparagus species may be capable of hybridizing with A. officinalis and that introducing wild characters through interspecific hybridization may offer advantages for breeding for novel traits. Takuro Ito and Toshinori Ochiai authors are contributed equally to this work     

A new gross morphological variation in the genus <Emphasis Type="Italic">Triticum</Emphasis> L.

The new character state “hairy neck” was found in Triticum dicoccoides from Lebanon differing morphologically from that which can be occasionally found as an introgressed character from rye into wheat. The material is newly described as botanical variety: T. dicoccoides var. johnsonii     

Dispersal of durum wheat [<Emphasis Type="Italic">Triticum turgidum</Emphasis> L. ssp. <Emphasis Type="Italic">turgidum</Emphasis> convar. <Emphasis Type="Italic">durum</Emphasis> (Desf.) MacKey] landraces across the Mediterranean basin assessed by AFLPs and microsatellites

A comprehensive characterization of crop germplasm is critical to the optimal improvement of the quality and productivity of crops. Genetic relationships and variability were evaluated among 63 durum wheat landraces from the Mediterranean basin using amplified fragment length polymorphisms (AFLPs) and microsatellites markers. The genetic diversity indices found were comparable to those of other crop species, with average polymorphism information content (PIC) values of 0.24 and 0.70 for AFLP and microsatellites, respectively. The mean number of alleles observed for the microsatellites loci was 9.15. Non-metric multi-dimensional scaling clustered the accessions according to their geographical origin with the landraces from the South shore of the Mediterranean Sea closely related. The results support two dispersal patterns of durum wheat in the Mediterranean basin, one through its north side and a second one through its south side.