High Molecular Weight (HMW) Glutenin Subunit Composition of Some Nordic and Middle European Wheats

A collection of 123 winter and 106 spring wheat (Triticum aestivum L.) cultivars and breeding lines commonly grown in Nordic and Middle European countries were characterised for the composition of high-molecular-weight (HMW) glutenin subunits on the ground of data from literature and experiments of author. HMW glutenin subunit composition was determined by one-dimensional sodium dodecyl sulphate- polyacrylamide gel electrophoresis (SDS-PAGE). The present database includes data for cultivars and breeding lines from Finland (FIN – 56), Estonia (EST – 27), Sweden (SW – 27), Germany (D – 24), Lithuania (LIT – 17), United Kingdom (UK – 6), France (F – 3), Poland (POL – 5), Italy (IT – 7), Netherlands (NL – 6), Norway (NOR – 12), Russian Federation (RUS – 14 items). The occurrence of individual alleles and corresponding HMW glutenin subunits in surveyed cultivars is demonstrated. Special attention has been paid to cultivars growing in neighbouring countries with the aim to apply them in Estonian plant breeding. The database consists of 4 tables with data of HMW glutenin subunits, encoding them Glu-1 alleles, quality score and sources of data. The database could be useful for varietal identification and for plant breeders to improve wheat quality and accelerate the breeding process.     

RAPD and AFLP assessment of genetic variation in a landrace of pepper (Capsicum annuum L.), grown in North-West Italy

In several regions of Italy as well as other parts of southern Europe, the heterogeneity of the land, the climate and the soil favour the survival in cultivation of a large number of landraces specifically adapted to local conditions. Knowledge on the level and distribution of their genetic variation can help to develop appropriate strategies, in order to suistainably manage in situ these germplasm resources at risk of genetic erosion. C. annuum is an herbaceous diploid species and is considered to be self-pollinating, although different rates of out-crossing have been recorded. We used random amplified polymorphic DNA (RAPD) and amplified fragment length polymorphism (AFLP) markers to assess genetic diversity within and between five populations of a landrace of Capsicum annuum L., grown in a limited area in north-west Italy and locally known as Cuneo pepper. Partitioning the genetic variation with Shannon's diversity index revealed that 41.6% occurred between and 58.4% within populations. Analogous results were obtained when the analysis was based only on RAPD or AFLP markers. However, AFLP was more reliable, since a lower range of variation was observed among primer combinations in detecting the two components of genetic variation. Notwithstanding the rather high level of within genetic variation detected, the five populations were clearly differentiated and differed in the frequency of alleles exclusive and/or present at very low frequencies. Our results show the need for accurate estimation of allele frequencies, in order to identify populations to which priority should be given for dynamic conservation of landraces.     

Genetic diversity of winter wheat in Shaanxi province, China, and other common wheat germplasm pools

Cultivated Chinese wheat germplasm has been a valuable genetic resourcein international plant breeding. Landraces endemic to China are a geneticresource that is distinct from other wheat germplasm. Patterns of geneticdiversity among cultivated Chinese accessions and relationship to othergermplasm pools are unknown, despite the proven value and potential novelty. Theobjective of this work was to determine the level of genetic diversity withinimproved Chinese germplasm in the context of several other wheat germplasmpools. We analyzed a set of improved accessions cultivated from the 1940s to the1990s in Shaanxi province, China, using six simple sequence repeat (SSR) primerpairs and 30 restriction fragment length polymorphism - probe enzymecombinations (RFLP-PEC) previously used to characterize 21 geographically basedgermplasm pools. Shaanxi germplasm consists of three groups based on foreignintroductions from Italy, Australia, Denmark, and Russia. There was a decreasein genetic diversity among Shaanxi accessions cultivated in the 1970s and 1980sto the 1990s, and accession classifications based on primary decade ofcultivation were found to be significantly undifferentiated. The analysis of themean genetic distance among 22 geographically based pools of germplasm suggestsseveral regions are significantly undifferentiated. A vast majority of the totalamount of variation was found within pools; therefore, pools appear to belargely differentiated based on small differences in band relative frequency andfew if any unique bands. Previous studies have identified some Chinese landracepools as morphologically and genetically unique. The Shaanxi pool does not havethe same unique morphological or genetic features, nor is it more similar to thelandrace pools than other improved germplasm pools.     

Genetic Diversity of Glutenin Protein Subunits Composition in Durum Wheat Landraces [Triticum turgidum ssp. turgidum Convar. durum (Desf.) MacKey] from the Mediterranean Basin

The genetic diversity of high and low molecular weight glutenin subunits of 63 durum wheat landraces from different geographical regions in the Mediterranean Basin was studied using SDS-PAGE. Great variability in glutenin composition was found, with 42 high and low molecular weight glutenin haplotypes, 20 allele combinations at the HMW-GS loci, and 18 at the LMW-GS. All five possible LMW models were detected in all Mediterranean regions. Rare alleles were found at Glu-B1 locus in high frequencies and a priori related alleles to grain quality were also observed. Global genetic diversity index was relatively high (0.67); it ranged from 0.33 to 0.66. Cluster analysis on the frequency patterns of origins grouped genotypes following a geographical structure. Rogers’ distance coefficient on frequency pattern for each region of origin showed two germplasm pools with distinct quality profiles, where South West Asian landraces were very different from the landraces of other Mediterranean areas. The relationship between different regions of origin is discussed and two possible ways of introduction of wheat in the Iberian Peninsula (N Africa and SE Europe) are hypothesized. The use of Mediterranean durum wheat landraces as source of genetic variability for grain quality improvement is highly recommended.     

Allelic diversity of HMW and LMW glutenin subunits and omega-gliadins in French bread wheat (Triticum aestivum L.)

Wheat endosperm storage proteins, namely gliadins and glutenins, are the major components of gluten. They play an important role in dough properties and in bread making quality in various wheat varieties. In the present study, the different alleles encoded at the 6 glutenin loci and at 3 -gliadin loci were identified from a set of 200 hexaploid wheat cultivars grown primarily in France using SDS PAGE. At Glu-A1, Glu-B1 and Glu-D1, encoding high molecular weight glutenin subunits (HMW-GS), 3, 8 and 5 alleles were observed respectively. Low molecular weight glutenin subunits (LMW-GS) displayed similar polymorphism, as 5 and 11 alleles were identified at loci Glu-A3 and Glu-B3 respectively. Four alleles were observed at Glu-D3 loci. Omega-gliadin diversity was also very high, as 7, 13 and 9 alleles were found at Gli-A1, Gli-B1 and Gli-D1, respectively. A total of 147 (or 149) patterns resulted from the genetic combination of the alleles encoding at the six glutenin loci (or Glu-1 and Gli-1 loci). Although Glu-1 and Glu-3 loci were located on different chromosome arms and were theoretically independent, some associations were revealed due to pedigree relatedness between some French wheat cultivars. The usefulness of allelic identification of LMW-GS together with HMW-GS and gliadins for future genetic and technological wheat improvement is discussed.     

Polymorphism of high molecular weight glutenin subunits in three species of Aegilops

A collection of 136 accessions of Aegilops umbellulata (39), Ae. comosa (75) and Ae. markgrafii (22) was analysed for high-molecular-weight (HMW) glutenin subunits composition. The homogeneity of the accessions was studied and 55.1% of the collection was homogeneous for HMW glutenin subunits (29 Ae. umbellulata, 33 Ae. comosa and 14 Ae. markgrafii). The HMW glutenin subunits of Ae. umbellulata are encoded by the Glu-U1 locus; in Ae. comosa results showed that this proteins are encoded at the 1M chromosome, and the locus was named Glu-M1. In Ae. markgrafii it was assumed that HMW glutenin subunits were encoded by an homoeologous locus and it was named Glu-C1. All the accessions of Ae. umbellulata and Ae. markgrafii expressed both, x-type and y-type subunits. Among the Ae. comosa accessions, only one expressed an x-type subunit alone. All the accessions of Ae. umbellulata and some of Ae. comosa had x-type glutenins of higher molecular weights than those commonly present in bread wheat. A total of 8 alleles were detected at the Glu-U1 locus, 11 at the Glu-M1 and 4 at the Glu-C1. The new HMW glutenin variation found in this work suggests their possible utilisation in breeding for wheat quality.     

Effect of farmers' seed selection on genetic variation of a landrace population of pepper (Capsicum annuum L.), grown in North-West Italy

Recognition by the European Community of élite landraces encourages farmers to grow these and earn more than is possible by growing modern varieties. However, farmers often exercise strong selection by collecting seed from a limited number of plants which best embody a few characters of prime interest. This, in the long run, may be responsible for considerable genetic erosion. In Southern Piedmont, North-West Italy, a local landrace of pepper (Capsicum annuum L.) known as Cuneo is grown; it deserves particular attention because of its hardiness, late production and fruit quality. We used random amplified polymorphic DNA (RAPD) and amplified fragment length polymorphism (AFLP) markers to assess the effect, on genetic composition of a population, of seed sampling carried out using the selection criteria adopted by the farmer. After two reproductive cycles using selection, it was already possible to detect loss in genetic variation and a change in allele frequencies, while no significant effect was found after two cycles of random sampling. Over this period, farmer selection pressure led to disappearance of eight low-frequency alleles, while only three alleles were lost in randomly sampled populations. Our results stress the need to adopt appropriate techniques for seed sampling in order to avoid genetic erosion of local landrace gene pools.     

Genetic variation of the durum wheat landrace Haurani from different agro-ecological regions

The durum wheat landrace Haurani (Triticum durum Desf.) is grown under contrasting climatic regions of Syria from Deir Ezzor, in the North-East (230 m altitude, 150 mm mean annual rainfall), to Qunaytra, in the South-West (1060 m altitude, 825 mm mean annual rainfall). In order to assess the genetic variation between and within Haurani populations, samples from eight provinces of Syria (Daraa, Damascus, Qunaytra, Deir Ezzor, Hassakeh, Aleppo, Homs and Hama) were analysed by RFLPs and seed storage proteins of glutenin subunits as markers. The analyses showed the presence of genetic polymorphisms in all populations with the highest values in those from Homs and Hassakeh. Moreover, the results point out differences in genetic distances between populations; some populations were further apart, such as Damascus and Aleppo, whereas others were closer to each other, for instance Homs and Hama. Cluster analysis identified two distinct groups of populations, characterized by geographical proximity, with similar rainfall and altitude. It is suggested that the similarity of landraces at locations close to each other might be the result of more frequent seed exchanges between farmers or of gene flow due to 5% estimated outcross rate of Haurani.     

Patterns of diversity in Cannabis

Summary A Cannabis germplasm collection was evaluated for traits relevant to the use of hemp as an arable source of paper pulp. The traits were related to stem yield and quality, psychoactive potency, resistance to root-knot nematodes and plant morphology. The present study is directed at diversity for these traits considered together. For the interpretation of diversity patterns, accessions were grouped on the basis of the presumed purpose and status of domestication resulting in the recognition of four plant-use groups: fibre cultivars, fibre landraces, drug strains, and truly wild or naturalized populations. Principal component analysis showed that stem quality, phenological development, psychoactive potency and host reactions to root-knot nematodes explained most of the total variation among accessions. The plant-use groups were reasonably well separated in the principal component space. Discriminant analysis showed that the a priori defined plant-use groups could be discriminated quite well on the basis of linear combinations of all agronomic and morphological traits. The contents of bark fibre and cannabinoids had the highest grouping value. Reallocations concerned mainly old fibre cultivars which were placed in the group of fibre landraces due to a low content of bark fibre.A genetic characterization of accessions was based on electrophoretic patterns of seed proteins. Isoelectric ocusing patterns of bulk seed extracts showed variation for the presence of six out of 45 protein bands in the H range from 4.75 to 6.40. Banding patterns were however independent from any grouping based on origin or gronomic and morphological traits and did not reflect expected common ancestry of accessions.     

Collection of Lupinus angustifolius L. Germplasm and Characterisation of Morphological and Molecular Diversity

Lupinus angustifolius L. is a Mediterranean species, domesticated in the 20th century, representing an important grain legume crop in Australia and other countries. This work is focused on the collection of wild germplasm and on the characterisation of morphological and molecular diversity of germplasm accessions. It reports the collection of 81 wild L. angustifolius accessions from the South and Centre of Portugal, available at the ‘Instituto Superior de Agronomia Gene Bank’, with subsequent morphological and molecular characterisation of a selection of these and other accessions. A multivariate analysis of morphological traits on 88 L. angustifolius accessions (including 59 wild Portuguese accessions, 15 cultivars and 14 breeding lines) showed a cline of variation on wild germplasm, with plants from Southern Portugal characterised by earlier flowering, higher vegetative development and larger seeds. AFLP and ISSR molecular markers grouped modern cultivars as sub-clusters within the wider diversity of wild germplasm, revealing the narrow pool of genetic diversity on which domesticated accessions are based. The importance of preserving, characterising and using wild genetic resources for L. angustifolius crop improvement is outlined by the results obtained.     

Sources of resistance to downy mildew in narbon (Vicia narbonensis) and common (Vicia sativa) vetches

Downy mildew (Peronospora viciae (Berk) de Bary) is an important disease of vetches (Vicia spp.) in the Mediterranean Region. Narbon and common vetch germplasm accessions originating from different countries, and advanced breeding narbon vetch genotypes, were evaluated for reaction to downy mildew for two seasons under field conditions. In narbon vetch, there were resistant sources from both germplasm accessions and advanced genotypes. From the germplasm accessions, 25 accessions were resistant while from the advanced breeding genotypes, 10 had resistance, with 2–3 ratings. In common vetch, many accessions were highly resistant to downy mildew. These were widely distributed in different countries, mainly Turkey, Italy, Syria and Iran. In general, common vetch had higher sources of resistance than the narbon vetch accessions. These new sources of resistance to downy mildew will be incorporated into the forage legume improvement project in a continuous effort to identify cultivars suitable for replacing fallow in the cereal-based cropping systems.     

Characterization of core collection of lentil germplasm for phenology, morphology, seed and straw yields

Increasing and maintaining crop residues in predominantly cereal-based rotations of the US Pacific Northwest is critical to controlling soil erosion. The core collection of lentil (Lens culinaris Medik.) germplasm comprising 287 accessions was evaluated for variation in phenological, morphological and growth parameters including seed yields and residue amounts over a two-year period under conventional tillage and no-till conditions. The objectives of this study were (i) assess lentil genetic variation in germplasm for variation in biomass production and seed yield, (ii) assess the relationship of phenological and morphological traits with biomass and seed yield and (iii) identify high biomass producing germplasm for use as parents in the breeding program. Days to flowering and days to maturity ranged from 31 to 78 and from 71 to 106 days, respectively. Time to flowering in terms of cumulative heat units was a more efficient measurement than days to flowering. Plant height and plant canopy width had a significant association with total biomass, seed yield and residue amounts. Total biomass ranged from 788–6389 kg ha^–1 under conventional tillage, while the range under no-till conditions was 1045–6195 kg ha^–1. Most of the lines with higher biomass also produced the highest seed yields and residue amounts. Overall, only one accession produced more residue than `Laird'. In the more favorable environment of 1997, six accessions exceeded the control cultivars, `Laird' and `Indianhead', for residue amounts, and seven and twenty-four accessions exceeded control cultivars, `Pardina' and `Brewer', for seed yield. Results indicated that plant height, canopy width at maturity and seed yield explained most of the variation in biomass and residue production. Large seeded germplasm consistently had a longer reproductive growth period than small seeded accessions and had 17%, 7% and 21% more biomass, seed yield and residue, respectively. Our data indicated significant variation in lentil germplasm for biomass, seed yield and residue amounts to warrant their use in the breeding program.     

Multivariate analysis of phenotypic diversity of Coffea arabica

Summary Eighteen different morphological and agronomical traits observed in a field collection of 148 accessions of Coffea arabica were analyzed using multivariate analysis. For the first time, a clear structure is observed within this species with the identification of two main groups. Accessions of group 1 have a more erect branching habit, narrower leaves and are on average more resistant to coffee leaf rust and coffee berry disease than accessions of group 2. Group 1 mostly contains Ethiopian accessions collected west of the Great Rift Valley, whereas group 2 mostly contain commonly cultivated varieties throughout the world and Ethiopian accessions collected east of the Great Rift Valley in Ethiopia. This phenotypic structure together with historical evidence suggest that group 1 has not been involved in the domestication of C. arabica. The present results confirm that sub-spontaneous genotypes west of the Rift Valley are highly valuable for enriching the genetic basis of cultivated C. arabica germplasm. The potential value of this material in breeding of C. arabica is discussed.     

Genetic diversity of cowpea [<Emphasis Type="Italic">Vigna unguiculata</Emphasis> (L.) Walp.] in four West African and USA breeding programs as determined by AFLP analysis

Cowpea is an important grain legume and hay crop of many tropical and subtropical regions, especially in the dry savanna region of West Africa. The cowpea gene pool may be narrow because of a genetic bottleneck during domestication. Genetic variation within specific breeding programs may be further restricted due to breeding methods, ‘founder effects’ and limited exchange of germplasm between breeding programs. Genetic relationships among 60 advanced breeding lines from six breeding programs in West Africa and USA, and 27 landrace accessions from Africa, Asia, and South America were examined using amplified fragment length polymorphism (AFLP) markers with six near infrared fluorescence labeled EcoRI + 3/1bases/MseI + 3/1bases primer sets. A total of 382 bands were scored among the accessions with 207 polymorphic bands (54.2%). Despite a diverse origin, the 87 cowpea accessions shared a minimum 86% genetic similarity. Principal coordinates analysis showed clustering of breeding lines by program origin, indicating lack of genetic diversity compared to potential diversity. Accessions from Asia and the Americas overlapped and were distinct from West African breeding lines, indicating that germplasm from Asia and the Americas have common origins outside West Africa. US and Asian breeding programs could increase genetic variability in their programs substantially by incorporating germplasm from West Africa, while national programs in West Africa should consider introgression of Asian germplasm and germplasm from other parts of Africa into their programs to ensure long-term gains from selection.     

Univariate and multivariate analysis performed on bio-agronomical traits of Cucumis melo L. germplasm

A large collection of melon (Cucumis melo L.) germplasm has been established at the Section of Genetics and Plant Breeding, Bari University (Italy). In the present work, data regarding the variation of 20 bio-agronomical traits recorded on entries and landraces collected in Albania and Apulia region (Southern Italy) are reported. The main objective of the study was to assess and describe, by means of univariate and multivariate analyses, the genetic diversity in the collection composed by genotypes classified in the Inodorus and Cantalupensis groups. The results obtained showed a large variation for all the traits examined. Furthermore, it was possible to identify valuable genotypes for future breeding programmes aimed at improving melon traits, particularly for the Inodorus group, which is an important crop in many Southern Italian sites. Genotypes of interest were especially selected for earliness and lateness, fruit shape, soluble solids content, storage time and fruit firmness.     

RAPD and seed coat morphology variation in annual and perennial species of the genus Cicer L.

Molecular and morphological variation of six perennial and five annual species including domesticated chickpea, C. arietinum, were inferred on the basis of RAPD and S.E.M. seed coat features using three outgroup taxa (Lens ervoides, Lathyrus japonica and Pisum sativum). Of the 66 decamer arbitrary primers tested, eight primers revealed 87 informative fragments. Neighbor-joining cluster analysis using Jaccard's coefficient of similarity on the basis of polymorphic fragments indicated a narrow variation in C. arietinum and recognized two major clusters in the genus Cicer. The first one included four species of sect. Monocicer: C. echinospermum, C. arietinum, C. reticulatum and Iranian material of C. bijugum. The second cluster contained annual and perennial species belonging to sections Chamaecicer, Polycicer and Acanthocicer. The character state of morphological and ecological traits on the RAPD phenogram indicated no monophyletic incision. Our results suggested that the high genetic difference between annual and perennial species might be regarded as a rapid speciation of section Monocicer. Reconsideration of traditional classification of sections Polycicer and Acanthocicer is necessary. The Desi and Kabuli types of C. arietinum could not be separately grouped at the DNA level, and the low level of genetic variation of C. arietinum may result from a bottleneck during the domestication process.     

Temporal variation of diversity in Italian durum wheat germplasm

The aim of this work is to analyse the temporal change of genetic diversity in Italian durum wheat germplasm. The germplasm deployed in this study (158 accessions), belonging to 5 different historical classes, was characterised for its microsatellite and gliadin markers. The level of genetic diversity (He), based on gliadin and SSR markers results – on average – greater in indigenous landraces present in Italy before 1915, with the exception of pure line material which had been selected from landraces (showing highest level of heterozigosity for gliadin markers). Genotypes obtained from crosses or mutagenesis (referring to the 1950–1960 period) along with those resulting from crosses between CIMMYT lines and old materials (1970s and beyond) were also genetically more diverse. Forty-nine percent of indigenous landraces were genetically heterogeneous. Nine out of 53 landrace accessions were able to capture 4 different SSR private alleles. It is speculated that the reduction of allele richness is an indicator of the genetic erosion of the pre-breeding germplasm and it is pointed out that the implementation of appropriate methods of genetic conservation of this germplasm is a priority for breeding and food safety.     

Physico-chemical characterisation of yam (Dioscorea alata L.) tubers from Vanuatu

The objectives of this study were: (1) to analyse the physico-chemical characteristics of 48 Dioscorea alata varieties representing a core sample of the Vanuatu national germplasm collection; (2) to relate those characteristics with the varieties eating quality; and (3) to assess the possibility of selecting varieties according to their chemotype. Overall, 331 accessions were collected from 15 different islands of Vanuatu, planted in an ex situ germplasm collection and described during 3 years. The 48 varieties included in the core sample were selected according to their island of origin, eating quality, tuber shape, tuber flesh colour and morphotype. Analyses of their tubers were made for percentage dry matter, starch, amylose, lipids, minerals, proteins, sugars and gelatinisation temperature range. Significant variation exists for each of these characteristics except for gelatinisation temperature. Varieties with good eating quality are characterised with high dry matter, starch and amylose contents. Chemotypes appear to be genetically controlled and further screening of germplasm and/or breeding will have to take into consideration these characteristics, important for farmers’ adoption.