Genetic diversity in Borago officinalis germplasm as revealed by seed oils and AFLP polymorphism

Ten accessions of wild Borago officinalis were collected during missions in the course of a research activity carried out on plants producing nutraceutical compounds. The aim was to identify populations that produce a higher content of γ-linolenic acid, as this compound has important therapeutic properties and has been used in medical tests to treat rheumatoid arthritis, eczema, and psoriasis with good therapeutical results. The present study reports on the characterization of both the polyunsaturated fatty acid composition and on genetic diversity estimated by means of AFLP markers in several B. officinalis accessions collected in Southern Italy and grown in isolation by means of screen houses specifically designed for each single population. Polyunsaturated fatty acid were found in different amounts in all investigated accessions. The populations were very variable at the molecular level, and the analysis of relationships based on AFLP data, showed that similarity based on Jaccard’s index ranged between 0.330 and 0.742. Our investigations allowed us to identify a population that may be considered as a good source of GLA.     

Digestive enzyme inhibitors and storage pest resistance in cowpea (Vigna unguiculata) seeds

Summary Amylase and trypsin inhibitors are proteins which inhibit digestive enzymes. The loss of the activity of these enzymes produces a reduction of starch and protein digestion. Amylase and trypsin inhibitor activity have been investigated in seeds of 20 cowpea lines in trying to establish their role in the storage pest resistance. A broad variation has been observed for both the inhibitors. Correlation analysis between inhibitor levels and extent of attack has shown that neither amylase nor trypsin inhibitors can separately explain the resistance to Callosobruchus maculatus (F.). Lines bruchid resistant in fact, have high levels of both inhibitors. Conversely lines with low levels of amylase and trypsin inhibitors are bruchid susceptible. The breeding for high contents of both amylase and trypsin inhibitors can be an effective way to obtain lines with resistance to storage pest.     

Assessment of genetic variation in a working collection of Vigna vexillata (L.) A. Rich. by isozyme and RAPD analyses

Genetic variation based on isozyme and RAPD analyses was investigated in 47 and 34 accessions respectively of Vigna vexillata from different geographical origins and belonging to three botanical varieties. A total of 9 enzyme systems were studied, accounting for 14 putative loci, 8 of which were polymorphic. The analysis of genetic diversity revealed a low level of within accession variation (H_S=0.013), while between accession diversity (D_ST) was 0.120. Coefficient of gene differentiation (G_ST) was 0.905, indicating that most variation was among accessions. Nei's genetic distances were calculated on the basis of allelic frequencies and a UPGMA dendrogram was constructed. Twenty arbitrary 10-mer oligonucleotides were used in RAPD analysis. Amplification profiles disclosed a higher level of polymorphism than isozymes. Based on amplification patterns, the similarity index of Jaccard was calculated and a dendrogram constructed on the basis of the similarity matrix. The final clustering based on RAPD data was similar to the one obtained using isozyme allelic frequencies. The classification in botanical varieties did not reflect the allelic constitution of the different samples. On the other hand, referring to geographical origin, most accessions from Africa and from Latin America were distributed respectively in two distinct clusters in the dendrogram. This grouping might also reflect the differences observed in the germination behaviour of V. vexillata from the two continents.     

Assessment of genetic variation in a collection of lentil using molecular tools

A collection of lentil landraces, mostly from Italy, but including Mediterranean and foreign germplasm as reference, was evaluated using RAPDs, microsatellite-primed PCR and ISSRs. A low level of useful polymorphic bands was detected with the former two markers, whereas ISSRs revealed a higher degree of variation. The UPGMA trees, based on Jaccard similarity index matrices, obtained with RAPD and ISSR respectively, did not produce similar clusters. However, in both cases, the accession which differentiated the most from the others was from Ethiopia, a country where also other cultivated species have shown a remarkable degree of variation. On the other hand, Italian accessions showed a trend to group together. ISSR markers proved to be useful for distinguishing closely related genotypes, and possibly for substantiating the genetic peculiarity of ecotypes applying for the obtainment of origin and quality marks. This revised version was published online in July 2006 with corrections to the Cover Date.     

On the origin of artichoke and cardoon from the <Emphasis Type="Italic">Cynara</Emphasis> gene pool as revealed by rDNA sequence variation

The evolutionary history of artichoke and cultivated cardoon and their relationships to wild allies of the genus Cynara are not fully understood yet. To try resolve the evolutionary patterns leading to the domestication of these two crops, a study of molecular evolution was undertaken. The species C. cardunculus, including artichoke, cultivated and wild cardoon, together with four wild Cynara species were taken into consideration. Internal (ITS) and external (ETS) rDNA transcribed spacers were used as markers of nuclear genome, the psbA-trnH spacer as a marker of chloroplast genome. Sequences were analysed using phylogenetic analysis packages. Molecular data indicate that the whole genus is quite recent and that the domestication of artichoke and cultivated cardoon, crops diverging for reproduction system and use, are independent events which diverge in time and space. As for wild Cynara species, an evolutionary pattern consistent with their present geographical distribution was hypothesized in relation to the climatic changes occurring in the Mediterranean during the last 20 millennia: C. humilis and C. cornigera appeared to have differentiated first, C. syriaca and C. baetica were differentiated in a second period, while C. cardunculus showed to be the most recent and plastic species. The high plasticity of C. cardunculus has not only allowed its nowadays wide distribution, but has also given the potential for domestication. Dedicated to the memory of Richard Neville Lester (1937–2006) who greatly contributed to the understanding of the evolution, domestication, and genetic resources of eggplants as well as to the biosystematics and taxonomy of allied species. IGV Publication N. 78 of the Institute of Plant Genetics, CNR     

Characterization of Italian chickpea (Cicer arietinum L.) germplasm by multidisciplinary approach

Characterization of 15 chickpea (Cicer arietinum L.) accessions (6 cultivars, 3 selected lines and 6 landraces) was performed using morphological, agronomic, and technological traits, together with biochemical (seed storage proteins) and molecular markers. Field trials, performed in two different geographical areas of Southern Italy, one in a plain area near Tyrrhenian sea and one in an hilly internal area, revealed a high influence of field location on yield. A wide variation was also registered for technological traits and molecular markers. Microsatellite (SSR) markers grouped chickpea accessions in different clusters, providing useful information on genetic variation and relationships among them, with polymorphic fragments useful to discriminate among all accessions. On the contrary, seed storage protein pattern showed scarce variation resulting very similar among all the accessions considered.     

Asparagine synthetase genes (<Emphasis Type="Italic">AsnS1</Emphasis> and <Emphasis Type="Italic">AsnS2</Emphasis>) in durum wheat: structural analysis and expression under nitrogen stress

Wheat is one of the most widely grown cereal crops based on the amount of calories it provides in the human diet. Durum wheat (Triticum turgidum ssp. durum) is largely used for production of pasta and other products. In order to use genetic knowledge to improve the understanding of N-use efficiency, we carried out, for the first time in durum wheat, the isolation and the characterization of four members of the asparagine synthetase (AsnS) gene family. Phylogenetic inference clustered the Ttu-AsnS1 (1.1 and 1.2) and Ttu-AsnS2 (2.1 and 2.2) genes in AsnS gene class I, which is present in monocots and dicots. Class I genes underwent a subsequent duplication leading to the formation of two subgroups. Plants of Svevo cultivar were grown under N-stress conditions and expression of the four AsnS genes was investigated at three developmental stages (seedling, booting, and late milk development), crucial for N absorption, assimilation and remobilization. AsnS1 genes were down-regulated in N-stressed roots, stems and leaves during seedling growth and booting, but seemed to play a role in N remobilization in flag leaves during grain filling. AsnS2 genes were scarcely expressed in roots, stems, and leaves. In N-stressed spikes there was no differential expression in any of the genes. The genes were mapped in silico using a durum wheat SNP map, assigning Ttu-AsnS1 genes to chromosome 5 and Ttu-AsnS2 to chromosome 3. These findings provide a better understanding of the role of ASN genes in response to N stress in durum wheat.     

Genetic variation in wild and cultivated artichoke revealedby RAPD markers

Determination of intraspecific genetic diversity is an important first step for the utilization of genetic resources and canprovide useful information on crop evolution. A living collection of Italian and foreign artichoke varieties and ecotypes is maintained at the Germplasm Institute, Bari, Italy. A total of 32 accessions of cultivated (Cynara cardunculus L. var.scolymus (L.) Fiori), three of wild (Cynara cardunculus var.cardunculus L.) artichoke and two ofcultivated cardoon (Cynara cardunculus var.altilis L.) were analysed in order to studygenetic variation and relationships within the species, using RAPDmarkers. Cultivated accessions were selected according tomorphological variation and geographical distribution available inthe collection. Fifty arbitrary decamer primers were initially tested, 18 ofwhich showed from 3 to 10 unambiguously interpretable fragments. Anintra-accession analysis using 4 varieties and 8 polymorphicprimers revealed that no RAPD variation was detected amongindividuals. Jaccard's similarity index (JSI) was comprisedamong 1 and 0.693 when all accessions were considered, on the otherhand, within the cultivated artichoke, JSI ranged between 1 and0.817. A UPGMA dendrogram based on the similarity matrix showed thatwild artichokes were clearly separated from cultivated accessions.Moreover, within cultivated artichokes, several groups could bedistinguished.     

Fertilization strategies on cultivars of globe artichoke: Effects on yield and quality performance

This paper refers on the yields and productive aspects of mineral and foliar nutrition of globe artichoke plants (cv. ‘Opal’ and ‘Madrigal’). Marketable yields were investigated in response to different nitrogen rates and foliar applications of biostimulating products. Field experiments were conducted over two seasons on an alluvial salty-clay soil in sub-humid climate conditions. Analyzed traits differed between years because of the age of plants while the lowest nitrogen rate and foliar spraying of biostimulants influenced significantly head weight, dry matter, and Soil Plant Analysis Division (SPAD) index. Biostimulants induced longer cultural cycle and heads lighter in weight. Cultivar ‘Madrigal’ was most productive than ‘Opal’, both in number (26.28% higher) and in weight (28.51%) of total heads. The two-year investigation showed that the synergic action was effective in improving some of the investigated productive traits of globe artichoke.     

Genetic diversity of some surviving on-farm Italian common bean (Phaseolus vulgaris L.) landraces

This study was undertaken to estimate the level of variation among and within 33 local populations belonging to seven Italian common bean landraces, by analysing the polymorphism of seed storage proteins, simple sequence repeat (SSR) and amplified fragment length polymorphism (AFLP) molecular markers. The nutritional seed quality of the landraces studied was also investigated. Results showed that the landraces retain a considerable level of heterogeneity. Use of both biochemical and molecular approaches provided a picture of the genetic diversity of each landrace. Cluster analyses based on Nei's genetic distances and Jaccard's similarity index as defined by SSR and AFLP markers, respectively, showed that all populations clustered into two groups corresponding to the Andean and the Mesoamerican gene pools. Knowledge of the genetic structure of a landrace is fundamental in elaborating strategies, which involve the local farmers, allowing us to improve and, at the same time, safeguard the genetic integrity of landrace genetic resources.