Characterization of dinucleotide and trinucleotide EST-derived microsatellites in the wheat genome

Over the past decade microsatellites or simple sequence repeats (SSRs) have attracted a considerable amount of attention from researchers. The aim of the present paper was to analyse expressed sequence tag-derived SSR (EST-SSR) marker variability in wheat and to investigate the relationships between the number and type of repeat units and the level of microsatellite polymorphism. Two hundred and forty-one new EST-SSR markers available in a public database () were characterized in eight durum wheat cultivars (Svevo, Ciccio, Primadur, Duilio, Meridiano, Claudio, Latino, Messapia), two accessions of Triticum turgidum var. dicoccoides (MG4343, MG29896), one accession of T. turgidum var. dicoccum (MG5323) and in the common wheat cv. Chinese Spring. Of these, 201 primer pairs (83.4%) amplified PCR products successfully, while the remaining 40 (16.6%) failed to amplify any product. Of the EST-SSRs analysed, 45.2% of the primer pairs amplified one or two PCR products. Multiple discrete PCR products were observed among both di- and trinucleotide EST-SSR markers (31.2 and 40.5%, respectively). Markers based on dinucleotide microsatellites were more polymorphic than those based on trinucleotide SSRs in the 12 wheat genotypes tested (68.9 and 52.7%, respectively). An average of 2.5 alleles for dinucleotide and 2.0 alleles for trinucleotide SSRs was observed. The data reported in the present work indicate the presence of a significant relationship between motif sequence types and polymorphism. The primer set based on the AG repeat motif showed the lowest percentage of polymorphism (55.0%), while the primer set based on the AC repeat motif showed t he highest percentage (85.0%). Among trinucleotide SSRs, the AGG microsatellite markers showed the highest percentage of polymorphism (70.0%), and the ACG motif the lowest value (25.0%). The characterization of these new EST-SSR markers and the results of our studyon the effect of repeat number and type of motifs could have important applications in the genetic analysis of agronomically important traits, quantitative trait locus discovery and marker-assisted selection.     

Characterization of flavonoids and pectins from bergamot (Citrus bergamia Risso) peel, a major byproduct of essential oil extraction

Bergamot peel is an underutilized byproduct of the essential oil and juice-processing industry. As with other Citrus peels, it still contains exploitable components, such as pectins and flavonoids. Commercial glycoside hydrolases, specifically a combination of pectolytic and cellulolytic enzymes, solubilized a high percentage of the material (81.94%). The flavonoid profile of the peel consisted of characteristic Citrus species flavanone rutinosides and neohesperosides derived from naringenin, eriodictyol, and hesperetin. In addition, a number of minor flavanone and flavone glycosides, not found in orange and lemon peels, were identified. The majority of flavonoids were extracted in the two 70% v/v EtOH extractions. Processing this material clearly has economic potential leading to low environmental impact.     

Diversity characterisation of broccoli (Brassica oleracea L. var. italica Plenck) landraces for their on-farm (in situ) safeguard and use in breeding programs

Brassica oleracea L. is an important species that originated in the eastern Mediterranean area. From there, B. oleracea L. subsp. capitata (L.) DC. convar. botrytis (L.) Alef. var. italica Plenck (broccoli) was introduced into Italy where considerable diversification took place. Several landraces of broccoli, which are highly appreciated by local people for their organoleptic traits, are presently cultivated in both large fields and home gardens. In this study, 10 landraces, 4 synthetics derived from landraces and 4 of the most used F1 hybrids were characterised for morpho-physiological and molecular traits with the principal aim of feeding information into the landrace protection schemes being developed in Italy and into breeding programs. The landraces and derived synthetics are well differentiated from each other and from the F1 hybrids for important agronomic and genetic traits. The principal component analysis based on analysis of morpho-physiological traits showed that overall the 2 main components accounted for 67.74 % of the total variation. The neighbour joining tree, based on 23 microsatellite markers, grouped accessions into 2 main clusters. One includes one synthetic variety and 2 F1 hybrids, while the other cluster includes all the other accessions. There were also several sub clusters. Most of the materials examined showed within-accession diversity. The information gathered in this study can be used to protect landraces through the schemes presently being developed in Italy and to register them in the European common catalogue of “conservation varieties”. In situ (on-farm) conservation of landrace diversity is discussed with reference to landrace use.     

Methyl bromide in natural fresh waters: Hydrolysis and volatilisation

The effect of some physical factors on the processes (hydrolysis, volatilisation) regulating the persistence of methyl bromide in fresh surface waters was investigated to complement previous research. It has been confirmed that electromagnetic radiation in the visible field has no influence on the hydrolysis kinetics of the fumigant. The removal of methyl bromide by volatilisation from stirred and ventilated solutions appears to be of greater importance in practice. It has been shown that under these conditions the half-life is directly proportional to the liquid depth. Without stirring volatilisation is greatly reduced, due to the low diffusivity of methyl bromide in the aqueous phase.     

Acylglycerol and fatty acid components of pulp,seed, and whole olive fruit oils. Their use to characterize fruit variety by chemometrics

The contents of triacylglycerols and diacylglycerols in three kinds of olive fruit oils (pulp, seed, and whole fruit) were determined. The fatty acid composition and the quality ratios 1,2-diacylglycerols/1,3-diacylglycerols and 1,2-diacylglycerols/total diacylglycerols were also assessed. Seven major Italian olive varieties were considered. Results of univariate statistical analyses indicated that the above analytical parameters (glyceridic ratios excepted) were effective in discriminating between pulp and seed oils. The seed oil fraction did not determine any change in the glyceridic indices and the acylglycerol or fatty acid composition concerning the whole fruit oil (mixture of pulp and seed oil fractions), the weight (%) of seed ( approximately 2%) being by far lower than the weight (%) of pulp ( approximately 85%) (fruit weight basis). Based on the data of triacylglycerol or fatty acid composition, and using appropriate parametric or nonparametric multivariate statistics, the genetic origins (olive variety) of the three fruit oil kinds were characterized.     

Landraces in Inland areas of the Basilicata region, Italy: monitoring and perspectives for on farm conservation

A collecting expedition has been carried out in the inland areas of the Basilicata Region, South Italy. Each of the 14 selected municipalities was subdivided in three strata based on the geographical distribution of population and orography: 1. city boundaries, 2. group of country houses, 3. isolated farm houses. Five sites were selected within each stratum/town for a total of 185 sites. In 101 sites crop species germplasm was collected and the farmer-maintainers were interviewed. Altogether 350 accessions were collected, the most common species were Capsicum annuum, Lycopersicon esculentum and Phaseolus vulgaris. A significant number of landraces are still preserved on farm, mostly in isolated farm houses (average age of farmers: 60?years) adopting traditional farming systems but a number of young farmers were also present. Perspectives for on farm conservation and management are discussed and public economic support for “farmer-maintainers” proposed.     

Leucettamols, Bifunctionalized Marine Sphingoids, Act as Modulators of TRPA1 and TRPM8 Channels

Leucettamols, bifunctionalized sphingoid-like compounds obtained from a marine sponge Leucetta sp., act as non-electrophilic activators of the TRPA1 channel and potent inhibitors of the icilin-mediated activation of the TRPM8 channel, while they are inactive on CB₁, CB₂ and TRPV1 receptors. Leucettamols represent the first compounds of marine origin to target TRPA1 and the first class of natural products to inhibit TRPM8 channels. The preparation of a small series of semi-synthetic derivatives revealed interesting details on the structure-activity relationships within this new chemotype of simple acyclic TRP modulators.     

Cadmium-Containing Carbonic Anhydrase CDCA1 in Marine Diatom Thalassiosira weissflogii

The Carbon Concentration Mechanism (CCM) allows phytoplakton species to accumulate the dissolved inorganic carbon (DIC) necessary for an efficient photosynthesis even under carbon dioxide limitation. In this mechanism of primary importance for diatoms, a key role is played by carbonic anhydrase (CA) enzymes which catalyze the reversible hydration of CO₂, thus taking part in the acquisition of inorganic carbon for photosynthesis. A novel CA, named CDCA1, has been recently discovered in the marine diatom Thalassiosira weissflogii. CDCA1 is a cambialistic enzyme since it naturally uses Cd²⁺ as catalytic metal ion, but if necessary can spontaneously exchange Cd²⁺ to Zn²⁺. Here, the biochemical and structural features of CDCA1 enzyme will be presented together with its putative biotechnological applications for the detection of metal ions in seawaters.     

Increased nitrogen-use efficiency of a short-rotation poplar plantation in elevated CO(2) concentration

We estimated nitrogen (N) use by trees of three poplar species exposed for 3 years to free air CO(2) enrichment (FACE) and determined whether the CO(2) treatment affected the future N availability of the plantation. Trees were harvested at the end of the first 3-year rotation and N concentration and content of woody tissues determined. Nitrogen uptake of fine roots and litter was measured throughout the first crop rotation. The results were related to previously published variations in soil N content during the same period. We estimated retranslocation from green leaves and processes determining N mobilization and immobilization, such as mineralization and nitrification, and N immobilization in litter and microbial biomass. In all species, elevated CO(2) concentration ([CO(2)]) significantly increased nitrogen-use efficiency (NUE; net primary productivity per unit of annual N uptake), decreased N concentration in most plant tissues, but did not significantly change cumulative N uptake by trees over the rotation. Total soil N was depleted more in elevated [CO(2)] than in ambient [CO(2)], although not significantly for all soil layers. The effect of elevated [CO(2)] was usually similar for all species, although differences among species were sometimes significant. During the first 3-year rotation, productivity of the plantation remained high in the elevated [CO(2)] treatment. However, we observed a potential reduction in N availability in response to elevated [CO(2)].