Effectiveness of microsatellite DNA markers in checking the identity of protected designation of origin extra virgin olive oil

Collina di Brindisi is an Italian extra virgin olive oil that obtained the mark of protected designation of origin (PDO) according to EC Regulation 2081/92. The varietal requirements of the official production protocol of this oil foresee that this oil is prepared from cultivar Ogliarola (minimum 70%) and other Olea europaea L. cultivars that are diffused in the production area, accounting for a maximum of 30%. The aim of this work was to verify the effectiveness of microsatellite analysis in verifying the identity of Collina di Brindisi PDO olive oil. A preliminary assessment of product's quality by means of chemical analyses was also carried out. Microsatellite analysis clarified that the generic name Ogliarola, indicated in the technical sheet of this PDO oil, actually corresponded to the Ogliarola salentina cultivar. Furthermore, the obtained results showed that the examination of a limited number of DNA microsatellites enables the identification of the Ogliarola salentina cultivar in this PDO oil.     

Effects of conventional tillage on biochemical properties of soils

Modification of soil environment by different farming practices can significantly affect crop growth. Tillage causes soil disturbance, altering the vertical distribution of soil organic matter and plant nutrient supplies in the soil surface, and it may affect the enzyme activity and microbial biomass which are responsible for transformation and cycling of organic matter and plant nutrients. In this study, the influence of three conventional tillage systems (shallow plowing, deep plowing and scarification) at different depths on the distribution and activity of enzymes, microbial biomass and nucleic acids in a cropped soil was investigated. Analysis of variance for depth and tillage showed the influence of the different tillage practices on the activity of some enzymes and on the nucleic acids. Glucosidase, galactosidase, nitrate reductase and dehydrogenase activity were significantly affected by the three tillage modalities. Activity in the upper layer (0–20 cm) was higher in the plots tilled by shallow plowing and scarification than in those tilled by deep plowing. Positive relationships were observed between the soil enzymes themselves, with the exception of urease and pyrophosphatase activity. Moreover, significant correlations were found between DNA and β-galactosidase, and between RNA and β-glucosidase, β-galactosidase, alkaline phosphatase and phosphodiesterase. α-Glucosidase, β-galactosidase, alkaline phosphatase and phosphodiesterase were highly correlated with biomass C determined by the fumigation-extraction method. Received: 27 June 1996     

Functional and molecular responses of soil microbial communities under differing soil management practices

The effects of soil management on some microbiological properties and soil bacterial community structure were evaluated. Two field sites with the same soil type, located on the same geographic area adjacent to one other, have received different soil management practices and cultivation. One site has been subjected for 20 years to intensive horticulture under conventional tillage and irrigation with low quality salt-rich water; the second field site has been uncultivated for a long period and was turned to organic farming practices over the last 5 years and is currently cultivated with fruit orchard. Total bacterial counts, microbial ATP, microbial community metabolic (BIOLOG^®) profiles, and DNA fingerprinting by PCR-DGGE were determined. Two-way ANOVA revealed that total bacterial counts were not significantly (P>0.3) affected by the two different management practices; ATP content was consistently and significantly (P<0.001) lower in salt-water irrigated soil than in organic soil at the three sampling times. The cluster analysis of community level physiological profiles indicated that microbial communities were much more uniform in organic soil than in irrigated one, suggesting that salt-water irrigation could have affected the size of the microbial population, its metabolic activities, as well as its composition. Molecular patterns fitted the BIOLOG^® profile diversity. In particular, at any sampling time, PCR-DGGE patterns of bacterial DNA, extracted by an indirect method, significantly discriminated irrigated from organic soil samples. The PCR-DGGE patterns of total soil DNA, extracted by a direct method, showed a moderate to significant variation among irrigated and organic soil samples. Biochemical, microbiological and molecular data contributed to evidence a significantly different response of indigenous microflora to soil management by using saline water or organic farming.     

Perturbation of the EphA2-EphrinA1 System in Human Prostate Cancer Cells by Colonic (Poly)phenol Catabolites

The Eph tyrosine kinase receptors and their ephrin ligands play a central role in human cancer as their deregulated expression induces tumorigenesis with aggressive phenotypes. To evaluate their potential contribution to EphA2-ephrinA1 modulation, several colonic catabolites of dietary (poly)phenolics, known to be generated in vivo, were screened using an ELISA-based binding assay. Some of the catabolites inhibited the binding in a dose-dependent manner (IC(50) values from 0.26 to 43 μM). Functional studies on prostate adenocarcinoma cells revealed that pyrogallol and protocatechuic acid specifically antagonized ephrinA1-Fc-induced EphA2 phosphorylation at concentrations that were not cytotoxic. The active concentrations of pyrogallol appear to be close to what can be reached in vivo under physiological conditions. Finally, because of the roles played by the Eph-ephrin system not only in cancer development but also in neurodegeneration and diabetes, pyrogallol and protocatechuic acid are candidates for more detailed functional studies to elucidate their role in these pathophysiological processes.     

Rhizospheric organic compounds in the soil–microorganism–plant system: their role in iron availability

Poor iron (Fe) availability in soil represents one of the most important limiting factors of agricultural production and is closely linked to physical, chemical and biological processes within the rhizosphere as a result of soil–microorganism–plant interactions. Iron shortage induces several mechanisms in soil organisms, resulting in an enhanced release of inorganic (such as protons) and organic (organic acids, carbohydrates, amino acids, phytosiderophores, siderophores, phenolics and enzymes) compounds to increase the solubility of poorly available Fe pools. However, rhizospheric organic compounds (ROCs) have short half‐lives because of the large microbial activity at the soil–root interface, which might limit their effects on Fe mobility and acquisition. In addition, ROCs also have a selective effect on the microbial community present in the rhizosphere. This review aims therefore to unravel these complex dynamics with the objective of providing an overview of the rhizosphere processes involved in Fe acquisition by soil organisms (plants and microorganisms). In particular, the review provides information on (i) Fe availability in soils, including mineral weathering and Fe mobilization from soil minerals, ligand and element competition and plant‐microbe competition; (ii) microbe–plant interactions, focusing on beneficial microbial communities and their association with plants, which in turn influences plant mineral nutrition; (iii) plant–soil interactions involving the metabolic changes triggered by Fe deficiency and the processes involved in exudate release from roots; and (iv) the influence of agrochemicals commonly used in agricultural production systems on rhizosphere processes related to Fe availability and acquisition by crops.     

Soil microbial diversity and activity in a Mediterranean olive orchard using sustainable agricultural practices

Sustainable soil management of orchards can have positive effects on both soils and crop yields due to increases in microbial biomass, activity and complexity. The aim of this study was to investigate medium‐term effects (12 yr) of two different management practices termed ‘sustainable’ (ST) and ‘conventional’ (CT) on soil microbial composition and metabolic diversity of a rainfed mature olive orchard in Southern Italy. ST included no‐till, self‐seeding weeds (mainly graminaceous and leguminosae), and mulch derived from olive tree prunings, whilst CT was managed by frequent tillage and included severe pruning with residues removed from the orchard. Microbial analyses were carried out by culture‐dependent methods (microbial cultures and Biolog^®). Molecular methods were used to confirm the identification by light microscopy of the isolates of fungi and Streptomyces. Significantly more culturable fungi and bacteria were found in ST than in CT. The number of fungal groups in ST was also significantly greater than in CT. Overall and substrate‐specific Biolog^® metabolic diversity indices of microbial communities and soil enzyme activities were greater in ST. The results demonstrate that soil micro‐organisms respond positively to sustainable orchard management characterized by periodic applications of locally derived organic matter. This study confirms the need to encourage farmers with orchards in the Mediterranean basin to practise soil management based on organic matter inputs associated with zero tillage to improve soil functionality.     

Detection and Identification of <Emphasis Type="Italic">Phytophthora</Emphasis> Species in Southern Italy by RFLP and Sequence Analysis of PCR-amplified Nuclear Ribosomal DNA

In four neighbouring regions of southern Italy, Basilicata, Campania, Apulia and Calabria, pepper and zucchini plants showing Phytophthora blight symptoms, tomato plants with either late blight or buckeye rot symptoms, plants of strawberry showing crown rot symptoms and declining clementine trees with root and fruit rot were examined for Phytophthora infections by means of polymerase chain reaction (PCR) assays, using primers directed to nuclear ribosomal DNA (rDNA) repeat sequences. All diseased plants and trees examined tested positive. The detected fungal-like organisms were differentiated and characterized on the basis of primer specificity as well as through extensive restriction fragment length polymorphism (RFLP) and sequence analysis of PCR-amplified rDNA. Phytophthora capsici was identified in diseased pepper and zucchini plants, P. infestans was identified in tomato with late blight symptoms whereas buckeye rot-affected tomatoes and diseased strawberry plants proved to be infected by P. nicotianae and P. cactorum, respectively. Declining clementine trees were infected with P. citrophthora and P. nicotianae in about the same proportion. Also, thirty-one pure culture-maintained isolates of Phytophthora which had previously been identified in southern Italy by traditional methods but were never examined molecularly, were examined by RFLP and sequence analysis of PCR-amplified nuclear rDNA. Among these, an isolate from gerbera which had previously been identified by traditional methods only at genus level, was assigned to P. tentaculata. For the remaining pure culture-maintained isolates examined, the molecular identification data obtained corresponded with those delineated by traditional methods. Most of the diseases examined were already known to occur in southern Italy but the pathogens were molecularly detected and fully characterized at nuclear rDNA repeat level only from other geographic areas, very often outside Italy. A new disease to southern Italy was the Phytophthora blight of zucchini. This is also the first report on the presence and molecular identification of P. tentaculata from Italy.     

Soil microbial dynamics and genetic diversity in soil under monoculture wheat grown in different long-term management systems

Organic matter incorporation into soil can increase nutrient availability to plants but it can affect soil microbial communities. These in turn influence soil fertility and plant growth. Soil biochemical and microbiological properties are indicators of soil quality, but there is still no consensus as to how these should be used. Recent developments in molecular biology have provided new tools to obtain a view of the whole microbial community. The long-term impact of crop residue management on the microbial biomass, and on the activity and community structure of soil bacteria was evaluated in a clay soil of Southern Italy, where a monoculture of durum wheat (Triticum durum Desf.) was grown in semiarid conditions, and burning or incorporation of post harvest plant residues were typical practices. The role of N-mineral fertilization, simultaneously with the ploughing in of crop residues and during the plant growth cycle was also investigated. Total bacterial counts of viable cells, biomass C, ATP content of soil microorganisms, genetic fingerprinting of the total eubacterial community and of ammonia oxidizers were evaluated. Burning and incorporation did not affect microbial biomass C, ATP content, and total bacterial counts of viable cells although statistically relevant changes were detected among rhizosphere and bulk soil samples regardless of the crop residue management used. Molecular fingerprinting confirmed that: no significant change in the composition and diversity of total bacteria, as well as of ammonia oxidizers was induced by the crop residue managements; that soil bacteria were more sensitive to N fertilizer application during the plant growth cycle; and that rhizosphere soil samples were significantly different from those of the bulk soil. As microbiological and genetic factors related to soil fertility were not affected significantly, the long-term incorporation of crop residues, under the field conditions investigated, is a sustainable practice to manage post-harvest residues.