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Development of an Efficient Real-Time Quantitative PCR Protocol for Detection of Xanthomonas arboricola pv. pruni in Prunus Species
No full textXanthomonas arboricola pv. pruni, the causal agent of bacterial spot disease of stone fruit, is considered a quarantine organism by the European Union and the European and Mediterranean Plant Protection Organization (EPPO). The bacterium can undergo an epiphytic phase and/or be latent and can be transmitted by plant material, but currently, only visual inspections are used to certify plants as being X. arboricola pv. pruni free. A novel and highly sensitive real-time TaqMan PCR detection protocol was designed based on a sequence of a gene for a putative protein related to an ABC transporter ATP-binding system in X. arboricola pv. pruni. Pathogen detection can be completed within a few hours with a sensitivity of 10(2) CFU ml(-1), thus surpassing the sensitivity of the existing conventional PCR. Specificity was assessed for X. arboricola pv. pruni strains from different origins as well as for closely related Xanthomonas species, non-Xanthomonas species, saprophytic bacteria, and healthy Prunus samples. The efficiency of the developed protocol was evaluated with field samples of 14 Prunus species and rootstocks. For symptomatic leaf samples, the protocol was very efficient even when washed tissues of the leaves were directly amplified without any previous DNA extraction. For samples of 117 asymptomatic leaves and 285 buds, the protocol was more efficient after a simple DNA extraction, and X. arboricola pv. pruni was detected in 9.4% and 9.1% of the 402 samples analyzed, respectively, demonstrating its frequent epiphytic or endophytic phase. This newly developed real-time PCR protocol can be used as a quantitative assay, offers a reliable and sensitive test for X. arboricola pv. pruni, and is suitable as a screening test for symptomatic as well as asymptomatic plant material
Effect of dsRNA on growth rate and reproductive potential of Monosporascus cannonballus
No full textThe effect of double stranded RNA (dsRNA) infection on growth rate and the reproductive potential of Monosporascus cannonballus was studied in 21 isolates collected in cucurbit growing areas of Spain and Tunisia. The isolates were incubated on potato dextrose agar (PDA) under different conditions of temperature, pH, and water potential (Psi(s)). They showed optimal growth temperatures over the range of 27-34 degrees C and perithecia formation was obtained mainly at 25 and 30 degrees C, although some isolates were able to produce perithecia at 35 degrees C. All isolates were able to produce perithecia in a broad range of pHs (4-8). Regarding the effect of Psi(s), the isolates were more tolerant to grow on KCl than on NaCl. For each solute, radial growth decreased progressively as Psi(s) decreased and was severely limited at -5.0 to -6.0 MPa. Perithecia formation was highest at -0.5 MPa, decreased at -1.0 MPa and occurred just in some isolates at -2.0 MPa. Nine of the M. cannonballus isolates harboured dsRNA with 2-6 bands each and a size range of 1.9-18.0 Kb. Phenotypical data were subjected to multivariate factorial analysis. Most of the isolates clustered in two groups corresponding with the presence/absence of dsRNA elements. Isolates without detectable dsRNA produced more perithecia. However, isolates with dsRNA produced lower number of perithecia depending on the pH, Psi(s) or solute used. These results improve our understanding of the behaviour and growth of this pathogen in soil, and can be useful to implement effective disease control. (C) 2011 The British Mycological Society. Published by Elsevier Ltd. All rights reserved
Antimicrobial Edible Films and Coatings for Fresh and Minimally Processed Fruits and Vegetables: A Review
No full textThe use of edible films and coatings is an environmentally friendly technology that offers substantial advantages for shelf-life increase of many food products including fruits and vegetables. The development of new natural edible films and coatings with the addition of antimicrobial compounds to preserve fresh and minimally processed fruits and vegetables is a technological challenge for the industry and a very active research field worldwide. Antimicrobial agents have been successfully added to edible composite films and coatings based on polysaccharides or proteins such as starch, cellulose derivatives, chitosan, alginate, fruit puree, whey protein isolated, soy protein, egg albumen, wheat gluten, or sodium caseinate. This paper reviews the development of edible films and coatings with antimicrobial activity, typically through the incorporation of antimicrobial food additives as ingredients, the effect of these edible films on the control of target microorganisms, the influence of antimicrobial agents on mechanical and barrier properties of stand-alone edible films, and the effect of the application of antimicrobial edible coatings on the quality of fresh and fresh-cut fruits and vegetables
SALTIRSOIL: a simulation model for the mid to long-term prediction of soil salinity in irrigated agriculture
No full textThe SALTIRSOIL model predicts soil salinity, sodicity and alkalinity in irrigated land using basic information on soil, climate, crop, irrigation management and water quality. It extends the concept of the WATSUIT model to include irrigation and crop management practices, advances in the calculation of evapotranspiration and new algorithms for the water stress coefficient and calculation of electrical conductivity. SALTIRSOIL calculates the soil water balance and soil solution concentration over the year. A second module, SALSOLCHEM, calculates the inorganic ion composition of the soil solution at equilibrium with soil calcite and gypsum at the soil’s CO2 partial pressure. Results from comparing predicted and experimentally determined concentrations, observations and predictions of pH, alkalinity and calcium concentration in calcite‐saturated solutions agree to the second significant figure; in gypsum‐saturated solutions the standard difference between observations and predictions is <3% in absolute values. The algorithms in SALTIRSOIL have been verified and SALSOLCHEM validated for the reliable calculation of soil salinity, sodicity and alkalinity at water saturation in well‐drained irrigated lands. In simulations for horticultural crops in southeast Spain, soil solution concentration factors at water saturation, quotients of electrical conductivity (EC25) at saturation to electrical conductivity in the irrigation water, and quotients of sodium adsorption ratio (SAR) are very similar to average measured values for the area
Occurrence, characterization and insecticidal activity of Bacillus thuringiensis strains isolated from argan fields in Morocco
No full textSoils collected from five locations in the argan forest (an endemic plant) in Morocco were used to form the first collection of Bacillus thuringiensis (Bt) strains from this area (58 strains). Here we found that the argan forest is a major source of Bt, as 90.62% of the samples contained Bt strains. These strains produced mainly spherical or irregular crystals that in some cases remained adhered to the spore after cell lysis. There was no strain producing bipyramidal crystals, suggesting the absence of strains bearing cry1 genes. This was confirmed by PCR analysis using eight primer pairs that can potentially detect 13 different groups of cry and cyt genes. Strains containing cry7/8 were the most abundant (25.53%), followed by strains harbouring cry9A (14.89%), cry11 (8.51%) and cry4 (4.25%). The mixtures of spores and crystals as well as culture supernatants were assayed for toxicity towards Ceratitis capitata (Medfly), showing up to 30% mortality. Our findings suggest that the argan region is a suitable target for future and wider screening programmes looking for strains bearing toxins or combinations of them to develop more efficient Bt-based formulates
Citrus-orchard ground harbours a diverse, well-established and abundant ground-dwelling spider fauna
Get PDFGround-dwelling spider assemblages comprise one of the most representative predatory groups to be found in many crops. There is some evidence of the role that ground-dwelling spiders play in controlling certain citrus pests; however, there are almost no studies about the abundance and composition of this predatory group in citrus orchards. A three-year survey conducted using pitfall traps in three citrus orchards in Eastern Spain yielded more than five-thousand ground-dwelling spiders belonging to more than 50 species and 20 families. Wandering families such as Lycosidae, Gnaphosidae and Zodariidae were the most numerous in terms of captures. The generalist predator Pardosa cribata Simon (Araneae: Lycosidae) was the most common species, representing a quarter of all captures, followed by Zodarion cesari Pekar. (Araneae: Zodariidae) and Trachyzelotes fuscipes (Koch) (Araneae: Gnaphosidae). Spiders were active throughout the year with a peak population in summer. The species abundance data for the three spider assemblages sampled fitted a log normal statistical model which is consistent with a well-established community. The presence of a cover crop provided higher abundance of alternative prey and consequently higher abundance and diversity of ground-dwelling spiders. This work demonstrates that the citrus-orchard ground harbours a diverse and abundant ground-dwelling spider fauna, which is also active throughout the year. A challenge for future studies will be to establish conservation management strategies for these predators, that will improve biological control of those citrus pests that inhabit or spend part of their life cycle on the orchard floor
Basic bio-ecological parameters of the invasive Red Palm Weevil, Rhynchophorus ferrugineus (Coleoptera: Curculionidae), in Phoenix canariensis under Mediterranean climate
No full textThe invasive red palm weevil, Rhynchophorus ferrugineus Olivier (Coleoptera, Curculionidae), is one of the most destructive pests of palms in the world. Since its detection in the Mediterranean Basin, the ornamental Phoenix canariensis Hort. ex Chabaud has become its main host. This study was aimed at determining the life cycle of R. ferrugineus in live P. canariensis palms. Egg lethal temperature threshold and thermal constant were determined in the laboratory and resulted in 13.1 degrees C and 40.4 degree days (DD), respectively. A semi field assay was carried out in a mesh enclosure where living P. canariensis palms were artificially infested with neonate larvae at one-month intervals from June 2008 to May 2009 under natural conditions. Infested palms were dissected at different time intervals. Maximum mortality rates for R. ferrugineus were observed for palms infested either in December or January (100%), whereas those infested from April through September showed maximum survival rates. Mean monthly temperatures below 10.3 degrees C were lethal for neonate larvae, as 4.5 degrees C were for older immature stages. All recovered larvae could be classed according to one of 13 instars. A thermal constant of 666.5 DD was estimated for complete larval development. Pupal develoment required an additional 282.5 DD. Based on these results and on the temperatures from 46 climatic stations selected in the Iberian Peninsula, less than one generation per year can be expected in areas with mean annual temperature below 15 degrees C and more than two where mean annual temperature is above 19 degrees C
Resistance to Plum pox virus in plants expressing cytosolic and nuclear single-chain antibodies against the viral RNA NIb replicase
No full textThe expression of engineered single-chain variable fragments specific to the NIb RNA replicase of Plum pox virus (PPV) (scFv2A) in transgenic plants was successfully used as a strategy to interfere with viral infection. Different scFv2A fusion proteins were constructed to target those subcellular compartments, such as the cytosol, endoplasmic reticulum (ER) membrane structures and the nucleus, where NIb protein presumably accumulates. Several transgenic lines of Nicotiana benthamiana plants expressing the scFv2A targeted to the cytosol (2A lines), ER (6K2 lines) and nucleus (NLS lines) were obtained. The protective effect of scFv expression was determined by mechanical virus inoculation in five 2A, three 6K2 and four NLS transgenic lines. The strongest resistance was afforded with the 2A-3 (six non-infected plants out of 10), 6K2-1 (17 out of 33) and NLS-11 (16 out of 19) transgenic lines. The success of this interference with PPV infection opens new possibilities for the control of this RNA virus and could be exploited not only to confer resistance in transgenic plants, but also to elucidate the role of the non-structural NIb protein in different cell compartments during viral infection
Water relations of field grown Pomegranate trees (Punica granatum) under different drip irrigation regimes
No full textPomegranate (Punica granatum L) is a deciduous fruit tree native of central Asia included in the so-called group of minor fruit tree species, not widely grown but of some importance in the south east of Spain. Fruit consumption interest is due to the organoleptic characteristics and to the beneficial effects on health. Pomegranate tree are considered as a culture tolerant to soil water deficit. However, very little is known about pomegranate orchard water management. The objective of this work was to characterize, for the first time in P. granatum, water relations aspects of applied significance for irrigation scheduling. Trees under different irrigation regimes were used and midday stem water potential (psi(stem)) and midday leaf gas exchange were periodically measured over the course of an entire season. During spring and autumn, psi(stem) did not show significant differences between irrigation treatments while there were considerable differences in leaf photosynthesis and stomatal conductance, suggesting a near-isohydric behaviour of pomegranate trees. This might explain why the signal intensity of psi(stem) was lower than those of gas exchange indicators during the experimental period. Thus, leaf photosynthesis rates and stomatal conductance might have a greater potential for irrigation scheduling of pomegranate trees than psi(stem) measured at solar noon. (C) 2010 Elsevier B.V. All rights reserved
Development and validation of an automatic thermal imaging process for assessing plant water status
Get PDFLeaf temperature is a physiological trait that can be used for monitoring plant water status. Nowadays, by means of thermography, canopy temperature can be remotely determined. In this sense, it is crucial to automatically process the images. In the present work, a methodology for the automatic analysis of frontal images taken on individual trees was developed. The procedure can be used when cameras take at the same time thermal and visible scenes, so it is not necessary to reference the images. In this way, during the processing in batch, no operator participated. The procedure was developed by means of a non supervised classification of the visible image from which the presence of sky and soil could be detected. In case of existence, a mask was performed for the extraction of intermediate pixels to calculate canopy temperature by means of the thermal image. At the same time, sunlit and shady leaves could be detected and isolated. Thus, the procedure allowed to separately determine canopy temperature either of the more exposed part of the canopy or of the shaded portion. The methodology developed was validated using images taken in several regulated deficit irrigation trials in Persimmon and two citrus cultivars (Clementina de Nules and Navel Line-Late). Overall, results indicated that similar canopy temperatures were calculated either by means of the automatic process or the manual procedure. The procedure developed allows to drastically reduce the time needed for image analysis also considering that no operator participation was required. This tool will facilitate further investigations in course for assessing the feasibility of thermography for detecting plant water status in woody perennial crops with discontinuous canopies. Preliminary results reported indicate that the type of crop evaluated has an important influence in the results obtained from thermographic imagery. Thus, in Persimmon trees there were good correlations between canopy temperature and plant water status while, in Clementina de Nules and Navel Lane-Late citrus cultivars canopy temperature differences among trees could not be related with tree-to-tree variations in plant water status. (C) 2011 Elsevier B.V. All rights reserved