Relationships of the Sardinian isolate of tomato yellow leaf curl geminivirus with its whitefly vectorBemisia tabaci Gen

Relationships of an Italian isolate of tomato yellow leaf curl geminivirus from Sardinia (TYLCV-S) with its whitefly vectorBemisia tabaci were studied by means of experimental transmissions from tomato to tomato plants. TYLCV-S was confirmed to be transmitted in a persistent, circulative manner. The minimum latent period in the vector was between 17 and 20 h from the beginning of the acquisition access period (AAP). The maximum retention of infectivity was 8 days from the end of the AAP. Both acquisition and inoculation feeding times influenced the detected proportion of infective insects, with patterns well described by an exponential model. Acquisition was more efficient than inoculation. Males were significantly less efficient vectors than females. Nymphs were as efficient as adults in acquiring the virus. The length of AAP influenced both the retention of infectivity, and the pattern of transmission in serial transfer transmission tests with individual females. No significant difference in transmission efficiency was detected between two colonies ofB. tabaci, one inducing typical silverleaf symptoms on squash, the other inducing only mild symptoms with more than 50 whiteflies per plant. The phenomenon of periodic acquisition was not unequivocally proved for TYLCV-S.Research supported by the National Research Council of Italy, Special Project RAISA, Sub-project N. 2. Paper N. 1961.Supported by a grant from the Istituto Agronomico Mediterraneo, Valenzano (Bari), Italy.     

Vitamin E Biochemistry and Function: A Case Study in Male Rabbit

Vitamin E supplementation has become a common procedure to promote growth and health and improve the qualitative characteristics of farm animals. It has been demonstrated to be an efficient strategy for improving their reproductive function. Germ cells are particularly vulnerable to oxidative damage and may thus require additional antioxidant protection. The aim of this review is to give a comprehensive overview of the current knowledge of the biochemistry and physiology of vitamin E; successively, the effect of this compound on the reproductive activity of rabbit buck is accurately described. In particular, this review examines studies on the effects of animal age, dose and duration of vitamin E supplementation, as well as the co-supplementation with selenium, vitamin C and polyunsaturated fatty acids. Several investigations have shown beneficial effects in bucks supplemented with vitamin E levels higher than the standard dietary requirement (50 mg/kg) particularly when the semen is stored. However, the exact dietary intake of vitamin E should be decided according to specific tissue needs for the individual vitamin E forms and the specific saturation markers.     

Acquisition capability of the grapevine Flavescence dorée by the leafhopper vector Scaphoideus titanus Ball correlates with phytoplasma titre in the source plant

Flavescence dorée (FD) is one of the most economically important grapevine diseases in Southern Europe, and it is associated with phytoplasmas, phloem-limited wall-less bacteria. Recovery from disease naturally occurs in infected grapevines during the following seasons after infection. The capability of the leafhopper vector Scaphoideus titanus to acquire FD phytoplasma (FDP) from recovered and infected grapevines of Barbera and Nebbiolo varieties was investigated in North-western Italy vineyards monitored from 2007 to 2011. Pathogen concentration was quantified by real-time PCR in FDP-infected grapevines and broad beans, also used as source plants under controlled conditions, to correlate acquisition capabilities and phytoplasma titre in source plants. S. titanus acquired FDP from infected, but not from recovered, grapevines. FDP titre was higher in Barbera than in Nebbiolo and higher in summer than in spring, and acquisition efficiency and pathogen titre in source plants were positively correlated, both in field and laboratory conditions. Recovered plants do not represent a source of inoculum for the vector and therefore do not contribute to FDP spread. The inability of recovered plants to serve as FDP acquisition sources for the vector as well as the effect of the season and of the two grapevine varieties on the FDP acquisition efficiency are relevant results to re-design disease management practices, especially since insecticide treatments against the vector are not fully effective, and newly designed successful control strategies are required.     

Inter-specific competition and management modify the morphology, nutrient content and resorption in Scots pine needles

Influence of tree-to-tree competition on nutrient resorption is still not well understood. To contribute filling this gap, we assessed the effects of thinning (0, 20 and 30 % extraction of basal area) and canopy type (beech–pine vs. pine subplots) on needle dry weight, needle length, nutrient content and nutrient resorption (N, P and K) in Scots pine (Pinus sylvestris) needles of different cohorts, 8–9 years after thinning. Thinning and canopy type often concurrently affected needle morphology (e.g. lighter and shorter needles in 30 % thinning, heavier and longer needles in pine canopy on the first year of study) and nutrient content (e.g. decrease in N, P and K in 30 %; N, P and K higher in pine canopy on the first year of study). However, effects of thinning appeared only in older cohorts for N and P but were found in old and new cohorts for needle dry weight, needle length and K, indicating that some thinning effects remained after 8–9 years. Canopy effects on morphology and nutrient content were more frequent in recent cohorts, in relation to an increase in beech cover over time. While no clear effects of thinning on the nutrient resorption were observed, higher values were observed in the pine than in the mixed canopy, which could be related to a higher Scots pine stem growth in those patches. The observed differences between treatments will likely increase as the stand develops, probably leading to beech trees being dominant, and as future thinnings are carried out.     

Pseudomonas syringae pv. coryli, the Causal Agent of Bacterial Twig Dieback of Corylus avellana

ABSTRACT Thirty-eight bacterial strains isolated from hazelnut (Corylus avellana) cv. Tonda Gentile delle Langhe showing a twig dieback in Piedmont and Sardinia, Italy, were studied by a polyphasic approach. All strains were assessed by fatty acids analysis and repetitive sequence-based polymerase chain reaction (PCR) fingerprinting using BOX and ERIC primer sets. Representative strains also were assessed by sequencing the 16S rDNA and hrpL genes, determining the presence of the syrB gene, testing their biochemical and nutritional characteristics, and determining their pathogenicity to hazelnut and other plants species or plant organs. Moreover, they were compared with reference strains of other phytopathogenic pseudomonads. The strains from hazelnut belong to Pseudomonas syringae (sensu latu), LOPAT group Ia. Both fatty acids and repetitive-sequence-based PCR clearly discriminate such strains from other Pseudomonas spp., including P. avellanae and other P. syringae pathovars as well as P. syringae pv. syringae strains from hazelnut. Also, the sequencing of 16S rDNA and hrpL genes differentiated them from P. avellanae and from P. syringae pv. syringae. They did not possess the syrB gene. Some nutritional tests also differentiated them from related P. syringae pathovars. Upon artificial inoculation, these strains incited severe twig diebacks only on hazelnut. Our results justify the creation of a new pathovar because the strains from hazelnut constitute a homogeneous group and a discrete phenon. The name of P. syringae pv. coryli is proposed and criteria for routine identification are presented.     

Enrichment and diversity of plant-probiotic microorganisms in the rhizosphere of hybrid maize during four growth cycles

The population dynamics of naturally-occurring antibiotic producing and nitrogen fixing rhizobacteria, as well as of arbuscular mycorrhizal fungi (AMF) was investigated for a hybrid of maize (Lo964×Lo1016) in comparison to its two parental lines (Lo964 and Lo1016), during four successive 5-week-long growth cycles in the same pot. Beneficial rhizobacteria, such as nitrogen fixers and 2,4-diacetylphloroglucinol (DAPG) and pyrrolnitrin (PRN) producers, as well as AMF were stimulated in the hybrid rhizosphere earlier than in those of the two parental lines. In fact, they were molecularly detected in all rhizospheric samples of the hybrid, independently of the cycle, whereas for the parental lines positive detections occurred only for samples collected after at least two growth cycles. Interestingly, a MPN-PCR approach on rhizospheric DNA samples indicated that, when detected, beneficial rhizobacteria reached similar density in all maize genotypes (2×10³ to 2.2×10⁴ target DNA sequences/g^?1 of root). Concerning the AMF, even if the three maize genotypes were cultivated in the same soil, it appears that each maize genotype stimulates the AMF population differently. Both the hybrid and the Lo964 line were able to select, in the observed time period, their own adapted phylogenetic AMF subgroups (Glomus A for the hybrid, Archeospora for Lo964), whereas the Lo1016 line was not.     

Separating the effects of habitat amount and fragmentation on invertebrate abundance using a multi-scale framework

Context

Herbicide treatments in viticulture can generate highly contrasting mosaics of vegetated and bare vineyards, of which vegetated fields often provide better conditions for biodiversity. In southern Switzerland, where herbicides are applied at large scales, vegetated vineyards are limited in extent and isolated from one another, potentially limiting the distribution and dispersal ability of organisms.

Objectives

We tested the separate and interactive effects of habitat amount and fragmentation on invertebrate abundance using a multi-scale framework, along with additional environmental factors. We identified which variables at which scales were most important in predicting patterns of invertebrate abundance.

Methods

We used a factorial design to sample across a gradient of habitat amount (area of vegetated vineyards, measured as percentage of landscape PLAND) and fragmentation (number of vegetated patches, measured as patch density PD). Using 10 different spatial scales, we identified the factors and scales that most strongly predicted invertebrate abundance and tested potential interactions between habitat amount and fragmentation.

Results

Habitat amount (PLAND index) was most important in predicting invertebrate numbers at a field scale (50 m radius). In contrast, we found a negative effect of fragmentation (PD) at a broad scale of 450 m radius, but no interactive effect between the two.

Conclusions

The spatial scales at which habitat amount and fragmentation affect invertebrates differ, underpinning the importance of spatially explicit study designs in disentangling the effects between habitat amount and configuration. We showed that the amount of vegetated vineyards has more influence on invertebrate abundance, but that fragmentation also contributed substantially. This suggests that efforts for augmenting the area of vegetated vineyards is more beneficial for invertebrate numbers than attempts to connect them.

     

Identification of mycobacterial infections in wild boars in Northern Sardinia, Italy

During a six-month period a region of Northern Sardinia was monitored to check the presence of mycobacterial infections in wild boars. Forty-eight serum and 229 biopsy samples were collected from different animals and examined by both traditional diagnostic techniques (culture, bacterioscopic and molecular tests) and enzyme-linked immunosorbent assay (ELISA). The latter was used to determine the antibody response against both methylated and nonmethylated Heparin-Binding Haemagglutinin (HBHA) protein. Nine mycobacterial strains were isolated: three M. avium ssp. paratuberculosis (Map), three M. avium, one M. interjectum and two M. scrofulaceum strains. By PCR, only one animal was positive for M. bovis, whereas 10 animals were positive for Map. Out of the 48 sera tested, 19 showed a good humoral response to methylated HBHA and 17 to nonmethylated HBHA. Our data provide new information on the prevalence of mycobacterial infection among wild boars in Northern Sardinia and suggest that a more effective program should be developed to monitor mycobacterial infections in the wild animal population.     

Quantitation over time of tomato yellow leaf curl geminivirus DNA in its whitefly vector

ABSTRACT The amount of tomato yellow leaf curl geminivirus (TYLCV) DNA that accumulated in the vector Bemisia tabaci was studied by quantitative chemiluminescent dot-blot assay, using digoxigenin-labeled specific DNA probes. Large groups of female whiteflies were allowed to feed for 4, 12, 24, or 48 h on TYLCV-infected tomato plants and then were transferred to TYLCV-immune cucumber plants. Insects were sampled at different times during and after acquisition access and tested for TYLCV-DNA content. TYLCV-DNA assays were done either on whole insects oron the head plus prothorax (to include salivary glands) and abdomen separately. The maximum amount of TYLCV DNA, averaging from 0.5 to 1.6 ng per insect, was always attained at the end of the acquisition period. The mean amount then decreased by about 1 to 2% per day, remaining clearly detectable up to 20 days after the end of the acquisition period. Only some whiteflies that were TYLCV-positive in the abdomen were positive for head plus prothorax. In both parts of the body, TYLCV DNA remained detectable up to 18 days after the end of the acquisition period, showing that TYLCV DNA remains in insect tissues much longer than infectivity indicates.     

Detection of cathepsin L in red cell membranes from fish blood

Cathepsin L, one of the cysteine proteases found in fish muscle, is considered to be the main cause of post-mortem autolysis of fish muscle. We have determined the presence of cathepsin L in the membranes of red blood cells of carp, amberjack, and red sea bream and measured its activity. Immunoblotting of an extract of the red blood cell membranes from these three fish species using human anti-cathepsin L antibody revealed the presence of cathepsin L of different molecular masses. The molecular masses of cathepsin L was estimated to be 120 and 85 kDa in the amberjack and 75 and 70 kDa in the carp. These proteins have higher molecular masses than the mature form of cathepsin L, suggesting that they are precursor forms. In contrast, the protein in the red sea bream was estimated to have a molecular mass of 30 kDa, suggesting that this cathepsin L is a mature form. The specific activity of cathepsin L was highest in the red blood cell membranes of the amberjack, followed by the carp and the red sea bream in descending order.