Suppression of conidial germination and appressorial formation by silicate treatment in powdery mildew of strawberry

The mode of action of soluble silicon against strawberry powdery mildew (Sphaerotheca aphanis var. aphanis) was investigated in four experiments. First, silicon-treated leaves from plants grown with silicate (Si+) and control leaves were excised, inoculated with conidia, and subsequent germination and formation of appressoria in a petri dish was assessed after 24 h. The germination rate was 49.7% on Si+ leaves, and was 67.2% on control leaves (t-test, P < 0.01). Second, we soaked cellulose membranes in various solvents and then placed the membranes on 4% water agar, dusted the membranes with conidia, and examined after 12 h. No difference was apparent between any treatment and the control (distilled water). Third, strawberries growing hydroponically with additional silicon in the medium were inoculated with conidia, and leaves were observed with a scanning electron microscope 1–2 days after inoculation. Germ tubes and secondary hyphae were shorter and had fewer branches on Si+ leaves than on the control. Moreover, penetration appeared to be inhibited. Fourth, the cuticle was separated from leaves from plants grown as in the third experiment, placed on water agar, and dusted with conidia. Germination of conidia, observed with a light microscope, on Si+ leaves was suppressed markedly to 40%–60% of that of the control. These results suggested that soluble silicon induced physiological changes in the cuticle layer after absorption by the plant. In addition, soluble silicate reduced germination of conidia, formation of appressoria, and possibly the penetration of powdery mildew.     

Fusariosis in rubber tree: pathogenic,morphological, and molecular characterization of the causal agent

Fusariosis, one of the phytosanitary problems found in rubber producing areas in the northwest of the state of São Paulo, is a disease that affects the bark of the adult plants, affecting the exploitation of latex. The typical symptoms appear as cracks in the bark that expand from the rootstock towards the tapping panel, causing a drying of the latex flux in the injured region, impeding latex tapping. Due to the recent incidence of this disease in rubber plantations, the goal of this study was to characterize the Fusarium associated with symptomatic rubber tissue in three different locations in the state of São Paulo. In order to identify Fusarium species, pathogenicity, morphological, cultural and molecular studies were carried out. A total of 51 isolates were obtained and separated into three groups based on macroconidium morphology, presence or absence of sporodochia, types of chlamydospores, formation of phialides and conidiogenesis of microconidia and mesoconidia, mycelial growth rate and coloring of the colonies. These groups were corroborated using DNA sequence information for five different genetic loci, and were subsequently recognized as Fusarium oxysporum, F. incarnatum and F. decemcellulare. Our results further showed that all 51 of the Fusarium isolates recovered were pathogenic to rubber tree seedlings of RRIM 600 standard clone. This study also reports for the presence of F. oxysporum and F. incarnatum in rubber plantations in the state of São Paulo and in Brazil.

     

Anti-Thy-1 Antibody-mediated Complement-dependent Cytotoxicity is Regulated by the Distribution of Antigen,Antibody and Membrane Complement Regulatory Proteins in Rats

Some therapeutic antibodies as anticancer agents exert their effects through the host immune system, but the factors that predict their cytotoxicity, including complement-dependent cytotoxicity (CDC), are unclear. In the present study, we attempted to elucidate some of these factors in a preclinical model. CDC-related mesangiolysis caused by administration of the anti-Thy-1.1 antibody can be studied in the rat anti-Thy-1 glomerulonephritis model, so the model was used in this study. Three animals each were sacrificed at 0.5, 1, 8, 24 and 48 hours after i.v. administration of the anti-Thy-1.1 antibody at 1mg/kg. The distribution of the Thy-1.1 antigen and 2 membrane complement regulatory proteins (mCRPs), Crry and CD55, in three non-treated animals and the distribution of the injected antibody and C3 in the model was studied by immunohistochemistry. In the mesangial cells of the kidney, both expression of the antigen and distribution of the antibody with C3 deposition were observed with weak expression of mCRPs. There was also antigen and antibody distribution in the medullary cells of the adrenal gland and in the lymphocytes of the thymus but no C3 deposition, which was thought to be related to high expression of mCRPs. The antigen was observed in several other organs and tissues without distribution of the antibody. Cell death was only observed in the mesangial cells. These results clearly demonstrate that activation of CDC is regulated by several factors, such as distribution of the target molecule, antibody distribution and the balance among the molecules of the CDC cascade and mCRPs.     

Development of an ELISA based on the baculovirus-expressed capsid protein of porcine circovirus type 2 as antigen

The genome of porcine circovirus type 2 (PCV2) contains two major open reading frames, which have been shown to encode the virus capsid and replication-associated proteins. The capsid protein is a major structural protein of the virus; it can be a suitable target antigen for detecting PCV2-specific antibodies to monitor PCV2 infection. To produce the antigen, the capsid protein coding sequence was cloned into a baculovirus transfer vector, and a recombinant capsid (rC) protein of PCV2 was expressed as a combined fusion protein in frame with a C-terminal peptide of six histidines. The affinity-purified rC protein was used as coating antigen to develop an ELISA for detecting the virus-specific antibodies in swine sera. The rC protein-based ELISA (rcELISA) was evaluated by examining a panel of 49 PCV2-positive and 49 PCV2-negative swine sera. In comparative experiments of immunoperoxidase monolayer assay (IPMA) using 102 field sera, there was 89.2% coincidence between data obtained by the rcELISA and IPMA. The rcELISA achieved 88.5% specificity and 89.4% sensitivity for detection of PCV2 antibody in the field sera. The assay showed no cross-reactivity with antibodies to PCV type 1, porcine reproductive and respiratory syndrome virus and porcine parvovirus. The results suggest that the rcELISA is suitable for routine serodiagnosis and epidemiological surveys of PCV2-associated diseases.