Factors affecting the virulence of Ralstonia solanacearum and its colonization on tobacco roots

Tobacco bacterial wilt caused by Ralstonia solanacearum is a serious disease affecting tobacco cultivation in southwest China. The response surface methodology was employed to evaluate the optimal conditions of tobacco bacterial wilt, and green fluorescent protein gene (gfp) labelling was applied to monitor the location and survival dynamics of R. solanacearum (Rs::gfp) on tobacco roots and in soil under these optimal conditions. The results showed that the highest wilt incidence was 91.13%, which occurred when the population reached 6.6 × 10⁶ CFU/g soil, the temperature was 30.55 °C, and the humidity was >81.42%. The Rs::gfp densely colonized the root tips and root hairs, and cells of Rs::gfp were observed intermittently in the elongation zone or at the point of the emerging lateral roots. The Rs::gfp number in the rhizosphere soil was 10.75‐, 73.13‐ and 74.86‐times higher than that in the bulk soil at 10, 15 and 20 days after transplantation, respectively. Increased colonization by Rs::gfp was related to the population of the pathogen, the environmental temperature and the humidity in the soil. These three conditions determined whether R. solanacearum would induce tobacco wilt. This is the first study to investigate factors affecting the virulence of a tobacco wilt bacterial pathogen, which is important for conducting field diagnosis and biocontrol of tobacco bacterial wilt.     

Distribution of Trace Metals and Organic Matter in the Sediments of Godavari Estuary of Kakinada Bay, East Coast of India

In Godavari estuary, the enhanced metal concentration in the sediment is localized, and is accompanied by marked enrichment in organic carbon, Kjeldahl nitrogen, and humic acid. High concentration of organic carbon was observed near the riverside sampling stations. A similar pattern was found for the humic acid concentrations. Relatively high organic carbon content was observed in pre-monsoon/monsoon compared to post-monsoon seasons which can be attributed to higher productivity due to upwelling in the former and landrunoff due to excessive rainfall in the later seasons. The trace metals in the sediments were found to be in the following order of their abundanceFe≥Mn≥Zn≥Pb≥Ni≥Cu≥Cd. In monsoon season relatively high levels of trace metals were reported in surface sediments which are transported by currents. The dominant process controllingthe metal enrichment may be either precipitation of metal-humic acid type of complexes. Our results indicate that in the Godavari estuary, the metal distribution is mainly influenced by sedimentarydynamics, while the contamination effect is very limited.     

Breeding maize lines for complete and partial resistance to maize streak virus (MSV)

Summary S₁ to S₅ inbred lines, derived from a maize population bred for its overall resistance to three tropical viruses, were screened for resistance to maize streak virus (MSV) by artificial plant infection using viruliferous leafhoppers. Symptoms were rated and intra-line frequency distributions studied for all pedigree inbred lines. Mortality due to MSV was very low among these inbreds. Symptoms appeared later, developed slower and were less severe than in the susceptible control hybrid. Results of a study of 500 S₁ and 93 S₂ lines suggested that resistance is under genetic control via a system involving loci with major genes (with dominance for resistance) controlling high to complete resistance, associated with a genetic system involving loci with minor genes controlling partial resistance. Lines expressing complete resistance to MSV were developed from 5 cycles of inbreeding and selection. The relevance of such complete and partial resistance is discussed.Abbreviations MRPS Mean Rating for Plants exhibiting Symptoms     

Detection of Bartonella spp. in wild rodents in Israel using HRM real-time PCR

The prevalence of Bartonella spp. in wild rodents was studied in 19 geographical locations in Israel. One hundred and twelve rodents belonging to five species (Mus musculus, Rattus rattus, Microtus socialis, Acomys cahirinus and Apodemus sylvaticus) were included in the survey. In addition, 156 ectoparasites were collected from the rodents. Spleen sample from each rodent and the ectoparasites were examined for the presence of Bartonella DNA using high resolution melt (HRM) real-time PCR. The method was designed for the simultaneous detection and differentiation of eight Bartonella spp. according to the nucleotide variation in each of two gene fragments (rpoB and gltA) and the 16S–23S intergenic spacer (ITS) locus, using the same PCR protocol which allowed the simultaneous amplification of the three different loci. Bartonella DNA was detected in spleen samples of 19 out of 79 (24%) black rats (R. rattus) and in 1 of 4 (25%) Cairo spiny mice (A. cahirinus). In addition, 15 of 34 (44%) flea pools harbored Bartonella DNA. Only rat flea (Xenopsyla cheopis) pools collected from black rats (R. rattus) were positive for Bartonella DNA. The Bartonella sp. detected in spleen samples from black rats (R. rattus) was closely related to both B. tribocorum and B. elizabethae. The species detected in the Cairo spiny mouse (A. cahirinus) spleen sample was closely related to the zoonotic pathogen, B. elizabethae. These results indicate that Bartonella species are highly prevalent in suburban rodent populations and their ectoparasites in Israel. Further investigation of the prevalence and zoonotic potential of the Bartonella species detected in the black rats and the Cairo spiny mouse is warranted.     

Autoclaved spent substrate of hatakeshimeji mushroom (Lyophyllum decastes Sing.) and its water extract protect cucumber from anthracnose

The protective effect of fresh spent mushroom substrate (SMS) of hatakeshimeji (Lyophyllum decastes Sing.), a popular culinary-medicinal mushroom, and its water extract against anthracnose of cucumber was investigated. Plants were treated with water extract from SMS or autoclaved water extract by spraying the whole plant or by dipping the first true leaf, and inoculated with Colletotrichum orbiculare seven days later. Plants treated with either of the extracts showed a significant reduction of necrotic lesions. On the other hand, when plants were grown in a mixture (1:2, v/v) of SMS or autoclaved SMS and soil, a disease reduction of over 70% was observed in autoclaved SMS. The water extract showed no antifungal activity against spore germination and mycelial growth of the pathogen. Real-time PCR analyses of chitinase and β-1,3-glucanase genes revealed a significant increase of expressions after 24 h of pathogen inoculation in water extract-treated plants compared with the control plants. These results suggest that water-soluble and heat-stable compounds in SMS enhance the state of systemic acquired resistance and protect cucumbers from anthracnose. Thus, the use of SMS for disease control may offer a new technology for the recycling and management of waste from mushroom cultivation.     

Antigenic variation of porcine transmissible gastroenteritis virus detected by monoclonal antibodies

Mouse myeloma cells (SP2/O) were fused with spleen cells from BALB/c mice immunized with detergent-solubilized antigen of purified virus, and 21 monoclonal (MC) antibodies reactive in enzyme-linked immunosorbent assay with the TO-163 strain of porcine transmissible gastroenteritis (TGE) virus were obtained. Of these MC antibodies, 14, 6 and 1 were IgG1, IgG2a and IgM, respectively. All of the MC antibodies contained light chains of the kappa type. Of these MC antibodies, 8 were found to have neutralization (NT) activity against the TO-163 strain. Comparison of 7 strains of TGE virus by NT tests using our panel of MC antibodies confirmed their close antigenic relationships, but also revealed the occurrence of distinct antigenic differences. These results suggest that there may be at least 6 different epitopes involved in NT reaction on the virion of the TO-163 strain. This notion was confirmed by the competitive binding assay.     

Rapid detection of canine parvovirus in feces using monoclonal antibodies and enzyme-linked immunosorbent assay

Monoclonal antibodies were used to develop a double antibody enzyme-linked immunosorbent assay for the detection of canine parvovirus (CPV) antigen in fecal samples. The assay was specific for the hemagglutinating protein of CPV and detected as little as 1.5 ng of virus within a 15-minute incubation period. The use of monoclonal antibodies against 2 epitopes on the CPV antigen permitted the simultaneous addition of test sample and enzyme-conjugated antibody, thus considerably simplifying the manipulations required for the assay. Results were visually determined without special instrumentation. Clinical studies revealed greater than 95% correlation between enzyme-linked immunosorbent assay results and hemagglutination titers.     

Dietary α‐linolenic acid affects lipid metabolism and tissue fatty acid profile and induces apoptosis in intraperitoneal adipose tissue of juvenile grass carp (Ctenopharyngodon idella)

A 56‐day feeding trial was conducted to elucidate the effects and mechanism action of dietary α‐linolenic acid (ALA, 18:3n‐3) on lipid accumulation and fatty acid profile of muscle, hepatopancreas and intraperitoneal fat (IPF) in juvenile grass carp using three isonitrogenous and isoenergetic semi‐purified diets containing 0.0% (control group), 1.0% and 2.0% ALA, respectively. The lowest intraperitoneal fat (IPF) ratio was found in 2.0% group. In the muscle, hepatopancreas and IPF, docosahexaenoic acid (DHA, 22:6n‐3) and eicosapentaenoic acid (EPA, 20:5n‐3) contents increased with the increase in dietary ALA. In the IPF, caspase 3, caspase 8 and caspase 9 showed the highest activities in 2.0% group, while the value of Bcl‐2/Bax (B‐cell leukaemia 2/Bcl‐2‐associated X protein) reached the lowest. Meanwhile, swelling of the IPF mitochondria was observed in 2.0% group. The gene expressions of fatty acid desaturase (FAD) and fatty acid elongase (ELO) in the hepatopancreas and muscle showed significantly higher levels in the treatment groups, whereas an opposite trend was existed in the IPF. Fatty acid synthase (FAS), sterol regulatory element binding protein‐1c (SREBP‐1c) in the IPF and hepatopancreas reached the lowest in 2.0% group. Overall, dietary ALA could promote n‐3 highly unsaturated fatty acids (HUFAs) synthesis and suppress the accumulation of lipid by decreasing the expression of related genes and promoting the apoptosis in IPF.