Virulence, resistance to magainin II, and expression of pectate lyase are controlled by the PhoP-PhoQ two-component regulatory system responding to pH and magnesium in Erwinia chrysanthemi 3937

Marker-exchanged mutants of phoP and phoQ of Erwinia chrysanthemi (Ech) strain 3937 became more sensitive to the cationic antimicrobial peptide (CAMP) magainin II than did the wild type at a low Mg²⁺ concentration and at either acidic or neutral pH. At high Mg²⁺ and acidic pH, only the phoQ mutant, but not the phoP mutant, became more sensitive to magainin II than did the wild type; both mutants were more sensitive at neutral pH. The hyperinduction of Pel synthesis in medium containing plant extracts and polygalacturonic acid (PGA) was confirmed in the wild type but not in the mutants at low Mg²⁺ and neutral pH. However, Pel was hyperinduced at high Mg²⁺ and neutral pH in these mutants but not in the wild type. Maceration was also greatly reduced by these mutants compared to the wild type when the inoculum was precultured and then resuspended in the medium with low Mg²⁺ at neutral pH. However, when bacteria were precultured and resuspended in the medium with high Mg²⁺ at neutral pH, severe maceration was observed in these mutants but not in the wild type. Thus, at low Mg²⁺, PhoP-PhoQ TCS seems to be stimulated for maceration and the hyperinduction of Pel synthesis. At high Mg²⁺, however, PhoP-PhoQ TCS may be repressed for these phenotypes, and PhoP may be controlled by a mechanism(s) other than PhoQ regulation.     

Characterization and genetic status of Banana bunchy top virus isolated from Okinawa, Japan

Isolates of Banana bunchy top virus (BBTV), which causes bunchy top disease in bananas, were collected in field surveys on seven islands in Okinawa Prefecture, Japan. From 44 banana samples, one isolate from each island was selected, and the DNA-1 and DNA-3 components were sequenced. Analysis of the major common region of DNA-1 showed that BBTV in Okinawa belongs to the Asian group of BBTV. DNA-1 and DNA-3 analysis revealed that Okinawan BBTV had a closer relationship with isolates from Taiwan and the Philippines than with some isolates from China and Vietnam. All the Okinawan BBTV isolates had high homology in the nucleotide sequences of DNA-1 and DNA-3 (%) because of a single, recent BBTV invasion of this area.The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under the accession numbers AB108449 – AB108458     

Virulence, accumulation of acetyl-coenzyme A, and pectate lyase synthesis are controlled by PhoP-PhoQ two-component regulatory system responding to organic acids of Erwinia chrysanthemi 3937

The PhoP-PhoQ two-components regulatory system is involved in the pathogenesis of animal, plant, and insect pathogenic bacteria in response to various environmental factors. To elucidate how this system contributes to the plant pathogenesis of Erwinia chrysanthemi 3937 (Ech 3937), marker-exchanged mutants of phoP and phoQ were constructed. Their role in the regulation of a major virulent factor, pectate lyase (Pel), in response to various organic acids was then tested. These mutants synthesized more Pel than did the wild type in the medium containing acetate or citrate as the sole source of carbon, but they synthesized less Pel than did the wild type in pyruvate or malate as the sole source of carbon. Synthesis of Pel did not differ in succinate, fumarate, or glycerol from the wild type. The phoP and phoQ mutants grown and resuspended in acetate or citrate also caused more maceration, and the wild type pretreated in pyruvate or malate caused more maceration than did the mutants. The level of intracellular acetyl-coenzyme A (acetyl-CoA) almost paralleled the synthesis of Pel in the wild type and in the mutants of the phoP and phoQ. These results suggested that acetyl-CoA may be involved in regulation of Pel synthesis through two-independent regulatory cascades via the PhoP-PhoQ system (in an opposite manner) in response to acetate/citrate and pyruvate/malate. However, ackA and pta genes, involved in the synthesis of acetyl-CoA in Escherichia coli, were not expressed as predicted on the basis of the level of acetyl-CoA. Thus there may be an additional regulation or pathway for the synthesis of acetyl-CoA in Ech 3937.     

Effects of inoculum density, leaf age, moisture, temperature, and wetness duration on black streak of edible burdock

Inoculum density, temperature, leaf age, and wetness duration were evaluated for their effects on the development of black streak (Itersonilia perplexans) on edible burdock (Arctium lappa L.) in a controlled environment. The effect of relative humidity (RH) on ballistospores production by I. perplexans was also evaluated. Symptoms of black streak on leaves increased in a linear fashion as the inoculum density of I. perplexans increased from 10² to 10⁶ ballistospores/ml. Rugose symptoms on young leaves were observed at densities of ≥10⁴ ballistospores/ml. Disease severity of I. perplexans in relation to leaf age followed a degradation curve when the leaves were inoculated with ballistospores. Disease severity was high in newly emerged leaves up to 5 days old, declined as leaf age increased to 29 days, and was zero when leaf age increased from 30 to 33 days. Disease development of edible burdock plants exposed to ballistospores of I. perplexans was evaluated at various combinations of temperature (10°, 15°, 20°, 25°C) and duration of leaf wetness (12, 24, 36, 48, and 72 h). Disease was most severe when plants were in contact with the ballistospore sources at 15° or 20°C. The least amount of disease occurred at 25°C regardless of wetness duration. Ballistospores required 24–36 h of continuous leaf wetness to cause visible symptoms by infection on edible burdock. Ballistospores production in infected lesions required at least 95.5% RH.     

Granulocyte colony-stimulating factor has no adverse effects on atherosclerotic lesions in high cholesterol-fed miniature Swine

Granulocyte colony-stimulating factor (G-CSF) is widely used to mobilize peripheral blood stem cells, and expected to restore cardiac function for patients with coronary artery diseases as a consequence of progression of atherosclerosis. Safety issues related to the administration of G-CSF to these patients, however, are still under study. The animal model for atherosclerosis was produced by feeding miniature swine a high-cholesterol diet for 3 months. G-CSF (5 or 10 microg/kg/day) was given to the animal model by daily subcutaneous injections for 10 days and 20 main arteries were evaluated pathologically. In addition, the general toxicological effects were studied on clinical signs, body weight, hematology, blood chemistry and pathology. In the G-CSF-treated groups, a variety of changes related to the major pharmacological activity of G-CSF including an increase in white blood cell (WBC) counts were observed. In many arteries, atherosclerotic lesions similar to Type I-V of the proposed classification by the American Heart Association were observed. No effects of the G-CSF treatment were seen on the histopathological findings, incidence, severity or distribution of atherosclerotic lesions. In addition, no infiltration of neutrophils to the lesions was observed. These findings suggest that the administration of G-CSF causes neither exacerbation or modification of atherosclerotic lesions nor adverse changes despite that a sufficient increase in WBC counts could be achieved in the peripheral blood.     

Effect of electric field in conditioned aversion response

The aim of the present study was to estimate whether rat sense exogenous electric field (EF) including one used in our previous studies. Employing a conditioned place aversion response paradigm based on an aversive behavior against light environment, alteration in both voluntary behavior of Wistar rat to a 50 Hz sinusoidal EF was examined. Following conditioning without EF, the times spent in white place in rats was significantly shortened (P<0.05). While, such changes were not shown in rats conditioned with EF. Thus, it was considered that the aversion response to light environment was interfered by exposure to EF. An interference in recognition of brightness via EF induced effect to visual system or in learning system via direct effect to central nerve system was considerable as a factor for EF-induced effect. In addition, it was remained that rat possibly sense exposure to EF as preferable. In order to confirm which factor functioned, further studies are needed.     

Expression of hedgehog family genes in the rat uterus during early pregnancy

Hedgehog (Hh) plays a pivotal role in various tissues during embryonic development, tissue homeostasis and tumorigenesis. In mammals, Hh exists in three homologs: Desert hedgehog (Dhh), Indian hedgehog (Ihh) and Sonic hedgehog (Shh). In this study, we cloned full-length cDNAs encoding Dhh and Ihh from the rat uterus. Their amino acid sequences have a high homology with those of the mouse and human. In addition, the changes of Hh gene expression in the rat uterus during early pregnancy were analyzed. The results showed that all three hedgehog mRNAs were detected in the rat uterus at the proestrus stage and during early pregnancy (1.5, 3.5, 5.5 and 7.5 days post coitus: dpc). Ihh mRNA expression varied and peaked at 3.5 dpc in the luminal and glandular epithelium. Expression was decreased on 5.5 dpc with the exception of sustained expression in the glandular epithelium. Despite such Ihh variability, the expressions of Dhh and Shh mRNA remained unchanged. This indicated that Ihh was mainly expressed in the rat uterus during early pregnancy. Moreover, the Hh target gene (glioma-associated oncogene homolog 1; Gli1) was also highly expressed at 3.5 dpc in the epithelium and periepithelial stroma in a manner similar to the temporal pattern of Ihh expression. This suggests that Ihh signaling axis play a role in the rat uterus during early pregnancy. In summary, our results elucidate that Ihh is a predominant Hh protein in the rat uterus during early pregnancy and that other Hhs have the potential to be expressed. This observation will help to elucidate the basic molecular mechanism of rat uterus during early pregnancy.     

Isolation and characterisation of Rhodococcus equi from submaxillary lymph nodes of wild boars (Sus scrofa)

Rhodococcus equi has been isolated from the submaxillary lymph nodes of domesticated pigs, but little is known about the presence of R. equi in wild boars. The aim of the study was the evaluation of the incidence of R. equi in wild boars and the characterisation of them. Of 482 submaxillary lymph nodes of wild boars shot in 39 settlements throughout Hungary, R. equi was isolated from 60 specimens, and plasmid types of 82 isolates were examined. The isolates were tested for the presence of 15-17-kDa (VapA) and 20-kDa virulence-associated protein antigen (VapB) genes by polymerase chain reaction (PCR). Plasmid DNAs were isolated and analysed by digestion with restriction endonucleases to estimate size and compare their polymorphisms. None of the 82 isolates contained vapA gene but 21 isolates (25.6%) were positive for vapB gene showing 827bp product of the expected size in the PCR amplification. Sixty-one strains (74.4%) did not contain plasmid. The 21 isolates of intermediate virulence contained virulence plasmids that were identified as types 1 (1 isolate), 5 (16 isolates), 21 (1 isolate), and three new distinct plasmid variants (1-1-1 isolate), respectively. On the basis of restriction digestion patterns of plasmid DNAs, we tentatively designated the new variants as types 25-27, respectively. The prevalence of R. equi strains of intermediate virulence among the isolates originated from the submaxillary lymph nodes of wild boars (25.6%) is very similar to those of domestic pigs (26.8%) in Hungary, and plasmid type 5 is the predominating one in both groups. This is the first report of isolation of VapB-positive R. equi from wild boars in the world.     

A genome‐wide association study for fat‐related traits computed by image analysis in Japanese Black cattle

The objective of this study was to identify genomic regions associated with fat‐related traits using a Japanese Black cattle population in Hyogo. From 1836 animals, those with high or low values were selected on the basis of corrected phenotype and then pooled into high and low groups (n = 100 each), respectively. DNA pool‐based genome‐wide association study (GWAS) was performed using Illumina BovineSNP50 BeadChip v2 with three replicate assays for each pooled sample. GWAS detected that two single nucleotide polymorphisms (SNPs) on BTA7 (ARS‐BFGL‐NGS‐35463 and Hapmap23838‐BTA‐163815) and one SNP on BTA12 (ARS‐BFGL‐NGS‐2915) significantly affected fat percentage (FAR). The significance of ARS‐BFGL‐NGS‐35463 on BTA7 was confirmed by individual genotyping in all pooled samples. Moreover, association analysis between SNP and FAR in 803 Japanese Black cattle revealed a significant effect of SNP on FAR. Thus, further investigation of these regions is required to identify FAR‐associated genes and mutations, which can lead to the development of DNA markers for marker‐assisted selection for the genetic improvement of beef quality.