Loop-mediated isothermal amplification of bacterial effector genes to detect Pseudomonas syringae pv. actinidiae biovars 1 and 3

Journal of General Plant Pathology - Loop-mediated isothermal amplification (LAMP) was designed to rapidly detect biovars 1 and 3 of Pseudomonas syringae pv. actinidiae (Psa), a serious, global...     

Phylogenetic and serological analyses reveal genetic diversity of Agrobacterium vitis strains in Japan

Twenty-eight strains of Agrobacterium vitis , including both tumorigenic and nonpathogenic phenotypes involving 26 isolated in Japan and strains NCPPB 3554^T and NCPPB 2562 isolated in Australia and Greece, respectively, were characterized by means of a slide agglutination test (SAT) using antisera prepared against somatic antigens. Phylogenetic analyses were also carried out, using the results of repetitive sequence-based polymerase chain reaction and the partial nucleotide sequences of pyrG , recA and rpoD . The A. vitis strains separated into four serogroups (A to D) based on the SAT reactivity. The phylogenetic analyses showed A. vitis strains separated into four tumorigenic groups (A to D) and one nonpathogenic group (E). Serogroups A to C corresponded exactly to genetic groups A to C, respectively, whereas serogroup D further divided into two distinct genetic groups, D and E. Genetic group E was isolated in Okayama Prefecture, Japan, and all strains of it have been found to coexist with tumorigenic strains belonging to the other groups within the same galled grapevine tissues. These results suggest that A. vitis strains are genetically heterogeneous and can be separated into several genetic groups. The differences between the nucleotide sequences of pyrG , recA and rpoD of the genetic groups will enable the development of a simple and convenient monitoring method, which will increase understanding of the dynamics of each genetic group of A. vitis in the natural environment.     

Gray mold of pearl lupine caused by <Emphasis Type="Italic">Botrytis cinerea</Emphasis>

Severe blight of stems, leaves and pods caused by Botrytis cinerea was found on pearl lupine (Lupinus mutabilis), a legume crop, grown in Kagawa Prefecture, Japan, in March–June 1996–2002. This disease was named “gray mold of pearl lupine” as a new disease. One of the fungal isolates obtained in this study was deposited in Genebank, National Institute of Agrobiological Sciences as accession MAFF238557.     

A method for predicting the occurrence of paralytic shellfish poisoning along the coast of Hokkaido in the Okhotsk Sea in summer

To design a method for predicting outbreaks of paralytic shellfish poisoning (PSP) in scallop fishing grounds, the relationship between the distribution of the toxic dinoflagellate Alexandrium tamarense and the dynamics of the Soya Warm Current (SWC) was examined in the Okhotsk Sea off Hokkaido. Surveys were conducted from May to June to clarify the transportation mechanism of A.?tamarense from the oceanic area to the coastal area. The sea-level difference (SLD) between Wakkanai and Abashiri was monitored as an index of the strength of the SWC southeastward flow in an alongshore belt to examine the possible occurrence of A.?tamarense in the coastal area during temporal weakening of the SWC. A bottom-mounted acoustic Doppler current profiler (ADCP) was used for direct observations of the SWC. The results indicated that PSP occurred when low-salinity water contaminated with A.?tamarense extended to the coast during temporal weakening of the SWC due to a decrease of the SLD. Our results strongly indicate that predictions can be realized by monitoring the decrease of SLD as an index of temporal weakening of the SWC after surveys of the distribution of A.?tamarense in the oceanic area before the period of PSP occurrence.     

Promotion of initial swimbladder inflation in Pacific bluefin tuna,Thunnus orientalis (Temminck and Schlegel), larvae

Initial swimbladder inflation (ISI) of Pacific bluefin tuna (PBT), Thunnus orientalis, larvae was studied to increase the survival of cultured larvae. Experiment 1 was conducted to explore promotion and inhibition of ISI under different water surface conditions; including the use of surface skimmer to remove autogenous surface substances (SS), covering the water surface with liquid‐paraffin‐layer (LP) and oil film (OF), and a control (non‐treatment, NT). Significantly higher inflation frequency was observed in SS (62.2%) than NT (11.9%), LP (2.7%) and OF (3.9%). This indicates that ISI in PBT larvae can be promoted by removal of surface substances on rearing water which inhibit larval air gulping. Experiment 2 aimed to elucidate proper day of larval age to start skimming for promoting ISI with four different periods of oil film removal: from 3 to 8 (SF3D), 4 to 8 (SF4D), 5 to 8 (SF5D), 6 to 8 (SF6D) days‐post‐hatch (dph). Significant improvement in ISI frequency was observed in SF3D (80.2%) but the frequency was very poor in SF4D, SF5D, and SF6D (17.8–7.5%). This implies the need of oil film removal without missing a narrow window, 1 day of 3 dph, to promote ISI in practical PBT larviculture.     

Effect of water surface condition on survival,growth and swim bladder inflation of yellowfin tuna,Thunnus albacares (Temminck and Schlegel), larvae

Early‐stage mortality due to surface water tension‐related death and due to sinking to the tank bottom was investigated for yellowfin tuna, Thunnus albacares (YFT), larvae. Different aeration rates and rearing water surface conditions were examined to evaluate the effect on larval survival, swim bladder inflation and growth. The percentage survival of yolk sac larvae was significantly higher when the rearing water surface was covered with fish oil at aeration rates of 0 and 50 mL min^?1. The highest mortality occurred at the highest aeration rate of 250 mL min^?1 regardless of surface water condition. A second experiment was conducted twice under different water surface conditions: the water surface was covered by fish oil (FO), skimmed of fish oil (SS), and was not treated (NC). The percentage survival was not significantly different between treatments after 7 days of feeding. In contrast to the survival, the proportion of larvae with inflated swim bladders was significantly higher for the NC and SS groups than that of the FO group. Results of these experiments indicate that the addition of oil to the rearing water surface without its removal interferes with the initial swim bladder inflation in YFT larvae. These results also indicate that YFT larvae need to obtain (gulp) air at the water surface for initial swim bladder inflation, and success of initial swim bladder inflation may be crucial for their survival.     

Effect of temperature on survival,growth and malformation of cultured larvae and juveniles of the seven-band grouper Epinephelus septemfasciatus

The effects of rearing temperature (23–29 °C) during the larval and juvenile stage on survival, growth and skeletal malformations in the seven-band grouper Epinephelus septemfasciatus were investigated. The survival rate of juveniles 30–40 mm in total length emerging from eggs was higher at 25 and 26 °C (0.1–1.3 %) than at 23 °C or 27–29 °C (0.004–1.5 %). Growth (increase in total length) was accelerated at higher temperatures. The frequency of malformed individuals was lower at 25–27 °C (36.0–61.5 %) than at 23, 28 or 29 °C (65.3–76.9 %). Specific incidences of spinal curvature and centrum fusion or defects in juveniles were not related to rearing temperature. However, incidences of twisted or compressed vertebrae (6.5–64.0 %) were higher at higher temperatures, while the incidence of bifurcated neural spine was significantly higher at 23 °C (43.6–54.4 %) than at other temperatures (3.3–22.7 %). The incidence rate of spinal curvature (23.3 %) was significantly higher in juveniles with a deflated swim bladder, regardless of rearing temperature. The results of this study suggest that the optimum culture temperature for seven-band grouper is 25–26 °C, collectively considering the survival, growth and incidences of abnormalities. Our results also demonstrate the significance of identifying the conditions for swim bladder inflation to prevent spinal curvature in seven-band grouper.     

Genetic population structure of the Pacific bluefin tuna Thunnus orientalis and the yellowfin tuna Thunnus albacares in the North Pacific Ocean

The genetic population structure of the Pacific bluefin tuna (PBF) Thunnus orientalis and the yellowfin tuna (YFT) T. albacares in the North Pacific Ocean was investigated. The polymorphism of microsatellite (SSR) loci and sequences of mitochondrial DNA control region (mtCR) were analyzed for 71 samples of PBF from Japan and Mexico and 45 samples of YFT from Japan and Panama. In the SSR analyses, both single-locus (?0.010 to 0.008 in PBF and ?0.023 to 0.020 in YFT) and global multilocus (0.003 in PBF and ?0.002 in YFT) F _ST values among the geographic populations were low and not significant in these species. In the mtCR analyses, neither the neighbor-joining tree nor the minimum spanning network showed genetic differentiation among the geographic populations in each species. The pairwise F _ST values among the geographic populations of them (?0.005 in PBF and ?0.020 to ?0.014 in YFT) were low and not significant. Our SSR and mtCR data suggested that genetic differentiations were not evident among the eastern and western populations in the North Pacific Ocean either in PBF or in YFT. Mismatch distributions, demographic parameters, and neutrality tests suggested that sudden population expansion of PBF and YFT in the North Pacific Ocean occurred 628,000–731,000 and 450,000–525,000 years ago, respectively.     

Model studies on precursor system generating blue pigment in onion and garlic

Reactions involved in blue-green discoloration in a mixture of onion (Allium cepa L.) and garlic (Allium sativum L.) were investigated. Vivid-blue color was successfully reproduced by using a defined model reaction system comprising only trans-(+)-S-(1-propenyl)-L-cysteine sulfoxide (1-PeCSO) from onion, S-allyl-L-cysteine sulfoxide (2-PeCSO) from garlic, purified alliinase (EC 4.4.1.4), and glycine (or some other amino acids). Four reaction steps identified and factors affecting the blue color formation were in good agreement with those suggested by earlier investigators. When crude onion alliinase was used in place of garlic alliinase, less pigment was formed. This result was explained by a difference in the amount of thiosulfinates, colorless intermediates termed color developers, yielded from 1-PeCSO by these enzymes.     

Antimicrobial resistance, virulence profiles, and phylogenetic groups of fecal Escherichia coli isolates: a comparative analysis between dogs and their owners in Japan

In this study, fecal Escherichia coli isolates (n=188) from 34 dog-owner pairs and 26 healthy control humans (2 isolates per individual) were tested for susceptibility to 6 antimicrobials and screened for virulence genes. Genetic diversity between canine and owner isolates was evaluated by pulsed-field gel electrophoresis (PFGE). Canine isolates exhibited significantly different rates of resistance to four and two antimicrobials, compared to control and owner isolates, respectively. Of the genes examined, the prevalence of sfa, hly, and cnf genes in canine isolates were higher than in control isolates, but not than in owner isolates. These results suggest that characteristics of owner isolates are somewhat similar to canine isolates, compared to isolates from non-dog owners. In addition, PFGE analysis revealed that transfer of E. coli between owners and their dogs had occurred within 3/34 (8.8%) households. Considering the effects of dog ownership on the population of E. coli isolates from owners, further epidemiological studies are required.