The advent of next generation sequencing has revolutionized research approaches to biology by making entire genome sequences available and marking a new age in biology that has the potential to open innovative research avenues in various... more
The advent of next generation sequencing has revolutionized research approaches to biology by making entire genome sequences available and marking a new age in biology that has the potential to open innovative research avenues in various fields. Genome sequencing is now being applied in the fields of forest ecology and forest pathology, which previously had limited access to molecular techniques. One of the most advanced areas of progress is the study of "pine wilt disease", which is caused by the parasitic nematode, Bursaphelenchus xylophilus. The entire genome sequence of B. xylophilus was determined in 2011, and since then, proteomic studies have been conducted to understand the molecular basis of the parasitism and pathogenicity of B. xylophilus. These postgenomic studies have provided numerous molecular insights and greatly changed our understanding of the pathogenesis of pine wilt disease. Here, we review the recent advances in genomic and proteomic approaches that address some of the longstanding questions behind the pathogenesis of pine wilt disease and have identified future questions and directions in this regard.
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BACKGROUND: Biocontrol achieved by a single biocontrol agent is generally inconsistent under field conditions. The aim of the present study was to increase the competitiveness and efficacy of biocontrol agents by using them together with... more
BACKGROUND: Biocontrol achieved by a single biocontrol agent is generally inconsistent under field conditions. The aim of the present study was to increase the competitiveness and efficacy of biocontrol agents by using them together with cattle manure.RESULTS: The effects of antagonistic fungi [Aspergillus niger v. Teigh., Paecilomyces lilacinus (Thom) Samson and Penicillium chrysogenum Thom] and plant‐growth‐promoting rhizobacteria (PGPR) [Azotobacter chroococcum Beijer., Bacillus subtilis (Ehrenberg) Cohn and Pseudomonas putida (Trev.) Mig.] were assessed with cattle manure on the growth of tomato and on the reproduction of Meloidogyne incognita (Kof. & White) Chitwood. Application of antagonistic fungi and PGPR alone and in combination with cattle manure resulted in a significant increase in the growth of nematode‐inoculated plants. The highest increase (79%) in the growth of nematode‐inoculated plants was observed when P. putida was used with cattle manure, followed by use of P. lilacinus plus cattle manure. Paecilomyces lilacinus resulted in a high reduction in galling and nematode multiplication, followed by P. putida, B. subtilis, A. niger, A. chroococcum and P. chrysogenum. The combined use of P. lilacinus with cattle manure resulted in a maximum reduction in galling and nematode multiplication.CONCLUSION: Application of P. lilacinus or P. putida with cattle manure was useful to achieve greater biocontrol of M. incognita on tomato. Copyright © 2009 Society of Chemical Industry
Research Interests: Horticulture, Biology, Fungi, Biological Control, Medicine, and 15 morePlant diseases, Animals, Biocontrol, Pseudomonas putida, Plant Roots, Cattle, Plant Growth Promoting Rhizobacteria, PEST, Manure, Biological Pest Control, Pest Management Science, Meloidogyne Incognita, Lycopersicon esculentum, ENVIRONMENTAL SCIENCE AND MANAGEMENT, and cattle manure
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Research Interests: Botany, Zoology, Nematology, Biology, Population, and 7 moreFlora, Parasite, Food Quality, VECTOR, PEST, Rhizoctonia, and Thallophyta
SummaryFor diagnosis of pine wilt disease, a simple PCR‐RFLP method was developed to identify and to differentiate two similar nematode species, based on a living or preserved single specimen. Pinewood nematodes, Bursaphelenchus... more
SummaryFor diagnosis of pine wilt disease, a simple PCR‐RFLP method was developed to identify and to differentiate two similar nematode species, based on a living or preserved single specimen. Pinewood nematodes, Bursaphelenchus xylophilus, and Bursaphelenchus mucronatus were examined. A single nematode in 1 µl of distilled water was put on a glass slide. When the water had almost dried the nematode was crushed with a filter paper chip, 1.5 mm × 1.5 mm, with the aid of forceps. The filter paper chip containing nematode remains was immediately placed into PCR buffer as the DNA template. The primer set used was to amplify ribosomal DNA containing the inter‐transcribed spacer (ITS) 1, 5.8S and ITS2 regions. The PCR product was consistently obtained from a single nematode, and digesting the product with restriction endonuclease, Hinf I, enabled discrimination between B. xylophilus and B. mucronatus. This method was simple, convenient and definitive, and could successfully determine the pathogen in the diagnosis of pine wilt disease. This method was applicable also to nematode specimens preserved under various conditions except in the case of those preserved in aldehyde‐containing fixatives.
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... Although the dominant fungi were constantly detected over the experimental period, the composi-tion of fungal species slightly varied among seasons as reported in previous studies (Kobayashi et al., 1974, 1975; Fukushige and Futai,... more
... Although the dominant fungi were constantly detected over the experimental period, the composi-tion of fungal species slightly varied among seasons as reported in previous studies (Kobayashi et al., 1974, 1975; Fukushige and Futai, 1987; Kuroda and Ito, 1992). ...
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ABSTRACT The inside of Pinus thunbergii could be a reliable screening source for a useful agent in controlling plant disease. Isolation of endophytic actinomycetes from P. thunbergii and their potential as biocontrol agents against the... more
ABSTRACT The inside of Pinus thunbergii could be a reliable screening source for a useful agent in controlling plant disease. Isolation of endophytic actinomycetes from P. thunbergii and their potential as biocontrol agents against the plant pathogen Cylindrocladium sp. were investigated. Two endophytic actinomycetes, Streptomyces sp. and Microbispora sp., were isolated from surface-sterilised root tissues of P. thunbergii seedlings. The recovery test of these two endophytic actinomycetes from pine seedling showed that Streptomyces sp. was isolated from only roots, but Microbispora sp. was isolated from both roots and leaves. Thus, Microbispora sp. is able to move to leaves from roots. Moreover, we evaluated the potential of both strains as biocontrol agents against Cylindrocladium sp. Two weeks after inoculation of Cylindrocladium sp. alone, pine seedlings showed a 50% mortality rate. Co-inoculation of Cylindrocladium sp. and Microbispora sp. did not affect seedling mortality rate. However, inoculation with both Cylindrocladium sp. and Streptomyces sp. reduced seedling mortality to 12%. Streptomyces sp. could be a useful agent in controlling pine disease caused by Cylindrocladium sp. Thus, it seems that Streptomyces sp. may induce a local host defence reaction and Microbispora sp. systemically spreads to aerial parts through the transpiration stream.
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Bursaphelenchus sinensis was isolated from a dead Japanese black pine, Pinus thunbergii in Japan. The Japanese population of B. sinensis was compared with the original descriptions of a B. sinensis population from China and of the close... more
Bursaphelenchus sinensis was isolated from a dead Japanese black pine, Pinus thunbergii in Japan. The Japanese population of B. sinensis was compared with the original descriptions of a B. sinensis population from China and of the close relative, B. aberrans from China. The molecular profiles of B. sinensis were determined by DNA sequencing and ITSRFLP analyses, and were compared with those of the other species in the genus. By morphological observations, several additional characters, such as a three-celled structure at the junction of the uterus and post-uterine sac, which has been reported in several Bursaphelenchus species belonging to various phylogenetic groups, were found in B. sinensis. The phylogenetic analysis of the small subunit of ribosomal DNA indicated that B. sinensis is included in an inner clade of the genus Bursaphelenchus; however, its precise phylogenetic position within the genus was not defined clearly.
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Research Interests: Zoology, Nematology, Biology, Virulence, Pathogen, and 5 moreNematode, Inoculation, Fungus, Wilt disease, and Host resistance
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Three- or 4-year-old Japanese black pine seedlings were exposed to simulated acid rain (SAR) at pH 3 for two months, then inoculated with a virulent isolate (S 10) of pinewood nematodes. The experiments were repeated three times in... more
Three- or 4-year-old Japanese black pine seedlings were exposed to simulated acid rain (SAR) at pH 3 for two months, then inoculated with a virulent isolate (S 10) of pinewood nematodes. The experiments were repeated three times in 1996–1998. The exposure to SAR killed no seedlings, and retarded the development of disease symptoms in the seedlings inoculated with nematodes. In
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Research Interests: Botany, Zoology, Nematology, Life history, Biology, and 7 moreDispersion, Parasite, VECTOR, PEST, Cerambycidae, Nematode, and Wilt disease
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Research Interests: Biology and Wilt disease
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The behavioral role of male ambrosia beetles, Platypus quercivorus , in subsocial colonies both field and laboratory has been investigated. The entrance tunnels, where the male beetles are staying, are short, with a mean 4 cm long and... more
The behavioral role of male ambrosia beetles, Platypus quercivorus , in subsocial colonies both field and laboratory has been investigated. The entrance tunnels, where the male beetles are staying, are short, with a mean 4 cm long and mostly incline upwards from outside to inside at an angle of around 20o. To examine the role of males, another male or female (as the invader) was placed into a tunnel. When inhabitant males stayed in the tunnel they quickly expelled the invaders, regardless of their sex of the invaders. However, when an inhabitant male was removed an introduced male or female could freely enter the tunnel and was accepted by the inhabitant female. Upon replacing an inhabitant male with an invader male or female and then putting another invader male or female into the tunnel, no rejection occurred, suggesting that invader males and females play no role in guarding the tunnel. Based on the results an inhabitant male seems to have three responsibilities; protection of th...
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Pine wilt disease, caused by the pinewood nematode, Bursaphelenchus xylophilus, is one of the world’s most serious tree diseases. Although the B. xylophilus whole-genome sequence and comprehensive secretome profile have been determined... more
Pine wilt disease, caused by the pinewood nematode, Bursaphelenchus xylophilus, is one of the world’s most serious tree diseases. Although the B. xylophilus whole-genome sequence and comprehensive secretome profile have been determined over the past decade, it remains unclear what molecules are critical in pine wilt disease and govern B. xylophilus virulence in host pine trees. Here, a comparative secretome analysis among four isolates of B. xylophilus with distinct virulence levels was performed to identify virulence determinants. The four candidate virulence determinants of B. xylophilus highly secreted in virulent isolates included lipase (Bx-lip1), glycoside hydrolase family 30 (Bx-GH30), and two C1A family cysteine peptidases (Bx-CAT1 and Bx-CAT2). To validate the quantitative differences in the four potential virulence determinants among virulence groups at the protein level, we used real-time reverse-transcription polymerase chain reaction analysis to investigate these determ...
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The advent of next generation sequencing has revolutionized research approaches to biology by making entire genome sequences available and marking a new age in biology that has the potential to open innovative research avenues in various... more
The advent of next generation sequencing has revolutionized research approaches to biology by making entire genome sequences available and marking a new age in biology that has the potential to open innovative research avenues in various fields. Genome sequencing is now being applied in the fields of forest ecology and forest pathology, which previously had limited access to molecular techniques. One of the most advanced areas of progress is the study of "pine wilt disease", which is caused by the parasitic nematode, Bursaphelenchus xylophilus. The entire genome sequence of B. xylophilus was determined in 2011, and since then, proteomic studies have been conducted to understand the molecular basis of the parasitism and pathogenicity of B. xylophilus. These postgenomic studies have provided numerous molecular insights and greatly changed our understanding of the pathogenesis of pine wilt disease. Here, we review the recent advances in genomic and proteomic approaches that address some of the longstanding questions behind the pathogenesis of pine wilt disease and have identified future questions and directions in this regard.