Project description:This study compared the different gene expression of Bursaphelenchus xylophilus in two growth conditions (growing on Botrytis cinerea and inoculating Pinus thunbergii). The goal was to analyze the specifically-expressed genes of the pine wood nematode involved in the early interaction between B. xylophilus and P. thunbergii and screen the pathogenesis related genes of B. xylophilus.
Project description:This study compared the different gene expression of Bursaphelenchus xylophilus in two growth conditions (growing on Botrytis cinerea and inoculating Pinus thunbergii). The goal was to analyze the specifically-expressed genes of the pine wood nematode involved in the early interaction between B. xylophilus and P. thunbergii and screen the pathogenesis related genes of B. xylophilus. Two-condition experiment, Growing on Botrytis cinerea vs. Inoculating Pinus thunbergii . Biological replicates: 3 replicates.
Project description:The pinewood nematode, Bursaphelenchus xylophilus, the pine wilt disease's causal agent, is a migratory endoparasitic nematode skilled to feed on pine tissues and on fungi that colonize the trees. In order to study B. xylophilus secretomes under the stimulus of pine species with different susceptibility to disease, nematodes were exposed to aqueous pine extracts from Pinus pinaster (high susceptible host) and P. pinea (low susceptible host). Sequential windowed acquisition of all theoretical mass spectra (SWATH-MS) was used to determine relative changes in protein amounts between B. xylophilus secretions, and a total of 776 secreted proteins were quantified in both secretomes.
Project description:Bursaphelenchus xylophilus is known as the causative agent of pine wilt disease with complex life cycles. In this research, newly published Bursaphelenchus xylophilus genome data were employed to annotate its miRNAs based on deep sequencing technologies. Four small RNA libraries derived from different infection stages of pine wilt disease were constructed and sequenced. Consequently, we obtained hundreds of evolutionarily conserved miRNAs as well as novel miRNA candidates. The analysis of miRNA expression patterns showed that most miRNAs were expressed at extraordinarily high levels during the middle stage of pine wilt disease. Subsequent stem-loop RT-PCR experiments were carried out to validate our results. Functional analysis proved that expression levels of miR-73 and miR-239 were mutually exclusive with their target GH45 cellulase genes., genes known to be responsible for the degradation of the pine cell walls. In addition, another set of atypical miRNAs, termed mirtrons, were identified from B. xylophilus introns. This discovery has expanded the current knowledgebase of such splicing-derived miRNAs into B. xylophilus. Thus, our research has provided detailed characterization of B. xylophilus miRNAs expression patterns during the pathological process of pine wilt disease. The findings will contribute to more in-depth understanding of this devastating plant disease. For the purposes of this study, we classified the pathogenic process associated with PWD into three stages in order to best characterize the expression patterns of microRNAs during the development of this devastating disease. The following describes the first stage (F): about seven days after pine trees are infected with PWNs, the tips of the pine needles begin to turn brown. Next, the middle stage (M) ensues approximately seven days later, when half of the needles on pine trees turn brown. The last stage (L) occurs another 10 days later and pine needles are complete browning. PWNs cultured on Botrytis cinerea grown on PDA medium served as the control stage (C).
Project description:to profile the adverse effects of an emamectin benzoate trunk-injection agent on pine wood nematode Bursaphelenchus xylophilus by analysing differential transcripts from the nematode whole genome through next-generation high-throughput sequencing.
Project description:DNA methylation is a pivotal process that regulates gene expression and facilitates rapid adaptation to challenging environments. The pine wood nematode (PWN; Bursaphelenchus xylophilus), the causative agent of pine wilt disease, survives and spreads at low temperatures through third-stage dispersal larvae, making it a major pathogen for pine wood in Asia. To comprehend the impact of DNA methylation on the formation and environmental adaptation of third-stage dispersal larvae, we conducted whole-genome bisulfite sequencing and transcriptional sequencing on both the third-stage dispersal larvae and three other stages propagative larvae of PWN.
Project description:Bursaphelenchus xylophilus is known as the causative agent of pine wilt disease with complex life cycles. In this research, newly published Bursaphelenchus xylophilus genome data were employed to annotate its miRNAs based on deep sequencing technologies. Four small RNA libraries derived from different infection stages of pine wilt disease were constructed and sequenced. Consequently, we obtained hundreds of evolutionarily conserved miRNAs as well as novel miRNA candidates. The analysis of miRNA expression patterns showed that most miRNAs were expressed at extraordinarily high levels during the middle stage of pine wilt disease. Subsequent stem-loop RT-PCR experiments were carried out to validate our results. Functional analysis proved that expression levels of miR-73 and miR-239 were mutually exclusive with their target GH45 cellulase genes., genes known to be responsible for the degradation of the pine cell walls. In addition, another set of atypical miRNAs, termed mirtrons, were identified from B. xylophilus introns. This discovery has expanded the current knowledgebase of such splicing-derived miRNAs into B. xylophilus. Thus, our research has provided detailed characterization of B. xylophilus miRNAs expression patterns during the pathological process of pine wilt disease. The findings will contribute to more in-depth understanding of this devastating plant disease.
Project description:DNA methylation is a pivotal process that regulates gene expression and facilitates rapid adaptation to challenging environments. The pine wood nematode (PWN; Bursaphelenchus xylophilus), the causative agent of pine wilt disease, survives and spreads at low temperatures through third-stage dispersal larvae, making it a major pathogen for pine wood in Asia. To comprehend the impact of DNA methylation on the formation and environmental adaptation of third-stage dispersal larvae, we conducted whole-genome bisulfite sequencing and transcriptional sequencing on both the third-stage dispersal larvae and three other stages propagative larvae of PWN.
Project description:Bursaphelenchus xylophilus (pinewood nematode, PWN) is a causal agent of pine wilt disease and results in economic and environmental losses in pine forests. The establishment of systemic acquired resistance (SAR) provides positive capacities to control PWN. We selected two SAR elicitors, acibenzolar-S-methyl (ASM) and methyl salicylic acid (MeSA), which effectively inhibited disease symptoms on PWN-infected pine trees. To understand dynamic interactions between pine host and PWN under SAR state, we characterized in vivo transcriptomes of pine trees infected by B. xylophilus according to the ASM and MeSA treatment. After distilled water treatment, pine trees infected by B. xylophilus was used as a negative control.
