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: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.
