Abstract
The pinewood nematode, Bursaphelenchus xylophilus, recognized as a worldwide major forest pest, is a migratory endoparasitic nematode with capacity to feed on pine tissues and also on fungi colonizing the trees. Bursaphelenchus mucronatus, the closest related species, differs from B. xylophilus on its pathogenicity, making this nematode a good candidate for comparative analyses. Secretome profiles of B. xylophilus and B. mucronatus were obtained and proteomic differences were evaluated by quantitative SWATH-MS. From the 681 proteins initially identified, 422 were quantified and compared between B. xylophilus and B. mucronatus secretomes and from these, 243 proteins were found differentially regulated: 158 and 85 proteins were increased in B. xylophilus and B. mucronatus secretomes, respectively. While increased proteins in B. xylophilus secretome revealed a strong enrichment in proteins with peptidase activity, the increased proteins in B. mucronatus secretome were mainly related to oxidative stress responses. The changes in peptidases were evaluated at the transcription level by RT-qPCR, revealing a correlation between the mRNA levels of four cysteine peptidases with secretion levels. The analysis presented expands our knowledge about molecular basis of B. xylophilus and B. mucronatus hosts interaction and supports the hypothesis of a key role of secreted peptidases in B. xylophilus pathogenicity.
Highlights
The pinewood nematode (PWN), Bursaphelenchus xylophilus, an European Plant Protection Organization A2 pest[1], is an invasive species responsible for the development of the pine wilt disease (PWD) and is recognized worldwide as a major forest pest
The entire reads set obtained for B. mucronatus transcriptome and used for the final assembly was submitted to the EMBL-EBI European Nucleotide Archive (ENA), under the study accession number PRJEB14884
A database of B. xylophilus and B. mucronatus transcriptomic data was produced and used for the identification of proteins secreted by these two nematodes under pine tree extract stimulation
Summary
The pinewood nematode (PWN), Bursaphelenchus xylophilus, an European Plant Protection Organization A2 pest[1], is an invasive species responsible for the development of the pine wilt disease (PWD) and is recognized worldwide as a major forest pest. Migration and pine cell destruction abilities are important factors affecting these two species different pathogenicity and recent studies showed that the number and area of dead epithelial cells in pine cuttings inoculated with B. mucronatus were smaller than in those inoculated with B. xylophilus, suggesting that the attacking ability of B. mucronatus is weaker than that of B. xylophilus[18]. SWATH-MS was used to determine changes in protein amounts between B. xylophilus and B. mucronatus secretions, bringing new insights into the molecular basis of these nematodes interaction with their hosts and PWN pathogenicity
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