Gene sequence and protein sequence of pinewood nematode Bursaphelenchus xylophilus expansin diagnostic composition for pine wilt disease and pharmaceutical composition for preventing and treating pine wilt disease

摘要

The present invention relates to a novel gene sequence, a protein sequence, a composition for the diagnosis of pine wilt disease, a diagnostic kit, and a medicament for the treatment or prevention of pine wilt disease in a pine nematode, Bursapelene cusylophilus exopancin. The present inventors identified two novel expansin genes in the expression sequence tag database of pine tree cynomolgus vulgaris cusylophilus and named them Bx-EXPB2 and Bx-EXPB3. The new Bx-EXPBs gene codes for 150 amino acids, and the similarity of the coding sequence was 70.7-84.0% as compared to EXPB1 of the previously reported Versafelen cuzylophilus. Bx-EXPB2 and Bx-EXPB3 were clustered with Bx-EXPB1 and Bm-EXPB1, respectively, and formed an independent lineage with other nematode EXP genes. All identified Bx-EXPBs contain signaling peptides and are only expressed during the proliferative phase and appear to be secreted to facilitate nematode migration through the host by loosening the cell wall during infection. Quantitative real-time PCR analysis showed that the relative accumulation of Bx-EXPB3 mRNA was highest among the three Bx-EXPs tested and the degree of mRNA accumulation was as follows: Bx-EXPB3 Bx-EXPB2 Bx-EXPB1. Similarity modeling of Bx-EXPBs showed that the optimal template was the Bx-EXPB1 and Bx-EXPB2 and residues essential for catalytic activity, except for Bx-EXPB3, which is an EXLX1 protein of Bacillus subtilis. Taken together, Bx-EXPB1 and Bx-EXPB2 are involved in migration through plant tissues and appear to play a role in virulence.