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Open AccessArticle

Cathepsin L-like Cysteine Proteinase Genes Are Associated with the Development and Pathogenicity of Pine Wood Nematode, Bursaphelenchus xylophilus

by Qi Xue 1,2, Xiao-Qin Wu 1,2,*, Wan-Jun Zhang 1,2, Li-Na Deng 3 and Miao-Miao Wu 1,2
1
Co-Innovation Center for Sustainable Forestry in Southern China, College of Forestry, Nanjing Forestry University, Nanjing 210037, China
2
Jiangsu Key Laboratory for Prevention and Management of Invasive Species, Nanjing Forestry University, Nanjing 210037, China
3
Yancheng Institute of Technology, School of Ocean and Biological Engineering, Yancheng 224051, China
*
Author to whom correspondence should be addressed.
Int. J. Mol. Sci. 2019, 20(1), 215; https://doi.org/10.3390/ijms20010215
Received: 16 December 2018 / Accepted: 1 January 2019 / Published: 8 January 2019
(This article belongs to the Section Biochemistry)
The pine wood nematode (PWN), Bursaphelenchus xylophilus, is the pathogen of pine wilt disease (PWD), resulting in huge losses in pine forests. However, its pathogenic mechanism remains unclear. The cathepsin L-like cysteine proteinase (CPL) genes are multifunctional genes related to the parasitic abilities of plant-parasitic nematodes, but their functions in PWN remain unclear. We cloned three cpl genes of PWN (Bx-cpls) by rapid amplification of cDNA ends (RACE) and analyzed their characteristics using bioinformatic methods. The tissue specificity of cpl gene of PWN (Bx-cpl) was studied using in situ mRNA hybridization (ISH). The functions of Bx-cpls in development and pathogenicity were investigated using real-time quantitative PCR (qPCR) and RNA interference (RNAi). The results showed that the full-length cDNAs of Bx-cpl-1, Bx-cpl-2, and Bx-cpl-3 were 1163 bp, 1305 bp, and 1302 bp, respectively. Bx-cpls could accumulate specifically in the egg, intestine, and genital system of PWN. During different developmental stages of PWN, the expression of Bx-cpls in the egg stage was highest. After infection, the expression levels of Bx-cpls increased and reached their highest at the initial stage of PWD, then declined gradually. The silencing of Bx-cpl could reduce the feeding, reproduction, and pathogenicity of PWN. These results revealed that Bx-cpls play multiple roles in the development and pathogenic processes of PWN. View Full-Text
Keywords: Bursaphelenchus xylophilus; cathepsin L; gene expression; development; pathogenicity Bursaphelenchus xylophilus; cathepsin L; gene expression; development; pathogenicity
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Xue, Q.; Wu, X.-Q.; Zhang, W.-J.; Deng, L.-N.; Wu, M.-M. Cathepsin L-like Cysteine Proteinase Genes Are Associated with the Development and Pathogenicity of Pine Wood Nematode, Bursaphelenchus xylophilus. Int. J. Mol. Sci. 2019, 20, 215.

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