Co-Innovation Center for Sustainable Forestry in Southern China, College of Forestry, Nanjing Forestry University, Nanjing 210037, China.
Jiangsu Key Laboratory for Prevention and Management of Invasive Species, Nanjing Forestry University, Nanjing 210037, China.
Int J Mol Sci. 2019 Jan 8;20(1):215. doi: 10.3390/ijms20010215.
The pine wood nematode (PWN), , 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 genes of PWN () by rapid amplification of cDNA ends (RACE) and analyzed their characteristics using bioinformatic methods. The tissue specificity of gene of PWN () was studied using in situ mRNA hybridization (ISH). The functions of s 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 , , and were 1163 bp, 1305 bp, and 1302 bp, respectively. s could accumulate specifically in the egg, intestine, and genital system of PWN. During different developmental stages of PWN, the expression of s in the egg stage was highest. After infection, the expression levels of s increased and reached their highest at the initial stage of PWD, then declined gradually. The silencing of could reduce the feeding, reproduction, and pathogenicity of PWN. These results revealed that s play multiple roles in the development and pathogenic processes of PWN.
松材线虫（PWN）是松材线虫病（PWD）的病原体，导致松林遭受巨大损失。然而，其致病机制尚不清楚。组织蛋白酶 L 样半胱氨酸蛋白酶（CPL）基因是与植物寄生线虫寄生能力相关的多功能基因，但它们在 PWN 中的功能尚不清楚。我们通过快速扩增 cDNA 末端（RACE）克隆了三个 PWN 的 CPL 基因（），并使用生物信息学方法分析了它们的特征。使用原位 mRNA 杂交（ISH）研究了 PWN 的基因的组织特异性。使用实时定量 PCR（qPCR）和 RNA 干扰（RNAi）研究了 s 在发育和致病性中的功能。结果表明，、和的全长 cDNA 分别为 1163bp、1305bp 和 1302bp。s 可以特异性地在 PWN 的卵、肠和生殖系统中积累。在 PWN 的不同发育阶段，s 在卵期的表达最高。感染后，s 的表达水平增加，并在 PWD 的初始阶段达到最高，然后逐渐下降。沉默可以降低 PWN 的取食、繁殖和致病性。这些结果表明 s 在 PWN 的发育和致病过程中发挥多种作用。
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