PDF(Pubmed)
Abstract：
Ditylenchus destructor is a migratory plant-parasitic nematode that severely harms many agriculturally important crops. The control of this pest is difficult, thus efficient strategies for its management in agricultural production are urgently required. Cathepsin L-like cysteine protease (CPL) is one important protease that has been shown to participate in various physiological and pathological processes. Here we decided to characterize the CPL gene (Dd-cpl-1) from D. destructor. Analysis of Dd-cpl-1 gene showed that Dd-cpl-1 gene contains a signal peptide, an I29 inhibitor domain with ERFNIN and GNFD motifs, and a peptidase C1 domain with four conserved active residues, showing evolutionary conservation with other nematode CPLs. RT-qPCR revealed that Dd-cpl-1 gene displayed high expression in third-stage juveniles (J3s) and female adults. In situ hybridization analysis demonstrated that Dd-cpl-1 was expressed in the digestive system and reproductive organs. Silencing Dd-cpl-1 in 1-cell stage eggs of D. destructor by RNAi resulted in a severely delay in development or even in abortive morphogenesis during embryogenesis. The RNAi-mediated silencing of Dd-cpl-1 in J2s and J3s resulted in a developmental arrest phenotype in J3 stage. In addition, silencing Dd-cpl-1 gene expression in female adults led to a 57.43% decrease in egg production. Finally, Dd-cpl-1 RNAi-treated nematodes showed a significant reduction in host colonization and infection. Overall, our results indicate that Dd-CPL-1 plays multiple roles in D. destructor ontogenesis and could serve as a new potential target for controlling D. destructor.
摘要：
Ditylenchus破坏因子是一种迁徙的植物寄生线虫，严重危害许多重要的农业作物。这种害虫的控制是困难的，因此，迫切需要有效的农业生产管理策略。组织蛋白酶L-样半胱氨酸蛋白酶（CPL）是一种重要的蛋白酶，已被证明参与各种生理和病理过程。在这里，我们决定表征来自D.destructor的CPL基因（Dd-cpl-1）。对Dd-cpl-1基因的分析表明,Dd-cpl-1基因含有一个信号肽,具有ERFNIN和GNFD基序的I29抑制剂结构域，和具有四个保守活性残基的肽酶C1结构域，显示与其他线虫CPL的进化保守性。RT-qPCR显示Dd-cpl-1基因在第三阶段青少年（J3s）和成年女性中显示出高表达。原位杂交分析表明Dd-cpl-1在消化系统和生殖器官中表达。通过RNAi在D.destructor的1细胞期卵中沉默Dd-cpl-1会导致发育严重延迟，甚至在胚胎发生过程中流产的形态发生。在J2s和J3s中RNAi介导的Dd-cpl-1沉默导致J3期的发育停滞表型。此外,在成年女性中沉默Dd-cpl-1基因表达导致产蛋量下降57.43％。最后,Dd-cpl-1RNAi处理的线虫显示宿主定植和感染的显著减少。总的来说,我们的结果表明，Dd-CPL-1在D.destructor本体发生中起着多种作用，可以作为控制D.destructor的新的潜在靶标。
