School of Forestry, Northeast Forestry University, Harbin 150000, China.
School of Biological Science and Medical Engineering, State Key Laboratory of Bioelectronics, Southeast University, Nanjing 210096, China.
Pestic Biochem Physiol. 2023 Feb;190:105334. doi: 10.1016/j.pestbp.2022.105334. Epub 2022 Dec 31.
Bursaphelenchus xylophilus is the causative agent of pine wilt disease. It has caused devastating damage to ecosystems worldwide, owing to the characteristic of being widely spread and uncontrollable. However, the current methods of control are mainly based on pesticides, which can cause irreversible damage to the ecosystem. Therefore, the search for new drug targets and the development of environmentally friendly nematicides is especially valuable. In this study, three key genes of the xenobiotic detoxification pathways were cloned from B. xylophilus, which were subsequently subjected to bioinformatic analysis. The bioassay experiment was carried out to determine the concentration of matrine required for further tests. Subsequently, enzyme activity detection and three gene expression pattern analysis were performed on matrine treated nematodes. Finally, RNA interference was conducted to verify the functions carried out by the three genes in combating matrine. The results indicated that cytochrome P450 and glutathione S-transferase of B. xylophilus were activated by matrine, which induced high expression of BxCYP33C4, BxGST1, and BxGST3. After RNA interference of three genes of B. xylophilus, the sensitivity of B. xylophilus to matrine was increased and the survival rate of nematodes was reduced to various degrees in comparison to the control group. Overall, the results fully demonstrated that BxCYP33C4, BxGST1, and BxGST3 are valuable drug targets for B. xylophilus. Furthermore, the results suggested that matrine has value for development and exploitation in the prevention and treatment of B. xylophilus.
松材线虫是松材线虫病的病原体。它具有分布广泛且不可控的特点，对全球生态系统造成了毁灭性的破坏。然而，目前的控制方法主要基于农药，这会对生态系统造成不可逆转的损害。因此，寻找新的药物靶点和开发环保型杀线虫剂尤为有价值。本研究从松材线虫中克隆了三个外来化合物解毒途径的关键基因，并对其进行了生物信息学分析。通过生物测定实验确定了进一步测试所需的苦参碱浓度。随后，对苦参碱处理的线虫进行了酶活性检测和三个基因表达模式分析。最后，通过 RNA 干扰验证了这三个基因在对抗苦参碱中的功能。结果表明，苦参碱激活了松材线虫的细胞色素 P450 和谷胱甘肽 S-转移酶，诱导了 BxCYP33C4、BxGST1 和 BxGST3 的高表达。松材线虫三个基因的 RNA 干扰后，与对照组相比，松材线虫对苦参碱的敏感性增加，线虫的存活率降低到不同程度。总体而言，这些结果充分表明 BxCYP33C4、BxGST1 和 BxGST3 是松材线虫有价值的药物靶点。此外，结果表明苦参碱在防治松材线虫方面具有开发和利用的价值。
