松材线虫(PWN)引起松树枯萎病(PWD),这是世界上最具破坏性的松树病之一。细胞色素P450(CYP)催化萜类化合物的生物合成代谢,在所有生物体的次级代谢产物修饰中起重要作用。我们研究了Bx-cyp29A3的分子特征和生物学功能。生物信息学分析结果表明,Bx-cyp29A3具有跨膜结构域,可以与L(-)-香芹酮对接。基因表达模式表明,Bx-cyp29A3在0.2、0.4、0.6、0.8和1.0mg/mLL(-)-香芹酮溶液中表达。当L(-)-香芹酮溶液浓度为0.8mg/mL时,Bx-cyp29A3表达以剂量依赖性方式增加,并在暴露24小时达到峰值。然而,基因表达在36小时后达到0.6mg/mL的峰值。此外,RNA干扰(RNAi)表明Bx-cyp29A3在对L(-)-香芹酮的反应中起重要作用。在暴露24小时或暴露36小时后,Bx-cyp29A3敲除组的死亡率高于对照组的0.4、0.6、0.8和1.0mg/mL香芹酮溶液。总之,生物信息学提供了Bx-cyp29A3及其编码蛋白的结构特征和保守序列性质,为木偶拟南芥P450家族的研究提供了靶基因。基因沉默实验阐明了Bx-cyp29A3在木蝇免疫防御中的功能。本研究为筛选新的分子靶标,为木线菌的防治提供了依据。
Bursaphelenchus xylophilus (PWN) causes pine wilt disease (PWD), which is one of the most devastating pine diseases worldwide. Cytochrome P450 (CYP) catalyzes the biosynthetic metabolism of terpenoids and plays an important role in the modification of secondary metabolites in all living organisms. We investigated the molecular characteristics and biological functions of Bx-cyp29A3 in B. xylophilus. The bioinformatics analysis results indicated that Bx-cyp29A3 has a transmembrane domain and could dock with L(-)-carvone. The gene expression pattern indicated that Bx-cyp29A3 was expressed in 0.2, 0.4, 0.6, 0.8, and 1.0 mg/mL L(-)-carvone solutions. The Bx-cyp29A3 expression increased in a dose-dependent manner and peaked at 24 h of exposure when the L(-)-carvone solution concentration was 0.8 mg/mL. However, the gene expression peaked at 0.6 mg/mL after 36 h. Furthermore, RNA interference (
RNAi) indicated that Bx-cyp29A3 played an essential role in the response to L(-)-carvone. The mortality rates of the Bx-cyp29A3 knockdown groups were higher than those of the control groups in the 0.4, 0.6, 0.8, and 1.0 mg/mL carvone solutions after 24 h of exposure or 36 h of exposure. In summary, bioinformatics provided the structural characteristics and conserved sequence properties of Bx-cyp29A3 and its encoded protein, which provided a target gene for the
study of the P450 family of B. xylophilus. Gene silencing experiments clarified the function of Bx-cyp29A3 in the immune defense of B. xylophilus. This
study provides a basis for the screening of new molecular targets for the prevention and management of B. xylophilus.