松材线虫谷胱甘肽S转移酶基因响应阿维菌素胁迫功能研究  

Functional study of the glutathione S-transferase gene of Bursaphelenchus xylophilus in response to abamectin exposion

作　　者：谭瑞娜 郝昕[1] 邓振 理永霞[3,4] 曹景鑫 苏光祥 陈洁 闵梦茹 马玲[1] TAN Ruina;HAO Xin;DENG Zhen;LI Yongxia;CAO Jingxin;SU Guangxiang;CHEN Jie;MIN Mengru;MA Ling(School of Forestry,Northeast Forestry University,Harbin 150040,Heilongjiang,China;College of Plant Protection,Nanjing Agricultural University,Nanjing 210095,Jiangsu,China;Laboratory of Forest Pathogen Integrated Biology Research Institute of Forestry New Technology,Chinese Academy of Forestry,Beijing 100091,China;Co-Innovation Center for Sustainable Forestry in Southern China,Nanjing Forestry University,Nanjing 210037,Jiangsu,China)

机构地区：[1]东北林业大学林学院,黑龙江哈尔滨150040 [2]南京农业大学植物保护学院,江苏南京210095 [3]中国林业科学研究院林业新技术研究所森林病原整合生物学研究室,北京100091 [4]南京林业大学南方现代林业协同创新中心,江苏南京210037

出　　处：《中南林业科技大学学报》2023年第8期159-168,共10页Journal of Central South University of Forestry & Technology

基　　金：中央高校基本科研业务费专项(2572022DP03,2572021AW25,2572021AW20),国家重点研发计划项目(2021YFD1400904)。

摘　　要：【目的】由松材线虫Bursaphelenchusxylophilus引起的松材线虫病严重破坏东亚和欧洲等多个国家和地区的森林生态系统。【方法】本研究从松材线虫基因组中获取Bx-gst12,随后对该基因的全长CDS区域进行基因克隆和序列分析、同时通过生物信息学方法对Bx-gst12所编码的蛋白质Bx-GST12的理化性质、亲疏水性、跨膜区、二级结构和三级结构进行了预测和分析。并通过基因干扰技术对Bx-gst12干扰后,分析该基因在松材线虫对阿维菌素敏感程度的影响。【结果】生物信息学结果显示Bx-GST12蛋白稳定系数为28.96,亲水系数为-0.412,三级结构预测Bx-GST12蛋白具有含有9个α螺旋和5个β折叠,但不具有跨膜结构域。应用基因干扰技术对Bx-gst12基因进行基因干扰,干扰后的Bx-gst12基因表达量变为原来的25.14%。阿维菌素溶液生物测定实验结果表明,Bx-gst12干扰后的松材线虫死亡率明显高于对照组,松材线虫死亡率平均提高了7.12%。【结论】本研究成功克隆Bx-gst12基因并对该基因的dsRNA进行了设计与合成。Bx-gst12基因干扰显著影响了松材线虫对阿维菌素的敏感性,在相同浓度的阿维菌素胁迫相同时间中,干扰组线虫死亡率明显高于对照组,表明Bx-gst12基因在松材线虫药物代谢过程中发挥着显著作用。【Objective】Pine wood disease caused by Bursaphelenchus xylophilus has devastated forest ecosystems in East Asia and Europe.【Method】In this study,Bx-gst12 was obtained from the genome,then this gene was cloned and analyzed,and bioinformatics were used to predict the physicochemical properties,hydrophilicity,transmembrane regions,secondary and tertiary structures of Bx-GST12.The effect of Bx-gst12 on the sensitivity to abamectin was also analyzed by RNAi.【Result】Bioinformatic analysis showed that Bx-GST12 has a stability factor of 28.96 and a hydrophilicity factor of-0.412.The tertiary structure prediction showed that Bx-GST12 has 9 alpha helices and 5 beta folds,but there is no transmembrane structural domain.RNAi was applied to the gene disruption of Bx-gst12,and the expression of Bx-gst12 was decreased to 25.14%of the original expression after RNAi.The results of the abamectin exposure experiments showed that the mortality of B.xylophilus after Bx-gst12 interference was significantly higher than the control group,with an average increase of 7.12%in mortality.【Conclusion】In this study,the Bx-gst12 was successfully cloned and the Bx-gst12-dsRNA was synthesized.Bx-gst12 interference significantly affected the susceptibility of B.xylophilus to abamectin.Under the same concentration of abamectin for the same time,the mortality rate in the interfered group was significantly higher than the control group,indicating that Bx-gst12 plays a significant role in the drug metabolism of B.xylophilus.

关 键 词：松材线虫 谷胱甘肽S转移酶基因 阿维菌素 生物信息学 基因干扰 

分 类 号：S763[农业科学—森林保护学]