【目的】细胞自噬在真核生物细胞应对不良环境中具有重要作用。探讨细胞自噬对松材线虫取食繁殖的影响,测定松材线虫与松树互作过程中松材线虫BxATG1和BxATG8两个自噬相关基因的表达。【方法】在前期研究发现松材线虫存在细胞自噬现象,并克隆了松材线虫BxATG1和BxATG8两个自噬相关基因的基础上,采用外源自噬诱导剂探讨了细胞自噬对松材线虫取食繁殖的影响,测定了松材线虫与松树互作早期自噬相关基因的表达。【结果】经雷帕霉素处理后,松材线虫的自噬启动基因BxATG1和自噬标志性基因BxATG8表达量显著上调,且松材线虫的取食速率及繁殖量也显著增加; 松材线虫在饥饿状态下的存活能力明显上升,脂肪含量的堆积也略有增加。松材线虫与松树互作早期(6、12、18、24 h)的qRT-PCR结果表明,松材线虫侵染松树12 h时,细胞自噬爆发达到最高峰。【结论】外源自噬诱导后促进了松材线虫细胞自噬的发生; 细胞自噬在促进松材线虫生长发育中具有一定作用; 在松材线虫侵染松树过程中,细胞自噬有助于线虫适应松树的早期抵抗反应。
Abstract
【Objective】Autophagy plays an important role in Eukaryotic cells adapting adversities. Nevertheless, there're few researches into autophagy and its functions in pine wood nematode(PWN), which is the pathogeny of pine wilt disease. 【Method】Our laboratory previously found that there existed autophagy in PWNs, thus, we cloned genes BxATG1 and BxATG8 of them, as the genes were related to autophagy. Based on that, this study discussed the impact of autophagy on PWNs' growth and development, as well as its roles during the interaction between PWNs and pines. 【Result】After being treated with Rapamycin, which was the autophagy inducer, the expression of BxATG1 and BxATG8 rose significantly, which demonstrated that the exogenous autophagy induction had promoted the occurrence of autophagy. Besides, the exogenous autophagy inducer notably helped with the boost in PWNs' feeding rate and reproduction. What's more, after autophagy induction, PWNs' viability under starving condition was stimulated obviously, and the accumulation of fats also improved. These facts showed that autophagy played an important role in the growth and development of PWNs. qRT-PCR results of early interaction between PWNs and pines showed that, the occurrence of autophagy reached its climax after infecting the pine seedlings for 12 hours, which implied that, during the course of PWNs infecting the pines, autophagy was helpful for PWNs to go through the early stage of plant's resistance response.【Conclusion】Induction of autophagy promoted the occurrence of PWN's autophagy, and autophagy played an important role in promoting the growth and development of Bursaphelenchus xylophilus. In the process of pine wood nematode infestation, autophagy can help the nematode adapt to the early resistance of pine.
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基金
收稿日期:2017-12-11 修回日期:2018-04-27
基金项目:国家重点研发计划(2016YFC1202100); 江苏省农业科技自主创新资助项目(CX(16)1005); 江苏高校优势学科建设工程资助项目(PAPD)
第一作者:吴凡(501888347@qq.com),硕士。*通信作者:吴小芹(xqwu@njfu.edu.cn),教授。
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