自噬拮抗GCRV的分子机制及育种潜力

草鱼出血病病毒（GCRV）引起的出血病是危害草鱼养殖业的重要病害，出血病防控以及抗病品系培育是亟待研究的课题。我们的前期研究结果表明，自噬相关通路及关键基因的表达量在对GCRV敏感性不同的草鱼中存在显著差异，暗示自噬水平与草鱼抗病力之间的相关性。进一步研究发现GCRV在机体和细胞水平上均能诱导自噬的发生，在细胞水平上利用药物促进自噬能够抑制GCRV复制，而抑制自噬却促进GCRV复制，但其分子机制尚不清楚。基于前期的工作基础，本项目拟在细胞和机体两个层次，通过自噬诱导或抑制技术并结合病毒感染实验，证实自噬对GCRV的拮抗作用以及对草鱼抗病力的影响；进一步通过检测自噬对宿主物质合成代谢、DNA修复损伤、免疫应答以及细胞凋亡等影响，阐明自噬拮抗GCRV的分子机制；同时利用关联分析等方法，评估利用自噬基因进行抗病育种的潜力。项目的研究结果将为出血病防控以及草鱼抗病品系培育提供重要指导作用。

Grass hemorrhagic disease that caused by grass carp reovirus (GCRV) is a serious disease of grass carp. Hemorrhagic disease prevention and control and disease-resistant breeding are urgent research projects. In our previous study, we found that the autophagy related pathways and key autophagy genes were differential expressed in grass carps that showed opposite susceptibility to GCRV, implying the relationship between autophagy and anti-disease ability of grass carp. Further study revealed that GCRV infection could trigger autophagy in both CIK cells and grass carp. Inducing autophagy by rapamycin in CIK cells repressed GCRV replication, while inhibited autophagy by 3-methladenine enhanced GCRV replication, whereas the molecular mechanism remain unclear. Based on our exciting preliminary data, we plan to verify the antagonistic effect of autophagy on GCRV proliferation and effect of autophagy on anti-disease ability of grass carp in cell and organism level by combining viral challenge experiment with autophagy induce or inhibit methods. Furthermore, the molecular mechanism of autophagy antagonizes GCRV will be illuminated by detecting the effects of autophagy on host biosynthesis and metabolism, DNA damage and repair, and immune response and cell apoptosis. In addition, the breeding potential of autophagy genes will be assessed by association analysis of death and survival samples from random mating grass carp population after GCRV infection. These results will provide important guidance for hemorrhagic disease prevention and control and disease-resistant breeding of grass carp.