镜鲤抗疱疹病毒(CyHV-3)F_(4)抗病品系病毒表达量评估

Research on expression of virus in Cyprinid herpesvirus 3 (CyHV-3) F_(4) disease resistant strains

以镜鲤抗疱疹病毒新品种F_(4)为实验材料,利用实时荧光定量PCR技术以TK(thymidine kinase)和ORF72基因为病毒标志基因,分析镜鲤抗疱疹病毒(CyHV-3)选育抗病品系F_(4)在病毒感染不同阶段中的表达量。结果显示:2个基因不同感染阶段脾、肾组织表达量趋势一致,24~144 h呈上升趋势,144~288 h呈下降趋势,且在144 h时脾、肾组织的病毒表达量均达到最高。脾组织2个基因表达量均低于肾组织。选育组2个基因在120 h均达到差异极显著,未选育组2个基因均在96~144 h差异极显著。选育组2个基因表达量低于未选育组。脾、肾组织TK基因均在96~168 h差异极显著;脾、肾组织ORF72基因均在96~192 h差异极显著,其中脾组织216 h也差异极显著。以上结果说明选育组抗病性能强于未选育组,其中肾组织抗病性能强于脾组织,且经选育肾组织抗病免疫性能得到提高。目前,未见其他鱼类的选育种在抗病性状上达到一个稳定阶段,对于病毒表达量的研究可为其他鱼类的研究提供重要的参考作用。

The new variety was bred to F_(4),and the disease resistance traits have reached stability. F_(4) was used as the experimental material, and TK and ORF72 were used as virus marker genes to analyze the expression of F_(4),a resistant strain of Cyprinus herpesvirus(CyHV-3),in different stages of viral infection, to investigate the effect of F_(4) on disease resistance and its virus expression pattern from the perspective of virus expression.The results showed that: The expression trends of spleen and kidney tissues of the two genes at different infection stages were consistent, with an upward trend from 24 h to 144 h and a downward trend from 144 h to 288 h.The spleen and kidney of the unselected group and the kidney of the selected group were significantly different at 144 h compared with other periods. The expression of both genes in spleen tissue was lower than that in kidney tissue.The two genes in the breeding group and the non-breeding group were significantly different between 96 h and 144 h, and the two genes in the breeding group were significantly different at 96 h and 144 h in the ORF72,and the other periods were significantly different. The expression levels of two genes in the breeding group were lower than those in the non-breeding group.The two genes in spleen tissue were significantly different from 96 h to 168 h, and the ORF72 was also significantly different at 192 h and 216 h;the two genes in kidney tissue were significantly different from 96 h to 168 h;ORF72 was also significantly different at 192 h.The above results show that the disease resistance of the selective group is stronger than that of the non-selected group, in which the kidney tissue has better disease resistance than the spleen, and the immune resistance of the selected kidney tissue is greatly improved.At present, no selective breeding of other fisher has reached a stable stage in disease resistance.This study can provide important guidance for other fishes in the study of viral expression.