›› 2010, Vol. 53 ›› Issue (2): 167-174.doi:

• 研究论文 • 上一篇    下一篇


郑桂玲, 李长友, 周洪旭, 李淑文, 李国勋, 薛明   

  • 出版日期:2010-03-25 发布日期:2010-02-20
  • 通讯作者: 李国勋, 薛明

Establishment of two new cell lines from the embryonic tissue of Helicoverpa armigera (Lepidoptera: Noctuidae)and their responses to baculovirus infection

ZHENG Gui-Ling, LI Chang-You, ZHOU Hong-Xu, LI Shu-Wen, LI Guo-Xun, XUE Ming   

  • Online:2010-03-25 Published:2010-02-20

摘要: 昆虫细胞系的建立在病毒学和昆虫学等领域的研究和应用中发挥着重要的作用。本研究由棉铃虫Helicoverpa armigera胚胎组织建立了两株细胞系, 分别命名为QB-Ha-E-1和QB-Ha-E-5, 在含10%胎牛血清的TNM-FH培养基中已传代60余代。两株细胞系均以圆形和短梭形细胞为主。DAF鉴定结果表明, 两株细胞系均来源于棉铃虫胚胎, 其扩增谱带与其他几种昆虫细胞系明显不同; QB-Ha-E-1和QB-Ha-E-5的第30代细胞群体倍增时间分别为63.7 h和66.9 h。两株细胞系均能被棉铃虫核型多角体病毒(HaSNPV)感染, 4 d的感染率分别为86.6%和56.5%, 对甘蓝夜蛾Mamestra brassicae核型多角体病毒(MbNPV)7 d的感染率均为15%左右, 但对苜蓿银纹夜蛾Autographa californica核型多角体病毒(AcMNPV)侵染的反应不同。DAPI染色和基因组DNA电泳结果表明, AcMNPV可诱导QB-Ha-E-5细胞发生凋亡, 极少数细胞内可形成多角体, 但不能诱导QB-Ha-E-1细胞发生凋亡, 其感染率为55.3%; 两株细胞系均可被1.25 μg/mL的放线菌素D诱导发生凋亡。两株细胞系具有相同的遗传背景, 但对AcMNPV侵染的反应不同, 可作为昆虫病毒和细胞之间相互关系以及细胞凋亡机制研究的理想材料。

关键词: 棉铃虫, 胚胎细胞系, 杆状病毒, 侵染, 细胞凋亡

Abstract: The development of insect cell lines plays a significant role in research and application of virology and entomology. Two new cell lines, i.e., QB-Ha-E-1 and QB-Ha-E-5, were established from the embryonic tissue of Helicoverpa armigera (Lepidoptera: Noctuidae). The cell lines had been subcultured over 60 passages in TNM-FH medium supplemented with 10% fetal bovine serum. Each cell line has two major morphological types, round cells and spindle-shaped cells. DNA amplification fingerprinting (DAF) analysis indicated that DNA profiles of the two cell lines were similar but distinctly different from cell lines of several other insects, suggesting that both originated from the embryonic tissue of H. armigera. The cell doubling time of the 30th passages of QB-Ha-E-1 and QB-Ha-E-5 were 63.7 h and 66.9 h, respectively. Both the cell lines could be infected by H. armigera single nucleopolyhedrovirus (HaSNPV) with the infection rates of 86.6% and 56.5%, respectively, 4 d post infection (p.i.). Approximately 15% of the two cell lines were infected by Mamestra brassicae nucleopolyhedrovirus (MbNPV) 7 d p.i. The two cell lines had differential responses to Autographa californica multiple nucleopolyhedrovirus (AcMNPV) infection. The results from the fluorescent staining with DAPI and electrophoresis of genomic DNA indicated that QB-Ha-E-5 cell line had typical apoptotic response following AcMNPV infection, but QB-Ha-E-1 cell line, with the AcMNPV infection rate of 55.3%, appeared anti-apoptosis. Actinomycin D could induce apoptosis of the two cell lines at the concentration of 1.25 μg/mL. Therefore, the two cell lines can be used as ideal materials to study interactions between insect viruses and insect cells.

Key words: Helicoverpa armigera, embryonic cell line, baculovirus, infection, apoptosis