Yeh Shih-Chia, Lee Song-Tay, Wu Chih-Yu, Wang Chung-Hsiung
Department of Entomology, National Taiwan University, Taipei, Taiwan, ROC.
J Invertebr Pathol. 2007 Oct;96(2):138-46. doi: 10.1016/j.jip.2007.04.004. Epub 2007 Apr 21.
Here we describe the establishment of a new cell line, NTU-MV, derived from pupal tissues of an economically important pest, the legume pod borer Maruca vitrata. This cell line contained four major cell types: polymorphic cells, round cells, spindle-shaped cells, and comma cells. The doubling time of MV cells in TNM-FH medium supplemented with 8% FBS at 28 degrees C was 27h. The chromosome numbers of MV cells varied widely from 16 to 268. Compared to other insect cell lines, the MV cell line produced distinct isozyme patterns with esterase, malate dehydrogenase (MDH), and lactate dehydrogenase (LDH). Confirmation that NTU-MV was derived from M. vitrata was demonstrated by showing that the sequence of the internal transcribed spacer regions (ITS) of the MV cells was 98% identical to that of M. vitrata larvae. Two NTU-MV cell strains, NTU-MV1 and NTU-MV56, were selected based on susceptibility to MaviMNPV (M. vitrata multiple nucleopolyhedrovirus). NTU-MV, MV1, and MV56 cells showed a high susceptibility to MaviMNPV and produced high yields of polyhedra (47-50OBs/cell, 4x10(7)-5.96x10(7)OBs/ml) after 2 weeks of MaviMNPV infection. We conclude that the NTU-MV cell line will be a useful tool for studying MaviMNPV as well as for the mass production of MaviMNPV polyhedra for the biocontrol of M. vitrata.
在此,我们描述了一种新细胞系NTU-MV的建立,该细胞系源自一种经济上重要的害虫——豆荚野螟Maruca vitrata的蛹组织。该细胞系包含四种主要细胞类型:多形细胞、圆形细胞、纺锤形细胞和逗号形细胞。在补充有8%胎牛血清的TNM-FH培养基中,28℃时MV细胞的倍增时间为27小时。MV细胞的染色体数目变化很大,从16到268条不等。与其他昆虫细胞系相比,MV细胞系在酯酶、苹果酸脱氢酶(MDH)和乳酸脱氢酶(LDH)方面产生了独特的同工酶模式。通过显示MV细胞的内部转录间隔区(ITS)序列与豆荚野螟幼虫的序列98%相同,证实了NTU-MV源自豆荚野螟。基于对MaviMNPV(豆荚野螟多角体病毒)的敏感性,选择了两个NTU-MV细胞株,NTU-MV1和NTU-MV56。NTU-MV、MV1和MV56细胞对MaviMNPV表现出高敏感性,在感染MaviMNPV 2周后产生了高产的多角体(47 - 50个多角体/细胞,4×10⁷ - 5.96×10⁷个多角体/毫升)。我们得出结论,NTU-MV细胞系将是研究MaviMNPV以及大规模生产用于生物防治豆荚野螟的MaviMNPV多角体的有用工具。