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作为一种改良农药的棉铃虫单粒包埋核多角体病毒的基因工程

Genetic engineering of Helicoverpa armigera single-nucleocapsid nucleopolyhedrovirus as an improved pesticide.

作者信息

Chen X, Sun X, Hu Z, Li M, O'Reilly D R, Zuidema D, Vlak J M

机构信息

Joint-lab of Invertebrate Virology, Chinese Academy of Sciences, Wuhan 430071, China.

出版信息

J Invertebr Pathol. 2000 Aug;76(2):140-6. doi: 10.1006/jipa.2000.4963.

Abstract

The Helicoverpa armigera single-nucleocapsid nucleopolyhedrovirus (HearNPV) has been registered and is commercially produced in China as a biopesticide to control the bollworm in cotton. However, the virus has a relatively slow speed of action. To improve its efficacy, recombinant HearNPVs were generated by deleting the ecdysteroid UDP-glucosyltransferase (egt) gene (HaCXW1 and HaLM2) or by inserting the insect-specific toxin gene AaIT in the egt locus (HaCXW2) of HearNPV using conventional recombination strategies in insect cell culture. The various recombinants remained genetically stable when cultured in HzAM1 insect cells. Bioassay data showed a significant reduction in the time required for all HearNPV recombinants to kill second instar H. armigera larvae. The LT(50) of the egt deletion recombinants HaCXW1 and HaLM2 was about 27% faster than that of wild-type HearNPV. The largest reduction in LT(50) was achieved by inserting the gene for the insect-specific neurotoxin, AaIT, in the egt locus, giving a reduction in LT(50) of 32% compared to wild-type HearNPV. The ability to genetically improve the properties of HearNPV as a biopesticide provides a further opportunity to develop this virus into a commercially viable product to control the bollworm in China.

摘要

棉铃虫单核衣壳核型多角体病毒(HearNPV)已在中国注册并作为生物农药进行商业化生产,用于防治棉花中的棉铃虫。然而,该病毒的作用速度相对较慢。为提高其效力,通过在昆虫细胞培养中使用常规重组策略,删除蜕皮甾类UDP-葡萄糖基转移酶(egt)基因(HaCXW1和HaLM2)或在HearNPV的egt基因座(HaCXW2)中插入昆虫特异性毒素基因AaIT,产生了重组HearNPV。当在HzAM1昆虫细胞中培养时,各种重组体在遗传上保持稳定。生物测定数据表明,所有HearNPV重组体杀死棉铃虫二龄幼虫所需的时间显著减少。egt缺失重组体HaCXW1和HaLM2的半数致死时间(LT(50))比野生型HearNPV快约27%。通过在egt基因座中插入昆虫特异性神经毒素AaIT基因,LT(50)减少幅度最大,与野生型HearNPV相比,LT(50)减少了32%。从基因层面改善HearNPV作为生物农药的特性,为将这种病毒开发成在中国控制棉铃虫的商业可行产品提供了进一步的机会。

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