Bianchi F J, Snoeijing I, van der Werf W, Mans R M, Smits P H, Vlak J M
Laboratory of Virology, Wageningen Agricultural University, Wageningen, 6709 PD, The Netherlands.
J Invertebr Pathol. 2000 Jan;75(1):28-35. doi: 10.1006/jipa.1999.4907.
Virulence and speed of action, as related to dose, are important effectiveness-determining properties of insect-pathogenic biocontrol agents. We used the droplet-feeding bioassay to compare dose responses between two wild-type baculoviruses, Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) and Spodoptera exigua MNPV (SeMNPV), and three deletion mutants of AcMNPV in S. exigua larvae. In each mutant one gene was deleted by genetic engineering: pp34, coding for the polyhedral membrane; egt, coding for ecdysteroid UDP-glucosyltransferase; or p10, coding for fibrillar structures in infected insect cells. SeMNPV had the lowest median lethal dose (LD(50)) as well as the highest speed of action (LT(50)) of all viruses investigated. In our comparative bioassays the only significant effect of gene deletions in AcMNPV was a slightly lower speed of action for the p10 deletion mutant. Otherwise, wild-type and recombinant AcMNPVs had similar biological activities. Our results suggest, in contrast to what is generally assumed, that gene deletions in AcMNPV for improved insecticidal activity should be critically assessed in each host system prior to further implementation as a control agent. Insertion of foreign genes coding for entomotoxins is less questionable and more promising in this respect.
毒力和作用速度与剂量相关,是昆虫病原生物防治剂重要的有效性决定特性。我们采用点滴饲喂生物测定法,比较了两种野生型杆状病毒,即苜蓿银纹夜蛾多粒包埋核型多角体病毒(AcMNPV)和甜菜夜蛾核型多角体病毒(SeMNPV),以及AcMNPV的三个缺失突变体在甜菜夜蛾幼虫中的剂量反应。在每个突变体中,通过基因工程删除了一个基因:编码多面体膜的pp34、编码蜕皮甾类UDP-葡萄糖基转移酶的egt或编码受感染昆虫细胞中纤维状结构的p10。在所研究的所有病毒中,SeMNPV的半数致死剂量(LD50)最低,作用速度(LT50)最高。在我们的比较生物测定中,AcMNPV基因缺失的唯一显著影响是p10缺失突变体的作用速度略低。否则,野生型和重组AcMNPV具有相似的生物活性。我们的结果表明,与通常的假设相反,在作为防治剂进一步应用之前,应在每个宿主系统中严格评估AcMNPV中为提高杀虫活性而进行的基因缺失。在这方面,插入编码昆虫毒素的外源基因问题较少且更有前景。