In Silico and in Vivo Investigations of Bioagent Helicoverpa Nucleopolyhedrovirus against Helicoverpa armigera in Chickpea

Ritu Srivastava, A. C. Shukla
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Abstract

During investigations; homology model of 3D-structure was built for sequence of polyhedrin protein of Helicoverpa armigera nucleopolyhedrovirus, containing 246 amino acids (Accession: ACI05106.1 GI: 205946055), and evaluated through multiple tools/ applications to judge extent of accuracy in light of existing crystal structure. Further, in vivo experiments were conducted and determined response of different adjuvants with HaNPV and their efficacy. The pooled mean mortality of larvae exposed to virus mixed with 5% green tea and 5% rice bran filtrates (8.3 larvae per 25 plants) was differ significantly from control (15.8 larvae per 25 plants), suggesting that UV protectants & diet enhancer (mannitol) has ability to protect stability of virulence of the virus, under field conditions. The minimum percent pod damage of 8.6% and maximum yield of 1604.8 Kg ha-1 at harvesting was recorded with formulation of indigenous BHA virus isolate @ 2.2 x 105 POBs mL-1 mixed with Roket @50 ppm; followed by formulation with mannitol (@ 1% + green tea 5% + 5% rice bran filtrates) with percent pod damage of 16.8 % and yield of 1045.8 Kg ha-1 of chickpea. Furthermore, in vitro toxicity of fresh virus suspension @ 250 mL ha-1 was recorded more toxic in terms of percent mortality and LT50 (5.65 days). However, three months stored HaNPV formulations [(A) mannitol @ 1%+ green tea@ 5% and (B) mannitol @ 1% + green tea 5% + 5% rice bran filtrates] were more effective in larval reduction with LT50 of 7.89 and 6.00 days, respectively. Virus mixed with 5% green tea and 5% rice bran filtrates gave stability to formulation up-to one year with LT50 of 7.64 days. Findings showed that HaNPV formulations with mannitol (B) have potential that can be used in integrated manner with other IPM practices, to reduce the use of toxic synthetic pesticides in chickpea.
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鹰嘴豆Helicoverpa核多角体病毒生物制剂对棉铃虫的体内和计算机研究
在调查;对含有246个氨基酸的棉铃虫多角体病毒多面蛋白序列(Accession: ACI05106.1 GI: 205946055)建立三维结构的同源性模型,并结合已有的晶体结构,通过多种工具/应用程序评估其准确性程度。进一步进行了体内实验,确定了不同佐剂对HaNPV的反应和疗效。与对照(15.8只/ 25株)相比,暴露于5%绿茶和5%米糠滤液混合的病毒的总平均死亡率(8.3只/ 25株)有显著差异,表明紫外线保护剂和日粮增强剂(甘露醇)在田间条件下具有保护病毒毒力稳定的能力。采用本地BHA病毒分离液(2.2 x 105 POBs mL-1)与Roket (50 ppm)混合配制,收获时荚果损失率最低为8.6%,产量最高为1604.8 Kg ha-1;其次是甘露醇(@ 1% +绿茶5% + 5%米糠滤液)配方,豆荚损坏率为16.8%,鹰嘴豆产量为1045.8 Kg hm -1。此外,新鲜病毒悬浮液@ 250 mL ha-1的体外毒性在死亡率百分比和LT50(5.65天)方面记录了更高的毒性。然而,储存3个月的HaNPV配方[(A)甘露醇@ 1%+绿茶@ 5%和(B)甘露醇@ 1%+绿茶5% + 5%米糠滤液]对幼虫的减少效果更好,LT50分别为7.89和6.00天。病毒与5%绿茶和5%米糠滤液混合后,配方稳定性长达一年,LT50为7.64天。研究结果表明,含有甘露醇(B)的HaNPV配方具有与其他IPM方法结合使用的潜力,可以减少鹰嘴豆中有毒合成农药的使用。
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