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Identification of Loci Associated with Enhanced Virulence in Spodoptera litura Nucleopolyhedrovirus Isolates Using Deep Sequencing

机译:使用深度测序鉴定斜纹夜蛾核多角体病毒分离株中与增强毒力相关的基因座

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Spodoptera litura is an emerging pest insect in cotton and arable crops in Central Asia. To explore the possibility of using baculoviruses as biological control agents instead of chemical pesticides, in a previous study we characterized a number of S. litura nucleopolyhedrovirus (SpltNPV) isolates from Pakistan. We found significant differences in speed of kill, an important property of a biological control agent. Here we set out to understand the genetic basis of these differences in speed of kill, by comparing the genome of the fast-killing SpltNPV-Pak-TAX1 isolate with that of the slow-killing SpltNPV-Pak-BNG isolate. These two isolates and the SpltNPV-G2 reference strain from China were deep sequenced with Illumina. As expected, the two Pakistani isolates were closely related with 99% sequence identity, whereas the Chinese isolate was more distantly related. We identified two loci that may be associated with the fast action of the SpltNPV-Pak-TAX1 isolate. First, an analysis of rates of synonymous and non-synonymous mutations identified neutral to positive selection on open reading frame (ORF) 122, encoding a viral fibroblast growth factor (vFGF) that is known to affect virulence in other baculoviruses. Second, the homologous repeat region hr17, a putative enhancer of transcription and origin of replication, is absent in SpltNPV-Pak-TAX1 suggesting it may also affect virulence. Additionally, we found there is little genetic variation within both Pakistani isolates, and we identified four genes under positive selection in both isolates that may have played a role in adaptation of SpltNPV to conditions in Central Asia. Our results contribute to the understanding of the enhanced activity of SpltNPV-Pak-TAX1, and may help to select better SpltNPV isolates for the control of S. litura in Pakistan and elsewhere.
机译:斜纹夜蛾是中亚棉花和耕作作物中的一种新兴害虫。为了探索使用杆状病毒代替生物农药作为生物防治剂的可能性,在先前的研究中,我们鉴定了巴基斯坦的许多斜纹夜蛾核多角体病毒(SpltNPV)分离株。我们发现杀灭速度是生物防治剂的重要特性,杀灭速度存在显着差异。在这里,我们通过比较快速杀灭的SpltNPV-Pak-TAX1分离株和慢杀性的SpltNPV-Pak-BNG分离株的基因组,了解杀灭速度差异的遗传基础。用Illumina对这两个分离株和来自中国的SpltNPV-G2参考菌株进行了深度测序。不出所料,这两个巴基斯坦分离株与> 99%的序列同一性密切相关,而中国分离株之间的亲缘关系更远。我们确定了两个可能与SpltNPV-Pak-TAX1分离物的快速作用有关的基因座。首先,对同义和非同义突变发生率的分析确定了在开放阅读框(ORF)122上对阳性选择是中性的,该开放阅读框编码已知会影响其他杆状病毒毒力的病毒成纤维细胞生长因子(vFGF)。其次,SpltNPV-Pak-TAX1中不存在同源的重复区hr17,它是转录和复制起点的公认增强子,表明它也可能影响毒力。此外,我们发现这两种巴基斯坦分离株之间几乎没有遗传变异,并且我们在这两种分离株中都鉴定出四个处于正选择状态的基因,这些基因可能在SpltNPV适应中亚条件方面发挥了作用。我们的结果有助于理解SpltNPV-Pak-TAX1的活性增强,并可能有助于选择更好的SpltNPV分离株来控制巴基斯坦和其他地方的斜纹夜蛾。

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