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Genetic Structure of a Spodoptera frugiperda Nucleopolyhedrovirus Population: High Prevalence of Deletion Genotypes

机译:草地贪夜蛾核多角体病毒种群的遗传结构:删除基因型的高流行。

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摘要

A Nicaraguan field isolate (SfNIC) of Spodoptera frugiperda nucleopolyhedrovirus was purified by plaque assay on Sf9 cells. Nine distinct genotypes, A to I, were identified by their restriction endonuclease profiles. Variant SfNIC-B was selected as the standard because its restriction profile corresponded to that of the wild-type isolate. Physical maps were generated for each of the variants. The differences between variants and the SfNIC-B standard were confined to the region between map units 9 and 32.5. This region included PstI-G, PstI-F, PstI-L, PstI-K and EcoRI-L fragments. Eight genotypes presented a deletion in their genome compared with SfNIC-B. Occlusion body-derived virions of SfNIC-C, -D and -G accounted for 41% of plaque-purified clones. These variants were not infectious per os but retained infectivity by injection into S. frugiperda larvae. Median 50% lethal concentration values for the other cloned genotypes were significantly higher than that of the wild type. The variants also differed in their speed of kill. Noninfectious variants SfNIC-C and -D lacked the pif and pif-2 genes. Infectivity was restored to these variants by plasmid rescue with a plasmid comprising both pif and pif-2. Transcription of an SfNIC-G gene was detected by reverse transcription-PCR in insects, but no fatal disease developed. Transcription was not detected in SfNIC-C or -D-inoculated larvae. We conclude that the SfNIC population presents high levels of genetic diversity, localized to a 17-kb region containing pif and pif-2, and that interactions among complete and deleted genotypic variants will likely influence the capacity of this virus to control insect pests.
机译:通过噬斑分析在Sf9细胞上纯化了斜纹夜蛾核多角体病毒的尼加拉瓜野外分离株(SfNIC)。通过限制性内切核酸酶谱鉴定了九种不同的基因型,A至I。选择变异SfNIC-B作为标准品,因为其限制性酶切图谱与野生型分离株相对应。为每个变体生成了物理图谱。变体与SfNIC-B标准之间的差异仅限于地图单元9和32.5之间的区域。该区域包括PstI-G,PstI-F,PstI-L,PstI-K和EcoRI-L片段。与SfNIC-B相比,八种基因型在其基因组中存在缺失。 SfNIC-C,-D和-G的咬合体病毒体占噬菌斑纯化克隆的41%。这些变种本身不是传染性的,但是通过注射到Srug frugiperda幼虫中而保留了传染性。其他克隆基因型的中值50%致死浓度值显着高于野生型。变体的杀死速度也不同。非感染性变体SfNIC-C和-D缺少pif和pif-2基因。通过用包含pif和pif-2的质粒进行质粒拯救来恢复对这些变体的感染性。通过反转录PCR检测昆虫中SfNIC-G基因的转录,但没有致命的疾病发生。在SfNIC-C或-D接种的幼虫中未检测到转录。我们得出的结论是,SfNIC人群呈现出高水平的遗传多样性,位于包含pif和pif-2的17kb区域,并且完整和缺失的基因型变体之间的相互作用将可能影响该病毒控制害虫的能力。

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