Munoz D, Castillejo JI, Caballero P
Laboratorio de Entomologia Agricola y Patologia de Insectos (LEAPI), Departamento de Produccion Agraria, Universidad Publica de Navarra, 31006 Pamplona, Spain.
Appl Environ Microbiol. 1998 Nov;64(11):4372-7. doi: 10.1128/AEM.64.11.4372-4377.1998.
A wild-type nucleopolyhedrovirus (NPV) isolate from Spodoptera exigua from Florida (Se-US2) is a variant of the SeMNPV type strain since it has a unique DNA profile but is closely related to other known geographical isolates of SeMNPV. It consists of several genotypic variants, of which seven were identified in a Se-US2 virus stock by a modification of the in vivo cloning method developed by Smith and Crook (Virology 166:240-244, 1988). The US2A variant was the most prevalent genotype, and it was designated the prototype Se-US2 variant, while four of the variants (US2B, US2D, US2F, and US2H) were found at low frequency. US2C and US2E were also very abundant, and their diagnostic bands were easily observed in wild-type isolate restriction endonuclease patterns. The analysis of each variant, compared to the prototype US2A, showed that US2B and US2H presented minor differences, while US2D and US2F contained slightly larger insertions or deletions. Variants US2C and US2E contained major deletions of 21.1 and 14 kb, respectively, mapping at the same genomic region (between 14.5 and 30.2 map units [m.u.] and between 12.8 and 23 m.u., respectively). This is the first report of such deletion mutants in a natural baculovirus population. Variants US2A, US2B, US2D, US2F, and US2H were isolated as pure genotypes, but we failed to clone US2C and US2E in vivo. When these two variants appeared without apparent contamination with any other variant, they lost their pathogenicity for Spodoptera exigua larvae. A further biological characterization showed evidence that these two naturally occurring deletion mutants act as parasitic genotypes in the virus population. Bioassay data also demonstrated that pure US2A is significantly more pathogenic against second-instar S. exigua larvae than the wild-type isolate. The need for precise genotypic characterization of a baculovirus prior to its development as a bioinsecticide is discussed.
从佛罗里达州的甜菜夜蛾中分离出的野生型核型多角体病毒(NPV)毒株(Se-US2)是SeMNPV标准毒株的一个变体,因为它具有独特的DNA图谱,但与其他已知的SeMNPV地理分离株密切相关。它由几个基因型变体组成,通过对Smith和Crook开发的体内克隆方法(《病毒学》166:240 - 244,1988年)进行改进,在一株Se-US2病毒株中鉴定出了7个变体。US2A变体是最常见的基因型,被指定为Se-US2原型变体,而其中4个变体(US2B、US2D、US2F和US2H)出现频率较低。US2C和US2E也非常丰富,它们的诊断条带在野生型分离株的限制性内切酶图谱中很容易观察到。与原型US2A相比,对每个变体的分析表明,US2B和US2H存在微小差异,而US2D和US2F包含稍大的插入或缺失。变体US2C和US2E分别包含21.1 kb和14 kb的大片段缺失,分别位于相同的基因组区域(分别在14.5至30.2个图谱单位[m.u.]之间和12.8至23 m.u.之间)。这是首次在天然杆状病毒群体中报道此类缺失突变体。变体US2A、US2B、US2D、US2F和US2H被分离为纯基因型,但我们未能在体内克隆出US2C和US2E。当这两个变体出现且没有明显受到任何其他变体污染时,它们对甜菜夜蛾幼虫失去了致病性。进一步的生物学特性分析表明,这两个天然存在的缺失突变体在病毒群体中表现为寄生基因型。生物测定数据还表明,纯US2A对二龄甜菜夜蛾幼虫的致病性明显高于野生型分离株。本文讨论了在杆状病毒开发为生物杀虫剂之前进行精确基因型鉴定的必要性。