摘要
【目的】从基因组序列角度进一步揭示自然界斜纹夜蛾核型多角体病毒(Spodoptera litura nucleopolyhedrovirus,SpltNPV)的基因型多态性。【方法】病毒克隆A5,F1,X3和X15分别以活体克隆法分离自SpltNPV埃及株、日本福冈株和日本小笠原株。根据SpltNPV基因组全序列(GenBank登录号:AF325155)和海灰翅夜蛾核型多角体病毒(S.littoralis NPV,SpliNPV)部分基因序列(GenBank登录号:X99377,X99376和X98924)设计引物,PCR扩增获得A5,F1,X3和X15的多角体蛋白(polyhedrin,polh)基因和ORF18～ORF23序列。【结果】根据多角体蛋白基因序列,X3和X15属于SpltNPV型,而A5和F1属于SpliNPV型。将A5,F1,X3和X15的ORF18～ORF23与SpltNPV和SpliNPV相应的基因序列进行同源性比较。结果发现,F1与SpliNPV以及X3与SpltNPV的核苷酸序列相似性高,但X3的ORF20在172～558nt处缺失387bp。尽管依据多角体蛋白基因序列X15属于SpltNPV型,但对于ORF18～ORF23序列,X15与SpliNPV的相似性高于与SpltNPV的相似性。同样,A5属于SpliNPV型,ORF18～ORF20与SpliNPV相应的核苷酸序列相似性高,但ORF21与SpltNPV相应的核苷酸序列一致性为100%,特别是ORF22,SpltNPV的特有序列出现在A5的基因组中,而且与SpltNPV的ORF22一致性为100%;反过来,ORF23又与SpliNPV相应的核苷酸序列相似性高。【结论】所有这些都表明,SpltNPV在自然界不仅存在基因型多态性,而且即使属于同一基因型,它们的基因组序列也有显著差异。可利用SpltNPV在自然界的这种异质性筛选适宜防治斜纹夜蛾幼虫的株系。
【Aim】 This study aims to further reveal genotypic polymorphism of Spodoptera litura nucleopolyhedrovirus （SpltNPV） in the wild based on genome sequences. 【Methods】 The virus clones A5, F1, X3 and X15 were isolated from SpltNPV Egypt stock, Fukuyama stock and Ogasawara stock by in vivo cloning technique, respectively. According to the complete genome sequence of SpltNPV （GenBank accession no.： NC_003102） and some SpliNPV gene sequences （GenBank accession no.： X99377, X99376 and X98924）, primers were designed. Polyhedrin and ORF18-ORF23 of A5, F1, X3 and X15 were obtained by PCR amplification. 【Results】 According to the polyhedrin sequence, A5 and F1 belong to SpliNPV type, while X3 and X15 belong to SpltNPV type. The homologous comparisons of ORF18-ORF23 were done between A5, F1, X3, X15 and SpltNPV or SpliNPV. The results showed that F1 shares high nucleotide identity with SpliNPV, and X3 shares high nucleotide identity with SpltNPV, but only 387 bp is deleted at 172-558 nt of ORF20. Though X15 belongs to SpltNPV type according to the polyhedrin sequence, ORF18-ORF23 all share high nucleotide identities to those of SpliNPV, and as a SpltNPV-unique ORF, ORF22 is deleted in X15 genome. A5 belongs to SpliNPV type, and the corresponding ORF18-ORF20 share high nucleotide identities with SpliNPV, but ORF21 shares 100% nucleotide identity with that of SpltNPV. As a SpltNPVunique ORF, ORF22, however, appears in A5 genome, and shares 100% nucleotide identity with that of SpltNPV. Furthermore, ORF23 shares high nucleotide identity with that of SpliNPV. 【Conclusion】 These results indicate that SpltNPV in the wild possesses genotypic polymorphisms, and moreover, the genomes of these virus clones with the same genotype have significant differences. The natural heterogeneity can be exploited to develop NPV strains suitable for the control of S. litura.
出处
《昆虫学报》
CAS
CSCD
北大核心
2013年第4期372-378,共7页
Acta Entomologica Sinica
基金
江苏省农业科技创新资金项目[cx(11)2046]
公益性行业科研专项(201203036)
关键词
斜纹夜蛾核型多角体病毒
病毒克隆
基因型
基因组
多态性
Spodoptera litura nucleopolyhedrovirus （SpltNPV） virus clones genotype genome
polymorphism
