Hubei Key Laboratory of Genetic Regulation and Integrative Biology, College of Life Sciences, Central China Normal University, Wuhan, China.
PLoS One. 2021 Jan 14;16(1):e0243143. doi: 10.1371/journal.pone.0243143. eCollection 2021.
In this study, the genomes of three Plutella xylostella granulovirus (PlxyGV) isolates, PlxyGV-W and PlxyGV-Wn from near Wuhan and PlxyGV-B from near Beijing, China were completely sequenced and comparatively analyzed to investigate genetic stability and diversity of PlxyGV. PlxyGV-W, PlxyGV-B and PlxyGV-Wn consist of 100,941bp, 100,972bp and 100,999bp in length with G + C compositions of 40.71-40.73%, respectively, and share nucleotide sequence identities of 99.5-99.8%. The three individual isolates contain 118 putative protein-encoding ORFs in common. PlxyGV-W, PlxyGV-B and PlxyGV-Wn have ten, nineteen and six nonsynonymous intra isolate nucleotide polymorphisms (NPs) in six, fourteen and five ORFs, respectively, including homologs of five DNA replication/late expression factors and two per os infectivity factors. There are seventeen nonsynonymous inter isolate NPs in seven ORFs between PlxyGV-W and PlxyGV-B, seventy three nonsynonymous NPs in forty seven ORFs between PlxyGV-W and PlxyGV-Wn, seventy seven nonsynonymous NPs in forty six ORFs between PlxyGV-B and PlxyGV-Wn. Alignment of the genome sequences of nine PlxyGV isolates sequenced up to date shows that the sequence homogeneity between the genomes are over 99.4%, with the exception of the genome of PlxyGV-SA from South Africa, which shares a sequence identity of 98.6-98.7% with the other ones. No events of gene gain/loss or translocations were observed. These results suggest that PlxyGV genome is fairly stable in nature. In addition, the transcription start sites and polyadenylation sites of thirteen PlxyGV-specific ORFs, conserved in all PlxyGV isolates, were identified by RACE analysis using mRNAs purified from larvae infected by PlxyGV-Wn, proving the PlxyGV-specific ORFs are all genuine genes.
在这项研究中,我们对来自中国武汉附近的 Plutella xylostella 颗粒体病毒(PlxyGV)分离株 PlxyGV-W 和 PlxyGV-Wn 以及来自北京附近的 PlxyGV-B 进行了全基因组测序和比较分析,以研究 PlxyGV 的遗传稳定性和多样性。PlxyGV-W、PlxyGV-B 和 PlxyGV-Wn 的基因组长度分别为 100,941bp、100,972bp 和 100,999bp,G+C 含量分别为 40.71-40.73%,核苷酸序列同一性为 99.5-99.8%。这三个分离株共含有 118 个推定的蛋白编码 ORF。PlxyGV-W、PlxyGV-B 和 PlxyGV-Wn 在六个、十四个和五个 ORF 中分别有十个、十九个和六个非同义的种内核苷酸多态性(NPs),包括五个 DNA 复制/晚期表达因子和两个经口感染性因子的同源物。PlxyGV-W 和 PlxyGV-B 之间的七个 ORF 中有十个非同义的种间核苷酸多态性,PlxyGV-W 和 PlxyGV-Wn 之间的四十七个 ORF 中有七十三个非同义核苷酸多态性,PlxyGV-B 和 PlxyGV-Wn 之间的四十六个 ORF 中有七十七个非同义核苷酸多态性。对迄今为止测序的 9 个 PlxyGV 分离株的基因组序列进行比对表明,除了来自南非的 PlxyGV-SA 基因组与其他基因组的序列同一性为 98.6-98.7%外,基因组之间的序列同源性均超过 99.4%。未观察到基因获得/缺失或易位事件。这些结果表明 PlxyGV 基因组在自然界中相当稳定。此外,通过使用 PlxyGV-Wn 感染幼虫纯化的 mRNA 进行 RACE 分析,鉴定了所有 PlxyGV 分离株中保守的 13 个 PlxyGV 特异性 ORF 的转录起始位点和聚腺苷酸化位点,证明 PlxyGV 特异性 ORF 均为真正的基因。