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The complete genome sequence of a third distinct baculovirus isolated from the true armyworm, Mythimna unipuncta, contains two copies of the lef-7 gene

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Abstract

A baculovirus isolate from a USDA Forest Service collection was characterized by electron microscopy and analysis of its genome sequence. The isolate, formerly referred to as Pseudoletia (Mythimna) sp. nucleopolyhedrovirus #7 (MyspNPV#7), was determined by barcoding PCR to derive from the host species Mythimna unipuncta (true armyworm) and was renamed Mythimna unipuncta nucleopolyhedrovirus #7 (MyunNPV#7). The occlusion bodies (OBs) and virions exhibited a size and morphology typical for OBs produced by the species of genus Alphabaculovirus, with occlusion-derived virions consisting of 2–5 nucleocapsids within a single envelope. The MyunNPV#7 genome was determined to be 148,482 bp with a 48.58% G+C nucleotide distribution. A total of 159 ORFs of 150 bp or larger were annotated in the genome sequence, including the 38 core genes of family Baculoviridae. The genome contained six homologous repeat regions (hrs) consisting of multiple copies of a 34-bp imperfect palindrome. Phylogenetic inference from concatenated baculovirus core gene amino acid sequence alignments placed MyunNPV#7 with group II alphabaculoviruses isolated from other armyworm and cutworm host species of lepidopteran family Noctuidae. MyunNPV#7 could be distinguished from other viruses in this group on the basis of differences in gene content and order. Pairwise nucleotide distances suggested that MyunNPV#7 represents a distinct species in Alphabaculovirus. The MyunNPV#7 genome was found to contain two copies of the late expression factor-7 (lef-7) gene, a feature not reported for any other baculovirus genome to date. Both copies of lef-7 encoded an F-box domain, which is required for the function of LEF-7 in baculovirus DNA replication.

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Acknowledgements

The authors would like to thank Pallavi Thapa, Daniel Barakh, and Anita Ghosh (USDA-ARS, Beltsille, MD) for assistance with dideoxy sequencing, and Doug Baldwin (AAFC) for assistance with assembly of 454 sequence reads and initial annotation of the genome. Mention of trade names or commercial products in this publication is solely for the purpose of providing specific information and does not imply recommendation or endorsement by the U.S. Department of Agriculture. USDA is an equal opportunity provider and employer.

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Conceived and designed the experiments: RLH, JDM, DLR, GRB, GFR, DAT, MAE. Performed the experiments: RLH, JDM, DLR, GRB, MAE. Analyzed the data: RLH, DLR. Wrote the paper: RLH, JDM, DLR, GRB, GFR, DAT, MAE.

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Correspondence to Robert L. Harrison.

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The research described in this paper does not use any human or animal subjects.

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Edited by A. Lorena Passarelli.

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Supplementary Fig. 1

Baculovirus core gene phylogenies carried out with MAFFT- or MUSCLE-aligned core gene amino acid sequences, using the same taxa as in Fig. 4 with (+) or without (-) sequences from the UrprNPV-Southern Brazil genome (PPTX 92 kb)

Supplementary Table 1

Names, abbreviations, and GenBank accession numbers of taxa used in phylogenetic inference (XLSX 13 kb)

Supplementary Table 2

MyunNPV#7 open reading frames (ORFs) and homologous repeat regions (hrs) (DOCX 42 kb)

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Harrison, R.L., Mowery, J.D., Rowley, D.L. et al. The complete genome sequence of a third distinct baculovirus isolated from the true armyworm, Mythimna unipuncta, contains two copies of the lef-7 gene. Virus Genes 54, 297–310 (2018). https://doi.org/10.1007/s11262-017-1525-0

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