Complete genome sequence of a novel rhabdo‐like virus from the Chinese black cutworm Agrotis ipsilon (Lepidoptera: Noctuidae) Su Chen1,2 · Haichao Li1 · Guohui Yuan1 · Ruobing Guan1 · Tong Li3

This work identified a novel rhabdo-like virus in a Chinese black cutworm (Agrotis ipsilon), which we tentatively named “Agrotis ipsilon virus” (AIpsV). The complete genome of AIpsV is 15,454 nucleotides in length and contains seven open reading frames, collectively encoding more than 160 amino acids. The AIpsV genome is predicted to encode three structural proteins, nucleoprotein (N), glycoprotein (G), and large polymerase protein (L), and four unknown proteins. Phylogenetic analysis revealed that the AIpsV clusters with Wuhan ant virus and Hubei rhabdo-like virus 1 within the rhabdo-like virus clade. The level of expression of AIpsV genes was found to be higher in the pupal and adult stages than in the egg and larval stages. Rhabdoviruses (family Rhabdoviridae) are negative-sense single-stranded RNA viruses that can infect a wide range of plants and animals [1]. To date, the family Rhabdoviridae contains 20 genera and one unassigned species [2]. In addition, using metagenomic methods, a number of unclassified rhabdo-like viruses have been found in arthropods [3, 4]. Agrotis ipsilon (Hufnagel) is a worldwide pest that causes considerable damage to vegetables and grains. However, the virome of A. ipsilon has not been well studied. Here, from an RNA-Seq database of Chinese black cutworm, we identified a novel rhabdo-like virus, which tentatively has been named “Agrotis ipsilon virus” (AIpsV). The complete genome sequence of AIpsV was determined. A. ipsilon caterpillars were collected in Shanghai, China. They were fed an artificial diet and raised at 25 °C (± 1 °C) under a 14 h light/10 h dark cycle. For RNA-Seq analysis, samples from the four stages, egg, larva, pupa, and adult, were collected. Total RNA was extracted using TRIzol Reagent (Invitrogen, USA). RNA purity and integrity were checked using a NanoPhotometer® spectrophotometer (Implen, CA, USA) and an Agilent 2100 Bioanalyzer (Agilent Technologies, USA), respectively. The concentration of total RNA in each sample was further measured using a Qubit® RNA Assay Kit in a Qubit® 2.0 fluorometer (Life Technologies, CA, USA). Library preparation was carried out by BGI (Shenzhen, China). The libraries were then sequenced on an Illumina Hiseq 2000 platform. De novo transcriptome assembly was performed in SOAPdenovo using default parameters [5]. In total, 62,933 unigenes, defined as the longest assembled transcript of a given gene, were obtained for subsequent analyses. To identify possible viral fragments in assembled unigenes, protein sequences of ssRNA viruses (txid 439488), dsRNA viruses (txid 35325), and members of the genus Deltavirus (txid 39759) were retrieved from the NCBI RefSeq database (reference sequences) and used as subjects in local BLASTx searches with an e-value threshold of 1 × 10−5. Non-virus-originating hits were filtered out by performing Handling Editor: T. K. Frey. Electronic supplementary material The online version of this article (https ://doi.org/10.1007/s0070 5-020-04559 -w) contains supplementary material, which is available to authorized users. * Ruobing Guan guanruobing@henau.edu. cn * Tong Li tongli84@hotmail.com 1 College of Plant Protection, Henan Agricultural University, Zhengzhou 450002, China 2 University of Chinese Academy of Sciences, Beijing, China 3 Institute of Plant Protection/Henan Key Laboratory of Crop Pest Control/Key Laboratory of Integrated Pest Management on Crops in Southern Region of North China, Henan Academy of Agricultural Sciences, Zhengzhou 450002, China