The use of Cydia pomonella granulovirus (CpGV) isolates as biological control agents of codling moth (CM) larvae is important in organic and integrated pome fruit production worldwide. The commercially available isolates CpGV-0006, CpGV-R5, and CpGV-V15 have been selected for the control of CpGV resistant CM populations in Europe. In infection experiments, CpGV-0006 and CpGV-R5 were able to break type I resistance and to a lower extent also type III resistance, whereas CpGV-V15 overcame type I and the rarely occurring type II and type III resistance. The genetic background of the three isolates was investigated with next generation sequencing (NGS) tools by comparing their nucleotide compositions to whole genome alignments of five CpGV isolates representing the known genetic diversity of the CpGV genome groups A to E. Based on the distribution of single nucleotide polymorphisms (SNPs) in Illumina sequencing reads, we found that the two isolates CpGV-0006 and CpGV-R5 have highly similar genome group compositions, consisting of about two thirds of the CpGV genome group E and one third of genome group A. In contrast, CpGV-V15 is composed of equal parts of CpGV genome group B and E. According to the identified genetic composition of these isolates, their efficacy towards different resistance types can be explained and predictions on the success of resistance management strategies in resistant CM populations can be made.

Six complete genome sequences of Cydia pomonella granulovirus (CpGV) isolates from Mexico (CpGV-M and CpGV-M1), England (CpGV-E2), Iran (CpGV-I07 and CpGV-I12), and Canada (CpGV-S) were aligned and analyzed for genetic diversity and evolutionary processes. The selected CpGV isolates represented recently identified phylogenetic lineages of CpGV, namely, the genome groups A to E. The genomes ranged from 120,816 bp to 124,269 bp. Several common differences between CpGV-M, -E2, -I07, -I12 and -S to CpGV-M1, the first sequenced and published CpGV isolate, were highlighted. Phylogenetic analysis based on the aligned genome sequences grouped CpGV-M and CpGV-I12 as the most derived lineages, followed by CpGV-E2, CpGV-S and CpGV-I07, which represent the most basal lineages. All of the genomes shared a high degree of co-linearity, with a common setup of 137 (CpGV-I07) to 142 (CpGV-M and -I12) open reading frames with no translocations. An overall trend of increasing genome size and a decrease in GC content was observed, from the most basal lineage (CpGV-I07) to the most derived (CpGV-I12). A total number of 788 positions of single nucleotide polymorphisms (SNPs) were determined and used to create a genome-wide SNP map of CpGV. Of the total amount of SNPs, 534 positions were specific for exactly one of either isolate CpGV-M, -E2, -I07, -I12 or -S, which allowed the SNP-based detection and identification of all known CpGV isolates.

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The genome of Agrotis segetum nucleopolyhedrovirus B (AgseNPV-B) was completely sequenced and compared with whole genome sequences of the Agrotis segetum nucleopolyhedrovirus A (AgseNPV-A) and Agrotis ipsilon nucleopolyhedrovirus (AgipNPV). The AgseNPV-B genome is 148,981 bp in length and encodes 150 putative open reading frames. AgseNPV-B contains two copies of the gene viral enhancing factor (vef), making the Agrotis nucleopolyhedroviruses and A. segetum granulovirus (AgseGV) very rich in vef in comparison to other baculoviruses. Genome alignments of AgseNPV-B, AgseNPV-A and AgipNPV showed a very high genome co-linearity interspersed with variable regions, which are considered as putative sites of genomic recombination. Phylogenetic analyses revealed that all three viruses are distinct. However, AgseNPV-B is more closely related to AgipNPV suggesting that both viruses are at an early stage of phylogenetic divergence. It is proposed that AgseNPV-B belongs to a third Alphabaculovirus species of the Agrotis baculovirus complex. The Agrotis exclamationis nucleopolyhedrovirus (AgexNPV) shared high nucleotide sequence identities with AgseNPV-B, suggesting it is actually an AgseNPV-B isolate.

Mixed infections of insect larvae with different baculoviruses are occasionally found. They are of interest from an evolutionary as well as from a practical point of view when baculoviruses are applied as biocontrol agents. Here, we report mixed-infection studies of neonate larvae of the common cutworm, Agrotis segetum, with two baculoviruses, Agrotis segetum nucleopolyhedrovirus B (AgseNPV-B) and Agrotis segetum granulovirus (AgseGV). By applying quantitative PCR (qPCR) analysis, coinfections of individual larvae were demonstrated, and occlusion body (OB) production within singly infected and coinfected larvae was determined in individual larvae. Mixtures of viruses did not lead to changes in mortality rates compared with rates of singlevirus treatments, indicating an independent action within host larvae under our experimental conditions. AgseNPV-B-infected larvae showed an increase in OB production during 2 weeks of infection, whereas the number of AgseGV OBs did not change from the first week to the second week. Fewer OBs of both viruses were produced in coinfections than in singly infected larvae, suggesting a competition of the two viruses for larval resources. Hence, no functional or economic advantage could be inferred from larval mortality and OB production from mixed infections of A. segetum larvae with AgseNPV-B and AgseGV.A large number of baculoviruses isolated from the larvae of species of the insect orders Lepidoptera, Diptera, and Hymenoptera have been described (1). According to their occlusion body (OB) morphology, baculoviruses are distinguished into two morphological groups: the granuloviruses (GVs) with a single virion in an ovocylindrical OB (granule) and the nucleopolyhedroviruses (NPVs) with a few to many virions in a polyhedral OB (polyhedron) (2). Taxonomically, the Baculoviridae family is subdivided into four genera, the Alpha-, Beta-, Gamma-, and Deltabaculovirus genera (2, 3). Alpha-and betabaculoviruses comprise lepidopteran-specific NPVs and GVs, respectively. Many lepidopteran species are susceptible to baculoviruses from different species. Even simultaneous infections, so-called coinfections or mixed infections, of two NPVs (4), two GVs (5), and NPVs and GVs (6, 7) have been observed.Larvae of the turnip moth, Agrotis segetum (Denis & Schiffermüller) (Lepidoptera: Noctuidae), so-called common cutworms, are important agricultural pests (8). Two different alphabaculovirus isolates, Agrotis segetum nucleopolyhedrovirus A (AgseNPV-A; also called the Polish isolate) and A. segetum nucleopolyhedrovirus B (AgseNPV-B; also termed the Oxford isolate) (9-11), as well as the betabaculovirus Agrotis segetum granulovirus (AgseGV) (12), were isolated and characterized from A. segetum larvae. AgseGV was tested extensively as a biocontrol agent for the control of A. segetum in the field (13-16), and AgseNPV-B has shown its potential as a biocontrol agent under laboratory conditions (13).Both viruses were found to infect A. segetum larvae simultaneously (17), but little is known about possible in...

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The mechanisms by which transposable elements (TEs) can be horizontally transferred between animals are unknown, but viruses are possible candidate vectors. Here, we surveyed the presence of host-derived TEs in viral genomes in 35 deep sequencing datasets produced from eleven host-virus systems, encompassing nine arthropod host species (five lepidopterans, two dipterans and two crustaceans) and six different double-stranded (ds) DNA viruses (four baculoviruses and two iridoviruses). We found evidence of viral-borne TEs in 14 datasets, with frequencies of viral genomes carrying a TE ranging from 0.01 to 26.33% for baculoviruses and from 0.45 to 7.36% for iridoviruses. The analysis of viral populations separated by a single replication cycle revealed that viral-borne TEs originating from an initial host species can be retrieved after viral replication in another host species, sometimes at higher frequencies. Furthermore, we detected a strong increase in the number of integrations in a viral population for a TE absent from the hosts’ genomes, indicating that this TE has undergone intense transposition within the viral population. Finally, we provide evidence that many TEs found integrated in viral genomes (15/41) have been horizontally transferred in insects. Altogether, our results indicate that multiple large dsDNA viruses have the capacity to shuttle TEs in insects and they underline the potential of viruses to act as vectors of horizontal transfer of TEs. Furthermore, the finding that TEs can transpose between viral genomes of a viral species sets viruses as possible new niches in which TEs can persist and evolve.

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