Native range assessment of classical biological control agents: impact of inundative releases as pre-introduction evaluation

Diadromus pulchellus Wesmael (Hymenoptera: Ichneumonidae) is a pupal parasitoid under consideration for introduction into Canada for the control of the invasive leek moth, Acrolepiopsis assectella (Zeller) (Lepidoptera: Acrolepiidae). Since study of the parasitoid outside of quarantine was not permitted in Canada at the time of this project, we assessed its efficacy via field trials in its native range in central Europe. This was done by simulating introductory releases that would eventually take place in Canada when a permit for release is obtained. In 2007 and 2008, experimental leek plots were artificially infested with pest larvae to mimic the higher pest densities common in Canada. Based on a preliminary experiment showing that leek moth pupae were suitable for parasitism up to 5–6 days after pupation, D. pulchellus adults were mass-released into the field plots when the first host cocoons were observed. The laboratory-reared agents reproduced successfully in all trials and radically reduced leek moth survival. Taking into account background parasitism caused by naturally occurring D. pulchellus, the released agents parasitized at least 15.8%, 43.9%, 48.1% and 58.8% of the available hosts in the four release trials. When this significant contribution to leek moth mortality is added to previously published life tables, in which pupal parasitism was absent, the total pupal mortality increases from 60.1% to 76.7%. This study demonstrates how field trials involving environmental manipulation in an agent's native range can yield predictions of the agent's field efficacy once introduced into a novel area.

Characterization of repetitive DNA regions and methylated DNA in ascovirus genomes

The accompanying phylogenetic study of large double-stranded DNA viruses based on their δ DNA polymerase genes suggests that ascoviruses (family Ascoviridae) and iridoviruses (family Iridoviridae) are closely related and may share a common ancestor. This relationship was unexpected because of marked differences between these viruses. Iridoviruses produce icosahedral virions and occur broadly among vertebrates and invertebrates, whereas ascoviruses typically produce reniform or bacilliform virions and are restricted to insect hosts, primarily lepidopterans. Detailed comparisons of these two virus types are not possible because fundamental information on the properties of the virions and their genomes is lacking, especially for ascoviruses. To facilitate further investigation of the putative evolutionary relationship between ascoviruses and iridoviruses, the genomes of representative viruses from each family were compared with respect to physical configuration, presence of DNA repeats and degree of DNA methylation. Genomes from Spodoptera frugiperda (SfAV1), Heliothis virescens (HvAV3) and Diadromus pulchellus (DpAV4) ascoviruses were all found to be circular and partially superhelical and to contain large interspersed repeats of 1–3 kbp. Mosquito (IV type 3), lepidopteran (IV type 6) and isopod (IV type 31) iridovirus genomes were all linear and lacked large regions of repetitive DNA. Ascovirus and iridovirus genomes were methylated and one, DpAV4, had the highest degree of methylation of any reported animal DNA virus. The major differences in the physical and biochemical characteristics of ascoviruses and iridoviruses reported here provide a foundation for further studies of their relatedness while making their possible close relationship and divergence during evolution of even greater interest.

Phylogenetic position of the Diadromus pulchellus ascovirus DNA polymerase among viruses with large double-stranded DNA genomes

The Ascoviridae is a family of large double-stranded (ds) DNA insect viruses that contains four species, the Spodoptera frugiperda (SfAV1), Trichoplusia ni (TnAV2), Heliothis virescens (HvAV3) and Diadromus pulchellus (DpAV4) ascoviruses. These are unique among insect viruses in that the primary means of transmission among their lepidopteran hosts is generally by being vectored mechanically by hymenopteran parasitoids. Ascoviruses are similar in virion structure, but their relationships with their parasitoid vectors vary from being opportunistic to obligate. Little is known, however, about the relatedness of these viruses to one another or to other large dsDNA viruses. We therefore cloned and sequenced the δ DNA polymerase gene of DpAV4, characterized it and compared it to 59 eukaryotic and viral δ and ϵ DNA polymerases. Phylogenetic analyses based on these genes revealed that the ascoviruses DpAV4 and SfAV1 formed a group of virus species distinct from, but closely related to, species of the family Iridoviridae. Detailed analyses of the relatedness of ascovirus species based on conserved δ DNA polymerase motifs showed two groups within the family Ascoviridae, one containing DpAV4 and the other containing SfAV1, TnAV2 and HvAV3, which was consistent with their host–vector relationships. Despite significant differences in capsid symmetry between ascoviruses and iridoviruses, these results suggest that these viruses may have originated from a common ancestral virus.