Abstract
Drosophila suzukii (Ds) is an invasive pest insect that infests ripening fruit, causing severe economic losses. Control measures based on chemical pesticides are inefficient and undesirable, so biological alternatives have been considered, including native Ds viruses. We previously isolated a strain of La Jolla virus (LJV-Ds-OS20) from Ds in Germany as a candidate biopesticide. Here we characterized the new strain in detail, focusing on the processing of its capsid proteins. We tested LJV growth during Ds development to optimize virus production, and established a laboratory production system using adult flies. This system was suitable for the preparation of virions for detailed analysis. The LJV-Ds-OS20 isolate was cloned by limiting dilution and the complete nucleotide sequence was determined as a basis for protein analysis. The terminal segments of the virus genome were completed by RACE-PCR. LJV virions were also purified by CsCl gradient centrifugation and analyzed by SDS-PAGE and electron microscopy. The capsid proteins of purified LJV virions were resolved by two-dimensional SDS-PAGE for N-terminal sequencing and peptide mass fingerprinting. The N-terminal sequences of VP1 and VP2, together with MS data representing several capsid proteins, allowed us to develop a model for the organization of the LJV structural protein region. This may facilitate the development of new viral strains as biopesticides.
Highlights
Drosophila suzukii (Ds; Matsumura, 1931) was originally native to East and Southeast Asia but has spread beyond its native range due to global trade and transport
We recently identified La Jolla virus (LJV, Iflaviridae) in Ds specimens captured in Germany [11], and others have detected the same virus species in Ds specimens from France, the UK and Japan [12]
LJV-Ds-OS20-open reading frame (ORF) sequences were aligned with 56 LJV sequences in GenBank using Geneious v10.2.6 and the ClustalW algorithm, and were manually edited to correct possible errors
Summary
Drosophila suzukii (Ds; Matsumura, 1931) was originally native to East and Southeast Asia but has spread beyond its native range due to global trade and transport This invasive species, known as the “spotted wing Drosophila”, is an economically devastating pest of fruit crops in the Americas and Europe [1,2,3,4]. The Iflavirus genome features a single open reading frame (ORF) that is translated into a polyprotein. The latter is processed into its functional subunits by a viral 3C-like protease [13]. We purified strain LJV-Ds-OS20 from Ds specimens captured in west-central Germany, allowing us to sequence the genome and investigate the processing of the capsid proteins, leading to a model of the structural protein region
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