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Published on: July 27, 2018
Genomic sequence analysis of a nucleopolyhedrovirus isolated from the diamondback moth, Plutella xylostella
Robert L Harrison1, Dwight E Lynn
1Insect Biocontrol Laboratory, USDA Agricultural Research Service, Plant Sciences Institute, Building 011A, Room 214, BARC-W, 10300 Baltimore Avenue, Beltsville, MD 20705, USA. Robert.L.Harrison@ars.usda.gov
Abstract:
The CL3 plaque isolate of Plutella xylostella multiple nucleopolyhedrovirus (PlxyMNPV-CL3) exhibits a high degree of genetic similarity with the Autographa californica MNPV (AcMNPV) but is significantly more virulent against the diamondback moth, P. xylostella, than AcMNPV. To identify genetic differences between PlxyMNPV-CL3 and AcMNPV that may account for the difference in virulence against P. xylostella, the genome sequence of the CL3 plaque isolate of PlxyMNPV was determined and compared to the genome sequence of AcMNPV isolate C6. The PlxyMNPV genome is 134,417 bp, 523 bp larger than the AcMNPV-C6 genome, and the nucleotide sequence is almost completely co-linear with that of AcMNPV-C6. Of the 153 open reading frames (ORFs) identified in PlxyMNPV, 151 had homologues in AcMNPV-C6, with a mean amino acid sequence identity of 98.5%. The PlxyMNPV genome possessed two features previously reported for other variants of AcMNPV: (1) an extra baculovirus repeated orf (bro) sequence located between the plxy29/ac30 and sod ORFs, and (2) the deletion of the AcMNPV pnk/pnl polynucleotide kinase/ligase gene. In addition, an 817 bp insert of unknown origin located between the fp25K and lef-9 genes was discovered. This insert contained two small ORFs and was detected in both tissue culture- and larvae-derived PlxyMNPV DNA by PCR. Finally, the PlxyMNPV-CL3 ie-2 gene encodes a product with a low level (37.3%) of amino acid sequence identity with the AcMNPV-C6 ie-2 product. PlxyMNPV-CL3 apparently acquired this variant ie2 gene by recombination with an undescribed nucleopolyhedrovirus.
Insights
Plutella xylostella multiple nucleopolyhedrovirus (PlxyMNPV-CL3) is more virulent than Autographa californica MNPV (AcMNPV) against diamondback moths. Genetic analysis revealed unique gene insertions and variations, including a distinct ie-2 gene, contributing to PlxyMNPV-CL3
Area of Science:
- Virology
- Insect Pathology
- Genomics
Background:
- Plutella xylostella multiple nucleopolyhedrovirus (PlxyMNPV-CL3) shows high genetic similarity to Autographa californica MNPV (AcMNPV).
- PlxyMNPV-CL3 exhibits significantly higher virulence against diamondback moth (P. xylostella) compared to AcMNPV.
Purpose of the Study:
- To identify genetic differences between PlxyMNPV-CL3 and AcMNPV.
- To understand the genetic basis for the enhanced virulence of PlxyMNPV-CL3 against P. xylostella.
Main Methods:
- Whole genome sequencing of PlxyMNPV-CL3.
- Comparative genomic analysis with AcMNPV isolate C6.
- Polymerase Chain Reaction (PCR) to detect specific genetic inserts.
Main Results:
- PlxyMNPV genome is 134,417 bp, slightly larger than AcMNPV-C6.
- 151 out of 153 identified open reading frames (ORFs) in PlxyMNPV have homologues in AcMNPV-C6 with 98.5% mean amino acid identity.
- Identified unique features: an additional baculovirus repeated orf (bro) sequence, deletion of the polynucleotide kinase/ligase gene, an 817 bp insert with two ORFs, and a divergent ie-2 gene.
Conclusions:
- The genetic variations, particularly the unique ie-2 gene acquired through recombination, likely contribute to the increased virulence of PlxyMNPV-CL3.
- The study elucidates key genomic distinctions driving differential virulence in related baculoviruses.
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