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Genomic sequence analysis of a nucleopolyhedrovirus isolated from the diamondback moth, Plutella xylostella

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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.

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Acknowledgements

The authors wish to thank Tad Sonstegard (USDA-ARS, Bovine Functional Genomics Laboratory) for the use of his GeneMachines Hydrohear, Wendy Sparks and Bryony Bonning (Iowa State University) for the gift of many oligonucleotides, Robert Farrar (USDA-ARS, Insect Biocontrol Laboratory) for larvae-derived polyhedra stocks of PlxyMNPV-CL3, Ellen Buckley (USDA-ARS, Insect Biocontrol Laboratory) for assistance with sequencing, and Stephen Rehner (USDA-ARS, Insect Biocontrol Laboratory) for helpful advice and discussion throughout the course of this study. Mention of trade names or commercial products in this publication is solely for the purpose of providing specific information and does not imply recommendation or endorsement by the U.S. Department of Agriculture

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Correspondence to Robert L. Harrison.

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The nucleotide sequence data reported in this paper have been submitted to the GenBank nucleotide sequence database and have been assigned the accession numbers DQ457003 (PlxyMNPV-CL3), DQ482644 (AcMNPV-C6 ac17), DQ482645 (AcMNPV-C6 ac52), DQ482646 (AcMNPV-C6 ac58/ac59), DQ482647 (AcMNPV-C6 ac106/ac107), DQ482648 (AcMNPV-C6 ac112/ac113), DQ482649 (AcMNPV-C6 odv-e18), DQ482650 (AcMNPV-C6 ac145), DQ482651 (AcMNPV-C6 arif-1), and DQ482652 (AcMNPV-C6 pep).

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Harrison, R.L., Lynn, D.E. Genomic sequence analysis of a nucleopolyhedrovirus isolated from the diamondback moth, Plutella xylostella . Virus Genes 35, 857–873 (2007). https://doi.org/10.1007/s11262-007-0136-6

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