Isolation, identification and molecular phylogenetic analysis of Hyblaea puera Nucleopolyhedrovirus
Author and article information
Abstract
Hyblaea puera (Lepidoptera: Hyblaeidae), is considered as a serious pest of teak in India and other tropical regions. It causes entire defoliation of teak trees and results in huge timber loss thereby decreasing forest productivity. Hyblaea puera Nucleopolyhedrovirus (HpNPV) is a baculovirus that has been employed in various parts of India as a bio-control agent against the pest H. puera. An unfeigned nucleopolyhedrovirus was isolated from the larvae of the moth, H. puera in Kerala, South India. Polh, lef-8, pif-2 gene sequences were amplified by PCR with degenerate primers and extracted for phylogenetic analysis. Hyblaea puera Nucleopolyhedrovirus appeared to be a distinct species of Group II NPV alphabaculovirus. Polyhedrin coding region was characterized by nucleotide sequence analysis. To date, Polyhedrin is the first isolated and characterized gene of HpNPV. It indicated the presence of ORF comprising 741 nucleotides which encode 246 amino acids with a predicted molecular mass of 28 KDa. Phylogeny based on three conserved baculovirus genes showed the highest homology of HpNPV to Helicoverpa armigera NPV. These findings were hardened by restriction endonuclease analysis, even though some differences in restriction pattern were observed. The current study will encourage future efforts to improve the efficacy of HpNPV against its natural host.
Cite this as
2018. Isolation, identification and molecular phylogenetic analysis of Hyblaea puera Nucleopolyhedrovirus. PeerJ Preprints 6:e27290v1 https://doi.org/10.7287/peerj.preprints.27290v1Author comment
This is a submission to PeerJ for review.
Sections
Supplemental Information
S1: Pairwise sequence comparisons of the deduced amino acid sequences of the concatenated lef-8 & pif-2 NPV genes used for phylogenetic analysis
S2: Pairwise comparison of the deduced amino acid sequences identity(%) of 36 polyhedrin genes of NPV used for phylogenetic analysis
Nucleotide sequence of the HpNPV polyhedrin gene and its flanking regions
The deduced amino acid sequences are indicated with one-letter code designation for polyhedrin. Putative baculovirus late promoter element ATAAG is shown in pink colour. Other potential eukaryotic promoter elements, TATA box and CAAT box are also shown, in green and blue respectively.
Nucleotide sequence of the HpNPV hypotetical protein
The deduced amino acids are indicated with one-letter code for hypothetical protein.
Comparison of nucleotide sequences of HpNPV and HearNPV
Dots indicate similarity and red indicates dissimilar nucleotide(s) of HearNPV.
Comparison of polyhedrin sequences of HpNPV and HearNPV
Dots in the HearNPV sequence indicates similar amino acids to HpNPV. Red indicates dissimilar amino acid(s).
Specific PCR amplification of HpNPV pol h and lef-8 genes
Agarose gel showing PCR amplification of full-length Polyhedrin gene
Specific PCR amplification of the HpNPV pif-2 gene.
Polyhedral Occlusion Bodies(POB) under light microscope(400x)
Additional Information
Competing Interests
The authors declare that they have no competing interests.
Author Contributions
Saranya Vijay Krishnan conceived and designed the experiments, performed the experiments, analyzed the data, contributed reagents/materials/analysis tools, prepared figures and/or tables, authored or reviewed drafts of the paper, approved the final draft.
Sajeev TV conceived and designed the experiments, analyzed the data, approved the final draft.
DNA Deposition
The following information was supplied regarding the deposition of DNA sequences:
HpNPV Sequences were deposited in GenBank and available under the following accession numbers: MH719085, MH2254886, MH254887 and MH362814.
Data Deposition
The following information was supplied regarding data availability:
Pairwise sequence comparisons of the deduced amino acid sequences of the concatenated lef-8 & pif-2 NPV genes after ClustalX used for phylogenetic analysis are provided in supplementary file 1.
Pairwise comparison of the deduced amino acid sequences identity(%) of 36 polyhedrin genes of NPV after ClustalX used for phylogenetic analysis are given in supplementary file 2.
Other data files regarding sequence features, and gel images are provided in the supplementary files from 3 to 10.
Funding
The authors received no funding for this work.
