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Published on: March 9, 2022
Characterization and polyhedrin gene cloning of Lymantria xylina multiple nucleopolyhedrovirus
Chih-Yu Wu1, Chung-Hsiung Wang
1Department of Entomology, National Taiwan University, Taipei, Taiwan, ROC.
Abstract:
A baculovirus has been isolated from infected larvae of the casuarina moth, Lymantria xylina Swinehoe (Lepidoptera: Lymantriidae) in Taiwan. Ultrastructural observation revealed that this virus is a multiple nucleopolyhedrovirus (MNPV), and the name L. xylina MNPV (LyxyMNPV) was proposed. Restriction endonuclease (BamHI, EcoRI, and EcoRV) profiles of LyxyMNPV genome differed from those of other known NPVs. The size of the LyxyMNPV genome was estimated to be 154+/-1.26 kbp (mean+/-SE). The polyhedrin gene is located in the BamHI-D, EcoRI-C, and EcoRV-K fragments of LyxyMNPV genome. The gene organization of the LyxyMNPV EcoRV-K fragment and the phylogenetic analysis based on the polyhedrin gene sequences showed that LyxyMNPV is closely related to the L. dispar MNPV (LdMNPV). Furthermore, a rapid assay method was developed to distinguish LyxyMNPV from LdMNPV based on PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism) analysis of polyhedrin genes.
Insights
A novel baculovirus, Lymantria xylina multiple nucleopolyhedrovirus (LyxyMNPV), was identified in Taiwan. This study characterizes its genome and develops a PCR-RFLP assay for distinguishing it from related viruses.
Area of Science:
- Entomology
- Virology
- Molecular Biology
Background:
- Baculoviruses are important insect pathogens.
- Nucleopolyhedroviruses (NPVs) are a significant group within baculoviruses.
- Identification and characterization of novel baculoviruses are crucial for understanding insect-pathogen interactions and developing biocontrol agents.
Purpose of the Study:
- To isolate and characterize a baculovirus from the casuarina moth, Lymantria xylina.
- To determine the genomic properties and phylogenetic relationships of the isolated virus.
- To develop a method for differentiating the new virus from related NPVs.
Main Methods:
- Isolation and ultrastructural observation of the baculovirus.
- Restriction endonuclease digestion (BamHI, EcoRI, EcoRV) of the viral genome.
- Estimation of viral genome size.
- Localization and sequencing of the polyhedrin gene.
- Phylogenetic analysis using polyhedrin gene sequences.
- Development of a PCR-RFLP assay.
Main Results:
- A multiple nucleopolyhedrovirus (MNPV), named L. xylina MNPV (LyxyMNPV), was isolated.
- LyxyMNPV genome size was estimated at 154+/-1.26 kbp.
- The polyhedrin gene was located on specific restriction fragments (BamHI-D, EcoRI-C, EcoRV-K).
- Genomic profiles differed from other known NPVs.
- Phylogenetic analysis indicated LyxyMNPV is closely related to L. dispar MNPV (LdMNPV).
- A rapid PCR-RFLP assay was successfully developed to distinguish LyxyMNPV from LdMNPV.
Conclusions:
- LyxyMNPV represents a distinct MNPV species infecting Lymantria xylina.
- Genomic and phylogenetic analyses provide insights into baculovirus evolution.
- The developed PCR-RFLP assay offers a practical tool for rapid identification and differentiation of LyxyMNPV.
