Morphological Characteristics of Erannis ankeraria Nucleopolyhedrovirus and Establishment of Molecular Detection Technique
[Objective]Erannis ankeraria nucleopolyhedrovirus(EranNPV)is a significant natural enemy microbial product that effectively regulates the population and quality of E.ankeraria.The elucidation of the ultramicromorphological characteristics of EranNPV and the establishment of a sensitive and convenient PCR-based rapid detection method will help to further investigate the epidemiology and transmission mechanisms of EranNPV in forest ecosystems.This,in turn,provides technical support for the continuous control of E.ankerariausing EranNPV.[Method]The external morphology and internal structural features of EranNPV polyhedra were observed using scanning electron microscopy and transmission electron microscopy.A pair of PCR detection primers,EaV,was designed based on the ac54 gene of EranNPV.The specificity of EaV was examined by PCR amplification results of various insect viruses.The sensitivity of PCR amplification using EaV primers was determined by testing different concentrations of EranNPV genomic DNA and polyhedrons.[Result]The electron microscopy results showed that the majority of the virus polyhedra were irregular polyhedra with smooth surfaces,and a few had scattered pore depressions with an average diameter of 1.43±0.20 μm.The virus particles within the polyhedra were distributed in a single bundle with an average length of 219.14±17.50 nm.When EaV is used as the primer,only EranNPV can be amplified to the target band.The PCR assay using EaV primers exhibited a sensitivity of 10 fg/mL for EranNPV genomic DNA and a sensitivity of 1×102 OBs/mL for polyhedrons.[Conclusion]EranNPV is a typical single nucleocapsid nucleopolyhedrovirus.The PCR assay using EaV primers demonstrates high specificity and sensitivity.It is expected to be further applied to the study of the diffusion and transmission mechanism of EranNPV in forest ecosystems.
Erannis ankerarianucleopolyhedrovirusscanning electron microscopetransmission electron microscopegraphPCR detection
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NSTL
万方数据
维普
