Abstract
Baculoviridae is a highly diverse family of rod-shaped viruses with double-stranded DNA. To date, almost 100 species have had their complete genomic sequences deposited in the GenBank database, a quarter of which comprises granuloviruses (GVs). Many of the genomes are sequenced using next-generation sequencing, which is currently considered the best method for characterizing new species, but it is time-consuming and expensive. Baculoviruses form a safe alternative to overused chemical pesticides and therefore there is a constant need for identifying new species that can be active components of novel biological insecticides. In this study, we have described a fast and reliable method for the detection of new and differentiation of previously analyzed granulovirus species based on a real-time polymerase chain reaction (PCR) technique with melting point curve analysis. The sequences of highly conserved baculovirus genes, such as granulin and late expression factors 8 and 9 (lef-8 and lef-9), derived from GVs available to date have been analyzed and used for degenerate primer design. The developed method was tested on a representative group of eight betabaculoviruses with comparisons of melting temperatures to allow for quick and preliminary granulovirus detection. The proposed real-time PCR procedure may be a very useful tool as an easily accessible screening method in a majority of laboratories.
Highlights
Insect pests pose a threat for crops and forests all over the world
There are 24 betabaculovirus genomes deposited in the GenBank database, all of which differ in the nucleotide sequence (Geneious Pro 7.1 (Biomatters, Auckland, New Zealand)-MAFFT
Real-time polymerase chain reaction (PCR) is commonly used for the detection of many different viruses, from various families, chosen
Summary
Insect pests pose a threat for crops and forests all over the world. To manage them, chemical control agents are usually applied, and are very often overused, because an immediate effect is sought.To reduce chemicals in the environment, there is a constant need for finding novel biologically safe alternatives. Insect pests pose a threat for crops and forests all over the world. Chemical control agents are usually applied, and are very often overused, because an immediate effect is sought. To reduce chemicals in the environment, there is a constant need for finding novel biologically safe alternatives. Some very promising candidates for biopesticides belong to the Baculoviridae family, which is currently the biggest insect virus family, comprising around 1000 described species of insect viruses [1,2,3]. Baculoviruses are known to naturally control many insect populations. They possess a very narrow host range and do not accumulate in the environment and, being selective, Viruses 2019, 11, 115; doi:10.3390/v11020115 www.mdpi.com/journal/viruses
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