Towards a molecular identification and classification system of lepidopteran-specific baculoviruses.
Virus genomics provides novel approaches for virus identification and classification. Based on the comparative analyses of sequenced lepidopteran-specific baculovirus genomes, degenerate oligonucleotides were developed that allow the specific amplification of several regions of the genome using polymerase chain reaction (PCR) followed by DNA sequencing. The DNA sequences within the coding regions of three highly conserved genes, namely polyhedrin/granulin (polh/gran), late expression factor 8 (lef-8), and late expression factor 9 (lef-9), were targeted for amplification. The oligonucleotides were tested on viral DNAs isolated from historical field samples, and amplification products were generated from 12 isolated nucleopolyhedrovirus (NPV) and 8 granulovirus (GV) DNAs. The PCR products were cloned or directly sequenced, and phylogenetic trees were inferred from individual and combined data sets of these three genes and compared to a phylogeny, which includes 22 baculoviruses using a combined data set of 30 core genes. This method allows a fast and reliable detection and identification of lepidopteran-specific NPVs and GVs. Furthermore, a strong correlation of the base composition of these three genome areas with that of the complete virus genome was observed and used to predict the base composition of uncharacterized baculovirus genomes. These analyses suggested that GVs have a significantly higher AT content than NPVs.
Lange
Martin
M
Laboratory for Biotechnological Crop Protection, Department of Phytopathology, Agricultural Service Center Palatinate, 67435 Neustadt an der Weinstrasse, Germany.
Wang
Hualin
H
Zhihong
Hu
H
Jehle
Johannes A
JA
D000818:Animals; D016367:Baculoviridae / Q000145:classification* / Q000235:genetics; D016679:Genome, Viral; D007915:Lepidoptera / Q000821:virology*; D000077183:Occlusion Body Matrix Proteins; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D014764:Viral Proteins / Q000235:genetics; D015678:Viral Structural Proteins
D000077183:Occlusion Body Matrix Proteins; D014764:Viral Proteins; D015678:Viral Structural Proteins; C087702:late expression factor 8, Baculovirus; C009829:polyhedrin protein, Nucleopolyhedrovirus