Molecular identification and phylogenetic analysis of baculoviruses from Lepidoptera.
PCR amplification of the highly conserved baculovirus genes late expression factor 8 (lef-8), late expression factor 9 (lef-9) and polyhedrin/granulin (polh/gran) combined with molecular phylogenetic analyses provide a powerful tool to identify lepidopteran-specific baculoviruses and to study their diversity. In the present investigation, we have improved the degenerate oligonucleotides and corroborated the approach that was recently described by Lange et al. (Lange, M., Wang, H., Zhihong, H., Jehle, J.A., 2004. Towards a molecular identification and classification system of lepidopteran-specific baculoviruses. Virology 325, 36-47.). Baculovirus DNA was isolated from 71 uncharacterized historic baculovirus samples, and partial gene sequences were amplified by using gene-specific degenerate PCR primers. The obtained PCR products were directly sequenced, and the deduced amino acid sequences were compiled and aligned with published sequences of these target genes. A phylogenetic tree of 117 baculoviruses was inferred using maximum parsimony and distance methods. Based on the comprehensive phylogenetic analysis of the partial lef-8, lef-9 and polh/gran genes, we propose a phylogenetic species criterion for lepidopteran-specific baculoviruses that uses the genetic distances of these genes for species demarcation.
Jehle
Johannes A
JA
Laboratory for Biotechnological Crop Protection, Department of Phytopathology, Agricultural Service Center Palatinate (DLR Rheinpfalz), Breitenweg 71, 67435 Neustadt an der Weinstrasse, Germany. Johannes.Jehle@dlr.rlp.de
Lange
Martin
M
Wang
Hualin
H
Hu
Zhihong
Z
Wang
Yongjie
Y
Hauschild
Rüdiger
R
D000818:Animals; D016367:Baculoviridae / Q000145:classification* / Q000235:genetics*; D004279:DNA, Viral / Q000032:analysis; D007915:Lepidoptera / Q000821:virology*; D008969:Molecular Sequence Data; D000077183:Occlusion Body Matrix Proteins; D010802:Phylogeny; D017422:Sequence Analysis, DNA; D014764:Viral Proteins / Q000235:genetics; D015678:Viral Structural Proteins
D004279:DNA, Viral; D000077183:Occlusion Body Matrix Proteins; D014764:Viral Proteins; D015678:Viral Structural Proteins; C087702:late expression factor 8, Baculovirus; C009829:polyhedrin protein, Nucleopolyhedrovirus