Genomic sequencing and analyses of HearMNPV--a new Multinucleocapsid nucleopolyhedrovirus isolated from Helicoverpa armigera.
BACKGROUND
HearMNPV, a nucleopolyhedrovirus (NPV), which infects the cotton bollworm, Helicoverpa armigera, comprises multiple rod-shaped nucleocapsids in virion(as detected by electron microscopy). HearMNPV shows a different host range compared with H. armigera single-nucleocapsid NPV (HearSNPV). To better understand HearMNPV, the HearMNPV genome was sequenced and analyzed.


METHODS
The morphology of HearMNPV was observed by electron microscope. The qPCR was used to determine the replication kinetics of HearMNPV infectious for H. armigera in vivo. A random genomic library of HearMNPV was constructed according to the "partial filling-in" method, the sequence and organization of the HearMNPV genome was analyzed and compared with sequence data from other baculoviruses.


RESULTS
Real time qPCR showed that HearMNPV DNA replication included a decreasing phase, latent phase, exponential phase, and a stationary phase during infection of H. armigera. The HearMNPV genome consists of 154,196 base pairs, with a G + C content of 40.07%. 162 putative ORFs were detected in the HearMNPV genome, which represented 90.16% of the genome. The remaining 9.84% constitute four homologous regions and other non-coding regions. The gene content and gene arrangement in HearMNPV were most similar to those of Mamestra configurata NPV-B (MacoNPV-B), but was different to HearSNPV. Comparison of the genome of HearMNPV and MacoNPV-B suggested that HearMNPV has a deletion of a 5.4-kb fragment containing five ORFs. In addition, HearMNPV orf66, bro genes, and hrs are different to the corresponding parts of the MacoNPV-B genome.


CONCLUSIONS
HearMNPV can replicate in vivo in H. armigera and in vitro, and is a new NPV isolate distinguished from HearSNPV. HearMNPV is most closely related to MacoNPV-B, but has a distinct genomic structure, content, and organization.
Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing, 100081, China.
Ping
P
Tang
Huan
H
Zhang
Yinü
Y
Li
Bin
B
Han
Guozeng
G
Wang
Qilian
Q
Qin
Zhifang
Z
Zhang
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003588:Cytopathogenic Effect, Viral; D023061:Gene Order; D016679:Genome, Viral; D007814:Larva / Q000821:virology; D008969:Molecular Sequence Data; D009036:Moths / Q000821:virology*; D017924:Nucleopolyhedroviruses / Q000145:classification / Q000235:genetics* / Q000648:ultrastructure; D016366:Open Reading Frames; D010802:Phylogeny; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D014779:Virus Replication