| Publication number |
CLPUB00722 |
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| Authors |
Hepat R.P., Erlandson M.A., Harrison R.L., Willis L.G., Theilmann D.A. |
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| Title |
Role of Plutella xylostella nucleopolyhedrovirus-CL3 ie2 in host range adaptation. |
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| Citation |
(In conference) 50th annual meeting and golden jubilee celebration of the Society for Invertebrate Pathology; pp.11-11; Society for Invertebrate Pathology; San Diego; USA (2017) |
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| Web pages |
http://www.sipweb.org/docs/SIP%202017_Complete%20Program.pdf |
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| Abstract |
The diamondback moth (Plutella xylostella) is one of the most important
destructive insect pests of cruciferous crops worldwide. Previous studies
identified both alpha-and betabaculoviruses that can infect and kill this
important lepidopteran pest. The genome of the alphabaculovirus isolate
Plutella xylostella nucleopolyhedrovirus-CL3 (PlxyNPV-CL3) was recently
sequenced and was found to be nearly identical (98.5%) to the genome of
Autographa californica multiple nucleopolyhedrovirus (AcMNPV).
Interestingly PlxyNPV-CL3 is highly lethal for P. xylostella, whereas
AcMNPV is not. Therefore, any differences in the genome of these two
viruses may identify genes that are required for host adaptation. The
dominant difference between the PlxyNPV-CL3 and AcMNPV-C6 genome is the
open reading frame of the ie2 gene. The predicted amino acid sequences of
the two IE2 proteins exhibit only 37.5% similarity. To determine if
PlxyNPV-CL3 IE2 is responsible for adaptation of AcMNPV to P. xylostella,
recombinant viruses were generated using bacmids. These viruses were an
ie2 knockout (vie2KO), and the ie2 knockout repaired with AcMNPV or
PlxyNPV-CL3 ie2 tagged with HA epitope (vie2KO-acie2HA and vie2KO-pxie2HA
respectively). Each of these viruses was tested for replication in various
cell lines which including Sf9, Tn5b1-4, and PxE-Po#583. Previous studies
have not analyzed the impact of a complete AcMNPV ie2 knockout.
Surprisingly, our results showed that deletion of ie2 led to a greater
than 90% reduction in budded virus (BV) production compared to wildtype
virus. In addition, occlusion body (OB) production was also greatly
reduced. Initial results with vie2KO-pxie2HA indicate that PlxyNPV-CL3 IE2
can rescue vie2KO producing BV titers nearly as high as AcMNPV IE2.
Further studies analyzing virus infection in P. xylotella larvae will be
reported.
|
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| Cell lines |
CVCL_C2VX; PxE-Po#583 |
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