A bacmid approach to the genetic manipulation of granuloviruses

Hilton, Sally; Kemp, E. Helen; Keane, Gary; Winstanley, Doreen

doi:10.1016/j.jviromet.2008.05.015

Cited by 26 publications

(22 citation statements)

References 27 publications

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“…To elucidate the potential role of pe38 in CpGV resistance, pe38 knockout and recovery mutants were constructed with the help of a CpGV bacmid (34). This bacmid, bacCpGV, is based on the genome of CpGV-M1, an in vivo cloned genotype of CpGV-M, and contains an 8.6-kbp BAC cassette, including an attachment site for 3 OB/mL of the isolates CpGV-M, -I12, -S, -E2, or -I07.…”

Section: Resultsmentioning

confidence: 99%

Baculovirus resistance in codling moth is virus isolate-dependent and the consequence of a mutation in viral genepe38

Gebhardt

Eberle

Radtke

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

103

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Significance Registered in 34 countries worldwide, Cydia pomonella granulovirus (CpGV) is a highly important biological agent to control the codling moth in pome fruit production. Since 2005, 38 codling moth field populations resistant to CpGV products containing the Mexican isolate (CpGV-M) were discovered in Europe, and this was the first demonstrated field resistance against commercial baculovirus products. By identifying and sequencing different resistance-breaking CpGV isolates, it was found that resistance of codling moth is directed toward CpGV-M only. As shown by mutation experiments, CpGV-M is prone to resistance because of a 24-nucleotide repeat within the viral gene pe38 . Thus, pe38 can be used as a genetic marker to identify resistance-breaking CpGV isolates for resistance management programs.

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Section: Resultsmentioning

confidence: 99%

Baculovirus resistance in codling moth is virus isolate-dependent and the consequence of a mutation in viral genepe38

Gebhardt

Eberle

Radtke

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

103

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“…CpGV is the only member of the betabaculoviruses whose replication has been studied in detail in vivo and in vitro (Winstanley & Crook, 1993;Hilton & Winstanley, 2007). Recently its genome has been genetically manipulated to generate a bacmid (Hilton et al, 2008).…”

Section: Introductionmentioning

confidence: 99%

Diversity and evolution of the Cydia pomonella granulovirus

Eberle

Sayed

Rezapanah³

et al. 2009

Journal of General Virology

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Eight new field isolates of Cydia pomonella granulovirus (CpGV) originating in Iran and Georgia and one English CpGV isolate were analysed for restriction fragment length polymorphisms (RFLPs) and by partial genome amplification and sequencing. According to the observed RFLPs, most of the predominant genotypes of these isolates could be assigned to those present in previously found isolates originating from Mexico (CpGV-M), England (CpGV-E) and Russia (CpGV-R). We suggest that these isolates should be designated genome A, B and C types, respectively. A fourth genome type was identified in three isolates and is designated D type. The isolates with A, B and D type genomes contained four open reading frames (ORFs) (ORF63-ORF66) not present in C type genomes. The lack of these ORFs in other granuloviruses suggests that the C type genome is evolutionarily ancestral to the other genome types. The B and D type genomes contained an additional insertion of a non-protein coding region of 0.7 kb, which was at different genome locations. Analysis of the partial gene sequences of late expression factor 8 (lef-8), lef-9 and polyhedrin/granulin (polh/gran) genes revealed single nucleotide polymorphisms (SNPs) that corresponded to the RFLP types. Phylogenetic analyses based on these SNPs corroborated the proposed ancestry of the C type genome. C type viruses were also less virulent to neonate codling moth larvae than the other virus types. In conclusion, the known diversity of CpGV isolates can be described by four major genome types, which appear to exist in different isolates as genotype mixtures.

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“…Helicoverpa armigera nucleopolyhedrovirus (Wang et al, 2003), Spodoptera exigua multiple nucleopolyhedrovirus (Pijlman et al, 2002) and Bombyx mori nucleopolyhedrovirus (Yao et al, 2012). Hilton et al (2008) constructed a bacmid of CpGV also using the unique Pac I site for inserting the transfer vector cassette that derived from the AcMNPV Bac-to-Bac system. In the present study, we used the CopyControl pCC1BAC cloning vector as the bacmid backbone (Wang et al, 2010).…”

Section: Discussionmentioning

confidence: 99%

Possible functional co-operation of palindromes hr3 and hr4 in the genome of Cydia pomonella granulovirus affects viral replication capacity

Elmenofy

Jehle

2015

Journal of General Virology

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After previous studies had shown that natural transposon insertion between the two homologous regions hr3 and hr4 of the genome of the Mexican (M) strain of Cydia pomonella granulovirus (CpGV-M) resulted in a loss of viral competitiveness, the function of these homologous regions was investigated. A CpGV-based bacmid (CpBAC) was constructed and mutants with deleted hr3 and hr4 palindromes (CpBAC hr3/hr4 KO ) and a construct (CpBAC hr3-kan-hr4 ) with physically separated hr3 and hr4 repeats were generated to investigate their involvement in in vivo replication. Based on median lethal concentration (LC 50 ) and median survival time (ST 50 ) of the mutant viruses vCpBAC hr3/hr4 KO and vCpBAC hr3-kan-hr4 it was found that the infectivity of both mutants for codling moth Cydia pomonella L. (Lep.: Tortricidae) larvae was not influenced compared with the parental virus vCpBAC. Co-infection experiments with vCpBAC hr3-kan-hr4 and vCpBAC using different virus ratios revealed that vCpBAC hr3-kan-hr4 was efficiently out-competed by vCpBAC during in vivo replication. These findings suggested that the separation of hr3 and hr4 resulted in a replication disadvantage of the mutant similar to the observation made in previous co-infection experiments using the transposon-carrying mutant CpGV-MCp5 and WT CpGV-M. It was concluded that the palindromes hr3 and hr4 may play a non-essential but co-functional role in the replication of CpGV-M.

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A bacmid approach to the genetic manipulation of granuloviruses

Cited by 26 publications

References 27 publications

Baculovirus resistance in codling moth is virus isolate-dependent and the consequence of a mutation in viral genepe38

Baculovirus resistance in codling moth is virus isolate-dependent and the consequence of a mutation in viral genepe38

Diversity and evolution of the Cydia pomonella granulovirus

Possible functional co-operation of palindromes hr3 and hr4 in the genome of Cydia pomonella granulovirus affects viral replication capacity

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