Morphological, genetic and biological characterisation of a novel alphabaculovirus isolated from Cryptophlebia peltastica (Lepidoptera: Tortricidae)
- Marsberg, Tamryn, Jukes, Michael, Krejmer-Rabalska, Martyna, Rabalski, Lukasz, Knox, Caroline, Moore, Sean D, Hill, Martin P, Szewczyk, Boguslaw
- Authors: Marsberg, Tamryn , Jukes, Michael , Krejmer-Rabalska, Martyna , Rabalski, Lukasz , Knox, Caroline , Moore, Sean D , Hill, Martin P , Szewczyk, Boguslaw
- Date: 2018
- Subjects: To be catalogued
- Language: English
- Type: text , article
- Identifier: http://hdl.handle.net/10962/419330 , vital:71635 , xlink:href="https://doi.org/10.1016/j.jip.2018.08.006"
- Description: Cryptophlebia peltastica is an agricultural pest of litchis and macadamias in South Africa with phytosanitary status for certain markets. Current control methods rely on chemical, cultural and classical biological control. However, a microbial control option has not been developed. An Alphabaculovirus from C. peltastica was recovered from a laboratory reared colony and morphologically characterised by transmission electron microscopy (TEM). Analysis of occlusion bodies indicated a single NPV (SNPV) varying in size from 421 to 1263 nm. PCR amplification and sequencing of the polh gene region using universal primers followed by BLAST analysis revealed a 93% similarity to a partial polh gene sequence from Epinotia granitalis NPV. Further genetic characterisation involving single restriction endonuclease (REN) digestion of genomic DNA was carried out to generate profiles for comparison against other baculovirus species and potential new isolates of the same virus. The complete genome of the virus was sequenced, assembled and analysed for a more comprehensive genetic analysis. The genome was 115 728 base pairs (bp) in length with a GC content of 37.2%. A total of 126 open reading frames (ORFs) were identified with minimal overlap and no preference in orientation. Bioassays were used to determine the virulence of the NPV against C. peltastica. The NPV was virulent against C. peltastica with an LC50 value of 6.46 × 103 OBs/ml and an LC90 value of 2.46 × 105 OBs/ml, and time mortality ranging between 76.32 h and 93.49 h. This is the first study to describe the isolation and genetic characterisation of a novel SNPV from C. peltastica, which has potential for development into a biopesticide for the control of this pest in South Africa.
- Full Text:
- Date Issued: 2018
- Authors: Marsberg, Tamryn , Jukes, Michael , Krejmer-Rabalska, Martyna , Rabalski, Lukasz , Knox, Caroline , Moore, Sean D , Hill, Martin P , Szewczyk, Boguslaw
- Date: 2018
- Subjects: To be catalogued
- Language: English
- Type: text , article
- Identifier: http://hdl.handle.net/10962/419330 , vital:71635 , xlink:href="https://doi.org/10.1016/j.jip.2018.08.006"
- Description: Cryptophlebia peltastica is an agricultural pest of litchis and macadamias in South Africa with phytosanitary status for certain markets. Current control methods rely on chemical, cultural and classical biological control. However, a microbial control option has not been developed. An Alphabaculovirus from C. peltastica was recovered from a laboratory reared colony and morphologically characterised by transmission electron microscopy (TEM). Analysis of occlusion bodies indicated a single NPV (SNPV) varying in size from 421 to 1263 nm. PCR amplification and sequencing of the polh gene region using universal primers followed by BLAST analysis revealed a 93% similarity to a partial polh gene sequence from Epinotia granitalis NPV. Further genetic characterisation involving single restriction endonuclease (REN) digestion of genomic DNA was carried out to generate profiles for comparison against other baculovirus species and potential new isolates of the same virus. The complete genome of the virus was sequenced, assembled and analysed for a more comprehensive genetic analysis. The genome was 115 728 base pairs (bp) in length with a GC content of 37.2%. A total of 126 open reading frames (ORFs) were identified with minimal overlap and no preference in orientation. Bioassays were used to determine the virulence of the NPV against C. peltastica. The NPV was virulent against C. peltastica with an LC50 value of 6.46 × 103 OBs/ml and an LC90 value of 2.46 × 105 OBs/ml, and time mortality ranging between 76.32 h and 93.49 h. This is the first study to describe the isolation and genetic characterisation of a novel SNPV from C. peltastica, which has potential for development into a biopesticide for the control of this pest in South Africa.
- Full Text:
- Date Issued: 2018
Characterisation of novel CrleGV isolates for false codling moth control-lessons learnt from codling moth resistance to CpGV. Characterisation of novel CrleGV isolates for false codling moth control-lessons learnt from codling moth resistance to CpGV
- Opoku-Debrah, John, Moore, Sean D, Hill, Martin P, Knox, Caroline
- Authors: Opoku-Debrah, John , Moore, Sean D , Hill, Martin P , Knox, Caroline
- Date: 2013
- Subjects: To be catalogued
- Language: English
- Type: text , article
- Identifier: http://hdl.handle.net/10962/425414 , vital:72237 , xlink:href="https://www.cabdirect.org/cabdirect/abstract/20133257674"
- Description: Recently some codling moth, Cydia pomonella, populations in Europe developed resistance to CpGV. In order to prepare for the possibility of a similar occurrence with the false codling moth, Thaumatotibia leucotreta, in South Africa, a search was conducted for novel CrleGV isolates. Through overcrowding, outbreaks of novel isolates were recorded from laboratory populations of five geographically distinct host populations. The genetic novelty of these and two commercially available isolates was confirmed through restriction enzyme analysis and sequence analysis of the granulin and egt genes. Phylogenetic analysis showed the existence of two CrleGV-SA genome types. Significant differences in virulence were also shown between certain isolates against certain host populations.
- Full Text:
- Date Issued: 2013
- Authors: Opoku-Debrah, John , Moore, Sean D , Hill, Martin P , Knox, Caroline
- Date: 2013
- Subjects: To be catalogued
- Language: English
- Type: text , article
- Identifier: http://hdl.handle.net/10962/425414 , vital:72237 , xlink:href="https://www.cabdirect.org/cabdirect/abstract/20133257674"
- Description: Recently some codling moth, Cydia pomonella, populations in Europe developed resistance to CpGV. In order to prepare for the possibility of a similar occurrence with the false codling moth, Thaumatotibia leucotreta, in South Africa, a search was conducted for novel CrleGV isolates. Through overcrowding, outbreaks of novel isolates were recorded from laboratory populations of five geographically distinct host populations. The genetic novelty of these and two commercially available isolates was confirmed through restriction enzyme analysis and sequence analysis of the granulin and egt genes. Phylogenetic analysis showed the existence of two CrleGV-SA genome types. Significant differences in virulence were also shown between certain isolates against certain host populations.
- Full Text:
- Date Issued: 2013
Overcrowding of false codling moth, Thaumatotibia leucotreta (Meyrick) leads to the isolation of five new Cryptophlebia leucotreta granulovirus (CrleGV-SA) isolates
- Opoku-Debrah, John K, Hill, Martin P, Knox, Caroline, Moore, Sean D
- Authors: Opoku-Debrah, John K , Hill, Martin P , Knox, Caroline , Moore, Sean D
- Date: 2013
- Subjects: To be catalogued
- Language: English
- Type: text , article
- Identifier: http://hdl.handle.net/10962/419360 , vital:71637 , xlink:href="https://doi.org/10.1016/j.jip.2012.12.008"
- Description: False codling moth, Thaumatotibia leucotreta (Meyrick) is a serious pest of economic importance to the South African fruit industry. As part of sustainable efforts to control this pest, biological control options that involve the application of baculovirus-based biopesticides such as Cryptogran and Cryptex (both formulated with a South African isolate of Cryptophlebia leucotreta granulovirus, CrleGV-SA) are popularly used by farmers. In order to safeguard the integrity of these biopesticides as well as protect against any future development of resistance in the host, we conducted a study to bioprospect for additional CrleGV isolates as alternatives to existing ones. Using overcrowding as an induction method for latent infection, we recovered five new CrleGV isolates (CrleGV-SA Ado, CrleGV-SA Mbl, CrleGV-SA Cit, CrleGV-SA MixC and CrleGV-SA Nels). Single restriction endonuclease (REN) analysis of viral genomic DNA extracted from purified occlusion bodies showed that isolates differed in their DNA profiles. Partial sequencing of granulin and egt genes from the different isolates and multiple alignments of nucleotide sequences revealed the presence of single nucleotide polymorphisms (SNPs), some of which resulted in amino acid substitutions in the protein sequence. Based on these findings as well as comparisons with other documented CrleGV isolates, we propose two phylogenetic groups for CrleGV-SA isolates recovered in this study.
- Full Text:
- Date Issued: 2013
- Authors: Opoku-Debrah, John K , Hill, Martin P , Knox, Caroline , Moore, Sean D
- Date: 2013
- Subjects: To be catalogued
- Language: English
- Type: text , article
- Identifier: http://hdl.handle.net/10962/419360 , vital:71637 , xlink:href="https://doi.org/10.1016/j.jip.2012.12.008"
- Description: False codling moth, Thaumatotibia leucotreta (Meyrick) is a serious pest of economic importance to the South African fruit industry. As part of sustainable efforts to control this pest, biological control options that involve the application of baculovirus-based biopesticides such as Cryptogran and Cryptex (both formulated with a South African isolate of Cryptophlebia leucotreta granulovirus, CrleGV-SA) are popularly used by farmers. In order to safeguard the integrity of these biopesticides as well as protect against any future development of resistance in the host, we conducted a study to bioprospect for additional CrleGV isolates as alternatives to existing ones. Using overcrowding as an induction method for latent infection, we recovered five new CrleGV isolates (CrleGV-SA Ado, CrleGV-SA Mbl, CrleGV-SA Cit, CrleGV-SA MixC and CrleGV-SA Nels). Single restriction endonuclease (REN) analysis of viral genomic DNA extracted from purified occlusion bodies showed that isolates differed in their DNA profiles. Partial sequencing of granulin and egt genes from the different isolates and multiple alignments of nucleotide sequences revealed the presence of single nucleotide polymorphisms (SNPs), some of which resulted in amino acid substitutions in the protein sequence. Based on these findings as well as comparisons with other documented CrleGV isolates, we propose two phylogenetic groups for CrleGV-SA isolates recovered in this study.
- Full Text:
- Date Issued: 2013
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