The establishment of a virus free laboratory colony of Cryptophlebia leucotreta (False Codling Moth) and characterisation of Cryptophlebia leucotreta Granulovirus (CrleGV) genes
- Authors: Ludewig, Michael Hans
- Date: 2003
- Subjects: Cryptophlebia leucotreta , Cryptophlebia leucotreta -- Control , Pests -- Biological control , DNA viruses , Agricultural pests -- Biological control , Baculoviruses
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3957 , http://hdl.handle.net/10962/d1004016 , Cryptophlebia leucotreta , Cryptophlebia leucotreta -- Control , Pests -- Biological control , DNA viruses , Agricultural pests -- Biological control , Baculoviruses
- Description: Cryptophlebia leucotreta is an economically important agricultural pest throughout Sub-Saharan Africa. CrleGV has been considered as an alternative to chemical control of this pest due to its host specificity and innocuous nature towards vertebrates. A CrleGV free laboratory colony of C. leucotreta would be useful for the isolation of genotypically pure strains of the CrleGV and for virulence comparisons between isolates. It is preferable to have a full characterisation of CrleGV prior to its registration and release into the environment as a biopesticide. A laboratory colony of C. leucotreta, set up at Rhodes University, containing a low level of infection indicated that CrleGV is vertically transmitted. To establish a virus free laboratory colony of C. leucotreta, a solution of 3.5% sodium hypochlorite and 1% Tween 20 was used to surface decontaminate C. leucotreta eggs for removal of transovum CrleGV from the laboratory colony. No apparent infection by CrleGV was induced by subjecting larvae to stress. PCR of DNA extracted from larvae using CTAB failed to detect virus in the laboratory colony. This detection protocol was able to detect down to 60 fg (480 genome copies of CrleGV). The possibility of low-level virus remaining in the colony requires monitoring of genotypic purity of virus manipulated in the colony. Sequencing of Bam HI/KpnI fragments produced a preliminary sequence of the granulin region of CrleGV. This preliminary sequence supports the trend that the gene organisation of the granulin region of the granuloviruses infecting the family Tortricidae is conserved.
- Full Text:
- Date Issued: 2003
- Authors: Ludewig, Michael Hans
- Date: 2003
- Subjects: Cryptophlebia leucotreta , Cryptophlebia leucotreta -- Control , Pests -- Biological control , DNA viruses , Agricultural pests -- Biological control , Baculoviruses
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:3957 , http://hdl.handle.net/10962/d1004016 , Cryptophlebia leucotreta , Cryptophlebia leucotreta -- Control , Pests -- Biological control , DNA viruses , Agricultural pests -- Biological control , Baculoviruses
- Description: Cryptophlebia leucotreta is an economically important agricultural pest throughout Sub-Saharan Africa. CrleGV has been considered as an alternative to chemical control of this pest due to its host specificity and innocuous nature towards vertebrates. A CrleGV free laboratory colony of C. leucotreta would be useful for the isolation of genotypically pure strains of the CrleGV and for virulence comparisons between isolates. It is preferable to have a full characterisation of CrleGV prior to its registration and release into the environment as a biopesticide. A laboratory colony of C. leucotreta, set up at Rhodes University, containing a low level of infection indicated that CrleGV is vertically transmitted. To establish a virus free laboratory colony of C. leucotreta, a solution of 3.5% sodium hypochlorite and 1% Tween 20 was used to surface decontaminate C. leucotreta eggs for removal of transovum CrleGV from the laboratory colony. No apparent infection by CrleGV was induced by subjecting larvae to stress. PCR of DNA extracted from larvae using CTAB failed to detect virus in the laboratory colony. This detection protocol was able to detect down to 60 fg (480 genome copies of CrleGV). The possibility of low-level virus remaining in the colony requires monitoring of genotypic purity of virus manipulated in the colony. Sequencing of Bam HI/KpnI fragments produced a preliminary sequence of the granulin region of CrleGV. This preliminary sequence supports the trend that the gene organisation of the granulin region of the granuloviruses infecting the family Tortricidae is conserved.
- Full Text:
- Date Issued: 2003
Investigation of the larval parasitoids of the false codling moth, Cryptophlebia Leucotreta (Meyrick) (Lepidoptera: Tortricidae), on citrus in South Africa
- Authors: Sishuba, Nomahlubi
- Date: 2004
- Subjects: Cryptophlebia leucotreta , Tortricidae , Cryptophlebia leucotreta -- Control , Pests -- Biological control , Parasitoids , Citrus -- Diseases and pests
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:5918 , http://hdl.handle.net/10962/d1016267
- Description: The study examined the larval parasitoids of Cryptophlebia leucotreta (Meyrick) on citrus in South Africa and aimed to improve the existing rearing techniques of C. leucotreta with a view to mass rearing of biological control agents. The biological characteristics of Agathis bishopi Nixon (Hymenoptera: Braconidae) were studied, with an emphasis on parasitism rates in the field, host stage preference, developmental rate, life span and offspring sex ratios. Two larval parasitoids, A. bishopi and Apophua leucotretae (Wilkinson) (Hymenoptera: Ichneumonidae), and an egg parasitoid, Trichogrammatoidea cryptophlebiae Nagaraja (Hymenoptera: Trichogrammatidae), were recorded on C. leucotreta on citrus. A. bishopi was the more abundant of the larval parasitoids and exhibited density dependent parasitism. The highest parasitism rates were observed in December with up to 38% in Sundays River Valley and 34% in Gamtoos River Valley, at a time when the highest false codling moth infestations were observed. Agathis bishopi was recorded only in the Eastern Cape Province. The sex ratio of A. bishopi was biased towards females throughout the study (77% in Gamtoos River Valley and 72% in Sundays River Valley). Agathis bishopi is a solitary, koinobiont, larval-pupal endoparasitoid. The species showed a preference for 1st and 2"d instar hosts. Females regulate the sex of their progeny according to the size and larval stage of the host, ovipositing unfertilised eggs in younger, smaller larvae (1st instars) and fertilised eggs in older, larger larvae (2nd instars). The developmental rate of A. bishopi is in synchrony with that of the moth and adults emerge when adult moths that have escaped parasitism emerge. Agathis bishopi and T. cryptophlebiae compliment each other because they have different niches and strategies of attack. Integrating A. bishopi and T. cryptophlebiae into the management of citrus orchards has potential to suppress false codling moth. Larger rearing containers seemed ideal for large-scale rearing of false codling moth. A higher percentage of adults was obtained from larvae reared in larger containers than in smaller ones. The width of the sponges used as stoppers prevented escape of the larvae. Media prepared in larger containers are easier and simpler to prepare than in smaller ones, thus eliminating many precautions otherwise necessary to prevent contamination. Moth production was greatly reduced by the high concentration of Sporekill used for egg decontamination.
- Full Text:
- Date Issued: 2004
- Authors: Sishuba, Nomahlubi
- Date: 2004
- Subjects: Cryptophlebia leucotreta , Tortricidae , Cryptophlebia leucotreta -- Control , Pests -- Biological control , Parasitoids , Citrus -- Diseases and pests
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:5918 , http://hdl.handle.net/10962/d1016267
- Description: The study examined the larval parasitoids of Cryptophlebia leucotreta (Meyrick) on citrus in South Africa and aimed to improve the existing rearing techniques of C. leucotreta with a view to mass rearing of biological control agents. The biological characteristics of Agathis bishopi Nixon (Hymenoptera: Braconidae) were studied, with an emphasis on parasitism rates in the field, host stage preference, developmental rate, life span and offspring sex ratios. Two larval parasitoids, A. bishopi and Apophua leucotretae (Wilkinson) (Hymenoptera: Ichneumonidae), and an egg parasitoid, Trichogrammatoidea cryptophlebiae Nagaraja (Hymenoptera: Trichogrammatidae), were recorded on C. leucotreta on citrus. A. bishopi was the more abundant of the larval parasitoids and exhibited density dependent parasitism. The highest parasitism rates were observed in December with up to 38% in Sundays River Valley and 34% in Gamtoos River Valley, at a time when the highest false codling moth infestations were observed. Agathis bishopi was recorded only in the Eastern Cape Province. The sex ratio of A. bishopi was biased towards females throughout the study (77% in Gamtoos River Valley and 72% in Sundays River Valley). Agathis bishopi is a solitary, koinobiont, larval-pupal endoparasitoid. The species showed a preference for 1st and 2"d instar hosts. Females regulate the sex of their progeny according to the size and larval stage of the host, ovipositing unfertilised eggs in younger, smaller larvae (1st instars) and fertilised eggs in older, larger larvae (2nd instars). The developmental rate of A. bishopi is in synchrony with that of the moth and adults emerge when adult moths that have escaped parasitism emerge. Agathis bishopi and T. cryptophlebiae compliment each other because they have different niches and strategies of attack. Integrating A. bishopi and T. cryptophlebiae into the management of citrus orchards has potential to suppress false codling moth. Larger rearing containers seemed ideal for large-scale rearing of false codling moth. A higher percentage of adults was obtained from larvae reared in larger containers than in smaller ones. The width of the sponges used as stoppers prevented escape of the larvae. Media prepared in larger containers are easier and simpler to prepare than in smaller ones, thus eliminating many precautions otherwise necessary to prevent contamination. Moth production was greatly reduced by the high concentration of Sporekill used for egg decontamination.
- Full Text:
- Date Issued: 2004
Geographic variation in the susceptibility of false colding Moth, Thaumatotibia Leucotreta, populations to a granulovirus (CrleGV-SA)
- Authors: Opoku-Debrah, John Kwadwo
- Date: 2008
- Subjects: Cryptophlebia leucotreta , Cryptophlebia leucotreta -- Control , Pests -- Biological control , Citrus -- Diseases and pests
- Language: English
- Type: Thesis , Masters , MTech
- Identifier: vital:10586 , http://hdl.handle.net/10948/984 , Cryptophlebia leucotreta , Cryptophlebia leucotreta -- Control , Pests -- Biological control , Citrus -- Diseases and pests
- Description: The false codling moth (FCM), Thaumatotibia (=Cryptophlebia) leucotreta (Meyrick) (Lepidoptera: Tortricidae) is a serious pest of citrus and other crops in Sub-Saharan Africa. The introduction of the Cryptophlebia leucotreta granulovirus (CrleGV-SA) Cryptogran and Cryptex (biopesticides) has proven to be very effective in the control of FCM. However, markedly lower susceptibility of some codling moth (CM), Cydia pomonella (L.) populations to Cydia pomonella granulovirus (CpGV-M), another granulovirus product used in the control of CM’s in Europe have been reported. Genetic differences between FCM populations in South Africa have also been established. It is therefore possible that differences in the susceptibility of these geographically distinct FCM populations to CrleGV-SA might also exist. To investigate this phenomenon, a benchmark for pathogenecity was established. In continuation of previous work with Cryptogran against the 1st and 5th instar FCM larvae, dose-response relationships were established for all five larval instars of FCM. In surface dose-response bioassays, the LC50 values for the 2nd, 3rd and 4th instars were calculated to be 4.516 x 104, 1.662 x 105 and 2.205 x 106 occlusion bodies (OBs)/ml, respectively. The LC90 values for the 2nd, 3rd and 4th instars were calculated to be 4.287 x 106, 9.992 x 106 and 1.661 x 108 OBs/ml, respectively. Susceptibility to CrleGV-SA was found to decline with larval stage and increase with time of exposure. The protocol was used in guiding bioassays with field collected FCM larvae. Laboratory assays conducted with Cryptogran (at 1.661 x 108 OBs/ml) against field collected FCM larvae from Addo, Kirkwood, Citrusdal and Clanwilliam as well as a standard laboratory colony, showed a significant difference in pathogenecity in only one case. This significant difference was observed between 5th instars from the Addo colony and 5th instars from the other populations. Four geographically distinct FCM colonies from Addo, Citrusdal, Marble Hall and Nelspruit were also established. Since Cryptogran and Cryptex are always targeted against 1st instar FCM larvae in the field, further comparative laboratory assays were conducted with the Addo colony and an old laboratory colony. Cryptogran was significantly more pathogenic than Cryptex against both the Addo and the old colony. However, a high level of heterogeneity was observed in responses within each population.
- Full Text:
- Date Issued: 2008
- Authors: Opoku-Debrah, John Kwadwo
- Date: 2008
- Subjects: Cryptophlebia leucotreta , Cryptophlebia leucotreta -- Control , Pests -- Biological control , Citrus -- Diseases and pests
- Language: English
- Type: Thesis , Masters , MTech
- Identifier: vital:10586 , http://hdl.handle.net/10948/984 , Cryptophlebia leucotreta , Cryptophlebia leucotreta -- Control , Pests -- Biological control , Citrus -- Diseases and pests
- Description: The false codling moth (FCM), Thaumatotibia (=Cryptophlebia) leucotreta (Meyrick) (Lepidoptera: Tortricidae) is a serious pest of citrus and other crops in Sub-Saharan Africa. The introduction of the Cryptophlebia leucotreta granulovirus (CrleGV-SA) Cryptogran and Cryptex (biopesticides) has proven to be very effective in the control of FCM. However, markedly lower susceptibility of some codling moth (CM), Cydia pomonella (L.) populations to Cydia pomonella granulovirus (CpGV-M), another granulovirus product used in the control of CM’s in Europe have been reported. Genetic differences between FCM populations in South Africa have also been established. It is therefore possible that differences in the susceptibility of these geographically distinct FCM populations to CrleGV-SA might also exist. To investigate this phenomenon, a benchmark for pathogenecity was established. In continuation of previous work with Cryptogran against the 1st and 5th instar FCM larvae, dose-response relationships were established for all five larval instars of FCM. In surface dose-response bioassays, the LC50 values for the 2nd, 3rd and 4th instars were calculated to be 4.516 x 104, 1.662 x 105 and 2.205 x 106 occlusion bodies (OBs)/ml, respectively. The LC90 values for the 2nd, 3rd and 4th instars were calculated to be 4.287 x 106, 9.992 x 106 and 1.661 x 108 OBs/ml, respectively. Susceptibility to CrleGV-SA was found to decline with larval stage and increase with time of exposure. The protocol was used in guiding bioassays with field collected FCM larvae. Laboratory assays conducted with Cryptogran (at 1.661 x 108 OBs/ml) against field collected FCM larvae from Addo, Kirkwood, Citrusdal and Clanwilliam as well as a standard laboratory colony, showed a significant difference in pathogenecity in only one case. This significant difference was observed between 5th instars from the Addo colony and 5th instars from the other populations. Four geographically distinct FCM colonies from Addo, Citrusdal, Marble Hall and Nelspruit were also established. Since Cryptogran and Cryptex are always targeted against 1st instar FCM larvae in the field, further comparative laboratory assays were conducted with the Addo colony and an old laboratory colony. Cryptogran was significantly more pathogenic than Cryptex against both the Addo and the old colony. However, a high level of heterogeneity was observed in responses within each population.
- Full Text:
- Date Issued: 2008
Understanding and improving the residual efficacy of the cryptophlebia leucotreta granulovirus (Cryptogran)
- Authors: Kirkman, Wayne
- Date: 2008
- Subjects: Cryptophlebia leucotreta , Cryptophlebia leucotreta -- Control , Pests -- Biological control , Citrus -- Diseases and pests
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:5794 , http://hdl.handle.net/10962/d1005482 , Cryptophlebia leucotreta , Cryptophlebia leucotreta -- Control , Pests -- Biological control , Citrus -- Diseases and pests
- Description: False codling moth (FCM), Thaumatotibia (=Cryptophlebia) leucotreta (Meyr) (Lepidoptera: Tortricidae), is one of the most important pests on citrus. The Cryptophlebia leucotreta granulovirus (CrleGV) has been developed into a successful biological control agent, registered under the name Cryptogran, and is currently the preferred product for the control of FCM on citrus in South Africa. A prerequisite to the continued success of Cryptogran as a means of controlling false codling moth is to understand the factors affecting field persistence of the virus, and to find ways to improve it. The aim of this study was to gain a clearer understanding of the product and the abiotic and biotic factors affecting its persistence in the field, and to investigate methods to improve this persistence. The effect of UV-irradiation on the virus was determined, and various products were tested as UV protectants in laboratory bioassays. Lignin was the most effective additive, and was tested in several field trials, where it also enhanced the efficacy of Cryptogran. Laboratory trials indicated that Cryptogran is rainfast. Cryptogran applications early in the season had a longer period of residual activity than sprays applied closer to harvest. Daytime applications were less effective that evening sprays. Sprays applied coinciding with peaks in pheromone moth trap catches were more effective than those applied between peaks. Biotic factors influencing persistence were investigated. Residual efficacy was longer when treatments were applied to blocks than as single tree treatments. Attempts were made to quantify the effect of the navel end of a navel orange on the field persistence of Cryptogran. Cryptogran was shown to be compatible with many agricultural chemicals used on citrus. Economic thresholds and various cost-benefit analyses are discussed. A list of practical recommendations to growers was drawn up, and possibilities for future research are presented.
- Full Text:
- Date Issued: 2008
- Authors: Kirkman, Wayne
- Date: 2008
- Subjects: Cryptophlebia leucotreta , Cryptophlebia leucotreta -- Control , Pests -- Biological control , Citrus -- Diseases and pests
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:5794 , http://hdl.handle.net/10962/d1005482 , Cryptophlebia leucotreta , Cryptophlebia leucotreta -- Control , Pests -- Biological control , Citrus -- Diseases and pests
- Description: False codling moth (FCM), Thaumatotibia (=Cryptophlebia) leucotreta (Meyr) (Lepidoptera: Tortricidae), is one of the most important pests on citrus. The Cryptophlebia leucotreta granulovirus (CrleGV) has been developed into a successful biological control agent, registered under the name Cryptogran, and is currently the preferred product for the control of FCM on citrus in South Africa. A prerequisite to the continued success of Cryptogran as a means of controlling false codling moth is to understand the factors affecting field persistence of the virus, and to find ways to improve it. The aim of this study was to gain a clearer understanding of the product and the abiotic and biotic factors affecting its persistence in the field, and to investigate methods to improve this persistence. The effect of UV-irradiation on the virus was determined, and various products were tested as UV protectants in laboratory bioassays. Lignin was the most effective additive, and was tested in several field trials, where it also enhanced the efficacy of Cryptogran. Laboratory trials indicated that Cryptogran is rainfast. Cryptogran applications early in the season had a longer period of residual activity than sprays applied closer to harvest. Daytime applications were less effective that evening sprays. Sprays applied coinciding with peaks in pheromone moth trap catches were more effective than those applied between peaks. Biotic factors influencing persistence were investigated. Residual efficacy was longer when treatments were applied to blocks than as single tree treatments. Attempts were made to quantify the effect of the navel end of a navel orange on the field persistence of Cryptogran. Cryptogran was shown to be compatible with many agricultural chemicals used on citrus. Economic thresholds and various cost-benefit analyses are discussed. A list of practical recommendations to growers was drawn up, and possibilities for future research are presented.
- Full Text:
- Date Issued: 2008
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