Assessment of pheromone specificity in Thaumatotibia leucotreta (Meyrick) populations with focus on pest monitoring and the regional rollout of the sterile insect technique in citrus
- Authors: Joubert, Francois D
- Date: 2018
- Subjects: Cryptophlebia leucotreta , Pheromone traps , Citrus -- Diseases and pests -- South Africa , Cryptophlebia leucotreta -- Contol , Cryptophlebia leucotreta -- Biological control
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/60665 , vital:27812
- Description: False codling moth (FCM), Thaumatotibia leucotreta (Meyrick) (Lepidoptera: Tortricidae) is considered the most important indigenous pest of citrus in southern Africa. It is recognized by several markets as a phytosanitary organism and the efficient control of this pest is now more important than ever. The pheromone communication between the male and female moths has been exploited in order to control FCM through the sterile insect technique (SIT). The sterilized males used for all SIT programmes across South Africa come from a colony that originates from wild material collected from the Citrusdal area of the Western Cape Province. The aim of this study was to determine if any differences in attractiveness of females to males exist between different geographical populations of FCM and if so what impact this would have on the male’s ability to locate females from other populations via the volatile sex pheromone released by the female. Laboratory trials with Y-tube olfactometers and flight tunnels tested the attraction of male moths to virgin females, but did not yield any consistent results. Field experiments were conducted with sterile male Citrusdal moths released and recaptured in yellow delta traps in two separate trials. For one trial, the traps were baited with live virgin females from five different geographical populations including Addo, Nelspruit, Marble Hall, Citrusdal and the Old colony, which is a mixture of several populations. For the other trial traps were baited with various synthetic pheromone blends including three regional blends which included South Africa, Ivory Coast and Malawi and three commercial blends including Pherolure, Isomate and Checkmate. For the virgin female trial the Citrusdal males showed a significant preference for females from their own population. There was also a significant difference in the recaptures from the different synthetic pheromones. The South African blend was the most attractive of all the regional and commercial blends. A cross-mating trial was also conducted under laboratory conditions in petri dishes with five different FCM populations including Citrusdal, Addo, Marble Hall, Nelspruit and Old (mixed origin). Females produced more eggs when mated with males from the same population for the Addo, Marble Hall, Nelspruit and Old (mixed origin) populations. The only case in which this was statistically significant was for the Marble Hall population. All the crosses produced viable eggs and the origin of the male or female did not influence egg hatch. The results from this study may lead to improvements in both the control and monitoring of FCM populations. The control methods include mating disruption, attract-and-kill and SIT. Tailoring these methods for a specific growing area with a pheromone blend originating from the area or releasing sterile moths from a colony that originates from the area may optimize the available monitoring and control options.
- Full Text:
- Date Issued: 2018
- Authors: Joubert, Francois D
- Date: 2018
- Subjects: Cryptophlebia leucotreta , Pheromone traps , Citrus -- Diseases and pests -- South Africa , Cryptophlebia leucotreta -- Contol , Cryptophlebia leucotreta -- Biological control
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/60665 , vital:27812
- Description: False codling moth (FCM), Thaumatotibia leucotreta (Meyrick) (Lepidoptera: Tortricidae) is considered the most important indigenous pest of citrus in southern Africa. It is recognized by several markets as a phytosanitary organism and the efficient control of this pest is now more important than ever. The pheromone communication between the male and female moths has been exploited in order to control FCM through the sterile insect technique (SIT). The sterilized males used for all SIT programmes across South Africa come from a colony that originates from wild material collected from the Citrusdal area of the Western Cape Province. The aim of this study was to determine if any differences in attractiveness of females to males exist between different geographical populations of FCM and if so what impact this would have on the male’s ability to locate females from other populations via the volatile sex pheromone released by the female. Laboratory trials with Y-tube olfactometers and flight tunnels tested the attraction of male moths to virgin females, but did not yield any consistent results. Field experiments were conducted with sterile male Citrusdal moths released and recaptured in yellow delta traps in two separate trials. For one trial, the traps were baited with live virgin females from five different geographical populations including Addo, Nelspruit, Marble Hall, Citrusdal and the Old colony, which is a mixture of several populations. For the other trial traps were baited with various synthetic pheromone blends including three regional blends which included South Africa, Ivory Coast and Malawi and three commercial blends including Pherolure, Isomate and Checkmate. For the virgin female trial the Citrusdal males showed a significant preference for females from their own population. There was also a significant difference in the recaptures from the different synthetic pheromones. The South African blend was the most attractive of all the regional and commercial blends. A cross-mating trial was also conducted under laboratory conditions in petri dishes with five different FCM populations including Citrusdal, Addo, Marble Hall, Nelspruit and Old (mixed origin). Females produced more eggs when mated with males from the same population for the Addo, Marble Hall, Nelspruit and Old (mixed origin) populations. The only case in which this was statistically significant was for the Marble Hall population. All the crosses produced viable eggs and the origin of the male or female did not influence egg hatch. The results from this study may lead to improvements in both the control and monitoring of FCM populations. The control methods include mating disruption, attract-and-kill and SIT. Tailoring these methods for a specific growing area with a pheromone blend originating from the area or releasing sterile moths from a colony that originates from the area may optimize the available monitoring and control options.
- Full Text:
- Date Issued: 2018
Thaumatotibia leucotreta (Meyrick) (Lepidoptera: Tortricidae) population ecology in citrus orchards: the influence of orchard age
- Authors: Albertyn, Sonnica
- Date: 2018
- Subjects: Cryptophlebia leucotreta , Population biology , Insect populations , Orchards , Insect nematodes , Entomopathogenic fungi
- Language: English
- Type: text , Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/62615 , vital:28213
- Description: Anecdotal reports in the South African citrus industry claim higher populations of false codling moth (FCM), Thaumatotibia (Cryptophlebia) leucotreta (Meyr) (Lepidoptera: Tortricidae), in orchards during the first three to five harvesting years of citrus planted in virgin soil, after which, FCM numbers seem to decrease and remain consistent. Various laboratory studies and field surveys were conducted to determine if, and why juvenile orchards (four to eight years old) experience higher FCM infestation than mature orchards (nine years and older). In laboratory trials, Washington Navel oranges and Nova Mandarins from juvenile trees were shown to be significantly more susceptible to FCM damage and significantly more attractive for oviposition in both choice and no-choice trials, than fruit from mature trees. Although fruit from juvenile Cambria Navel trees were significantly more attractive than mature orchards for oviposition, they were not more susceptible to FCM damage. In contrast, fruit from juvenile and mature Midnight Valencia orchards were equally attractive for oviposition, but fruit from juvenile trees were significantly more susceptible to FCM damage than fruit from mature trees. Artificial diets were augmented with powder from fruit from juvenile or mature Washington Navel orchards at 5%, 10%, 15% or 30%. Higher larval survival of 76%, 63%, 50% and 34%, respectively, was recorded on diets containing fruit powder from the juvenile trees than on diets containing fruit powder from the mature trees, at 69%, 57%, 44% and 27% larval survival, respectively. Bioassays were conducted to determine if differences in plant chemistry between fruit from juvenile and mature trees will have an impact on the susceptibility FCM to entomopathogenic nematodes (EPN), entomopathogenic fungi (EPF) and Cryptophlebia leucotreta granulovirus (CrleGV). No significant differences in the susceptibility of larvae reared on diets containing 15% fruit powder from juvenile and mature trees to EPN and EPF were recorded. Mortality of neonate larvae was significantly lower when placed on diets containing 15% fruit powder from mature trees (45% mortality) than diets containing 15% fruit powder from juvenile trees (61% mortality), after larvae ingested the lowest virus concentration tested, being 2 x104 OBs/ml. Data collected from field surveys showed significantly lower egg parasitism, virus infection of larvae and EPF occurrence in juvenile orchards than mature orchards. Egg parasitism was between 11% and 54% higher in mature orchards than juvenile orchards, with the exception of Mandarins during 2015, where egg parasitism was slightly higher in juvenile orchards, but not significantly so. A significantly higher proportion of larvae retrieved from mature orchards (7% of larvae) were infected with CrleGV than larvae retrieved from juvenile orchards (4% of larvae). A significantly higher occurrence of EPF was recorded in non-bearing and mature orchards, with 40% and 37% occurrence respectively, than in juvenile orchards, with 25% occurrence recorded. EPF occurrence in juvenile orchards increased significantly by 16% to 32% from the first to the third year of sampling. In contrast to results recorded in laboratory trials, similar or higher pest pressure in juvenile orchards than mature orchards did not always result in significantly higher levels of FCM damage under field conditions. FCM damage in juvenile orchards may have been lower than expected, as greater extremes of temperature and lower humidity were recorded in juvenile orchards, which would increase larval mortality. Results of this study showed that juvenile and mature orchards are significantly different and should be managed differently.
- Full Text:
- Date Issued: 2018
- Authors: Albertyn, Sonnica
- Date: 2018
- Subjects: Cryptophlebia leucotreta , Population biology , Insect populations , Orchards , Insect nematodes , Entomopathogenic fungi
- Language: English
- Type: text , Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10962/62615 , vital:28213
- Description: Anecdotal reports in the South African citrus industry claim higher populations of false codling moth (FCM), Thaumatotibia (Cryptophlebia) leucotreta (Meyr) (Lepidoptera: Tortricidae), in orchards during the first three to five harvesting years of citrus planted in virgin soil, after which, FCM numbers seem to decrease and remain consistent. Various laboratory studies and field surveys were conducted to determine if, and why juvenile orchards (four to eight years old) experience higher FCM infestation than mature orchards (nine years and older). In laboratory trials, Washington Navel oranges and Nova Mandarins from juvenile trees were shown to be significantly more susceptible to FCM damage and significantly more attractive for oviposition in both choice and no-choice trials, than fruit from mature trees. Although fruit from juvenile Cambria Navel trees were significantly more attractive than mature orchards for oviposition, they were not more susceptible to FCM damage. In contrast, fruit from juvenile and mature Midnight Valencia orchards were equally attractive for oviposition, but fruit from juvenile trees were significantly more susceptible to FCM damage than fruit from mature trees. Artificial diets were augmented with powder from fruit from juvenile or mature Washington Navel orchards at 5%, 10%, 15% or 30%. Higher larval survival of 76%, 63%, 50% and 34%, respectively, was recorded on diets containing fruit powder from the juvenile trees than on diets containing fruit powder from the mature trees, at 69%, 57%, 44% and 27% larval survival, respectively. Bioassays were conducted to determine if differences in plant chemistry between fruit from juvenile and mature trees will have an impact on the susceptibility FCM to entomopathogenic nematodes (EPN), entomopathogenic fungi (EPF) and Cryptophlebia leucotreta granulovirus (CrleGV). No significant differences in the susceptibility of larvae reared on diets containing 15% fruit powder from juvenile and mature trees to EPN and EPF were recorded. Mortality of neonate larvae was significantly lower when placed on diets containing 15% fruit powder from mature trees (45% mortality) than diets containing 15% fruit powder from juvenile trees (61% mortality), after larvae ingested the lowest virus concentration tested, being 2 x104 OBs/ml. Data collected from field surveys showed significantly lower egg parasitism, virus infection of larvae and EPF occurrence in juvenile orchards than mature orchards. Egg parasitism was between 11% and 54% higher in mature orchards than juvenile orchards, with the exception of Mandarins during 2015, where egg parasitism was slightly higher in juvenile orchards, but not significantly so. A significantly higher proportion of larvae retrieved from mature orchards (7% of larvae) were infected with CrleGV than larvae retrieved from juvenile orchards (4% of larvae). A significantly higher occurrence of EPF was recorded in non-bearing and mature orchards, with 40% and 37% occurrence respectively, than in juvenile orchards, with 25% occurrence recorded. EPF occurrence in juvenile orchards increased significantly by 16% to 32% from the first to the third year of sampling. In contrast to results recorded in laboratory trials, similar or higher pest pressure in juvenile orchards than mature orchards did not always result in significantly higher levels of FCM damage under field conditions. FCM damage in juvenile orchards may have been lower than expected, as greater extremes of temperature and lower humidity were recorded in juvenile orchards, which would increase larval mortality. Results of this study showed that juvenile and mature orchards are significantly different and should be managed differently.
- Full Text:
- Date Issued: 2018
Yeast-baculovirus synergism: investigating mixed infections for improved management of the false codling moth, Thaumatotibia leucotreta
- Authors: Van der Merwe, Marcel
- Date: 2018
- Subjects: Cryptophlebia leucotreta , Baculoviruses , Yeast , Citrus Diseases and pests , Biological pest control agents , Pests Integrated control
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10962/62963 , vital:28347
- Description: Thaumatotibia leucotreta (Meyrick) (Lepidoptera: Tortricidae) or otherwise commonly known as the false codling moth is an indigenous pest of the citrus industry in southern Africa. The pest is highly significant as it impacts negatively on the export of fresh citrus fruits from South Africa to international markets. To control T. leucotreta in South Africa, an integrated pest management (IPM) programme has been implemented. One component of this programme is the baculovirus Cryptophlebia leucotreta granulovirus (CrleGV-SA) which has been formulated into the products Cryptogran™ and Cryptex®. It has previously been reported that there is a mutualistic association between Cydia pomonella (L.) (Lepidoptera: Tortricidae) also known as codling moth, and epiphytic yeasts. Cydia pomonella larval feeding galleries were colonised by yeasts and this, in turn, reduced larval mortality and enhanced larval development. It has been demonstrated in laboratory assays and field trials that combining yeast and brown cane sugar with Cydia pomonella granulovirus (CpGV) significantly increased larval mortality and lowered the proportion of injured apple fruit. This suggests that yeasts can enhance the effectiveness of an insect virus in managing pest larvae. In this study, we proposed to determine which species of yeast occur naturally in the digestive tract, frass and on the epidermis of T. leucotreta larvae and to examine whether any of these yeasts, when combined with the CrleGV-SA, have a synergistic effect in increasing mortality of T. leucotreta larvae. Firstly, Navel oranges infested with T. leucotreta larvae were collected from orchards in Sundays River Valley in Eastern Cape of South Africa. Larvae were extracted and analysed for the presence of yeast on their surface, or in their gut and frass. Four yeasts were isolated from T. leucotreta larvae and identified down to species level via PCR amplification and sequencing of internal transcribed spacer (ITS) region and D1/D2 domain of the large subunit (LSU) of rDNA region. These yeasts were isolated from the frass, epidermis and digestive tract of T. leucotreta larvae. The yeast isolates were identified as Meyerozyma caribbica, Pichia kluyveri, Pichia kudriavzevii and Hanseniaspora opuntiae. A yeast preference assay was conducted on female T. leucotreta moths to examine whether any of the isolated yeast species affected their oviposition preference. Navel oranges were inoculated with the isolated yeast species at a concentration of 6 × 108 cells.ml-1. The assay also included a Brewer’s yeast and distilled water control. Pichia kudriavzevii was shown to be the preferred yeast species for oviposition, as significantly more eggs were deposited on Navel oranges inoculated with this yeast compared to the other treatments. Lastly, a detached fruit bioassay was performed to evaluate the efficacy of mixing P. kudriavzevii with CrleGV-SA to enhance T. leucotreta larvae mortality. Pichia kudriavzevii was selected as it was demonstrated as having an effect on the oviposition preference of female T. leucotreta moths. The concentration at which P. kudriavzevii was applied remained the same as in the preference assay while CrleGV-SA was applied at lethal concentration required to kill 50 % of the population (9.31 × 107 OBs.ml-1). Although an increase in larval mortality was observed between CrleGV-SA being applied alone and the yeast/virus mixture, this result was determined not to be statistically significant. The experiments performed in this study provide a platform for further research into the application of a yeast-virus combination as a novel control option for T. leucotreta in the field. , Thesis (MSc) -- Faculty of Science, Biochemistry and Microbiology, 2018
- Full Text:
- Date Issued: 2018
- Authors: Van der Merwe, Marcel
- Date: 2018
- Subjects: Cryptophlebia leucotreta , Baculoviruses , Yeast , Citrus Diseases and pests , Biological pest control agents , Pests Integrated control
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10962/62963 , vital:28347
- Description: Thaumatotibia leucotreta (Meyrick) (Lepidoptera: Tortricidae) or otherwise commonly known as the false codling moth is an indigenous pest of the citrus industry in southern Africa. The pest is highly significant as it impacts negatively on the export of fresh citrus fruits from South Africa to international markets. To control T. leucotreta in South Africa, an integrated pest management (IPM) programme has been implemented. One component of this programme is the baculovirus Cryptophlebia leucotreta granulovirus (CrleGV-SA) which has been formulated into the products Cryptogran™ and Cryptex®. It has previously been reported that there is a mutualistic association between Cydia pomonella (L.) (Lepidoptera: Tortricidae) also known as codling moth, and epiphytic yeasts. Cydia pomonella larval feeding galleries were colonised by yeasts and this, in turn, reduced larval mortality and enhanced larval development. It has been demonstrated in laboratory assays and field trials that combining yeast and brown cane sugar with Cydia pomonella granulovirus (CpGV) significantly increased larval mortality and lowered the proportion of injured apple fruit. This suggests that yeasts can enhance the effectiveness of an insect virus in managing pest larvae. In this study, we proposed to determine which species of yeast occur naturally in the digestive tract, frass and on the epidermis of T. leucotreta larvae and to examine whether any of these yeasts, when combined with the CrleGV-SA, have a synergistic effect in increasing mortality of T. leucotreta larvae. Firstly, Navel oranges infested with T. leucotreta larvae were collected from orchards in Sundays River Valley in Eastern Cape of South Africa. Larvae were extracted and analysed for the presence of yeast on their surface, or in their gut and frass. Four yeasts were isolated from T. leucotreta larvae and identified down to species level via PCR amplification and sequencing of internal transcribed spacer (ITS) region and D1/D2 domain of the large subunit (LSU) of rDNA region. These yeasts were isolated from the frass, epidermis and digestive tract of T. leucotreta larvae. The yeast isolates were identified as Meyerozyma caribbica, Pichia kluyveri, Pichia kudriavzevii and Hanseniaspora opuntiae. A yeast preference assay was conducted on female T. leucotreta moths to examine whether any of the isolated yeast species affected their oviposition preference. Navel oranges were inoculated with the isolated yeast species at a concentration of 6 × 108 cells.ml-1. The assay also included a Brewer’s yeast and distilled water control. Pichia kudriavzevii was shown to be the preferred yeast species for oviposition, as significantly more eggs were deposited on Navel oranges inoculated with this yeast compared to the other treatments. Lastly, a detached fruit bioassay was performed to evaluate the efficacy of mixing P. kudriavzevii with CrleGV-SA to enhance T. leucotreta larvae mortality. Pichia kudriavzevii was selected as it was demonstrated as having an effect on the oviposition preference of female T. leucotreta moths. The concentration at which P. kudriavzevii was applied remained the same as in the preference assay while CrleGV-SA was applied at lethal concentration required to kill 50 % of the population (9.31 × 107 OBs.ml-1). Although an increase in larval mortality was observed between CrleGV-SA being applied alone and the yeast/virus mixture, this result was determined not to be statistically significant. The experiments performed in this study provide a platform for further research into the application of a yeast-virus combination as a novel control option for T. leucotreta in the field. , Thesis (MSc) -- Faculty of Science, Biochemistry and Microbiology, 2018
- Full Text:
- Date Issued: 2018
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