Developing an attractant for monitoring fruit-feeding moths in citrus orchards
- Authors: Goddard, Mathew Keith
- Date: 2016
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/2981 , vital:20349
- Description: Fruit-piercing moths are a sporadic pest of citrus, especially in the Eastern Cape Province of South Africa, where the adults can cause significant damage in outbreak years. Currently the only way in which to successfully control fruit-feeding moths within the orchards is the use of repellent lights. However, growers confuse fruit-piercing moths with fruit-sucking moths that don‘t cause primary damage, and there is no way of monitoring which moth species are attacking the fruit in the orchards during the night. In a previous study, banana was shown to be the most attractive bait for a variety of fruit-feeding moth species. Therefore the aim of this study was to determine the population dynamics of fruit-feeding moths develop a cost-effective alternative to the use of fresh banana as a bait for fruit-piercing moths. Fresh banana was compared to nine alternative synthetic attractants, frozen banana and a control under field conditions in several orchards in the Eastern Cape Province. Once again, banana was shown to be the most attractive bait. Some 23 species of fruit-feeding moth species were sampled in the traps, but there was only two fruit-piercing species, Serrodes partita (Fabricius) (Lepidoptera: Noctuidae) and Eudocima sp. Surprisingly S. partita, which was thought to be the main pest, comprised only 6.9% of trap catches. Serrodes partita, is a sporadic pest, only becoming problematic every five to 10 years after good rainfall in the Little Karoo region that causes flushes of their larval host, wild plum, Pappea capensis (Ecklon and Zeyher). During these outbreaks, damage to fruit can range from 70 to 90% and this is especially so for soft skinned citrus. A study on the morphology of the proboscis confirmed that only two species of fruit-piercing moths were present. Trap catches over three citrus growing seasons was linked to fruit damage found within several orchards. Once again fruit-piercing moth damage was relatively low in comparison to other types of damage such as mechanical and undefined damage. There was a very weak correlation between S. partita trap catches and damage, but generally damage was recorded two to three weeks after a peak in S. partita trap catches. Climatic conditions were also recorded and compared to weekly trap catches of S. partita, and while temperature and wind direction had no influence on moth populations, precipitation in the orchards was weakly correlated with trap catches. This study has shown that in non-outbreak seasons, the main fruit-piercing moth, S. partita comprises a small percentage of fruit-feeding moths in citrus orchards, but that growers are unable to determine the difference between fruit-piercing species and the harmless fruit-sucking species. Further fresh banana remains the best method for attracting fruit-piecing moths to traps, but this is not cost effective and thus a commercially viable protocol for monitoring these species remains elusive.
- Full Text:
- Authors: Goddard, Mathew Keith
- Date: 2016
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/2981 , vital:20349
- Description: Fruit-piercing moths are a sporadic pest of citrus, especially in the Eastern Cape Province of South Africa, where the adults can cause significant damage in outbreak years. Currently the only way in which to successfully control fruit-feeding moths within the orchards is the use of repellent lights. However, growers confuse fruit-piercing moths with fruit-sucking moths that don‘t cause primary damage, and there is no way of monitoring which moth species are attacking the fruit in the orchards during the night. In a previous study, banana was shown to be the most attractive bait for a variety of fruit-feeding moth species. Therefore the aim of this study was to determine the population dynamics of fruit-feeding moths develop a cost-effective alternative to the use of fresh banana as a bait for fruit-piercing moths. Fresh banana was compared to nine alternative synthetic attractants, frozen banana and a control under field conditions in several orchards in the Eastern Cape Province. Once again, banana was shown to be the most attractive bait. Some 23 species of fruit-feeding moth species were sampled in the traps, but there was only two fruit-piercing species, Serrodes partita (Fabricius) (Lepidoptera: Noctuidae) and Eudocima sp. Surprisingly S. partita, which was thought to be the main pest, comprised only 6.9% of trap catches. Serrodes partita, is a sporadic pest, only becoming problematic every five to 10 years after good rainfall in the Little Karoo region that causes flushes of their larval host, wild plum, Pappea capensis (Ecklon and Zeyher). During these outbreaks, damage to fruit can range from 70 to 90% and this is especially so for soft skinned citrus. A study on the morphology of the proboscis confirmed that only two species of fruit-piercing moths were present. Trap catches over three citrus growing seasons was linked to fruit damage found within several orchards. Once again fruit-piercing moth damage was relatively low in comparison to other types of damage such as mechanical and undefined damage. There was a very weak correlation between S. partita trap catches and damage, but generally damage was recorded two to three weeks after a peak in S. partita trap catches. Climatic conditions were also recorded and compared to weekly trap catches of S. partita, and while temperature and wind direction had no influence on moth populations, precipitation in the orchards was weakly correlated with trap catches. This study has shown that in non-outbreak seasons, the main fruit-piercing moth, S. partita comprises a small percentage of fruit-feeding moths in citrus orchards, but that growers are unable to determine the difference between fruit-piercing species and the harmless fruit-sucking species. Further fresh banana remains the best method for attracting fruit-piecing moths to traps, but this is not cost effective and thus a commercially viable protocol for monitoring these species remains elusive.
- Full Text:
Genetic and biological characterisation of a novel South African Cydia pomonella granulovirus (CpGV-SA) isolate
- Motsoeneng, Boitumelo Madika
- Authors: Motsoeneng, Boitumelo Madika
- Date: 2016
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:20503 , http://hdl.handle.net/10962/d1021266
- Description: The codling moth, Cydia pomonella (L.) (Lepidoptera: Tortricidae), is the primary pest of pome fruit cultivated worldwide. The control of this insect pest has been dependent on the frequent use of broad-spectrum chemical pesticides, which has led to the development of resistance in pest populations and negative effects on human health and the environment. The Betabaculovirus of C. pomonella has successfully been applied as a biological control agent in integrated pest management (IPM) programmes for the suppression of pest populations worldwide. Previously, all Cydia pomonella granulovirus (CpGV) biopesticides were based on a Mexican isolate (CpGV-M) and although these products are highly efficient at controlling C. pomonella, resistance cases have been reported across Europe. The identification of novel CpGV isolates as additional or alternative control agents to manage resistance is therefore necessary. This study aimed to genetically and biologically characterise a novel South African C. pomonella granulovirus isolate and to test its virulence against neonate larvae. Based on the morphology of the occlusion bodies observed using transmission electron microscopy, granuloviruses were recovered from diseased and dead larvae collected from an orchard in South Africa where no virus applications had been made. DNA was extracted and the identification of the isolated granulovirus was achieved through the PCR amplification and sequencing of the lef-8, lef-9, granulin and egt genes. Submission of the gene sequences to BLAST revealed high percentage identities to sequences from various CpGV isolates, resulting in the naming of the isolate in this study as the South African Cydia pomonella granulovirus (CpGV-SA) isolate. Phylogenetic analysis based on the single nucleotide polymorphisms (SNPs) detected in the lef-8, lef-9 and granulin nucleotide sequences grouped the South African isolate with CpGV-E2 (genome type B) and CpGV-S (genome type E). The CpGV-SA isolate was further genetically characterised by restriction endonuclease analysis and complete sequencing of the genomic DNA. Differences were observed for the BamHI, EcoRI, PstI and XhoI profiles of CpGV-SA in comparison to the respective profiles generated for CpGV-M extracted from a biopesticide, Carpovirusine® (Arysta Lifescience, France). Several genetic variations between the complete genome sequence of CpGV-SA and the reference isolate, CpGV-M1, as well as a recent genome submission of CpGV-M, both representing genome type A were observed. The complete genome analysis confirmed that CpGV-SA is genetically different from the Mexican CpGV isolate, used in thedevelopment of most biopesticides. In silico restriction profiles of the genome sequence obtained for CpGV-SA and genome sequences of genetically different CpGV isolates originating from Mexico (M1 and M), England (E2), Canada (S) and Iran (I12 and I07), available on the NCBI’s GenBank database confirmed that CpGV-SA is of mixed genotypes. Furthermore, the South African isolate shared the single common difference found in the pe38 gene of resistance overcoming isolates, which was the absence of an internal 24 nucleotide repeat present in CpGV-M1. In addition to the common difference, SNPs detected in the pe38 gene grouped the isolate with the CpGV-S isolate, suggesting that the CpGV-SA isolate is predominantly of genome type E. To determine the biological activity of CpGV-SA against neonate C. pomonella larvae, surface bioassays were conducted alongside CpGV-M (Carpovirusine®) bioassays. The LC50 and LC90 values for the South African isolate were 1.6 × 103 and 1.2 × 105 OBs/ml respectively. The LT50 was determined to be 135 hours. These values were similar to the values obtained for CpGV-M (Carpovirusine®). The results in this study suggest that a novel South African CpGV isolate of mixed genotypes, potentially able to overcome resistance in C. pomonella, with biological activity similar to CpGV-M (Carpovirusine®) and important for the control of C. pomonella was recovered. The CpGV-SA isolate could therefore potentially be developed into a biopesticide for use in resistance management strategies against C. pomonella populations in South Africa.
- Full Text:
- Authors: Motsoeneng, Boitumelo Madika
- Date: 2016
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:20503 , http://hdl.handle.net/10962/d1021266
- Description: The codling moth, Cydia pomonella (L.) (Lepidoptera: Tortricidae), is the primary pest of pome fruit cultivated worldwide. The control of this insect pest has been dependent on the frequent use of broad-spectrum chemical pesticides, which has led to the development of resistance in pest populations and negative effects on human health and the environment. The Betabaculovirus of C. pomonella has successfully been applied as a biological control agent in integrated pest management (IPM) programmes for the suppression of pest populations worldwide. Previously, all Cydia pomonella granulovirus (CpGV) biopesticides were based on a Mexican isolate (CpGV-M) and although these products are highly efficient at controlling C. pomonella, resistance cases have been reported across Europe. The identification of novel CpGV isolates as additional or alternative control agents to manage resistance is therefore necessary. This study aimed to genetically and biologically characterise a novel South African C. pomonella granulovirus isolate and to test its virulence against neonate larvae. Based on the morphology of the occlusion bodies observed using transmission electron microscopy, granuloviruses were recovered from diseased and dead larvae collected from an orchard in South Africa where no virus applications had been made. DNA was extracted and the identification of the isolated granulovirus was achieved through the PCR amplification and sequencing of the lef-8, lef-9, granulin and egt genes. Submission of the gene sequences to BLAST revealed high percentage identities to sequences from various CpGV isolates, resulting in the naming of the isolate in this study as the South African Cydia pomonella granulovirus (CpGV-SA) isolate. Phylogenetic analysis based on the single nucleotide polymorphisms (SNPs) detected in the lef-8, lef-9 and granulin nucleotide sequences grouped the South African isolate with CpGV-E2 (genome type B) and CpGV-S (genome type E). The CpGV-SA isolate was further genetically characterised by restriction endonuclease analysis and complete sequencing of the genomic DNA. Differences were observed for the BamHI, EcoRI, PstI and XhoI profiles of CpGV-SA in comparison to the respective profiles generated for CpGV-M extracted from a biopesticide, Carpovirusine® (Arysta Lifescience, France). Several genetic variations between the complete genome sequence of CpGV-SA and the reference isolate, CpGV-M1, as well as a recent genome submission of CpGV-M, both representing genome type A were observed. The complete genome analysis confirmed that CpGV-SA is genetically different from the Mexican CpGV isolate, used in thedevelopment of most biopesticides. In silico restriction profiles of the genome sequence obtained for CpGV-SA and genome sequences of genetically different CpGV isolates originating from Mexico (M1 and M), England (E2), Canada (S) and Iran (I12 and I07), available on the NCBI’s GenBank database confirmed that CpGV-SA is of mixed genotypes. Furthermore, the South African isolate shared the single common difference found in the pe38 gene of resistance overcoming isolates, which was the absence of an internal 24 nucleotide repeat present in CpGV-M1. In addition to the common difference, SNPs detected in the pe38 gene grouped the isolate with the CpGV-S isolate, suggesting that the CpGV-SA isolate is predominantly of genome type E. To determine the biological activity of CpGV-SA against neonate C. pomonella larvae, surface bioassays were conducted alongside CpGV-M (Carpovirusine®) bioassays. The LC50 and LC90 values for the South African isolate were 1.6 × 103 and 1.2 × 105 OBs/ml respectively. The LT50 was determined to be 135 hours. These values were similar to the values obtained for CpGV-M (Carpovirusine®). The results in this study suggest that a novel South African CpGV isolate of mixed genotypes, potentially able to overcome resistance in C. pomonella, with biological activity similar to CpGV-M (Carpovirusine®) and important for the control of C. pomonella was recovered. The CpGV-SA isolate could therefore potentially be developed into a biopesticide for use in resistance management strategies against C. pomonella populations in South Africa.
- Full Text:
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