Improving the Cold Tolerance of False Codling Moth
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IMPROVING THE COLD TOLERANCE OF FALSE CODLING MOTH, THAUMATOTIBIA LEUCOTRETA, FOR BETTER PERFORMANCE IN A STERILE INSECT RELEASE PROGRAMME A thesis submitted in the fulfilment of the requirements for the degree of MASTER OF SCIENCE of RHODES UNIVERSITY by CLAIRE ASHLEIGH DANIEL June 2016 i| Page ABSTRACT The false codling moth (FCM), Thaumatotibia leucotreta (Meyrick) (Lepidoptera: Tortricidae) is a major pest of citrus and other important crops in Sub-Saharan Africa. The introduction of a sterile insect technique (SIT) programme for FCM in South Africa has proven to be very effective in the control of FCM. The objective was to flood citrus orchards with large numbers of sterile males resulting in a ratio of at least 10 sterile to 1 wild moth, increasing the probability of a female moth mating with a sterile male. This target is often achieved and the programme is generally successful, however there are some challenges regarding this programme. The mass rearing envir onment, artificial diet, handling methods and irradiation have an impact on sterile insect quality as environmental differences between the rearing facility and field, influences the insect phenotype and competitiveness. This is evident as wild male moths can theoretically actively fly at a temperature of 12°C and laboratory-reared sterilized moths, due to the radiation treatment, appear unable to fly below 20°C. As a consequence, sterile males are out-competed by wild males during the cooler months of the year. This is detrimental to the SIT programme as FCM do not undergo diapause, meaning they are active during winter and will still reproduce. Therefore, to maximize the effect of the SIT programme, it is vital to increase the flight ability of mass reared sterile males at lower temperatures. Various studies have shown that by adding cryoprotectants to the basic laboratory diets increases the cold tolerance of certain insects and thus may allow them to be mobile at lower temperatures, however it imperative that any chemical used to augment the commercial diet of the insect has no negative effects on the insect physiology and development. To investigate this detail for FCM, five generations of FCM were reared on diets augmented with various known insect cryoprotectants. These augmented FCM were subsequently used in experiments designed to determine firstly, if the cryoprotectants had a positive result on the cold tolerance of the FCM, and secondly, if they had any adverse effects on other physiological aspects such as duration of development. Laboratory trials indicated that the flight ability of male FCM was improved when larvae were reared on diets augmented with trehalose and cholesterol (with an average of 40 % of cholesterol and trehalose augmented males that flew at 15 °C where 0 % of the control flew). Results obtained during the field trials support the laboratory results as there was a significant increase in the number of trehalose augmented moths caught in the field during March and July (winter). Results also showed potential for cholesterol to be used as an additive. Other ii| Page important findings show that both cholesterol and trehalose have no negative impacts on developmental rate, pupal size, and egg production and viability. Trehalose was found to increase the pupal mass of male and female FCM, as well as the number of eggs laid per female. Cholesterol was found to increase developmental rate and the number of eggs laid. The main findings of this study were that diet additives could improve the mass rearing of FCM for SIT and the competitiveness of the males, especially at lower temperatures. However, the additives were expensive and cost could well be a constraint to the wide scale implementation of the new technology. iii| Page To my family, Henry, Jenny and Liam Thank you all for your unconditional love and support iv| Page DECLARATION The following thesis has not been submitted to any university other than Rhodes University, Grahamstown, South Africa. The work presented here is that of the author. 29/06/2015 Date: v| Page ACKNOWLEDGEMENTS • My supervisor, Martin Hill and co- supervisor Sean Moore for their guidance, patience and constructive criticism. I take responsibility for a few of your grey hairs. • Martin Villet for your statistical assistance as well as being a friend when times were tough. • Julie Coetzee, for all your help with my statistics- I would be very lost without your guidance with this. • Mellissa Peyper and Tanya Pretorius for all their help in the lab. • Jeanne Van der Merwe for all those times I had no idea what I needed to do with paperwork and deliveries, you were very patient, thank you. • Claire Love and Wayne Kirkman from CRI for all their help in the field. • Craig Chambers from River Bioscience for all the times I bugged you for anything from egg sheets to email addresses, I really appreciate your help. • Eugene Nepgen for help with papers and advice. • Nevill Boersma, you have helped me so much with the sterilization part of my project and I know you took on a great deal of extra work helping me, I cannot say thank you enough. • Kobus, the farm manager working on the citrus farm where my project was based. Thank you for your help and awesome 4X4 rides. • A special thank you goes to my bursars, CRI, citrus academy THRIP and Rhodes University as without the funding I have received, I would not have been able to do my honours and master’s degrees. • Granny, you helped out so much and I know you would be proud, I wish you could enjoy this with me. • Liam Richter- My love/ assistant. All those late nights when you helped me and never complained once! You are amazing and I doubt I would have been able to accomplish this without your help and support, thank you so much. • To my parents, without you and your hard work I would never have been able to attend varsity. I thank you for all your guidance and support through the tough times, and laughter and celebration during the good times. I appreciate you guys every day. • Finally I would like to give thanks to my heavenly father. vi| Page TABLE OF CONTENTS Abstract................................................................................................................................... ii Declaration............................................................................................................................... v Acknowledgements................................................................................................................. vi Table of Contents................................................................................................................... vii List of Tables............................................................................................................................ x List of Figures......................................................................................................................... xi List of Abbreviations............................................................................................................ xiii CHAPTER I-GENERAL INTRODUCTION 1.1 FALSE CODLING MOTH, THAUMATOTIBIA LEUCOTRETA................................ 1 1.1.1 Taxonomy and geographic distribution................................................................1 1.1.2 Life History and host plants................................................................................ 2 1.1.3 Economic damage due to false codling moth......................................................8 1.1.4 Current control tactics..........................................................................................9 1.1.5 Resistance to chemical control and the evolution of IPM strategies.................12 1.2 STERILE INSECT TECHNIQUE...............................................................................13 1.2.1 Development of the sterile insect technique.......................................................13 1.2.2 Sterile insect technique programmes worldwide................................................14 1.2.3 Sterile insect technique programme for false codling moth in South Africa....18 1.2.4 Integrated pest management and SIT.................................................................19 1.2.5 Disadvantages.................................................................................................... 20 1.2.6 Female sterility and its role in SIT, particularly for FCM ................................22 1.3 PHYSIOLOGY OF INSECT COLD SENSITIVITY................................................ 23 1.3.1 The effects of low temperatures on insects....................................................... 23 1.3.2 Mechanisms promoting insect survival at low temperatures............................ 24 1.3.3 Cryoprotectants found in insects....................................................................... 26 1.3.4 Improving the cold tolerance of an insect using cryoprotectants......................26 1.4 INSECT FLIGHT....................................................................................................... 27 1.4.1 Fuel utilization in energy production................................................................ 27 1.4.2 Carbohydrates and insect flight......................................................................... 28 1.4.3 Lipids and insect flight...................................................................................... 28 1.4.4 Proline and insect flight..................................................................................... 29 vii| Page 1.5 PROJECT