Predation of Cabbage Pests

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Predation of Cabbage Pests PREDATION OF CABBAGE PESTS: A COMBINED ECOLOGICAL AND MOLECULAR APPROACH Rini Murtiningsih A thesis submitted for the degree of Doctor of Philosophy at The University of Queensland in 2014 School of Biological Sciences Abstract Plutella xylostella L (Lepidoptera: Plutellidae) and Crocidolomia pavonana F (Lepidoptera: Crambidae) are serious pests of Brassica crops in the highlands of West Java, Indonesia, where they co-occur, and in Southeast Queensland, Australia where C. pavonana is a serious pest early in the crop season (February–May) and P. xylostella is a pest later in the year (June-November). Introductions of parasitoids of P. xylostella, especially Diadegma semiclausum Hellén (Hymenoptera: Ichneumonidae), have had considerable success in both countries, but no effective larval or pupal parasitoids of C. pavonana are known. The use of insecticides to manage C. pavonana disrupts P. xylostella parasitoids, leading to pest outbreaks. Thus, an integrated pest management (IPM) strategy, that takes account of both pests, is needed in both regions. Although P. xylostella parasitoids have been the centre of much research over the past 50-60 years, the impact of predatory arthropods on pest populations has received much less attention. Similarly, little is known about the impact of predators on C. pavonana populations and it appears to be attacked by a particularly sparse parasitoid fauna. In recent years, considerable advances have been made in the development of DNA-based molecular techniques to detect the remains of insect prey within the guts of predatory arthropods. However, the methods have rarely been applied in conjunction with field experiments that measure the impact of natural enemies on pest/ prey populations. In this research, field studies used a combination of ecological (natural enemy exclusion experiments and the construction of life tables) and DNA-based molecular methods to quantify the impact of predatory arthropods on P. xylostella and C. pavonana populations and to identify the most important predatory groups preying on these insect pests in the different agro-ecosystems of Queensland and West Java. The specificity of previously designed primer sequences for P. xylostella, C. pavonana and Pieris rapae L. (Lepidoptera: Pieridae) mtCO1 DNA was confirmed by testing them against a wide range of herbivores and predatory arthropods collected from the field in both West Java and Queensland. Based on field studies, key groups of predators were selected to determine the detectability half-life of target P. xylostella and C. pavonana mtCO1 DNA in their guts following consumption of single prey items. For the P. xylostella mtCO1 DNA, when tested at 25 °C the detectability half-life (DHL) was lowest in Clubionidae (18 h) < Linyphiidae (42 h) ≤ Lycosidae (61 h) < Theridiidae (135 h). Similarly when tested at this temperature, the DHL for C. pavonana mtCO1 DNA was lower in Lycosidae (93 h) than in Theridiidae (193 h). Despite having a longer base pair sequence, the target C. pavonana fragment (276 bp) typically persisted for longer in the guts of predators than ii the P. xylostella fragment (165 bp) and in Theridiidae, the DHL of target DNA of both species decreased with increasing temperature when tested at 20, 25 and 30 °C Field studies in Australia showed that the impact of the endemic natural enemy complex could suppress P. xylostella and C. pavonana populations, but that predators had a greater impact on P. xylostella populations than on C. pavonana populations. Based on their relative abundance, the proportion sampled that tested positive for target prey DNA and the DHL for target prey DNA in gut-contents analysis, Clubionidae and Linyphiidae were identified as the major predators of P. xylostella, although Lycosidae and Theridiidae, and probably the coccinelid H. varieagata, also contributed to P. xylostella suppression. The endolarval parasitoid D. semiclausum also contributed significantly to P. xylostella mortality but overall, the combined action of predators caused greater mortality. Similar analysis identified Araneidae, Clubionidae, Lycosidae, Linyphiidae, Oxyopidae and Salticidae as predators of C. pavonana, although more limited determination of DHL for C. pavonana DNA for these groups makes it difficult to rank predators in order of importance. In Queensland, C. pavonana did not suffer significant mortality due to parasitoids. Studies in West Java investigated the impact of natural enemies on P. xylostella and C. pavonana populations simultaneously. The endemic predator complex and D. semiclausum suppressed P. xylostella populations, but predators had a greater impact than the parasitoid. Crocidolomia pavonana was not attacked by parasitoids in West Java and although predators could reduce the pest population, they had a lower and more variable effect on C. pavonana than on P. xylostella. Most foliar dwelling spiders (Araneidae, Clubionidae, Gnaphosidae, Linyphiidae, Salticidae and Tetragnathidae) consumed P. xylostella but only a small number of Araneidae and Gnaphosidae tested positive for C. pavonana DNA. When sampled at the soil surface, Gnaphosidae and Lycosidae were positive for C. pavonana DNA and only Lycosidae were positive for P. xylostella DNA. The predatory insect fauna was more diverse in West Java than in Queensland and many groups, but particularly larval Syrphidae, adult and larval Coccinellidae and Staphylinidae tested positive for both pests. iii Declaration by author This thesis is composed of my original work, and contains no material previously published or written by another person except where due reference has been made in the text. I have clearly stated the contribution by others to jointly-authored works that I have included in my thesis. I have clearly stated the contribution of others to my thesis as a whole, including statistical assistance, survey design, data analysis, significant technical procedures, professional editorial advice, and any other original research work used or reported in my thesis. The content of my thesis is the result of work I have carried out since the commencement of my research higher degree candidature and does not include a substantial part of work that has been submitted to qualify for the award of any other degree or diploma in any university or other tertiary institution. I have clearly stated which parts of my thesis, if any, have been submitted to qualify for another award. I acknowledge that an electronic copy of my thesis must be lodged with the University Library and, subject to the General Award Rules of The University of Queensland, immediately made available for research and study in accordance with the Copyright Act 1968. I acknowledge that copyright of all material contained in my thesis resides with the copyright holder(s) of that material. Where appropriate I have obtained copyright permission from the copyright holder to reproduce material in this thesis. iv Publications during candidature Furlong, Michael J., Rowley, Daniel L., Murtiningsih, Rini and Greenstone, Matthew H. (2014). Combining ecological methods and molecular gut-content analysis to investigate predation of a lepidopteran pest complex of Brassica crops. Entomologia Experimentalis et Applicata, 153(2), 128-141. Publications included in this thesis No publications included Contributions by others to the thesis None Statement of parts of the thesis submitted to qualify for the award of another degree None v Acknowledgements Thanks and praises be to Jesus Christ, God Almighty for His grace, guidance and wisdom. I would like to express my deep gratitude to my research supervisors: Dr. Mike Furlong, Dr. Peter Ridland, and Dr. Lyn Cook for their patience, guidance, enthusiastic encouragements and constructive critiques and comments in this research project. I wish to acknowledge JAF for providing scholarship which enabling me to pursuing the post graduate education at University of Queensland, Australia. My grateful thanks are also extended to Dr. Eri Sofiari (former director of IVEGRI) and Dr. Terry Hill for encouraging me to apply for a JAF scholarship and providing the letter of recommendation to support my scholarship application. I am particularly grateful for the supports and encouragements from UQ academic staffs, particularly Prof. Meron Zalucky and A/Prof. Gimme Walter. I would also like to thank to A/Prof. Sassan Asgari and Dr. Bronwen Cribb for their constructive suggestions in each of milestone stage of my study. I also would like to offer my thanks to Ms Gail Walter for her assistance to keeping my study progress on schedule. My grateful thanks are also extended to the staff from Gatton Research Facility, DAFF, and Queensland: Mr. Stephen S, Chris, Vince, Ken, and Mr. Gavin for their enthusiastic and careful help during the Gatton field work. I would also like to extend my thanks to IVEGRI staff for their help during the Lembang field works in 2012. My grateful thank are also extended to my laboratory mates: Paul Lyn, Penny, Alicia, James, Jude, James, for their continuing support and advice during the difficult laboratory work. I am particularly grateful for the friendship and supports provided by my office mates Barry Ale, Rehan Silva, Gurion Ang, Praise Tadle, Tuyet Thi, Komal Gudarsani, and Alana Dunne. I wish to thank the warm friendships offered by many friends of mine: Yani, Maya, Agnes, Ivone, Andy, Anita and many others. Finally, I wish to thanks my mother for her continuing prayers, support and encouragements throughout my study, my father, who
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