CAPTIVE REARING AND SEMIOCHEMCIAL ECOLOGY OF TRICHOGRAMMA PAPILIONIS (HYMENOPTERA: TRICHOGRAMMATIDAE) A DISSERTATION SUBMITTED TO THE GRADUTE DIVISION OF THE UNIVERSITY OF HAWAIʻI AT MĀNOA IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF DOCTOR IN PHILOSOPHY IN ENTOMOLOGY MAY 2020 BY ABDULLA N. ALI DISSERTATION COMMITTEE: MARK G. WRIGHT, CHAIRPERSON JON- PAUL BINGHAM (UNIVERSITY REP) LEYLA KAUFMAN PETER FOLLETT GORDON BENNETT i Ó copyright 2020 By Abdulla N. AlI i DEDICATION At first dedicatIng thiS diSSertatIon to AlmIghty Allah, wIthout hiS mercy and sympathy, I waS not able to accomplISh thiS study. AlmIghty Allah gave me the power and confidence to done project work and alSo holy prophet Muhammed and hiS famIly (Peace Be Upon Them) who are a lIght for my lIfe. I alSo dedicate thiS diSSertatIon to my lovely parentS wIth the deepeSt gratItude Whose love and prayers have alWays been a source of strength for me. To my father who did not lIve long enough to see me complete thiS succeSSful mISSIon. My dedicated alSo goeS to my lovely wIfe Maha and my children WISam and TaIm Who fill my heart wIth hope and happineSS. To my sIblIngs and friends, theIr contInuous support and advice have helped me to pursue my goalS. ii ACKNOWLEDGMENTS Apart from my effortS, the succeSS of my project depends largely on the encouragement and guidelIneS of many others. I take thiS opportunity to expreSS my gratItude towards them. FIrst of all, I would lIke to thank my major profeSSor Dr. Mark G. Wright for hiS contInual and valuable guidance, huge support that I belIeve wIthout hiS superviSIon I would not finiSh any of my reSearch. It iS not enough to expreSS myself for all the great things he haS done to me. Dr. Wright iS more than an adviSor. I have to be humble wIth all the things I learned from him in hiS laboratory. I learned from him how to be a profeSSIonal; how do I think critIcally and lInk ideaS. I would lIke to expreSS my serious thank you to my diSSertatIon commIttee members, Drs. Leyla Kaufman, Jon-Paul Bingham, Peter Follett, and Gordon Bennett. I apprecIate all thoughtful feedback and critIcISm of my chapters that were very delIberate and directed me through my Ph.D. Study. I am truly thankful for all useful InputS. My famIly (Mom, Dad, brothers and SISters). To my SWeetheart WIfe Maha A. Najm and my kids (WISam and TaIm AlI), my depth thank you to you all for your prayers and immersIve support. Thank you Maha for beIng very penitent and genuinely supportIve to me. Thank you for giving me such a beautIful gift (WISam and TaIm). I would alSo lIke to thank the MIniStry of HIgher EducatIon & ScIentIfic ReSearch, UniversIty of Kufa, Iraq for grantIng me a scholarship and an opportunity to study abroad. Iraqi cultural office in WaShington D.C., they are apprecIated for theIr genuine support and financIal aId through five and a half year. To my all Student colleagueS, it WaS such a bleSSIng tIme to be wIth you. I am indebted to David Honsberger for hiS aSSIStance wIth fields work, critIcal reading and editIng on the first draftS of my theSIS manuscriptS. FInally, It iS my pleaSure to be indebted to all people who belIeved in me and Supported me even wIthout my aWareneSS. There are definitely many people iii Whose nameS have not mentIoned, helped me to accomplISh thiS important mISSIon in my lIfe. ThiS work waS funded by Hatch Project 919-H, admIniStered by CTAHR, and the MIniStry of HIgher EducatIon & ScIentIfic ReSearch, UniversIty of Kufa, provided support to me. iv LIST OF PUBLICATIONS AlI, A.N. and Wright, M.G. 2020. Behavior reSponse of Trichogramma papilionis In reSponse to host eggs, host plantS, and induced plant cueS. Biological Control. (Accepted) AlI, A.N. and Wright, M.G. FItneSS effectS of founder female number of Trichogramma papilionis reared on a factItuouS host Ephestia kuehniella (Zeller). Proceedings of the HaWaIIan Entomological SocIety (In revieW) AlI, A.N. and Wright, M.G. 2018. ReSponse of Trichogramma papilionis to plant volatIleS aSSocIated wIth Lepidoptera oviposItIon. Biological Control in PeSt management SystemS of PlantS Annual MeetIng. WhitefiSh, Montana, October 2018. (Oral preSentatIon) Papers in Preparation: AlI, A.N. and Wright, M.G. ReSponse of Trichogramma papilionis WaSps to blends of synthetIc SemIochemIcalS. AlI, A.N. and Wright, M.G. The effect of plant derived semIochemIcalS on searching behavior of Trichogramma papilionis In different environmentS. v ABSTRACT ThiS study addreSSed aSpectS of maSS rearing of Trichogramma papilionis (Hymenoptera; TrichogrammatIdae), including the effectS of varied colony founder sIze on waSp fitneSS, and the exploitatIon of the waSps to locate egg hostS in which to deposIt thereof progeny. EffectS of InitIal founder female number of T. papilionis Were inveStIgated usIng fItneSS parameters (emergency rate, sex ratIo and fecundity) to quantIfy the effectS of a severe bottleneck (sIngle founder female) on 10 subsequent generatIons. ReSultS Showed that no sIgnificant difference for eggs laId per female over ten generatIons, suggeStIng that the imposed bottleneck did not reSult In reduced female fecundity for any founder populatIon sIze. However, founder numbers did affect both the emergence rate and sex ratIo of T. papilionis. Further inveStIgatIon of the impactS of inbreeding on field performance of the waSps waS diScontInued aS extremely lImIted host finding abilIty of the waSps waS observed in some habitatS. The emphaSIS of the work waS thus Shifted to elucIdatIng the searching behaviors of T. papilionis In relatIon to chemIcal cueS. The hypotheSIS that T. papilionis are attracted to host habitat by host plant or egg-aSSocIated volatIle chemIcalS waS teSted. The reSponse of T. papilionis femaleS to olfactory cueS from host eggs, host plantS and Induced plant volatIleS were studied. The reSponse of T. papilionis femaleS to different info- chemIcal cueS waS teSted in Y-tube olfactory aSSayS. WaSps made a posItIve reSponse to odors from corn earworm (CEW) eggs Helicoverpa zea (Lepidoptera: Noctuidae) compared wIth blank aIr, while there waS a negatIve reSponse to Ephestia kuehniella eggs (Lepidoptera: PyralIdae) compared blank aIr: T. papilionis femaleS thus preferred odors from corn earworm eggs over the Mediterranean flour moth eggs. Further, the WaSps were attracted to volatIle emISSIons from Sunn hemp Crotalaria juncea (L.) over maize Zea mays (L.), deSpite both plantS InfeSted wIth H. zea vi eggs. No preference waS observed for plantS not infeSted wIth H. zea eggs, suggeStIng T. papilionis Showed a posItIve reSponse to stImulI from sunn hemp plantS that mIght be induced by H. zea oviposItIon. ChemIcal volatIle collectIon and headspace analysIS WaS conducted. Headspace analysIS and thermal deSorptIon and gaS chromatography–maSS spectrometry (TD- GCMS) was used to qualItatIvely and semI-quantItatIvely determIne the difference in plant volatIle organic componentS (VOCs) from Helicoverpa zea egg infeSted Sunn hemp plantS compared wIth intact sunn hemp plantS and H. zea eggs only. TD iS used aS a preconcentratIon technique of VOCs for gaS chromatography-maSS spectrometry (GC–MS), making it useful to detect low-concentratIon analyteS that would otherwISe be undetectable. ReSultS demonstrated that sunn hemp plantS releaSed 55 chemIcal volatIleS wIth five compounds that Were unique, or Were emItted in higher concentratIons, for plantS infeSted wIth CEW eggs. TheSe volatIle compounds Were consIStent wIth lInear alkaneS, aldehydeS, aromatIcS, polyterpene-related compounds, naphthalene derivatIveS, and eSter-related compounds. HIgh concentratIons of aniSole, β-myrcene, cIS-butyric acId, trans-ISoeugenol, and biS(2-ethylhexyl) phthalate Were found in infeSted Sunn hemp. The majority of GCMS peaks detected from H. zea eggs were consIStent wIth phosphateS, pheromone-related compounds, various natural productS, a serieS of glycol-related compounds, and a serieS of fatty acId eSter-related compounds. Several compounds were shared in sunn hemp sampleS and corn earworm eggs: aniSole, β-myrcene, and bIS (2-ethylhexyl) phthalate, but were detected in higher concentratIons from the plantS wIth H. zea eggs. EvaluatIon of the reSponse and the performance of T. papilionis femaleS In y-tube olfactory bioaSSays to SIngle compounds, and blends of synthetIc chemIcal showed that the WaSps were sIgnificantly attracted to only tWo of the aSSayed chemIcal volatIleS (anISole and vii biS(2-ethylhexyl) phthalate). Some concentratIons of aniSole and biS (2-ethylhexyl) phthalate Were attractant to the waSps, whereaS Some concentratIons of the other teSted chemIcal compounds repelled the waSps. WaSps were attracted to a blend of aniSole and biS(2-ethylhexyl) phthalate (25μL /100μL ratIo) which iS sImIlar to the ratIo of aniSole to biS(2-ethylhexyl) phthalate detected In the (GC-DMS) chromatograph for C. juncea plantS infeSted wIth H. zea eggs. No sIgnificant attractIon to any other blend ratIos of aniSole and biS(2-ethylhexyl) phthalate WaS observed. Greenhouse and field experimentS were conducted to determIne whether the patterns observed in the y-olfactometer were consIStent under leSS constraIned conditIons. The optImal blend identIfied above waS initIally teSted in a greenhouse, and later in closed-canopy environmentS (under treeS) and open habitat wIth no treeS. The paraSItISm rate by T. papilionis WaSps waS sIgnificantly increaSed When the waSps were exposed to aniSole and biS(2-ethylhexyl) phthalate blend In both greenhouse and outdoor trialS (covered habitat), at leaSt over short diStance (up to 2m from the volatIle sourceS). The findings preSented in thiS diSSertatIon underscore the importance of improving our understanding of how tri-trophic interactIons (natural enemIeS- herbivoreS and host plantS) Interact to influence insect behavior, aS well aS the impact of variable environmentS, impact paraSItoid waSps. The reSultS may alSo contribute to finding a way to improve natural enemy efficacy in augmentatIve and conservatIon biocontrol effortS. SemIochemIcal cueS can posItIvely or negatIvely affect the reSponse of paraSItIc waSps.