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ARS Home » Southeast Area » Gainesville, Florida » Center for Medical, Agricultural and Veterinary Entomology » Insect Behavior and Biocontrol Research » Research » Research Project #430081

Research Project: Improved Biologically-Based Methods for Insect Pest Management of Crop Insect Pests

Location: Insect Behavior and Biocontrol Research

2016 Annual Report

Objective 1. Develop new transgenic conditional lethal strains for sexing and sterility in tephritid and drosophilid fruit flies to be used in the sterile insect technique, produce redundant lethality systems for ecological safety, and transgenic technology for emerging pest species such as the Asian citrus psyllid. Objective 2. Develop paratransgenic strains that eliminate the ability of host populations to vector plant disease by using Wolbachia cytoplasmic incompatibility to drive pathogen immunity throughout populations of key pests such as Asian citrus psyllid, glassy-winged sharpshooter, and potato/tomato psyllid. Objective 3. Develop automated acoustic methods for improved surveillance and detection of hidden and invasive pests such as red palm and citrus root weevils and Asian long-horned beetle that will facilitate more rapid information collection/processing by use of big data technologies. Objective 4. Develop improved visual-cue trap systems for surveillance of invasive and outbreak insect pests such as Asian citrus psyllid and corn silk fly, and improve strategies for detecting and predicting the dispersal of these pests by understanding the role of visual and other stimuli in affecting their behavior. Objective 5. Develop predictive models for fall armyworm migration pathways that are shifting due to climate change, and improve area-wide landscape management tactics for these pests by developing cover crop and biological control strategies to control them. Sub-objective 5.A. Develop genetic methods to monitor fall armyworm population behavior and air transport models to describe and predict its migration pathways and potential changes in infestation patterns due to climate change. Sub-objective 5.B. Improve area-wide landscape management tactics by developing cover crop and other strategies to mitigate pest populations such as fall armyworm, and attract or support natural enemies and pollinators.

The experimental approaches to achieve these objectives is multidisciplinary and integrates genetics, ecology, behavior, and engineering to address various stages of control, from molecular genetics leading to autocidal strain development to predicting changes in pest migration in response to global climate change. These approaches will apply, initially, to high priority invasive fruit flies, beetles, psyllids, fall armyworm and corn silk flies, and will include studies for development of molecular genetics methods for gene discovery and manipulation to develop genetically-modified pest strains to suppress wild populations, or eliminate their ability to vector pathogens of plant disease; development of detection and surveillance methods for optimization of acoustic, chemical and visual-cue detectors for detection and surveillance of hidden, invasive and outbreak pests; and biological control studies to develop predictive models to target shifting migrations of noctuid pests in response to climate change, and development of improved area-wide landscape management tactics to mitigate pest populations and attract natural enemies and pollinators.

Progress Report
The efforts of the Insect Behavior and Biocontrol Research Unit, IBBRU, resulted in significant progress towards all five objectives and subobjectives for this project. Under Objective 1 in collaboration with ARS researchers in Hilo, Hawaii, and an international group of academic colleagues, we completed the sequencing and annotation of the Mediterranean fruit fly genome. This information will be utilized to enhance all phases of insect pest management of the medfly, including development of transgenic strains that will enhance control by Sterile Insect Technique (SIT). Under Objective 2 lipid biosynthesis was examined in the obligate intracellular endosymbiotic bacterium, Wolbachia. We demonstrated that lipid biosynthesis precursors are not derived from the host, but are produced in the rickettsial cell. A novel inhibitor of this enzyme, itaconic acid was used effectively as an antibiotic for Wolbachia to cure insect lines. We also developed an artificial feeding protocol for the delivery of bioactive agents to the Varroa mite, Varroa destructor. This Varroa mite feeding protocol offers a simple reliable means of testing biologically active materials to disrupt reproduction or development and provide control of the pest Varroa mites. Significant progress was made in Objective 3, we tested a vibration trap and mating disruption system for Asian citrus psyllids (ACP). We recently developed a device that mimics female­ produced courtship vibrations to trap male ACP searching in citrus tree branches for females. In addition, methods of using vibrational signals to disrupt mating were assessed. Under Objective 4, we identified several chemical attractants for use in current traps for collection of several species of corn silk flies. Use of these attractants can enhance sensitivity of current surveillance traps to better guide optimal timing of pesticide applications for sweet corn production. We also assessed the impact of feeding on a wide range of citrus plants and found clear differences in both size and shape of wings from ACP reared on different citrus cultivars. Such differences in wing size and shape may be associated with greater dispersal capability of ACP reared on specific plants and is of importance in predicting the potential risk for spreading of citrus greening into uninfected citrus. Under Objective 5A, we developed new genetic tools to monitor migration of fall armyworm populations. A screen of intron sequences which have higher genetic variability than exon sequences was examined. The intron sequences of a nuclear gene was developed as a marker for identifying subgroups of fall armyworm that will facilitate ongoing studies investigating the distribution of host-specific strains and pesticide resistance in this species. Under Objective 5B, the summer cover crop plant sunn hemp was shown to attract pollinators and insect predators and the use sweet alyssum, provides nutrition to egg and larval parasitoids that attack noctuid pests.

1. Development of a vibration trap and mating disruption systems for Asian citrus psyllids (ACP). Asian citrus psyllids (ACP) are an invasive insect pest in Florida citrus groves and the arthropod vector of the bacterium that causes citrus greening. Unlike most insects, male and female ACP use wing-buzzing vibrations instead of pheromones to locate and court one another in citrus trees. Researchers at ARS, Gainesville, Florida, developed and tested a highly efficient device that mimics female-produced courtship vibrations to trap male ACP. As an extension of this research, they have also used vibrational signals that mimic females to disrupt mating. Tests in the laboratory have proven successful and devices are being engineered for the transfer of this technology to field environments to improve detection and control of ACP populations.

2. Completed whole genome sequencing, assembly and annotation of the Mediterranean fruit fly. The Med Fly is one of the most serious agricultural pests to U.S. agriculture and throughout the world. The Med fly genome was sequenced and annotated in order to obtain extensive knowledge of its genetic pathways essential to development, behavior and reproduction. Researchers at ARS, Gainesville, Florida, and at Hilo, Hawaii, along with an international assemblage of academic colleagues, collaborated to conduct the sequencing and an in-depth manual curation of the genome and transcriptome. They identified specific genetic expansions that can be related to invasiveness and host adaptation, including gene families for chemoreception, toxin and insecticide metabolism, immunity, cuticle proteins, and aquaporins among others. This new knowledge, along with genetic tools available for functional and applied studies, should significantly advance the means of controlling the size and invasive potential of medfly populations and to enhance control by Sterile Insect Technique (SIT).

3. Modeling fall armyworm migration. Fall armyworm is a pest of vegetable, row, and turf crops in the eastern and central U.S. This species annually migrates north from southern Florida and southern Texas, although the pathways and destination of these two populations was unknown. Using genetic analyses of moths collected throughout the U.S., researchers at ARS, IBBRU, Gainesville, Florida, and collaborators have been able to identify the pathways Florida and Texas moths use to migrate north. Computer model simulations of migration that include population densities, wind patterns, and corn acreage relate well with field data obtained from pheromone traps. These results contribute to the knowledge of fall armyworm population ecology on a continental scale and will aid in the prediction and interpretation of migration patterns as they correspond to climate and pest management changes.

4. Alternative management tactics for fall armyworm. Cover crop and weedy refuge plants in vegetable and row crop agroecosystems offer a potential to attract and maintain populations of beneficial insects. Researchers at ARS, Gainesville, Florida, found that the summer cover crop plant, sunn hemp, cultivar AU Golden, provides many of the benefits of traditional cover crops. However, because of its ability to produce lots of flowers, sunn hemp plantings attract pollinators and insect predators. The use of cool season weedy plants, such as sweet alyssum will augment and extend the duration of control by providing nutrition to egg and larval parasitoids that attack noctuid pests. Utilization of sunn hemp as a management tool against fall armyworm is currently being published which will be followed by placement in extension-related publications for adoption by farmers and recommendation by seed suppliers.

5. Identified chemical attractants for corn silk flies.  Several species of corn silk flies have emerged as the primary insect pests threatening production of sweet corn in the southeastern U.S. Production relies on numerous applications of pesticides, however current surveillance methods for corn silk flies are poor and do not provide adequate information for optimally timed pesticide applications. Researchers at ARS, Gainesville, Florida, identified attractants for the corn silk flies. Use of these attractants can enhance sensitivity of current surveillance traps to better guide optimal timing of pesticide applications for sweet corn production.

6. Control of Rickettsial bacteria through lipid biosynthesis genes. Lipids are critical molecules especially in membrane production. Using bioinformatics, it was found that obligate intracellular reproductive-parasite alpha-proteobacterium infecting many arthropods, Wolbachia and other Rickettsia, lack the genes recognized to produce the building blocks of lipids. Utilizing a conditionally lethal strain of E. coli that is lipid-biosynthesis deficient, researchers at ARS, Gainesville, Florida, determined that alternate genes from Rickettsial bacteria can genetically complement for lipid building block biosynthesis. This discovery demonstrates that lipid biosynthesis precursors are not derived from the host, but in fact are produced in the rickettsial cell. A novel inhibitor of this enzyme, itaconic acid was used effectively as an antibiotic for Rickettsia.

7. Artificial feeding protocol for the Varroa mites. An artificial feeding protocol for the delivery of bioactive agents to the Varroa mites was developed by researchers at ARS, Gainesville, Florida. Various diet components were assessed in the medium and it was established that honey bee blood was necessary but feeding could be enhanced by addition of cultured insect cells. Feeding with Tebufenozide, an insect steroid hormone mimic was included in the medium, the levels for the last gene in the steroid pathway increased. This Varroa mite feeding protocol offers a simple reliable means of testing biologically active materials to disrupt reproduction or development and provide control of the pest Varroa mites.

1. Served as Board member for Center for Biological Control at College of Agriculture and Food Sciences Florida A&M University (FAMU). 2. Served as a member of graduate committees for 1 M.S. students (one male minority student) at FAMU; appointed member of the College of Agriculture and Food Sciences (CAFS)-FAMU graduate committee, member of the search committee for 2 faculty positions at the Center for Biological Control at CAFS-FAMU. 3. Mentored and provided experiential research training to one volunteer female undergraduate student (minority student) at University of Florida. 4. Mentored two female minority graduate students. 5. Mentored three female hispanic undergraduate students. 6. Served on the committee for a male M.S. graduate student at Florida A&M. 7. Hosted minority female and male undergraduate students from the Louis A. Stokes Minority Alliance program for conducting research on corn silk fly behavior. 8. Hired minority male undergraduate study in Pathways program and provided mentoring for research. 9. Mentored and hosted visiting minority male graduate student for several weeks to complete research project.

Review Publications
Alvarenga, C.D., Diaz, V., Stuhl, C.J., Sivinski, J.M. 2015. Contrasting brood-sex ratio flexibility in two opiine (Hymenoptera: Braconidae) parasitoids of tephritid (Diptera) fruit files. Journal of Insect Behavior. 29(1):25-36.
Peng, W., Wenping, Z., Handler, A.M., Zhang, H. 2015. The role of the transformer gene in sex determination and reproduction in the tephritid fruit fly, Bactrocera dorsalis (Hendel). Genetica. 143(6):717-727.
Shirk, P.D., Furlong, R.B. 2016. Insect cell transformation vectors that support high level expression and promoter assessment in insect cell culture. Plasmid Journal. 83:12-19. Available:
Mankin, R.W., Al-Ayedh, H.Y., Aldryhim, Y., Rohde, B. 2016. Acoustic detection of Rhynchophorus ferrugineus (Coleoptera: Dryophthoridae) and Oryctes elegans (Coleoptera: Scarabaeidae) in Phoenix dactylifera (Arecales: Arecacae) trees and offshoots in Saudi Arabian orchards. Journal of Economic Entomology. 109(2):622-628.
Meagher Jr, R.L., Nuessly, G.S., Nagoshi, R.N., Hay-Roe, M.M. 2016. Parasitoids attacking fall armyworm (Lepidoptera: Noctuidae) in sweet corn habitats. Biological Control. 95:66-72.
Mura, M.G., Nagoshi, R.N., Dos Santos, D.A., Hay-Roe, M.M., Meagher Jr, R.L., Vilardi, J.C. 2015. Demonstration using field collections that Argentina fall armyworm populations exhibit strain-specific host plant preference. Journal of Economic Entomology. 108(5):2305-2315.
Genc, H., Schetelig, M.F., Xavier, N., Handler, A.M. 2015. Germline transformation of the olive fruit fly, Bactrocera oleae (Rossi)(Diptera:Tephritidae) with a piggyBac transposon vector. Turkish Journal of Biology. 40:845-855.
Coates, B.S., Poelchau, M.F., Childers, C., Evans, J.D., Handler, A.M., Guerrero, F., Skoda, S.R., Hopper, K.R., Wintermantel, W.M., Ling, K., Hunter, W.B., Oppert, B.S., Perez De Leon, A.A., Hackett, K.J., Shoemaker, D.D. 2015. Arthropod genomics research in the United States Department of Agriculture-Agricultural Research Service: Current impacts and future prospects. Trends in Entomology. 11(1):1-27.
Schetelig, M.F., Targovska, A., Meza, J.S., Bourtzis, K., Handler, A.M. 2016. Tetracycline-suppressible female lethality and sterility in the Mexican fruit fly, Anastrepha ludens. Insect Molecular Biology. 25(4):500-508.
Shirk, P.D., Perera, O.P., Shelby, K., Furlong, R.B., LoVullo, E.D., Popham, H.J. 2015. Unique synteny and alternate splicing of the chitin synthases in closely related heliothine moths. Gene. 574(1):121-139.
Machtinger, E.T., Geden, C.J., Lovullo, E.D., Shirk, P.D. 2015. Impacts of extended laboratory rearing on female fitness in Florida colonies of the parasitoid spalangia cameroni (Hymenoptera: Pteromalidae) with an analysis of wolbachia strains. Annals of the Entomological Society of America. 109(2):176-182.
Zagvazdina, N.Y., Paris, T.M., Udell, B.J., Stanislauskas, M., Mcneill, S., Allan, S.A., Mankin, R.W. 2015. Effects of atmospheric pressure trends on calling, mate-seeking, and phototaxis of Diaphorina citri (Hemiptera: Liviidae). Annals of the Entomological Society of America. 108(5):762-770.
Njoroge, A.W., Affognon, H., Mutungi, C., Rohde, B., Richter, U., Hensel, O., Mankin, R.W. 2016. Frequency and time pattern differences in acoustic signals produced by Prostephanus truncatus (Horn) (Coleoptera: Bostrichidae) and Sitophilus zeamais (Motschulsky) (Coleoptera: Curculionidae) in stored maize. Journal of Stored Products Research. 69:31-40.
Snyder, D., Cernicciaro, N., Allan, S.A., Cohnstaedt, L.W. 2016. Insecticidal sugar baits for adult biting midges. Medical and Veterinary Entomology. doi: 10.1111/mve.12158.
Ruktanonchai, N., Lounibos, L.P., Smith, D.L., Allan, S.A. 2015. Experience- and egg-mediated oviposition behaviour in the yellow fever mosquito Stegomyia aegypti (=Aedes aegypti). Medical and Veterinary Entomology. 29(3):255-262.
Burroughs, J.E., Thomasson, J.A., Marsella, R., Greiner, E.C., Allan, S.A. 2016. Ticks associated with domestic dogs and cats in Florida, USA. Experimental and Applied Acarology. 69(1):87-95.
Junnila, A., Revay, E.E., Muller, G.C., Kravchenko, V., Qualls, W.A., Xue, R., Allan, S.A., Beier, J.C., Schlein, Y. 2015. Efficacy of attractive toxic sugar baits (ATSB) against Aedes albopictus with garlic oil encapsulated in beta-Cyclodextrin as the active ingredient. ACTA TROPICA. 152(1):195-200.
Eiden, A.L., Kaufman, P.E., Allan, S.A., Oi, F. 2016. Establishing the discriminating concentration for permethrin and fipronil resistance in Rhipicephalus sanguineus (Latreille) (Acari:Ixodidae), the brown dog tick. Pest Management Science. 72(7):1390-1395.
Hay-Roe, M., Nagoshi, R.N., Meagher Jr, R.L., Arias De Lopez, M., Trabanino, R. 2015. Isolation and DNA barcode characterization of a permanent Telenomus (Hymenoptera: Platygastridae) population in Florida that targets fall armyworm (Lepidoptera: Noctuidae). Annals of the Entomological Society of America. 108(5): 729-735.
Nagoshi, R.N., Meagher Jr, R.L. 2016. Using intron sequence comparisons in the triose-phosphate isomerase gene to study the divergence of the fall armyworm host strains. Insect Molecular Biology. 25(3):324-337.
Hay-Roe, M.M., Meagher Jr, R.L., Nagoshi, R.N., Newman, Y. 2016. Distributional patterns of fall armyworm parasitoids in a corn field and pasture field in Florida. Biological Control. 96:48-56.
Hay-Roe, M.M., Nagoshi, R.N., Meagher Jr, R.L., Lopez, M., Trabanino, R. 2015. Isolation and DNA barcode characterization of a permanent Telenomus (Hymenoptera: Platygastridae) population in Florida that targets fall armyworm (Lepidoptera: Noctuidae). Annals of the Entomological Society of America. 108(5):729-735.
Boucias, D., Liu, S., Meagher Jr, R.L., Baniszewski, J. 2016. Fungal dimorphism in the entomopathogenic fungus Metarhizium rileyi: detection of an in vivo quorum-sensing system. Journal of Invertebrate Pathology. 136:100-108.
Niu, Y., Qureshi, J.A., Ni, X., Head, G.P., Price, P.A., Meagher Jr, R.L., Kerns, D., Levy, R., Yang, X., Huang, F. 2016. F2 screen for resistance to Bacillus thuringiensis Cry2Ab2-maize in field populations of Spodoptera frugiperda (Lepidoptera: Noctuidae) from the southern United States. Journal of Invertebrate Pathology. 138(2016): 66-72. doi.10.1016/j.jip.2016.06.005.
Westbrook, J.K., Nagoshi, R.N., Meagher Jr, R.L., Fleischer, S.J., Jairam, S. 2015. Modeling seasonal migration of fall armyworm moths. International Journal of Biometeorology. 60:255-267.
Mankin, R.W., Rohde, B., Mcneill, S. 2016. Vibrational duetting mimics to trap and disrupt mating of the devasting Asian citrus psyllid insect pest. Proceedings of Meetings on Acoustics (POMA). 25,010006. doi: 10.1121/2.0000185.
Pregmon, E.A., Lujo, S., Norton, K.R., Hartman, E.N., Barukh, R., Zagvazdina, N.Y., Mankin, R.W. 2016. A “walker” tool to place Diaphorina citri (Hemiptera: Liviidae) adults at predetermined sites for bioassays of behavior in citrus (Sapindales: Rutacease) trees. Florida Entomologist. 99(2): 308-310.