2007 Annual Report 1a.Objectives (from AD-416) To determine chemical and biochemical factors involved in feeding, development, dormancy and interactions with hosts of economically important insects, both pest and beneficial. To characterize and evaluate the function of cuticular lipids, internal lipids, intermediary metabolites and recognition chemicals for pest and beneficial insects, and the effect of lipids on interactions of natural enemies with their insect hosts. Use this information to enhance the efficiency of insect pest control programs and to improve management of beneficial insects. 1b.Approach (from AD-416) Microscopic techniques and biochemical technology will be used to characterize stylet feeding for adult and nymph stages of economically important homopteran insects. The physiological, biochemical, behavioral, anatomical, and ultrastructural mechanism(s) of stylet feeding, stylet-sheath formation, plant tissue damage by sheath material will be determined. Chemical and biochemical approaches will be used to document some specific and new functions of lipids, such as determining which chemical cues specific parasitic wasps use to mark and identify homopteran hosts that have already been parasitized and identifying lipids and determining their roles in several biological processes of beneficial solitary bees that includes: functioning as an important source of stored energy and intracellular communication during diapause-related processes; functioning as chemical cues in the process of nesting site recognition for female pollinator bees; providing a matrix and possibly nutrition for the larval-gut germination and growth of a pathogen fungus that causes chalkbrood disease in solitary bees. 3.Progress Report For Project 5442-22000-041-00D: Progress has been made on using microscopic techniques for studying the physiology and morphology of the Dufour’s gland from female alfalfa leafcutting and blue orchard bees in regard to the cuticular site(s) for glandular secretion release. Dufour’s glands (associated with the stinger apparatus) were dissected and analyzed by light microcopy and by transmission electron microscopy of embedded and ultra-sectioned tissues. Results for both species of pollinator bees indicate that posterior end of Dufour’s gland enters the base of the stinger apparatus. Microscopic analysis continues on determining the actual site(s) of release for Dufour’s gland secretion. These findings are important in regard to determining if Dufour’s gland secretory material plays a role in providing nesting attractants for these bees. This research related to Component III: Bees and Pollination, Problem Statement B. Bee Management and Pollination of the NP (305) Action Plan. Subordinate Project Report:SCA 58-5442-1-0331: Specific cooperative agreement (58-5442-1-0331 “Xylem and Phloem penetration by Plant Feeding Insects”) between ARS and North Dakota State University-Department of Plant Pathology was terminated 09-14-2006 and final report by P.I. was submitted at that time. The results of the collaborative research provided a better understanding of how homopteran insects (whiteflies, leafhoppers) can cause damage to vital host plant tissues. This research related to Component III: Plant, Pest, and Natural Enemy Interactions and Ecology, Problem Statement A. Understanding the Complex Interactions of the NP (304) Action Plan. 4.Accomplishments Identifying Nesting Compounds for Alfalfa Leafcutting Bees The use of olfactory cues for nest recognition by the solitary bee, alfalfa leafcutting bee (ALCB) Megachile rotundata (Hymenoptera: Megachilidae) was studied in a greenhouse environment at the USDA-ARS Bee Biology Laboratory, Logan, UT and chemical/biochemical analyses at the USDA-ARS Biosciences Research Laboratory, Fargo, ND. Glass tube segments (two) were provided for each nesting cavity of eight test female bees so that in-nest behavior could be observed and tube segment contents analyzed. Organic solvent extracts of glass tube contents were analyzed for composition and chemical structure by chromatography (TLC, GC) and GC-mass spectrometry. Similarly, the identity and composition of compounds from surfaces and contents of the test female ALCBs, ALCB Dufour’s gland contents and alfalfa leaves and alfalfa pollen were determined and compared to the identified compounds from the various glass nesting tube segments. Analyses revealed the presence of saturated and unsaturated hydrocarbons and wax esters, aldehydes and free fatty acids. The composition and distribution of those segment compounds from tube segments that were found to be common to compounds from test bees will be tested by bioassay techniques for nesting attractant activity. A better understanding of nesting attractants will contribute to improved management of this important pollinating bee (and other pollinator bee species) by scientists, commercial bee providers and growers of agricultural crops that depend on bee pollinators. This research relates to Component III: Bees and Pollination, Problem Statement B. Bee Management and Pollination of the NP (305) Action Plan. 5.Significant Activities that Support Special Target Populations None. 6.Technology Transfer Number of new CRADAs and MTAs 1 Number of non-peer reviewed presentations and proceedings 2 Review Publications Eltahlawy, H., Buckner, J.S., Foster, S.P. 2007. Evidence for two-step regulation of pheromone biosynthesis by the pheromone biosynthesis-activating neuropeptide in the moth Heliothis virescens. Archives of Insect Biochemistry and Physiology. 64(3):120-130. Eltahlawy, H., Buckner, J.S., Foster, S.P. 2007. Regulation of pheromone biosynthesis in the "Z strain" of the European corn borer, Ostrinia nubilalis. Archives of Insect Biochemistry and Physiology. 65(1):29-38.