2007 Annual Report 1a.Objectives (from AD-416) Objective 1. Develop new knowledge of life history, behavior, genetics, physiology, and ecology of CM, leafrollers, pear psylla, cherry fruit fly, apple maggot, secondary pests, and their natural enemies that provide opportunities for new and improved bio-rational management of tree fruit insect pests. Objective 2. Develop monitoring methods and techniques that provide information to growers on the presence and threat level from pear psylla and CM. Objective 3. Develop biorational and sustainable methods for control of CM, leafrollers, pear psylla, and fruit fly pests of tree fruits. 1b.Approach (from AD-416) Objective 1. Determine foods used by fruit flies over nature. Determine local movement patterns of fruit flies. Evaluate nutritional requirements of fruit flies. Characterize pear psylla behavioral responses to psylla and pear odors. Determine impact of insecticides on codling moth development by looking at mating and oviposition following exposures. Anthocoroid predators of psylla will be studied to determine behavioral, morphological, and molecular differences among species. Monoclonal antibodies and PCR techniques will be used to identify pear psylla consumed by predators. New methods for moving and stabilizing genes will be evaluated for use against codling moth. Objective 2. Formulations and chemicals for feeding attractant and kairomone lures will be tested for fruit flies. Arena and flight tunnel assays, GC-EAD and GC-MS will be used to isolate and identify new kairomonal attractans and pheromones for psylla and codling moth. Objective 3. New formulations will be developed for virus and nematode pathogens of insects. Baits and insecticides will be modified and formulated for killing fruit flies. Dispensers, formulations, and strategies for codling moth mating disruption will be evaluated. Formerly 5352-22000-015-00D (3/05). 4.Accomplishments Tree Fruit- 5352-22000-017-00D. One method for controlling codling moth, which is a major pest of pome and stone fruits, is the sterile insect technique. To improve effectiveness and economics of this method it is desirous to develop genetic sexing lines of this pest to allow for the release of sterile males. Yakima Agricultural Research Laboratory, Wapato, WA, has developed a method to determine the sex of an egg using real-time PCR (DNA amplifications) with a greater than 70% efficiency. This will allow for the identification and isolation of sex specific genes to be used in transformation cassettes for the development of genetic sexing lines. Improvement of transformation cassettes will facilitate the development of genetic sexing lines to be used in codling moth sterile insect technique. NP 304 Component II: Biology of Pests & Natural Enemies (Microbes), Problem Statement A: Basic Biology; Component V: Pest Control Technologies, Problem Statement D: Other Biologically-Based Control Management of Insect Pests of Temperate Tree Fruit Crops. The efficacy of using sex pheromones to manage codling moth needs further improvements. In addition, the use of many of the new insecticides are causing problems with a number of secondary pests such as phytophagous mites. A new management approach has been developed at the Yakima Agricultural Research Laboratory, Wapato, WA, which combines the use of reduced rates of insecticides and a sprayable sex pheromone formulation. Seasonal trials demonstrated that this integrated approach significantly improved the management of this pest with only pheromones; and integrated mite management was improved when compared with similar plots treated with the conventional air blast insecticide applications. NP 304 Component V: Pest Control Technologies, Problem Statement D: Other Biologically-Based Control Effects of Several Newer Insecticides and Kaolin on Oviposition and Adult Mortality in Western Cherry Fruit Fly. Cherry fruit fly is a serious quarantine pest of cherry in central Washington. Knowledge about the effects of newer and safer insecticides is needed to control the fly. Studies of insecticide effects and kaolin clay (a non-insecticide)at the Yakima Agricultural Research Laboratory, Wapato, WA, on reducing or preventing egg laying by this fly into cherries reveled that when sprayed onto cherries, all insecticides (azinphos-methyl, spinosad bait, imidacloprid, thiamethoxam, and indoxacarb) reduced the numbers of eggs flies laid into the cherries. Some insecticides did not need to be highly toxic or cause high mortality to reduce the numbers of eggs laid. When kaolin clay was sprayed on cherries, the numbers of eggs laid were also reduced. The results of this study are important because they show that eggs laid by cherry fruit flies can be reduced to similar levels by materials with different toxicities, the insecticides tested can reduce damage to cherries, and that in residential trees where no insecticides is desirable, kaolin may be useful in reducing egg laying and buildup of fly populations. NP 304 Pest Control Technologies, Problem Statement E: Chemical Control New Kairomonal Lure for Codling Moth, a Pest of Apple. Codling moth is the key pest of apple and pear, and new methods are needed to manage it that are environmentally compatible, safe, economical and effective. Chemical attractants are used for monitoring, for mating disruption, and for lure and kill or baiting techniques. Yakima Agricultural Research Laboratory, Wapato, WA, discovered a novel chemical combination from the odor of fruits that is more strongly attractive to female codling moths compared to any other lures. Attracting females of a pest species suggests potential to directly reduce oviposition and reproduction. This new attractant holds promise for use in baits to attract and kill codling moths as a management strategy that uses much less pesticide, and with no pesticide contact with the environment or the crop. NP 304 Component II: Biology of Pests & Natural Enemies (Microbes), Problem Statement A: Basic Biology; Component V: Pest Control Technologies, Problem Statement D: Other Biologically-Based Control Molecular gut content studies of codling moth and leafroller predators. Predation is an important component of the natural regulation of insect pests but it is difficult to quantify due to the destruction of the prey by the predator and the secretive nature of predators. At the Yakima Agricultural Research Laboratory, Wapato, WA, primers for polymerase chain reactions specific to codling moth and the oblique banded leafroller, were developed and used to detect predation by multiple species of insect and spider predators and one bat species. A salt-based DNA extraction method was refined for this high through-put application. Predator species considered of minor importance were shown to be consistent predators of codling moth. This information may lead to conservation and enhancement of specific predators to improve insect control in orchards. NP 304 Component II: Biology of Pests & Natural Enemies (Microbes), Problem Statement A: Basic Biology; Component III: Plant, Pest, & Natural Enemy Interactions & Ecology, Problem Statement A: Understanding the Complex Interactions Two viruses that produce little cherry diseases found in Washington. Little cherry disease is a serious concern for sweet cherry growers in Washington and accounted for near destruction of the cherry industry in British Columbia several decades ago. Knowledge of the presence of the disease in Washington can support remediation procedures. A diagnostic test is needed to identify the virus that causes this disease. At the Yakima Agricultural Research Laboratory, Wapato, WA, the two viruses that produce little cherry disease were both shown to occur in Washington using newly refined reverse transcriptase methods. Also significant variation with the little cherry virus 1 was found in Washington, British Columbia, California, and Pennsylvania, which were similar but distinct to isolates from Germany and Turkey. Reliable methods to validate little cherry viruses as causes of little cherry symptoms represent an important step in managing this disease in orchards and understanding the potential of various insect vectors in transmitting the disease. NP 304 Component II: Biology of Pests & Natural Enemies (Microbes), Problem Statement A: Basic Biology; Component III: Plant, Pest, & Natural Enemy Interactions & Ecology, Problem Statement A: Understanding the Complex Interactions Formulation of virus and nematode pathogens for managing codling moth. Codling moth is the most significant pest of apple in the United States and control of the moth by conventional orchardists primarily relies on broad spectrum insecticides such as azinphos-methyl, which may have deleterious effects on beneficial insects and the environment. Microbial control agents of CM offer safe alternative means of control with minimal impact on the environment and food supply. Yakima Agricultural Research Laboratory, Wapato, WA, combined the CM granulovirus with the solar protectant lignin, particulate clays and others and evaluated of these adjuvants in both laboratory and field studies. Our results indicate that the most effective means of protecting the virus from ultra-violet radiation is encapsulation within lignin-sulfate micro-granules. Nematode activity against cocooned larvae in fruit bins and orchards was improved by formulating nematodes with wetting agent and humectants, and the use of mulching in orchards also improved persistence and activity of nematodes against CM larvae. Micontrol agents provide effective means of CM control without the risks associated with chemical pesticides, where as,b formulation of the CM granulovirus and insect specific nematodes provides good control of CM and better persistence than unformatted virus and nematodes. Use of alternatives to broad spectrum pesticides will provide control, have little or no effect beneficial insects, applicators, the environment and the food supply. NP 304 Component V: Pest Control Technologies, Problem Statement A: Traditional Biological Control 5.Significant Activities that Support Special Target Populations None. 6.Technology Transfer Number of invention disclosures submitted 1 Number of patent granted 1 Number of web sites managed 1 Number of non-peer reviewed presentations and proceedings 24 Number of newspaper articles and other presentations for non-science audiences 14 Review Publications Horton, D.R., Unruh, T.R., Lewis, T.M., Thomsen Archer, K.L. 2007. Morphological and genetic divergence in three populations of anthocoris antevolens (hemiptera: heteroptera: anthocoridae). Annuals of the Entomological Society of America 100(3):403-412. Miliczky, E., Horton, D.R. 2007. Natural Enemy Fauna (Insecta, Araneae) Found on Native Sagebrush Steppe Plants in Eastern Washington with Reference to Species Also Found in Adjacent Apple and Pear Orchards. Pan Pacific Entomology 83(1):50-65. Horton, D.R., Guedot, C.N., Landolt, P.J. 2007. Diapause status of females affects attraction of male pear psylla, cacopsylla pyricola, to volatiles from female-infested pear shoots. Entomologia Experimentalis et Applicata. 123:185-192. Horton, D.R., Landolt, P.J. 2007. Attraction of male pear psylla, cacopsylla pyricola, to female-infested pear shoots. Entomologia Experimentalis et Applicata. 123:177-183. Knight, A.L. 2006. Multiple-mating of male and female codling moth (lepidoptera: tortricidae) in apple orchards treated with sex pheromone. Environmental Entomology. 36:157-164. Knight, A.L. 2007. Disruption of mating in codling moth (lepidoptera: tortricidae) by an anthranilamid insecticide. Pest Management Science. 63:180-189. Knight, A.L., Fisher, J. 2007. Increased catch of male codling moth (lepidoptera: tortricidae)in orange plastic delta-shaped traps. Environmental Entomology. 35:1597-1602. Knight, A.L. 2006. Assessing the mating status of female codling moth (lepidoptera:tortricidae) in orchards treated with sex pheromone using traps baited with ethyl (e, z)-2,4-decadienoate. Environmental Entomology 35:894-900. Arthurs, S.P., Lacey, L.A., Miliczky, E. 2007. Evaluation of the codling mont granulovirus and spinosad for codling moth control and impact on non-target species in pear orchards. Biological Control 41:99-109. Dolinski, C., Lacey, L.A. 2007. Microbial control of arthropod pests of tropical tree fruit. Neotropical Entomology. 36:161-179. Lacey, L.A., Unruh, T.R. 2007. Biological control of codling moth (cydia pomonella, tortricidae: lepidoptera) and its role in integrated pest management, with emphasis on entomopathogens. Vedalia 12:33-60. Riga, E., Lacey, L.A., Guerra, N., Headrick, H.L. 2006. Control of the oriental fruit moth, Grapholita molesta, using entomopathogenic nematodes in laboratory and fruit bin assays. Journal of Nematology. 38(1): 168-171. Arthurs, S.P., Lacey, L.A., Behle, R.W. 2006. Evaluation of spray-dried lignin-based formulations and adjuvants as solar protectants for the granulovirus of the codling moth, Cydia pomonella (L). Journal of Invertebrate Pathology. 93: 88-95. Landolt, P.J., Toth, M., Josvai, J. 2007. First European Report of Social Wasps Trapped in Response to Acetic acid, Isobutanol, 2-Methyl-2-propanol, and Heptyl butyrate in Tests Conducted in Hungary. Bulletin of Insectology 60(1):7-11. Dvorak, L., Landolt, P.J. 2006. Social wasps trapped in the Czech Republic with syrup and fermented fruit and comparison with similar studies (Hymenoptera: Vespidae). Bulletin of Insectology 59(2):115-120. Mackenzie, J.K., Landolt, P.J., Zack, R.S. 2006. Attraction to peony (Paeonia, Paeoniaceae) by Polistes dominulus (Hympenoptera: Vespidae) demonstrated using Y-tube and parallel tube olfactometeters. Journal of the Kansas Entomological Society. 79(3): 231-238. Yee, W.L., Oriki, J., Nash, M. 2007. Mortality of Rhagoletis pomonella (Diptera: Tephritidae) exposed to field-aged spinetoram, GF-120, and azinphos-methyl in Washington state. Florida Entomologist 90:335-342. Yee, W.L. 2006. Feeding history effects on feeding responses of rhagoletis indifferens (dipt., tephritidae) to gf-120 and nulure. Journal of Applied Entomology. 130(9-10), 538-550. Yee, W.L., Alston, D.G. 2006. Effects of spinosad, spinosad bait, and chloronicotinyl insecticides on mortality and control of adult and larval western cherry fruit fly, rhagoletis indifferens (diptera: tephritidae). Journal of Economic Entomology 99(5):1722-1732. Yee, W.L., Landolt, P.J., Darnell, T.J. 2006. Attraction of rhagoletis pomonella (diptera: tephritidae) and non-target flies to traps baited with ammonium carbonate and fruit volatile lures in washington and oregon. Journal of Agricultural and Urban Entomology p. 133-149. Knight, A.L. 2007. Influence of within-orchard trap placement on catch of codling moth (lepidoptera: tortricidae) in sex pheromone-treated orchards. Environmental Entomology 36:425-432. Arthurs, S.P., Hilton, R., Knight, A.L., Lacey, L.A. 2007. Evaluation of the Pear Ester Kairomone as a Formulation Additive for the Granulovirus of Codling Moth, Cydia pomonella (L.) in Pome Fruits. Journal of Economic Entomology 100:702-709.