2006 Annual Report 1.What major problem or issue is being resolved and how are you resolving it (summarize project aims and objectives)? How serious is the problem? Why does it matter? Cotton yield losses by tarnished plant bugs (TPB) and their western cousins are second only to the bollworm/budworm complex. Percent of yield lost to plant bugs has been reasonably consistent since 1990. In recent years economic losses from tarnished plant bugs in US cotton alone have totaled approximately $100 million in control costs and yield losses. Current control practices rely primarily on a few chemical insecticide classes and resistance development is a growing concern. Alternative control options including entomopathogenic fungi and sterile TPB are being developed by this project to strategically target populations developing on alternate hosts before migrating to cotton. Highly pathogenic and selective isolates that are adapted to the target environment and formulations that improve field persistence will improve mycoinsecticide efficacy. Additional research is being conducted on TPB attraction, trapping efficiency, diapause, and the potential of augmenting natural epizootics of the cotton aphid fungus. 2.List by year the currently approved milestones (indicators of research progress) Year 1 (FY 2006) 1. Small scale biphasic production of B. bassiana. 2. Field trials of isolates in wild host plants and cotton (MS) and alfalfa (CA) 3. Initial field trials of UV protective formulations. 4. Establishment of radiation doses to produce sterility in F1 progeny of treated TPB males. 5. Design and construct an effective non-sticky field trap for TPB. 6. Develop laboratory and field tests that will give consistent measures of mating propensity, attractiveness of females, and responsiveness of males to determine the effectiveness of sterile TPB. 7. Determine interactions between cotton development and cotton aphid fungus epizootiology. Year 2 (FY 2007) 1. Scale up of biphasic production of B. bassiana. 2. Design and construction of an effective olfactometer will depend on being able to test TPB under conditions conducive to female pheromone production and male response. 3. Compare the competitiveness of sterile and normal insects. 4. Initiate epizootics by inoculation of the cotton aphid fungus. Year 3 (FY 2008) 1. Additional scale-up of UV protective formulations. 2. Field tests with sterile TPB releases. 3. Preservation of cotton aphid fungus inocula. Year 4 (FY 2009) 1. Effects of B. bassiana on beneficial insects during field trials. 2. Establish diapause models based on dynamic photoperiods and temperatures. 4a.List the single most significant research accomplishment during FY 2006. Mass production of a B. bassiana isolate from Lygus spp. was demonstrated on a scale practical for areawide management demonstrations. This production capacity greatly improved our ability to conduct multiple field trials both in MS and with collaborators to determine application rates, compare isolates, evaluate formulations, and potential for use with trap crops. 4b.List other significant research accomplishment(s), if any. The feasibility of the sterile TPB project depends largely on the ability to mass-produce the bugs. The source of a serious, intermittent diet-contamination problem that has occurred at both our laboratory and the mass-rearing laboratory formerly located at Mississippi State University was identified and corrected. 4c.List significant activities that support special target populations. None. 4d.Progress report. None. 5.Describe the major accomplishments to date and their predicted or actual impact. Beauveria bassiana isolates from Lygus spp. were discovered and described using a wide range of characteristics relevant to microbial biopesticide development. Improvement in formulation and mass production technology and twelve field trials have been conducted either in MS or through collaborators in CA, AR, and WA to evaluate efficacy, compare isolates, determine application rates, and evaluate formulations. Data on infection rates for target Lygus spp. and beneficial insects and populations impacts were determined from these field trials. The first effective, non-sticky TPB trap was developed. Attractiveness of females, responsiveness of males, flight propensity, and mating propensity of irradiated TPB and their F1 progeny were determined, and rearing methods were improved. These accomplishments have stimulated additional research to determine the feasibility of using irradiated TPB for suppressing native populations. 6.What science and/or technologies have been transferred and to whom? When is the science and/or technology likely to become available to the end-user (industry, farmer, other scientists)? What are the constraints, if known, to the adoption and durability of the technology products? No science or technology transferred yet. Commercial interest in entomopathogenic fungi for Lygus control will first require demonstration of successful field trials. Field efficacy may be improved by selection of highly pathogenic isolates and improvements in persistence through formulation. Isolates have been selected that are highly pathogenic and formulations that significantly improve UV tolerance. Other technical factors critical to commercial development, such as in vitro production capacity and mycotoxin production, have been investigated and results are favorable. Market factors, including the size and penetration of the market, will need to be considered. Regarding potential market size, Lygus is a significant pest of many high value crops. Control of Lygus and other susceptible insect pests in a range of crops will need to be investigated through collaborations to expand market size. Regarding potential market penetration, entomopathogenic fungi could fill niches not currently targeted with chemical insecticides (e.g. populations in wild host plants), and a relatively narrow spectrum of insecticide chemistry is currently available for Lygus control, which may be threatened by regulatory actions (e.g. FQPA) and resistance development. Information on the economic impact of area-wide management of TPB in wild host plants through wild host plant destruction is currently being gathered in another project. This information may help to justify the investment control strategies targeting populations in wild host plants. Ultimately the projected returns based on market size and penetration will need to justify registration costs and investment. Some data currently being collected in the development of the isolates could potentially be referred to in the registration package differing some of the registration costs. Commercial development of the cotton aphid fungus would not follow the typical microbial biopesticide development paradigm as long as it cannot be produced in vitro. However, epizootics of this fungus have tremendous potential to spread once initiated. Therefore, relatively little investment in inoculation could have large returns in control. Potentially, grower cooperative groups or small companies could provide sufficient infrastructure for the collection, storage, and inoculation of fields with N. fresenii infected cadavers. A model for grower cooperative investment in the cotton aphid fungus is currently in place through the Cotton Aphid Fungus Sampling Service funded by Cotton Incorporated. This program has provided very significant returns to the growers from saved aphicide applications. We are still in the proof-of-concept phase of this project, demonstrating that epizootics may be initiated prior to natural epizootics and, if so, how early and under what conditions. If these results are promising we will investigate the rate of spread of initiated epizootics and improve efficiencies of collection, storage, and inoculation. Futher investigation will need to be done to determine if such an approach to control would be exempt from registration requirements, a major hurdle to small startup companies, since the fungus is highly specific and is naturally prevalent over a wide geographic area. There should also be additional research into the potential of using N. fresenii for inoculative biocontrol of soybean aphids. 7.List your most important publications in the popular press and presentations to organizations and articles written about your work. (NOTE: List your peer reviewed publications below). None. Review Publications Leland, J.E., Snodgrass, G.L. 2004. Prevalence of naturally occurring beauveria bassiana in lygus lineolaris populations on wild host plants in the delta and hill country regions of mississippi. Journal of Agricultural and Urban Entomology. Leland, J.E. 2005. Characteristics of beauveria bassiana isolates from lygus lineolaris populations of mississippi. Journal of Agricultural and Urban Entomology. Villavaso, E.J. 2004. A non-sicky trap for tarnished plant bug (heteroptera: miridae). Journal of Entomological Science.