Progress 04/02/07 to 11/07/10
Outputs
Progress Report Objectives (from AD-416) Development of effective biological control technology for selected pests, including importation and culture of appropriate natural enemies and microbial control agents of insects and development and testing of technology for production, processing, storage, packaging, distribution, and release of natural enemies of insects and weed pests. Effective augmentative and classical biological control strategies for key pests are developed. Production of biological control agents for use in field evaluations in cooperation with other labs. Improvements in mosquito biological control agent formulation, delivery, specificity and storage stability will be achieved by modifying the physical and chemical properties of the formulations. Approach (from AD-416) Research and development activities are directed, primarily, at the development of automated mass rearing technologies for important biological control agents, including the development of artificial diets. Agents for study are selected on the basis of the needs of cooperators; the potential for success in the development of mass rearing technology; and the availability of basic rearing procedures, artificial diets, etc. Quality and process control criteria are developed. Techniques for packaging, distribution, and release of biological control agents are developed and tested. Biological control strategies are developed and tested through the mass rearing and field evaluation of the released agents. This is the final report for project 6402-22000-057-00D terminated in October 2010. Substantial results were realized over the 5 years of this project. A stable red-eyed strain of tarnish plant bug (Lygus lineolaris) was established to supplement insect genetics efforts. A new mutation involving deformed limbs (�Pirate�) was identified and a colony was established. Specialized needle preparation, microinjection, and arthropod handling protocols were established for embryos, nymphs, and adults of the tarnish plant bug and embryonic lacewings (Chrysoperla rufilabris). A method was invented for microencapsulating conidia (spores) of an insect-infecting fungus with sucrose by spray drying at higher temperatures. We achieved 68.5% survival of conidia using this method. This invention has significantly increased the amount of living spores in dried preparations. This invention can provide formulations for a variety of applications, from seed coatings to sprayable conidia. This invention can be used to dry other microbial biological control agents. Efforts to improve the mass rearing technology of predatory mites (Phytoseiulus persimilis) culminated in the development of a caged rearing system. This system allows the continuous production of predatory mites with minimal loss and consistent quality. Prototypes were shipped to Syngenta Bioline in Oxnard, California, for testing, and a patent application was filed. Laboratory studies on the effects of temperature on growth, development, and reproduction of the predatory mites culminated in the determination of an optimal range of temperatures for mass production of these natural enemies of the worldwide pest called the two-spotted spider mite. New artificial diets for the predatory mite and encapsulation methods were invented. For the first time, predatory mites were induced to lay eggs in artificial diet in absence of their prey using ethanol-water extracts from spider mite prey. This new discovery opens the door for the development of a complete artificial based rearing system eliminating the need for greenhouses. Studies on the nutritional ecology of the predatory mite yielded the discovery of alternate food sources for them. It was discovered that predatory mites regularly feed in extra-floral nectaries of bean plants. This discovery will now allow for the development of new techniques to extend the life of the predators in the field. Studies on the bio-ecological interactions between plant chemicals, the two-spotted spider mites, and its predators, showed that plant chemicals such as linamarin accumulate in the spider mites and impact negatively on the survival, development time, growth, and reproduction of two of their predators, the predatory mite (Phytoseiulus persimilis) and a lady beetle (Stethorus punctillum). Based on these results, it was concluded that the use of lima bean varieties with low content of linamarin were better suited to mass produce these 2 natural enemies. Accomplishments 01 A True Artificial Diet for the Predatory Mite Phytoseiulus Persimilis. Previous reports reported the development of a new artificial diet for t predatory mite (Phytoseiulus persimilis). Predatory mites completed development in the diet; however, mites reared exclusively in artificial diet failed to produce progeny. Recent improvements in diet encapsulatio and oviposition substrates impregnated with chemicals extracted from the prey (the two-spotted spider mite) have yielded reproduction of the predatory mite entirely in artificial diet. 02 Spore Separation of a Beneficial Fungus from a Solid Substrate. Biologic control products based on fungi (such as Trichoderma harzianum) that are parasitic on plant disease-causing microbes are currently marketed in th U.S. and many other countries to control soil-born plant diseases. Large quantities of conidia spores of these fungi are produced through two-pha solid fermentation. Because these conidial spores tend to absolve water, it is difficult to separate them from the solid substrate by sieving. A method of microencapsulating conidia spores with sucrose through spray drying at elevated temperatures was invented. Trace amount of sucrose microencapsulation protected conidia during the drying process. The survival of the conidial spores achieved 68.5%. This invention has significantly increased the viability levels of dried conidia preparatio The dried preparations consisted of more than 99% of pure conidia. Therefore, this invention can provide formulations for a variety of applications, from seed coating to sprayable conidia for foliage disease controls. Advanced Biological Marketing, Inc. conducted field experiment using microencapsulated conidia for seed treatment in spring; all feedba received from these trials were positive. This invention can also be use to dry other microbial biological control agents of insect pests (such a Metarhizium anisopliae, Penicillium and Beauveria bassiana), greatly expanding the potential for widespread application by commercial compani and thus reducing reliance on harsh chemicals. 03 Sequencing Lacewing Genes. An additional set of sequences (85) was obtained from the green lacewing (Chrysoperla rufilabris) and used to establish quantitative expression standards for this insect. These 85 sequences were deposited in GenBank. These first lacewing expressed gene can be used to compare gene expression under varied conditions. 04 A Rearing System for the Mass Production of Predatory Mites. A third generation cage system was developed to mass rear a beneficial mite (Phytoseiulus persimilis), key predator of the worldwide plant pest, the two-spotted spider mite. This system is an encaged and continuous method of producing predatory mites with minimal loss and consistent quality. Four prototypes have been constructed, two of which have been sent to ou cooperators for testing. Tests of the system in a commercial environment concluded that the system requires significant development in mechanization before becoming viable for commercial use. However, this system showed promise as a method of implementing artificial diet rearin at a commercial level. The potential of developing a functional encaged system for large scale production of predatory mites on artificial diets is very significant for the biological control industry. This system is the first step in accomplishing a fully mechanized commercial system. A patent disclosure has been submitted, approved and a patent application has been filed. 05 Optimal Environmental Conditions for Mass-rearing Predatory Mites. Previous studies on temperature dependent development rate and fecundity of predatory mites (Phytoseiulus persimilis) showed that the most favorable temperatures were between 25 and 27�C. New studies were done a 24, 25, 26, and 27�C to more closely determine the optimal temperature f mass propagation of this predator of the two spotted spider mite. The previous study showed that temperature affected development time, surviv and fecundity in different ways. The life and fertility table analysis allowed for a better analysis of fitness and population growth. Doubling time is a function of intrinsic rate of population increase (r) and represents the time required (in days) to double the population. Bases o these results it was concluded that optimal rearing temperature for the predatory mite can fluctuate between 25 and 26�C. These results provide critical information about the optimal conditions of mass propagation of these predatory mites. 06 Plant Chemicals Impact Lady Beetle Predators of the Two-spotted Spider Mite. Research has been conducted on differential effects of plant varie on the life history/rearing parameters of a mite-eating lady beetle (Stethorus punctillum). Research has revealed impact of lima bean cultivar on predator development time, growth, and size through the consumption of their spider mite prey. Leaves of lima bean (Phaseolus lunatus) cultivar Henderson Bush contain plant defensive compounds (allelochemicals, such as linamarin) that appear to block the absorption of protein by spider mites. This resulted in negative effects on development time, growth, and body size of the predator fed spider mites of all stages reared on foliage from this cultivar. In contrast, foliag from the Fordhook cultivar contains comparatively low levels of these defensive compounds and supports the development and growth of the predator. Predators consuming prey of reduced nutritional quality may reduce predator reproductive output (egg production). Another possible explanation is that toxicity of plant defensive chemicals accumulated in the spider mite bodies may be impacting the reproductive capacity of the predatory lady beetles. The next step will be to use artificial diets to test these 2 hypotheses. This discovery indicates that the lima bean Henderson Bush cultivar is not a suitable substrate for mass production the predator probably due to high levels of defensive chemicals present the foliage. The Fordhook cultivar is more suitable for mass production of this lady beetle.
Impacts
(N/A)
Publications
- Allen, M.L., Walker III, W.B. 2010. Expressed Sequence Reference Standards for Evaluating Stage-specific Gene Expression in Southern Green Lacewings, Chrysoperla rufilabris. Biocontrol Science and Technology. 20: 6, 589- 593, doi: 10.1080/09853151003657969.
- Chen, J., Shang, H., Jin, X. 2010. Interspecific Variation of delta-1,6- piperideines in Imported Fire Ants. Toxicon. 55(2010)1181-1187.
- Riddick, E.W., Cottrell, T.E. 2010. Is the Prevalence and Intensity of the Ectooparasitic Fungus Hesperomyces virescens Related to the Abundance of Entomophagous Coccinellids?. Bulletin of Insectology. 63 (1):71-78, 2010.
- Riddick, E.W. 2010. Ectoparasitic mite and fungus on an invasive lady beetle: parasite coexistence and influence on host survival. Bulletin of Insectology. 63 (1):13-20, 2010.
- Morales Ramos, J.A., Rojas, M.G. 2009. Some Organic Acids Acting as Stimulants of Recruitment and Feeding for the Formosan Subterranean Termite (Isoptera: Rhinotermitidae). Sociobiology. 54 (3):861-871.
- Morales Ramos, J.A., Rojas, M.G., Shapiro Ilan, D.I., Tedders, W.L. 2009. Developmental plasticity in Tenebrio molitor (Coleoptera: Tenebrionidae): Analysis of Instar Variation in Number and Development Time under Different Diets. J. Entomol. Sci. 45(2):75-90.
- Blanco, C. A., Rojas, M. G., Groot, A., Morales Ramos, J. A., Abel, C. A. 2009. Size and Chemical Composition of Heliothis virescens (Lepidoptera: Noctuidae) Spermatophores. Ann. Entomol. Soc. Am. 102(4): 629-637.
- Shapiro Ilan, D.I., Morales Ramos, J.A., Rojas, M.G., Tedders, W.L. 2010. Effects of a novel entomopathogenic nematode-infected host formulation on cadaver integrity, nematode yield, and suppression of Diaprepes abbreviatus and Aethina tumida under controlled conditions. Journal of Invertebrate Pathology. Journal of Invertebrate Pathology. 103:103-108.
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Progress 10/01/08 to 09/30/09
Outputs
Progress Report Objectives (from AD-416) Development of effective biological control technology for selected pests, including importation and culture of appropriate natural enemies and microbial control agents of insects and development and testing of technology for production, processing, storage, packaging, distribution, and release of natural enemies of insects and weed pests. Effective augmentative and classical biological control strategies for key pests are developed. Production of biological control agents for use in field evaluations in cooperation with other labs. Improvements in mosquito biological control agent formulation, delivery, specificity and storage stability will be achieved by modifying the physical and chemical properties of the formulations. Approach (from AD-416) Research and development activities are directed, primarily, at the development of automated mass rearing technologies for important biological control agents, including the development of artificial diets. Agents for study are selected on the basis of the needs of cooperators; the potential for success in the development of mass rearing technology; and the availability of basic rearing procedures, artificial diets, etc. Quality and process control criteria are developed. Techniques for packaging, distribution, and release of biological control agents are developed and tested. Biological control strategies are developed and tested through the mass rearing and field evaluation of the released agents. Significant Activities that Support Special Target Populations Broad goal of project is development of in vivo and in vitro mass-rearing methods and technology, including technology for harvesting, packaging, storage, and distribution of quality-assured natural enemies (including microbials). In area of functional genomics, continuous colonies of two arthropod species are still (yr 2) maintained in the Quarantine Facility. Specialized needle preparation, micro-injection, and arthropod handling protocols are established for embryos, nymphs, and adults of Lygus lineolaris and embryonic Chrysoperla (C.) rufilabris. Primary focus is on L. lineolaris with baseline autofluorescence was reported for both insects. A first survivor generation (G0) of embryo injected Lygus has been produced, but the first potentially transgenic generation (G1) has not yet been screened. A red-eyed strain of Lygus is being established to supplement insect transformation efforts. Additional set of sequences (85) were obtained from C. rufilabris and used to establish quantitative expression standards for this insect. These 85 sequences were deposited in GenBank. In area of formulation, progress has been made in applying hydrophilic-lipophilic balance (HLB) number in wetting agent selection for formulation development of hydrophobic conidia of Metarhizium anisopliae and Beauveria bassiana. Focus has been placed on identifying the hydrophilic-lypophilic balance (HLB) numbers of surfactants that can reduce the wetting time required by the dried conidia of these two entomopathogenic fungi. These studies introduced the concept of �wettability� that means how fast the hydrophobic conidia can be wetted by wetting agents that have different HLB numbers. Progress was also made in applying HLB number system to optimize surfactant selection that improved bio-herbicidal efficacy of Myrothecium verrucaria to control sicklepod seedlings. In area of investigating the biology, behavior, and ecology of natural enemies for improvement of mass rearing systems and for managing predator populations, research has revealed that host plant (lima bean, Henderson Bush variety) resistance can have a negative effect on a mite-eating predator, Stethorus punctillum. Our remedy to this problem is to use filter paper rather than leaves as a rearing substrate for predator larvae. This observation and remedy has greatly increased our ability to rear S. punctillum and should help the biocontrol industry increase their production. Team research is underway to determine if another possible form of host plant resistance in lima bean (Henderson Bush variety), such as allelochemicals (e.g., linamarin and lotaustralin) in the leaves, also negatively affects the development of S. punctillum larvae. These molecules may have little or no demonstrable effect on spider mites but have negative effect on the development of predator larvae. In area of bio-ecological interactions of parasites (mites and fungi) and their impact on the multicolored Asian lady beetle (Harmonia axyridis) and other lady beetles, research has revealed that the presence of the mite (Coccipolipus hippodamiae) affects the location intensity of the fungus (Hesperomyces virescens). Technology Transfer Number of Active CRADAS: 4 Number of Invention Disclosures submitted: 5 Number of Web Sites managed: 1
Impacts
(N/A)
Publications
- Celorio-Mancera, M., Allen, M.L., Powell, A.L., Ahmadi, H., Salemi, M.R., Phinney, B.S., Shackel, K.A., Greve, L., Teuber, L.R., Labavitch, J.M. 2008. Polygalacturonase Causes Lygus-like Damage on Plants: Cloning and Identification of Western Tarnished Plant Bug (Lygus hesperus) Polygalacturonases Secreted During Feeding. Arthropod-Plant Interactions. 2:215-225.
- Vogt, J.T., Allen, M.L., Wallet, B., Boykin, D.L., Smith, W.A. 2009. Distribution patterns of imported fire ants (Hymenoptera: Formicidae) on a sheep and goat farm in Oklahoma. Environmental Entomology. 38(3):551-560
- Rojas, M.G., Morales Ramos, J.A. 2008. Phytoseilus persimilis (Mesostigmata: Phytoseiidae) Feeding on Extrafloral Nectar: Reproductive Impact of Sugar Sources in Presence of Prey. Biopesticides International. 4(1): 1-5 (2008).
- Chen, J., Cantrell, C.L., Shang, H., Rojas, M.G. 2009. Piperideine Alkaloids from the Poison Gland of the Red Imported Fire Ant (Hymenoptera: Formicidae). Journal of Agricultural and Food Chemistry. Available: http://pubs.acs.org
- Jin, X., Ugine, T. A., Chen, J., Streett, D. A. 2009. Method for Determining the best Hydrophilic-lipophilic Balance (HLB) Number of a Compatible Non-ionic Surfactant in Formulation Development for Areal Conidia of Metarhizium anisopliae (Deuteromycotina: Hyphomycetes). Biocontrol Science and Technology. 19(3): 341-347.
- Handler, A.M., Allen, M.L., Skoda, S.R. 2009. Development and utilization of transgenic New World screwworm, Cochliomyia hominivorax. Medical and Veterinary Entomology. 23(s1):98-105.
- Shapiro Ilan, D.I., Rojas, M.G., Morales Ramos, J.A., Lewis, E.E., Tedders, W.L. 2008. Effects of host nutrition on virulence and fitness of entomopathogenic nematodes: lipid and protein based supplements in Tenebrio molitor diets. Journal of Nematology. 40:13-19.
- Riddick, E. W., Cottrell, T. E., Kidd, K. A. 2009. Natural Enemies of the Coccinellidae: Parasites, Pathogens, and Parasitoids. Biological Control 51 (2009) 306-312.
- Rojas, M., Morales-Ramos, J., Lockwood, M., Etheridge, L., Carroll, J., Coker, C., Knight, P. 2008. Area-wide management of subterranean termites using baits in South Mississippi. Agricultural Research Service Station Bulletin pp. 1-42.
- Weaver, M. A., Jin, X., Hoagland, R. E., Boyette, C. D. 2009. Improved bioherbicidal efficacy by Myrothecium verrucaria via spray adjuvants or herbicide mixtures. Biocontrol. 50: 150-156.
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Progress 10/01/07 to 09/30/08
Outputs
Progress Report Objectives (from AD-416) Development of effective biological control technology for selected pests, including importation and culture of appropriate natural enemies and microbial control agents of insects and development and testing of technology for production, processing, storage, packaging, distribution, and release of natural enemies of insects and weed pests. Effective augmentative and classical biological control strategies for key pests are developed. Production of biological control agents for use in field evaluations in cooperation with other labs. Improvements in mosquito biological control agent formulation, delivery, specificity and storage stability will be achieved by modifying the physical and chemical properties of the formulations. Approach (from AD-416) Research and development activities are directed, primarily, at the development of automated mass rearing technologies for important biological control agents, including the development of artificial diets. Agents for study are selected on the basis of the needs of cooperators; the potential for success in the development of mass rearing technology; and the availability of basic rearing procedures, artificial diets, etc. Quality and process control criteria are developed. Techniques for packaging, distribution, and release of biological control agents are developed and tested. Biological control strategies are developed and tested through the mass rearing and field evaluation of the released agents. Significant Activities that Support Special Target Populations Broad goal of project is development of in vivo and in vitro mass-rearing methods and technology, including technology for harvesting, packaging, storage, and distribution of quality-assured natural enemies (including microbials). In the area of functional genomics, continuous colonies of two arthropod species are being maintained in the Quarantine Facility. The microinjection laboratory is in operation, with capabilities including needle preparation, embryo and nymph injection, and fluorescence screening. Routine arthropod handling protocols are well established for the species Lygus lineolaris and Chrysoperla rufilabris. Baseline autofluorescence has been observed in both insects, and microinjection conditions are under continuous investigation. The laboratory is now capable of performing many molecular genetic protocols. High quality genomic DNA (gDNA) can be routinely extracted from a variety of insects including Solenopsis invicta, S. richteri, L. lineolaris, L. hesperus, and C. rufilabris. Substantial quantities of total ribonucleic acid (RNA) have been extracted from L. lineolaris, and C. rufilabris. Complementary DNA (cDNA) can be prepared from L. lineolaris either after further purifying polyadenylated (polyA) message RNA (mRNA), as for cDNA library construction, or without the polyA purification step for plymerase chan reaction (PCR) expression analysis. Double stranded RNA has been prepared for gene knock-down studies. Reverse transcriptase polymerase chain reaction (RT-PCR) has been tested successfully using the polygalacturonase 2 & 3 sequences. Libraries have been constructed from embryos and salivary glands, but not yet sequenced. A library has been prepared and is being sequenced from L. lineolaris exposed to the fungal pathogen, Beauveria bassiana. The current total L. lineolaris submissions from our lab is 381 expressed sequence tag (ESTs), 31 CoreNucleotides and 26 Protein accessions. Progress has been made in understanding the nutrition and ecological relationships among plant-pest- predator (beans-two-spotted spider mites-predatory mites). This approach has proven the most effective method for developing mite artificial diets. Industry is using this approach to improve in vivo mass production systems for mites. In the area of developing a mass production system for P. persimilis, several rearing cage designs have been evaluated. Current cage system provides for the continuous rearing and scale-up to 15,000 mites per week. An improved large scale production system is undergoing development and will be evaluated by industry. Finally, an efficient system for rearing the mite predator, Stethorus punctillum, is underway and involves the development of an artificial diet for this predator. Development of an artificial diet would eliminate need of using large numbers of the 2-spotted spider mite for production of this predator. NP304,Comp2,Prob b. Technology Transfer Number of Active CRADAS: 4 Number of Invention Disclosures submitted: 3 Number of Web Sites managed: 3
Impacts
(N/A)
Publications
- Allen, M.L., Mertens, J.A. 2008. Molecular cloning and expression of three polygalacturonase cDNAs from the tarnished plant bug, Lygus lineolaris. Journal of Insect Science 8:27, available online: insectscience.org/8.27.
- Riddick, E.W., Brown, A.E., Chauhan, K.R. 2008. Harmonia Axyridis Adults Avoid Catnip and Grapefruit-derived Terpenoids in Laboratory Bioassays. Bulletin of Insectology. 61(1):81-90.
- Riddick, E.W. 2009. Benefits and Limitations of Factitious Prey and Artificial Diets on Life Parameters of Predatory Beetles, Bugs, and Lacewings: A Mini-Review. Biocontrol. 54:325-339.
- Morales Ramos, J.A., Rojas, M.G. 2007. Importance of Lipids for Queen Fecundity and Colony Growth of Coptotermes formosanus (Isoptera: Rhinotermitidae). Environmental Entomology 36(5):1014-1017.
- Streett, D. A., Ni, Xinzhi, and Lawrence, A. M. 2008. Effect of DNA Gyrase Inhibitors in the NI diet on Biological Fitness of the Western Tarnished Plant Bug (Heteroptera: Miridae). J. Entomol. Sci. 43: 86-94.
- Jin, X., Streett, D. A., Dunlap, C. A., Lyn, M. E. 2008. Application of hydrophilic-lypophilic balance (HLB) number to optimize a compatible non- ionic surfactant for dried aerial conidia of Beauveria bassiana. Biological Control. 46: 226-233.
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Progress 10/01/06 to 09/30/07
Outputs
Progress Report Objectives (from AD-416) Development of effective biological control technology for selected pests, including importation and culture of appropriate natural enemies and microbial control agents of insects and development and testing of technology for production, processing, storage, packaging, distribution, and release of natural enemies of insects and weed pests. Effective augmentative and classical biological control strategies for key pests are developed. Production of biological control agents for use in field evaluations in cooperation with other labs. Improvements in mosquito biological control agent formulation, delivery, specificity and storage stability will be achieved by modifying the physical and chemical properties of the formulations. Approach (from AD-416) Research and development activities are directed, primarily, at the development of automated mass rearing technologies for important biological control agents, including the development of artificial diets. Agents for study are selected on the basis of the needs of cooperators; the potential for success in the development of mass rearing technology; and the availability of basic rearing procedures, artificial diets, etc. Quality and process control criteria are developed. Techniques for packaging, distribution, and release of biological control agents are developed and tested. Biological control strategies are developed and tested through the mass rearing and field evaluation of the released agents. Significant Activities that Support Special Target Populations Deployed Warfighter Protection Research Program. -- Collaborative research project was conducted under a Reimbursable Agreement between ARS and the Department of Defense (6402-22000-057-07R). For deployment settings, the Department of Defense is in need of innovative vector control technologies that reduce or eliminate exposure of U.S. forces and civilians to traditional pesticides and other chemicals. The overall goal of this project is to develop new formulations for mosquito control. Our focus will be on the development of ultra low volume (ULV) formulations for adult mosquito control using a commercially available active ingredient. A Cooperative Research and Development Agreement (CRADA) with a mosquito control industry partner interested in developing a ULV formulation for mosquito control is currently undergoing development. Ongoing monitoring of this project is accomplished through emails and phone conversations. Several chemicals will be evaluated as potential solvents, carriers, or diluents for the active ingredient in the development of a new ULV formulation. Effectiveness of test formulations against mosquitoes will be determined in laboratory bioassays using a Potter spray tower. A Malvern laser diffraction analysis system will be used to characterize spray properties of test formulations. Test formulations with the best overall characteristics will advance to field trials. Formulation activity in small scale field assays will involve adult mosquitoes and a range of adult mosquito species. Mortality and knockdown times will be determined to characterize formulation activity. A Non-Funded Cooperative Agreement between ARS and the University of California, Davis (6402-22000-057-06N) was established to conduct research on three sequences that bore distinctive similarity to polygalacturonase (PG) genes were identified in a library of expressed sequences from the tarnished plant bug, Lygus lineolaris. PG enzymes are key to plant damage inflicted by tarnished plant bugs (TPBs), as described by our collaborators. PGs degrade the plant�s cell walls and lead to wilting and a drop of flowers and fruit. Our collaborators have been purifying PGs from TPB salivary glands, and have accumulated evidence that several active PGs are present in the saliva. Our genomic evidence of multiple expressed PG geness supports the conclusion that multiple PGs are used by TPB to attack plants. Therefore, we fully sequenced the genes for all three TPB genes from Lygus lineolaris, and are looking for similar genes in Lygus hesperus (the Western tarnished plant bug). We hope to clarify the roles of the different PGs and identify genetic mitigation strategies. The three gene sequences were deposited the the National Center for Biotechnology Information (NCBI) GenBank, numbers DQ399525, DQ399526, DQ399527. Accomplishments Mechanization of in-vivo production of entomopathogenic nematodes in Tenebrio molitor. -- This work was done under Cooperative Research and Development Agreement (CRADA) with Southeastern Insectaries. This project has also been founded by a Small Business Initiative Research (SBIR) grant and it is currently in its second phase which was approved this year. Improved diets for T. molitor are being developed by systematically testing food supplements containing three basic nutrients including complex carbohydrates, protein, and lipids. During the first phase we determined that adding 10% protein in the form of dry egg whites to a supplement consisting of dry potato significantly shorten developmental time and increase the pupal weight. We are currently working on testing and comparing three different protein sources including wheat gluten, soy protein, and egg white protein. An experiment is in process to test the optimal ratio of dry potato to wheat bran for immature survival, developmental time, and pupal weight. A poster was presented in the 2006 ESA Annual meeting Indianapolis, IN, titled �Effect of Nutrition on the Life Cycle of Tenebrio molitor (Coleoptera: Tenebrionidae).� A poster presentation has been submitted for the 2007 ESA Annual meeting, San Diego, CA, titled �Effect of Adult Age of Tenebrio molitor (Coleoptera: Tenebrionidae) on Progeny Survival, Growth, and Development.� Significant improvements of mass rearing of T. molitor have been accomplished. A new rearing tray design has been developed, which allows first instar larvae to drop to a second tray by passing through a screen. This method reduces egg cannibalism and increases progeny survival. A similar design has been implemented to grow the larvae and allows for continuous frass separation from the food source. This new design is currently being evaluated to determine the optimal mesh size of the separation screen for both, first instar separation from adults, and for frass separation from food source. No presentation has been done or submitted at present. A new method for sorting different larval stages consisting of a multiple screen separator has been adapted and is currently being evaluated. A mechanized system to pack T. molitor larvae infected by entomopathogenic nematodes between two strips of masking tape has been developed. A prototype has been constructed and is currently under evaluation. A patent disclosure has been submitted titled �Mechanized Packing and Delivery System for Entomopathogenic Nematodes in Infected Mealworm Cadavers� (Docket No. 140.07). This project contributes to the National Program 304 �Crop Protection and Quarantine� and specifically addresses Component I: A. Insects and Mites, Problem Area: Identification and Classification of Insects and Mites, and Problem Area: Biology of Pests and Natural Enemies (includes microbes). New tarnished plant bug genes. -- We identified and sequenced muscle actin and three polygalacturonase (PG) genes completely by a process called rapid amplification of cDNA ends (RACE). We submitted the completed cDNA sequences and protein translations to GenBank. The PG sequences will be used to study feeding damage and may be instrumental in designing feeding disruption mechanisms. This project contributes to the National Program 304 �Crop Protection and Quarantine� and specifically addresses Component I: A. Insects and Mites, Problem Area: Identification and Classification of Insects and Mites, and Problem Area: Biology of Pests and Natural Enemies (includes microbes) and National Program 301 �Plant Genetic Resources, Genomics and Genetic Improvement� and specifically addresses Components I: Genetic Resource Management, and Component III: Genome Databases and Bioinformatics. Mass production of spider mites. -- The two spotted spider mite is an important economic pest that has an economic impact on U.S. crops of over $100 million in control costs and yield loss. Optimization of its in-vivo mass rearing all year long is key for the production of its predators. This pest can be reared on bean leaves in greenhouses for the purpose of developing and evaluating improved mass rearing systems of its predator P. persimilis. Several bean varieties and environmental conditions were tested and it was found that the right combination of plant variety, humidity, temperature, and lighting were enough to maintain a constant and good quality production of spider mites for the whole year. These findings will allow to reduce costs by eliminating waste of excessive production of predatory mites and maintain their quality by reducing their storage time. This project contributes to the National Program 304 �Crop Protection and Quarantine� and specifically addresses Component II: Biology of Pests and Natural Enemies (Microbes) and Problem Area IIB: Rearing of Insects and Mites. Technology Transfer Number of New CRADAS and MTAS: 1 Number of Active CRADAS and MTAS: 4 Number of Invention Disclosures submitted: 2 Number of Web Sites managed: 1 Number of Non-Peer Reviewed Presentations and Proceedings: 10 Number of Newspaper Articles,Presentations for NonScience Audiences: 1
Impacts
(N/A)
Publications
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