Progress 10/01/99 to 09/30/05
Outputs
Progress was made in the following areas of research: (1) We developed in the laboratory the first insect strain resistant to the new biological insecticide, Spinosad, and provided the lead information in the field on the genetics and likely mechanism of resistance for this insecticide. Spinosad is a new generation insecticide chemistry with a novel mode of action. Our pioneering studies were used for the development of strategies for resistance management for this important pesticide technology. (2) We developed a novel feeding disruption test (FDT) for monitoring larval insect resistance to Bt and chemical insecticides. This assay can also be used for the diagnosis of species in the budworm/bollworm complex, important in cotton pest management. The assay involves a hydrateable meal pad with a diagnostic dose of insecticide and a novel assay delivery system. The technology has resulted in multiple patents, the technology has been licensed, and is in commercial
production. (3) We have demonstrated feasibility of an adult assay to monitor caterpillar resistance to transgenic crops modified for insect resistance. The technology provides the opportunnity to produce crop and product specific, built-in diagnostics for monitoring pesticide resistance in the easily collectable adult stage for moth species. The technologies have been submitted to NCSU for commercial development. (4) We have discovered a new insect repellent as a replacement for DEET. This new insect repellent will be all natural, potentially can be organically certified and will be safe for use on children. Several US and foreign patents have been issued; the technology has been licensed and is in the advanced stages of commercial development. The submission of the EPA registration is expected early in 2006. Details on the repellent, referred to as BioUD, can be found at www.bioud.com. BioUD is as effective as DEET against mosquitoes and is also highly active against ticks and other
species of insects. (5) We have discovered a new, non-chemical method for insect control using atmospheric plasma. Our mode of action studies show that plasma disrupts the insect nervous system causing rapid death in seconds. There are many potential uses for this technology in the control of stored product insects and insects of public health importance, in the development of mosquito traps and for quarantine appplications. This work has been accepted by NCSU as intellectual property and is being commercilized in cooperation with private industry. (6) We have developed a novel method for the syntheisis of insecticides from nicotine. Our invention (patents pending) involves both novel synthetic chemistry and novel composition of matter. Current efforts for commercialization are ongoing with a major Ag company in the US. (7) We have discovered a novel method for moving proteins intact across the insect digestive system. This technology will permit the use of toxic proteins other than
Bt as insecticides. The technology has been submitted for patenting and is in commercial development.
Impacts
Our research with the highist potential commercial value and impact are as follows: (1) My insect repellent, BioUD, which is in the advance stages of commercialization is a potential replacement for DEET. It is as effective as DEET against mosquitoes and is highly active against ticks and other insects. It is all natural, can be organically certified, is non-flammable, is safe for use on children and can be bound to cloth and used in many other commercial applications. (2) We have developed novel technologies for monitoring caterpillar resistance to any transgenic crop engineered for insect resistance; the method assays the adult moth. This technology will allow industry to construct a built-in resistance monitoring system for their specific plant technologies as well as monitor for resistance with their current commercial technologies. (3) We have developed a non-chemical method for insect control that has a number of important applications in human health and
agriculture. (4) We have discovered a new chemistry to facilitate the movement of protein insecticides across the insect digestive system. This will permit the use of proteins other than Bt for insect control.
Publications
- Donohue, K. V., B. L. Bures, M. A. Bourham and R. M. Roe. 2005. Investigation of the mode of action of a non-equilibrium discharge on arthropod pests. Proceedings, 2005 Beltwide Cotton Conferences, New Orleans, LA, Jan. 4-7, 2005, pp. 1453-1462. National Cotton Council, Memphis, TN.
- Roe, R. M., J. Van Kretschmar, D. M. Thompson, K. V. Donohue and C. E. Sorenson. 2005. Larval feeding disruption tests (FDT) for monitoring insect resistance to Cry1Ac, Cry1f and Cry1ab. Proceedings, 2005 Beltwide Cotton Conferences, New Orleans, LA, Jan. 4-7, 2005, pp. 1651-1661. National Cotton Council, Memphhis, TN.
- Bures, B. L., K. V. Donohue, R. M. Roe and M. A. Bourham. 2005. Visualization of helium dielectric barrier discharge treatment of green peach aphids on tobacco leaves. IEEE Transactions on Plasma Sciences 33, 290-291.
- Vanderherchen, M. B., M. Isherwood, D. M. Thompson, R. J. Linderman and R. M. Roe. 2005. Toxicity of novel aromatic and aliphatic organic acid and ester analogs of trypsin modulating oostatic factor to larvae of the northern house mosquito, Culex pipiens complex, and the tobacco hornworm, Manduca sexta. Pesticide Biochemistry and Physiology 81, 71-84.
- Anspaugh, D. D., S. M. Khalil, M. D. Tomalski, D. M. Thompson and R. M. Roe. 2005. Molecular characterization of epoxide hydrolase cDNAs from the cabbage looper, Trichoplusia ni. Proceedings, 2005 Beltwide Cotton Conferences, New Orleans, LA, Jan. 4-7, pp. 1463-1474. National Cotton Council, Memphis, TN.
- Roe, R. M., V. Kallapur, R. J. Linderman and F. Viviani. 2005. Organic synthesis and bioassay of novel inhibitors of JH III epoxide hydrolase activity from fifth stadium cabbage loopers, Trichoplusia ni. Pesticide Biochemistry and Physiology. 83, 140-154.
- Anspaugh, D. D. and R. M. Roe. 2005. Regulation of JH epoxide hydrolase versus JH esterase activity in the cabbage looper, Trichoplusia ni, by juvenile hormone and xenobiotics. Journal of Insect Phyiology 51, 523-535.
- Jeffers, L. A., D. M. Thompson, D. Ben-Yakir and R. M. Roe. 2005. Movement of proteins across the digestive system of the tobacco budworm, Heliothis virescens. Entomologia Experimentalis et Applicata. 117, 135-146.
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Progress 10/01/03 to 09/30/04
Outputs
Progress has been made in the following areas: (1) We have developed a new generation of feeding disruption assay kits for monitoring insect resistance to the latest Bt toxins found in commercially available transgenic Bt cotton. The kits contain a diagnostic dose of Cry1Ac, 1Ab or 1F that has been incorporated into our patented hydreatable meal pads and assay devices. The assays were successfully tested for resistance diagnosis on laboratory-reared Bt susceptible tobacco budworms and cotton bollworms, laboratory-reared budworms resistant to Bt and field collected budworms from North Carolina. The kits were also field tested successfully by cooperators in the US and abroad during the 2004 growing season. These feeding disruption assay kits are being developed as a standard method for monitoring Bt resistance in larval lepidoptera and also are applicable to chemical and other biological insecticides. (2) Methodology was developed for the use of atmospheric plasma as a
non-chemical method of insect control for applications in agriculture and medical entomology. A license agreement was issued for one application of the technology, and a USDA SBIR Phase I grant was submitted to determine commericial feasibility. Research has continued on the determination of the mode of action of this control method on insects. (3) Novel neonicotinoid insecticides are being synthesized from nicotine. The compounds are also synthesized by novel methods (patents pending on structure and method of synthesis). Receptor studies indicate the mode of action is binding to the acetylcholine receptor producing classical poisoning symptomology as would be expected for a neonicotinoid. The compounds demonstrated significant insecticidal activity against agriculural pests and could potentially represent a new chemical class of neonicotinoid insecticides. One advantage of the chemistry is synthesis directly from nicotine, reducing the cost of production. (4) A US patent was obtain
on a novel insect repellent and potential replacement for DEET. The technology was licensed and is in commercial development for use as an animal and human mosquito and tick repellent, as a cloth treatment to repel insects, and for area wide control of biting insects.
Impacts
Feeding disruption assays (two US patents issued) have been successfully tested as a standard method for monitoring caterpillar resistance to Bt transgenic crops, as a high throughput-automated screening method for insecticide discovery, and for routine insect rearing. A device was constructed that generates atomospheric plasma discharges which can be used as a non-chemical method of insect control in a variety of applications in agriculture and medical entomology (license agreement awarded on one application). A novel class of neonicotinoid insecticides have been developed (US patents pending). The compounds are made directly from nicotine which may reduce the cost of insecticide production. A novel insect repellent was discovered as a possible replacement for DEET (multiple US and foreign patents awarded and pending). The technology has been licensed for applications on animals and humans to repel mosquitoes and ticks, for cloth treatments and area wide insect
control.
Publications
- T. Iwasa, N. Motoyama, J.T. Ambrose, and R.M. Roe. 2004. Mechanism for the differential toxicity of neonicotinoid insecticides in the honey bee, Apis mellifera. Crop Protecton. 23: 371-378.
- R.M. Roe, D.M. Thompson, J.L. Rhein, D.V. Donohue, S. Long, J.S. Bacheler , C.E. Sorenson and C.L. Sutula. 2004. Feeding disuption bioassay for field monitoring of insect susceptibility to Bt-transgenic crops and traditional pesticies. Proceedings, 2004 Beltwide Cotton Conferences, San Antonio, TX--January 5-9. P. 326.
- B.L. Bures, M.A. Bourham, K.V. Donohue, S. Long and R.M. Roe. 2004. Mortality of insects on the surface of plants using an atmospheric pressure plasma discharge. Proceedings, 2004 Beltwide Cotton Conferences, San Antonio, TX--January 5-9. Pp. 1518-1522.
- R.M. Roe. 2004. Method of repelling insects. US Patent Number: 6,800,662 B2, October 5, 2004.
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Progress 10/01/01 to 09/30/02
Outputs
An intellectual property disclosure was accepted by North Carolina State University on the use of atmospheric plasms as a novel method for insect control. The invention includes a device and its application along with proof of concept. Atmospheric plasma was successfully used for the control of lepidopteran neonates, aphids and thrips on and off host plants. Applications for this new technology include using atmospheric plasma to remove insects from plants before transportation and for routine insect control in green houses, the home and row crops. In some cases, 100% insect control can be achieved on plants in a few seconds of exposure. The device is inexpensive to manufacture and minimal electrical current is needed to generate the plasma. The plasma should be safe to humans. The environmental impact needs to be studied but is expected to be minimal. Atmospheric plasma may be especially relevant to applicatons in organic farming, and could eliminate or reduce the
need for insecticide use for some pest species. The mechanism of action is unknow.
Impacts
The invention is the use of atmospheric plasma as a new method of insect control. The application of this technology could be used as a replacement for traditional methods of insect control, i.e., insecticides and methy bromide, for examples. Atomospheric plasma could be used in organic farming and allow for agricultural production at a lower cost. The plasma also has applications in green house production, row crops, global transport of agriculture products and home land secruity related to bio/agro-terriorism.
Publications
- Roe, R.M., S. Long, S. Cawsey, J.S. Bacheler, C.E. Sorenson, N. Hoffman and C.L. Sutula. 2002. New commercial feeding disruption bioassay kit for species and insecticide resistance diagnosis in the tobacco budworm and cotton bollworm in cotton. Proceedings of the Beltwide Cotton Conferences. National Cotton Council, Memphis, TN.
- Vanderherchen, M.B., D.M. Thompson, R.M. Roe, M. Isherwood and R.J. Linderman. 2002. Evaluation of novel nonpeptidic chemical analogs of trypsin modulating oostatic factor (TMOF): A new biorational approach for managing lepidopteran pests of cotton. Proceedings of the Beltwide Cotton Conferences. National Cotton Council, Memphis, TN.
- Khalil, S., S. Long, H. Young and R.M. Roe. 2002. Development of a laboratory strain of the tobacco budworm resistant to Denim and a field kit for resistance monitoring. Proceedings of the Beltwide Cotton Conferences. National Cotton Council, Memphis, TN.
- Thompson, D.M., R.M. Roe, H.P. Young, F.W. Edens, A.W. Olmstead, G.A. LeBlanc and E. Hodgson. 2002. Trypsin modulating oostatic factor (TMOF): an environmentally safe insecticide. Proceedings of the Beltwide Cotton Conferences. National Cotton Council, Memphis, TN.
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Progress 10/01/00 to 09/30/01
Outputs
A consumer version of our feeding disruption bioassay technology for pesticide resistance detection and species diagnosis is currently in the commercialization phase of development. The kit measures the potential effectiveness of pesticide treatments against lepidopteran pests. This past year our research was aimed mostly toward the tobacco budworm (TBW)-cotton bollworm (CBW) complex as it occurs in cotton crops. The kit consists of a specially designed white plastic 16-well plate with recessed hydrateable meal pads (patent pending) containing an insecticide diagnostic dose and a blue feeding indicator dye. The appearance of blue feces easily seen on the background of the white plate is a marker for feeding. Insects that produce blue feces are resistant to the diagnostic dose of the pesticide. The assay kit can be stored at room temperature until needed, the test is performed in 24h on individual neonates or older insects, and the results are read in as little as 2 h
on groups of insects. Resistant assay kits were developed for the TBW and/or the CBW for the following insecticides: Bt, spinosyn, permethrin, indoxacarb and emamectin benzoate. A species diagnosis kit which distinguishes TBW from CBW was also developed based on the same principle and using a diagnostic dose of Bt. The species diagnostic kit was successfully field tested with insects from North Carolina and Louisiana. Finally, the utility of our new assay approach has been demonstrated by the development of a kit for spinosyn in the cabbage looper. We also developed a strain of the tobacco budworm resistant to emamectin benzoate.
Impacts
The technology being developed can be used to monitor insect populations for resistance to pesticides like Bt, pyrethroids, indoxacarb, and others. This should result in overall reductions in pesticide usage, maintain the efficacy of current insecticide technology, reduce crop losses, increase farming productivity and profits, and reduce the risk to the environment of the over-use of insecticides.
Publications
- Wyss, C. F., H. P. Young, J. Shukla and R. M. Roe. 2001. Inheritance and stability of spinosad resistance in a laboratory strain of the tobacco budworm. Proceedings of the Beltwide Cotton Conferences 2:1163-1166.
- H. P. Young, W. D. Bailey, R. M. Roe and T. Iwasa. 2001. Mechanism of resistance and cross resistance in a laboratory, spinosad-selected strain of the tobacco budworm and resistance in laboratory-selected cotton bollworms. 2001. Proceedings of the Beltwide Cotton Conferences 2:1167-1170.
- Bailey, W. D., C. Brownie, J. S. Bacheler, F. Gould, G. G. Kennedy, C. E. Sorenson and R. M. Roe. 2001. Species diagnosis and Bacillus thuringiensis resistance monitoring of Heliothis virescens and Helicoverpa zea (Lepidoptera: Noctuidae) field strains from the southern United States using feeding disruption bioassays. Journal of Economic Entomology 94:76-85.
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Progress 10/01/99 to 09/30/00
Outputs
We were issued a US Patent for a novel feeding disruption assay technology for monitoring insect resistance to pesticides and for species diagnosis of the tobacco budworm and cotton bollworm in cotton. A licensing agreement is now in place between North Carolina State University and the private company, Agdia Inc. (Elkhart, Indiana), to commercialize our invention. In this regard, we have received a USDA SBIR Phase II grant to conduct research leading to a prototype for test marketing by the company. Associated with our feeding disruption assay technology, the University has submitted a second patent which is a continuation in part of the first. This pending patent includes our invention of hydrateable meal pads which will allow for the production of ready-to-use, off-the-shelf assays for resistance and species diagnosis, high through-put automated in vivo screening for insecticide discovery, routine insect rearing, and devices for housing insects and meal pads to
facilitate their routine use. Research has continued on a novel strain of the tobacco budworm developed by this laboratory which is over 1000-fold resistant to the topical application of spinosad. This is especially significant because this is the first insect strain developed to be resistant to spinosad, and spinosad represents a new insecticide chemistry with a novel mode of action against the economically important Lepidoptera. In our research this year, no cross resistance was found between the spinosad resistant strain and the other insecticide classes used for the control of caterpillars in cotton and other crops. Electrophysiological studies indicate possible target site-based resistance for our spinosad-resistant tobacco budworms. The spinosad resistant strain is currently being offered for licensing to private industry as a screening tool for the discovery of new compounds with the same mechanism of action as spinosad and as a method to isolate the spinosad receptor.
Confidentiality agreements are currently in place with two companies in this regard.
Impacts
Our feeding disruption assay technology combined with meal pads provide a new method for resistance and species diagnosis, high through-put insecticide screening, and routine insect rearing; and is currenty under commercial development by a private company. This invention has the potential of changing insect rearing as we know it today. Our spinosad resistant tobacco budworm strain provides a method for the discovery of a novel insecticide target in the insect nervous system. This could have wide and significant commercial value.
Publications
- ROE, R.M., BAILEY, W., GOULD, F., AND KENNEDY, G.G. 2000. Insecticide Resistance Assay. US Patent Number: 6,060,039. Date of Patent: May 9, 2000.
- ROE, R.M., BAILEY, W.D., GOULD, F., SORENSON, C.E., KENNEDY, G.G., BACHELER, J.S., ROSE, R.L., HODGSON, E., AND SUTULA, C.L. 2000. Detection of resistant insects and IPM, In: Emerging Technologies for Integrated Pest Management: Concepts, Research, and Implementation (G.G. Kennedy and T.B. Sutton, Eds.), pp. 67-84. APS Press, St. Paul, Minnesota.
- ROE, R.M., BAILEY, W.D., YOUNG, H.P., AND WYSS, C.F. 2000. Characterization of spinosad (Tracer) resistance in a laboratory strain of the tobacco budworm and development of novel diagnostics for resistance monitoring in the field. In: 2000 Beltwide Cotton Conferences, pp. 926-929. National Cotton Council, Memphis, TN.
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Progress 10/01/98 to 09/30/99
Outputs
A proprietary co-technology to our feeding disruption assay was discovered that greatly enhances the shelf-life of our Bt resistance and species diagnositic kit for cotton. Multiple strains of the tobacco budworm and cotton bollworm were collected from the field in the southeastern U.S. and established in the laboratory at NC State University. Using these field strains, a predictive model was developed for the use of the feeding disruption assay for distinguishing budworm from bollworm and for the diagnosis of Bt resistance in both species. A feeding disruption assay kit was also developed for resistance monitoring of larval tobacco budworms to spinosad. We developed and are maintaining a laboratory strain of the tobacco budworm resistant to this new biological insecticide. This is the first insect strain resistant to spinosad. This is especially significant since spinosad represents a novel insecticide chemistry with an unknown binding site in the insect nervous
system. Studies were initiated to determine the mechanism of resistance. Resistance when compared to laboratory and field susceptible strains was measured when spinosad was applied topically, orally and by injection. No differences in the rate of penetration or metabolism of spinoad were found between the resistance and the parental strain. These studies suggest that spinosad resistance is not the result of reduced penetration, increased metabolism or changes in behavior.
Impacts
Our feedings disruption assay technology is the only field assay method for monitoring insect resistance to Bt transgenic crops. A proprietary co-technology extends its use to high through put screening and general insect rearing. Our spinosad-resistant tobacco budworm strain can be used for studies in spinosad resistance management and to screen for novel compounds with the same mode of action as spinosad, and gives us a clear path to the identification of the novel binding site of spinosad.
Publications
- DEVORSHAK, C., AND ROE, R.M. 1998. The role of esterases in insecticide resistance. Reviews in Toxicology 2:501-537.
- ROSE, R.L., HODGSON, E., AND ROE, R.M. 1999. Pesticides. In: Toxicology, Marquardt, H., Schafer, S.G., McClellan, R., and Welsch, F. (Eds.). Academic Press, New York. pp. 663-697.
- ROE, R.M., BAILEY, W.D., ZHAO, G., YOUNG, H.P., CARTER, L.M., GOULD, F., SORENSON, C.E., KENNEDY, G.G., AND BACHELER, J.S. 1999. Assay kit for species and insecticide resistance diagnosis for tobacco budworm and bollworm in cotton. Beltwide Cotton Conferences, pp. 926-930.
- BAILEY, W.D., YOUNG, H.P., AND ROE, R.M. 1999. Laboratory selection of a Tracer-resistant strain of the tobacco budworm and comparisons with field strains from the southeastern US. Beltwide Cotton Conferences, pp. 1221-1224.
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