Progress 10/01/19 to 09/30/20
Outputs
Progress Report Objectives (from AD-416): Objective 1: Identify, synthesize, and develop semiochemicals (pheromones, kairomones, plant volatiles, and other attractants and repellants) of significant insect crop pests (such as harlequin bug, bagrada bug, spotted wing drosophila, striped cucumber beetle, kudzu bug) and their natural enemies for use in integrated pest management. Subobjective 1a: Develop attractant volatiles and trap designs to monitor and manage the harlequin bug. Subobjective 1b: Assess known stink bug attractants for value in monitoring and/or managing bagrada bug. Subobjective 1c: Identify plant volatiles that attract squash bugs. Subobjective 1d: Identify plant and/or insect volatiles for detection and attraction of kudzu bug. Subobjective 1e: Improve lures for spotted wing drosophila by identification of fruit-based attractants. Subobjective 1f: Synthesize and test aggregation pheromone in combination with baits and traps for striped cucumber beetle management. Objective 2: Develop arthropod biological controls for managing key vegetable pests such as stink bugs, squash bug, and cucumber beetles, including integration of natural enemies with other tactics such as microbial control, semiochemicals, and cultural pest controls. Subobjective 2a: Evaluate the efficacy of the squash bug natural enemy complex for use in pest management and identify the most effective biological control agents. Subobjective 2b: Assess the value of native egg parasitoids to suppress BMSB. Objective 3: Advance effective microbial controls for key crop pests such as stink bugs, noctuid moths, and other seedling pests, including discovery of molecular and ecological mechanisms for sustained field reproduction and persistence, using diverse Bacillus thuringiensis (Bt) strains, Chromobacterium spp., baculoviruses, and other entomopathogens. Subobjective 3a. Discover additional environmental isolates of C. subtsugae and related species, and characterize their insecticidal properties. Subobjective 3b. Determine ability of Bt strains to persist in common vegetable mulches. Subobjective 3c. Develop effective baculoviruses for key pest diamondback moth. Objective 4: Discover naturally occurring biopesticides (such as botanical compounds and/or RNAis) targeting key vegetable pests such as stink bugs and cucumber beetles. Objective 5: Determine and strengthen the genetic basis of plant defense mechanisms (e.g., pest aversion and resistance) for protection of high- value crops such as cole crops on small farms or gardens against destructive insects, and determine the influence of the microflora of pest insects and/or plants on the plant defense response. Subobjective 5a: Determine the effect of knocking out or over-expressing the infestation responsive TF StZFP2 in its native species, and in Arabidopsis. Subobjective 5b: Determine how infestation inducible Q-type C2H2 TFs affect resistance to pest insects in Arabidopsis and B. oleracea. Subobjective 5c: Determine the influence of plant and insect microflora on the infestation response of Arabidopsis to T. ni. Approach (from AD-416): The project brings together a research team with diverse expertise for multiple approaches to insect management. The proposed project will focus on control of key insect pests in small farms and urban gardens, both organic and non-organic. Bio-based integrated pest management approaches to be developed will include: (i) discovery and deployment of natural insect attractants and repellents; (ii) conservation and augmentation of beneficial insects including use of their semiochemicals; (iii) pest-specific microbial controls [bacteria (including Bacillus thuringiensis strains and Chromobacterium spp.) and baculoviruses]; and, (iv) crop genetic resistance using molecular-based gene discovery. Research will target insect pests that cause major damage to key crops such as cucurbits and cole crops, although other important crops such as small fruit, beans, and potatoes may receive attention for specific problems. The combination of semiochemical approaches, biological controls, molecular techniques, and crop resistance, will offer a range of non-chemical tactics useful to integrated pest management strategies for major crop pests in urban small farms and gardens. Objective 1: Identify, synthesize, and develop semiochemicals (pheromones, kairomones, plant volatiles, and other attractants and repellants) of significant insect crop pests and natural enemies. ARS scientists in Beltsville, Maryland, continued to develop the controlled-release dispenser for optimization of spotted-winged drosophila (SWD) attraction and application of attract-and-kill strategy to manage SWD populations. As a result of chemical syntheses initiated by Invasive Insect Biocontrol and Behavior Laboratory (IIBBL), the aggregation pheromones of beetle vegetable pests (striped cucumber beetle, western striped cucumber beetle, crucifer flea beetle, and striped crucifer flea beetle) are now being tested in the field in a total of ten states (Maryland, Virginia, Pennsylvania, New York, Vermont, New Hampshire, Maine, Indiana, North Dakota, California) with university collaborators. In each case, ARS scientists have discovered that plant compounds are very important in enhancing the attraction and feeding of the beetles, making attract-and- kill techniques potentially more effective. Also, for the cucumber beetles, field trials have already shown attraction to other cucurbit pests, an unexpected benefit. Both aggregation pheromones will be deployed together and separate from the plant compounds, to determine the most effective pest management approach for monitoring and possible suppression. ARS scientists in Beltsville, Maryland, have found that a border trap crop of mustard protected a main crop of collards from its key pest, the harlequin bug, but only if the bordering mustards were separated by two rows rather than adjacent to the main crop. If there was no separation, female harlequin bugs, although they prefer to feed on mustard plants, ⿿commute to the collards to lay their eggs, thereby defeating the trap crop tactic. Sufficient separation demonstrates that small modifications in trap cropping can make this a valuable tactic for crop protection among small vegetable growers. Addition of the harlequin bug aggregation pheromone, murgantiol, which discoveries in IIBBL made commercially available, may allow further improvement in trap cropping tactics for this key pest. ARS scientists have confirmed that the superpest Colorado potato beetle (CPB), in addition to possessing a male-produced aggregation pheromone discovered earlier at Beltsville, also have a female-produced pheromone that prompts mate-seeking and mating behavior by males. Further research will attempt to identify and to synthesize this sex pheromone for use in combination with the aggregation pheromone for management of CPB. Objective 2: Develop arthropod biological controls for managing key vegetable pests. Flowering borders did not have a significant effect on parasitism by either egg or adult parasitoids of two squash bug species, Anasa tristis and Anasa armigera. Egg parasitism in the early season was significantly lower than egg parasitism in the late season crop; it is likely that the use of augmentative releases early in the season could improve the efficacy of Gryon pennsylvanicum as a biological control agent, especially for the early planting. Further research on the use of augmentative releases is needed. Measurements taken using transmission electron microscopy determined that the average combined width of the epicuticle and exocuticle of the egg chorion was significantly greater for A. tristis eggs than for A. armigera eggs. This difference may account for the lower rates of parasitism and parasitoid emergence and for the increased time spent drilling into A. tristis eggs compared with A. armigera eggs. ARS scientists in Beltsville, Maryland, and Newark, Delaware, continued collaboration in identifying volatiles emitted by brown marmorated stink bug (BMSB) that could be used as attractants or repellents for the parasitoid wasp Trissolcus japonicus. Two compounds, tridecane, and (E)-2- decenal, in the BMSB volatiles emission were behaviorally active against this known egg parasitoid. While the former chemical seemed to attract the egg parasitoid, the latter displayed a repellent activity when tested in laboratory bioassay. For brown marmorated stink bug, implementation research is supported by USDA NIFA Specialty Crop Research Initiative (SCRI), Coordinated Agricultural Programs (CAP) grant, ⿿Management of BMSB in U.S. Specialty Crops, through North Carolina State University in cooperation with several other universities and ARS labs. Objective 3: Advance effective microbial controls for key crop pests, using diverse Bacillus thuringiensis (Bt) strains, Chromobacterium spp., baculoviruses, and other entomopathogens. With respect to Objective 3, bioassays were conducted with larvae of a Bacillus thuringiensis (Bt) toxin-resistant strain of diamondback moth to see if simultaneous ingestion of baculovirus and Bt toxin would increase the susceptibility of the larvae to baculovirus infection. With a concentration of Bt toxin that normally kills 100% of wild-type/Bt- susceptible larvae but has no effect on Bt-resistant larvae, no increase was seen in virus-induced mortality compared to a treatment consisting of baculovirus alone. This result indicates that Bt toxin does not increase baculovirus susceptibility in Bt-resistant diamondback moth larvae. The newly discovered microsporidian pathogen of stink bugs, Nosema maddoxi, appears to have caused catastrophic declines of laboratory colonies of brown marmorated stink bug (BMSB), and possibly to have also caused declines in the field, where it infects BMSB and at least two species of native stink bugs. Its possible value in stink bug management is being investigated under the USDA NIFA Specialty Crop Research Initiative (SPRI) Coordinated Agricultural Projects (CAP) grant, ⿿Management of Brown Marmorated Stink Bug (BMSB) in U.S. Specialty Crops, through North Carolina State University in cooperation with Cornell University and ARS labs. Research continues as to the host range of this pathogen, which may be useful in suppressing other invasive stink bugs Objective 4: Discover natural biopesticides (such as botanical compounds and/or RNAi⿿s) targeting key vegetable pests. ARS scientists in Beltsville, Maryland, also continued to evaluate methyl benzoate and its analogs as insecticides, with efficacy against bed bugs, mosquitoes, thrips, white flies, nematodes, and some stored product insect pests demonstrated under laboratory conditions. Candidate gene targets for RNAi were identified in squash bug and used to design dsRNAs targeting these genes. Thirteen dsRNAs were and generated and tested against squashbug in preliminary assays. Potential gene targets for dsRNA/RNAi experiments were also identified in bagrada bug. Objective 5: Determine and strengthen the genetic basis of plant defense mechanisms (e.g., pest aversion and resistance) for protection of high value cole crops. To study microbes within the pest cabbage looper (CL) that might affect the defense response in Arabidopsis or Brassica, we began by identifying what microbes were found in CL using next generation sequencing to identify the variation in microbes found in different organs of CL that could affect the plant response during infestation. In the first study of its type, we characterized bacteria associated with different digestive tract organs of cabbage looper feeding on collards. They found that the bacteria inhabiting the mandibular glands, Malpighian tubules, and midgut of the caterpillar were broadly similar, but that the bacterial species associated with the salivary glands were substantially different. These results suggest that bacteria inhabiting the salivary glands, and their metabolic products, should be examined separately for their effect on plant defensive responses. Results of those studies may provide methods of interfering with insect countermeasures to plant defenses, reducing damage to crops. Accomplishments 01 Development of new trapping system for spotted wing drosophila pest management. ARS scientists in Beltsville, Maryland, collaborated with scientist in Towson University, as well as scientists in France, Germany, Spain, and Italy, to develop a dry trap baited with an optimized controlled-release attractant dispenser for spotted wing drosophila (SWD). The new trapping system demonstrated earlier SWD detection compared to the conventional liquid traps that have been currently used in orchards/farms. Due to the simplicity of the dry trap design with controlled release dispenser, it has provided growers/ farmers with an efficient, convenient, and easy processing tool for SWD infestation detection and control.
Impacts
(N/A)
Publications
- Boyle, S.M., Weber, D.C., Hough-Goldstein, J., Hoelmer, K.A. 2019. Host kairomones influence searching behavior of Trissolcus japonicus (Hymenoptera: Scelionidae), a parasitoid of Halyomorpha halys (Heteroptera: Pentatomidae). Environmental Entomology. 49(1):15-20.
- Dos Santos, E.R., Ecker, A., Trentin, L.B., Da Silva, L.A., Borges, M., Ribeiro, B.M., Mowery, J.D., Harrison, R.L., Ardisson-Araujo, D.M. 2019. An iflavirus found in stink bugs (Hemiptera: Pentatomidae) of four different species. Virology. 534:72-79.
- Blassiolli-Moraes, M., Khrimian, A., Michereff, M., Magalhaes, D., Hickel, E., Freitas, T., Barrigossi, A., Laumann, R., Telles, A., Guggilapu, S.D., Silva, C., Sant'Ana, J., Borges, M. 2019. Male-produced sex pheromone of Tibraca limbativentris revisited: absolute configurations of zingiberenol stereoisomers and their influence on chemotaxis behaviour of conspecific females. Journal of Chemical Ecology. 46:1-9.
- Dou, X., Zhang, A., Jurenka, R. 2019. Functional identification of fatty acyl reductases in female pheromone gland and tarsi of the corn earworm, Helicoverpa zea. Insect Biochemistry and Molecular Biology.
- Farrar, R.R., Gundersen, D.E., Kuhar, D.J., Blackburn, M.B. 2020. Insecticidal activity of Chromobacterium phragmitis, a recently described bacterium from tidal marshes. Journal of Entomological Science. 55(1):98- 104.
- Harrison, R.L., Rowley, D.L., Popham, H.R. 2019. A novel alphabaculovirus from the soybean looper, Chrysodeixis includens, that produces tetrahedral occlusion bodies and encodes two copies of he65. Viruses. 11:579.
- Harrison, R.L., Herniou, E.A., Bezier, A., Jehle, J.A., Burand, J.P., Theilmann, D.A., Krell, P.J., Van Oers, M.M., Nakai, M. 2020. ICTV virus taxonomy profile: Nudiviridae. Journal of General Virology. 101:3-4.
- Larson, N.R., Strickland, J.A., Shields, V., Zhang, A. 2020. Controlled- release dispenser and dry trap developments for Drosophila suzukii detection. Frontiers in Ecology and Evolution.
- Larson, N., Zhang, A., Feldlaufer, M.F. 2019. Fumigation activities of methyl benzoate and its derivatives against the common bed bug (Hemiptera: cimicidae). Journal of Medical Entomology.
- Lawrence, S.D., Novak, N.G., Shao, J.Y., Ghosh, S.B., Blackburn, M.B. 2020. Cabbage looper (Trichoplusia ni) salivary glands contain unique bacterial flora in contrast with their alimentary canal, mandibular gland and Malpighian tubules . MicrobiologyOpen. 9(4):e994.
- Morrison III, W.R., Larson, N.R., Brabec, D.L., Zhang, A. 2019. Methyl benzoate as a putative alternative, environmentally-friendly fumigant for the control of stored product insects. Journal of Economic Entomology. 112(5):2458-2468.
- Sparks, M., Gundersen, D.E., Bansal, R., Oppert, B.S., Poelchau, M.F., Childers, C., Rhoades, J.H., Velamuri, A.S., Benoit, J.B., Chao, H., Blackburn, M.B., Johnston, J. 2020. Brown marmorated stink bug, Halyomorpha halys (Stål), genome: underpinnings of polyphagy, insecticide resistance potential and biology of a top worldwide pest. BMC Genomics.
- Su, J., Zhao, B., Zhang, A., Bu, X., Chen, J., Yan, Z., Wang, S. 2019. Pore-ridged nanostructures on the surface of trichoid sensilla of the male Bombyx mori: Aerodynamic trapping, sensing, and transporting of the pheromone molecules. Arthropod Structure and Development. S1467-8039(18) 30167-1.
- Weber, D.C., Duan, J.J., Haber, A.I. 2020. Male Colorado potato beetles alter search behavior in response to prior female presence on potato plants. Journal of Pest Science. 93:595-604.
- Weber, D.C., Morrison III, W.R., Khrimian, A., Rice, K.B., Short, B.D., Herlihy, M.V., Leskey, T.C. 2019. Attractiveness of pheromone components with and without the synergist, methyl (2E,4E,6Z)-decatrienoate, to brown marmorated stink bug, Halyomorpha halys (Stål) [Hemiptera: Pentatomidae]. Journal of Economic Entomology. 113:712-719.
- Guo, T., Mehan, S., Gitau, M.W., Wang, Q., Kuczek, T., Flanagan, D.C. 2017. Impact of number of realizations on the suitability of simulated weather data for hydrologic and environmental applications. Stochastic Environmental Research and Risk Assessment (SERRA). 32(8):2405-2421.
- Blackburn, M.B., Farrar, R.R., Sparks, M., Kuhar, D.J., Mowery, J.D., Mitchell, A.D., Gundersen, D.E. 2019. Chromobacterium phragmitis sp. nov., an insecticidal bacterium isolated from estuarine marshes. International Journal of Systematic and Evolutionary Microbiology.
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Progress 10/01/18 to 09/30/19
Outputs
Progress Report Objectives (from AD-416): Objective 1: Identify, synthesize, and develop semiochemicals (pheromones, kairomones, plant volatiles, and other attractants and repellants) of significant insect crop pests (such as harlequin bug, bagrada bug, spotted wing drosophila, striped cucumber beetle, kudzu bug) and their natural enemies for use in integrated pest management. Subobjective 1a: Develop attractant volatiles and trap designs to monitor and manage the harlequin bug. Subobjective 1b: Assess known stink bug attractants for value in monitoring and/or managing bagrada bug. Subobjective 1c: Identify plant volatiles that attract squash bugs. Subobjective 1d: Identify plant and/or insect volatiles for detection and attraction of kudzu bug. Subobjective 1e: Improve lures for spotted wing drosophila by identification of fruit-based attractants. Subobjective 1f: Synthesize and test aggregation pheromone in combination with baits and traps for striped cucumber beetle management. Objective 2: Develop arthropod biological controls for managing key vegetable pests such as stink bugs, squash bug, and cucumber beetles, including integration of natural enemies with other tactics such as microbial control, semiochemicals, and cultural pest controls. Subobjective 2a: Evaluate the efficacy of the squash bug natural enemy complex for use in pest management and identify the most effective biological control agents. Subobjective 2b: Assess the value of native egg parasitoids to suppress BMSB. Objective 3: Advance effective microbial controls for key crop pests such as stink bugs, noctuid moths, and other seedling pests, including discovery of molecular and ecological mechanisms for sustained field reproduction and persistence, using diverse Bacillus thuringiensis (Bt) strains, Chromobacterium spp., baculoviruses, and other entomopathogens. Subobjective 3a. Discover additional environmental isolates of C. subtsugae and related species, and characterize their insecticidal properties. Subobjective 3b. Determine ability of Bt strains to persist in common vegetable mulches. Subobjective 3c. Develop effective baculoviruses for key pest diamondback moth. Objective 4: Discover naturally occurring biopesticides (such as botanical compounds and/or RNAis) targeting key vegetable pests such as stink bugs and cucumber beetles. Objective 5: Determine and strengthen the genetic basis of plant defense mechanisms (e.g., pest aversion and resistance) for protection of high- value crops such as cole crops on small farms or gardens against destructive insects, and determine the influence of the microflora of pest insects and/or plants on the plant defense response. Subobjective 5a: Determine the effect of knocking out or over-expressing the infestation responsive TF StZFP2 in its native species, and in Arabidopsis. Subobjective 5b: Determine how infestation inducible Q-type C2H2 TFs affect resistance to pest insects in Arabidopsis and B. oleracea. Subobjective 5c: Determine the influence of plant and insect microflora on the infestation response of Arabidopsis to T. ni. Approach (from AD-416): The project brings together a research team with diverse expertise for multiple approaches to insect management. The proposed project will focus on control of key insect pests in small farms and urban gardens, both organic and non-organic. Bio-based integrated pest management approaches to be developed will include: (i) discovery and deployment of natural insect attractants and repellents; (ii) conservation and augmentation of beneficial insects including use of their semiochemicals; (iii) pest-specific microbial controls [bacteria (including Bacillus thuringiensis strains and Chromobacterium spp.) and baculoviruses]; and, (iv) crop genetic resistance using molecular-based gene discovery. Research will target insect pests that cause major damage to key crops such as cucurbits and cole crops, although other important crops such as small fruit, beans, and potatoes may receive attention for specific problems. The combination of semiochemical approaches, biological controls, molecular techniques, and crop resistance, will offer a range of non-chemical tactics useful to integrated pest management strategies for major crop pests in urban small farms and gardens. Objective 1: Identify, synthesize, and develop semiochemicals (pheromones, kairomones, plant volatiles, and other attractants and repellants) of significant insect crop pests and natural enemies. As a result of syntheses initiated by the Invasive Insect Biocontrol and Behavior Laboratory (IIBBL), pheromones are now available in quantity for field deployment of aggregation pheromones for striped cucumber beetle, a key pest of cucurbit crops in North America, and for two species of flea beetles that are key pests of brassica vegetable crops like canola, cabbage, broccoli, kale, collards, and mustard greens. In each case, ARS scientists have discovered that plant compounds are very important in enhancing the attraction and feeding of the beetles, making attract-and- kill techniques potentially more effective. Therefore, the aggregation pheromones will be deployed together and separate from the plant compounds, to determine the most effective pest management approach. Research cooperators are in New York, New Hampshire, Virginia, North Dakota, and California. IIBBL has also been investigating biosynthesis of pheromones and possible pest management applications. RNA samples from male and female striped cucumber beetles collected in the field were sequenced to obtain global male- and female-specific transcriptomes. Data from this analysis are currently being used to assess differential gene expression between male and female adults. Homologs to known pheromone biosynthesis-related genes previously identified in stink bug species have been identified and are being considered as targets of future functional interrogations. Within the stink bug family, ARS researchers at Beltsville, Maryland, as part of the USDA NIFA Foundational grant, ⿿Defining Molecular Mechanisms of Terpene Aggregation Pheromone Biosynthesis in Stink Bugs to Engineer Trap Crops, have discovered key enzymes and intermediates in the biosynthetic pathways that allow the male insects to make their pheromones. Results include transcriptomic and biochemical discoveries for three pest species (harlequin bug, brown marmorated stink bug, and southern green stink bug), with more species underway. Results offer added targets to disrupt pest reproduction and/or divert pest populations with future incorporation into pheromone-producing trap plants. Objective 2: Develop arthropod biological controls for managing key vegetable pests. ARS scientists in Beltsville, Maryland, and Newark, Delaware, collaborated in identifying volatiles emitted by brown marmorated stink bug (BMSB) that could be used as attractants or repellents for the parasitoid wasp Trissolcus japonicus. Two compounds, tridecane, and (E)-2-decenal, in the BMSB volatiles emission were behaviorally active against this known egg parasitoid. While the former chemical seemed to attract the egg parasitoid, the latter displayed a repellent activity when tested in laboratory bioassay. For brown marmorated stink bug, implementation research is supported by USDA NIFA Specialty Crop Research Initiative (SCRI), Coordinated Agricultural Programs (CAPS), grant, ⿿Management of BMSB in U.S. Specialty Crops, through North Carolina State University in cooperation with several other universities and ARS labs. A two-year study has been initiated to evaluate parasitism rates and species composition of parasitoids attacking sentinel egg masses of BMSB and two native stink bug species, Euschistus servus and Podisus maculiventris. In the first year of the study, only 4.5% of BMSB eggs were parasitized compared with 31.4% of E. servus eggs and 28.1% of P. maculiventris eggs. Five parasitoid species were identified emerging from BMSB, with Trissolcus brochymenae being the most prevalent. Seven parasitoid species were identified emerging from E. servus and six from P. maculiventris. Results already demonstrate the higher level of parasitism of native species (both herbivore and predator) of stink bugs, showing the necessity of added biological control for the invasive BMSB. The exotic species T. japonicus parasitized a single egg mass of BMSB and a single egg mass of E. servus. The newly discovered microsporidian pathogen of stink bugs, Nosema maddoxi, appears to have caused catastrophic declines of laboratory colonies of brown marmorated stink bug (BMSB), and possibly to have also caused declines in the field, where it infects BMSB and at least two species of native stink bugs. This pathogen is apparently native both to Asia and North America. Its possible value in stink bug management is being investigated under the USDA NIFA SCRI CAP grant, ⿿Management of BMSB in US Specialty Crops, through North Carolina State University in cooperation with Cornell University and ARS labs. Research continues as to the host range of this pathogen, which may be useful in suppressing other invasive stink bugs such as Southern green stink bug (Nezara viridula) and Bagrada bug (Bagrada hilaris). Research is being conducted to evaluate the efficacy of the egg parasitoid Gryon pennsyvanicum as a biological control agent of two species of squash bugs, Anasa tristis and Anasa armigera and to compare ovipositional behavior of the parasitoid on the two squash bug species. Laboratory bioassays are being conducted to evaluate the effects of increased parasitoid pressure on parasitism rates and successful emergence when a single egg mass is exposed to either one, two, or three parasitoids. Laboratory bioassays are also being conducted to determine if parasitism rates on large egg masses could be increased if parasitoids were continuously provided with nutrition. These studies will provide information on factors influencing parasitism rates on squash bug egg masses and result in better biological control strategies for managing squash bug populations. Objective 3: Advance effective microbial controls for key crop pests, using diverse Bacillus thuringiensis (Bt) strains, Chromobacterium spp., baculoviruses, and other entomopathogens. More progress was made towards identifying a suitable alternative production host for the diamondback moth alphabaculovirus. The amount of diamondback moth virus needed to kill 50% of corn earworm larvae in a bioassay was 1,000-fold higher than that required to kill beet armyworm. Also, this baculovirus was unable to infect and cause mortality of larvae of the fall armyworm. The results suggest that, of the lepidopteran species test, beet armyworm may be the most suitable alternative host for the diamondback moth alphabaculovirus. Objective 4: Discover natural biopesticides (such as botanical compounds and/or RNAi⿿s) targeting key vegetable pests. ARS scientists in Beltsville, Maryland, continued to evaluate methyl benzoate and its analogs as insecticides, with efficacy against bed bugs, mosquitoes, thrips, nematodes, and some stored product insect pests demonstrated under laboratory conditions. Objective 5: Determine and strengthen the genetic basis of plant defense mechanisms (e.g., pest aversion and resistance) for protection of high value cole crops. The publication of several Brassica whole genome sequences allows mining for genes in cole crops. These encompass several nutritional powerhouses such as broccoli, collards, and kale along with two other similar species Chinese cabbage and canola. To broaden IIBBL investigations of C2H2 transcription factors (TF), we identified 146 C2H2 TF genes in three plant species. The identification and analysis of this gene family helps to focus on genes with similar protein structures to those in the potato (C2H2 TF, StZFP2), which upon over-expression can increase resistance to potato late blight and inhibit growth of the insect pest tobacco hornworm. This advance will allow work directly in these target cole crops. The publication of the biography of Charles Valentine Riley, Founder of Modern Entomology. An ARS entomologist in Beltsville, Maryland, has co- authored this definitive biography, which is the first full account of a fascinating American scientist whose leadership created the modern science of entomology which recognizes both the essential role of insects in natural ecosystems and their challenge to the agricultural food supply that sustains humankind. The book, published by University of Alabama Press, tells the story of how Riley (1843⿿1895), a young British immigrant to America, came to play a key role in the reorientation of entomology from the collection and arrangement of specimens to a scientific approach to insect evolution, diversity, ecology, and applied management of insect pests. Riley was USDA Entomologist from 1877 until his death in 1895. Among other achievements, he founded the National Insect Collection, vanquished the Grape Phylloxera that ravaged French vineyards through use of pest-resistant rootstocks, initiated the first successful classical biological control project through importation of the Vedalia Beetle from Australia, and discovered the Viceroy-Monarch mimicry as well as the yucca moth system of coevolution of plants and insects. This publication will be of interest to entomologists and other scientists and members of the general public interested in agricultural, demonstrating the value of government-funded science to solve agricultural challenges. Accomplishments 01 Development of attract & kill strategy for cocoa pod borer pest management. The cocoa pod borer (CPB) is one of the most important insects affecting cocoa production in Southeast Asia. This pest is responsible for more than 50% of cocoa crop losses in the Indo-Malayan archipelago. Until now, management of CPB has heavily relied on pesticide applications, which is neither environmentally sustainable nor economically effective. ARS scientists in Beltsville, Maryland, have developed a more economically productive method to synthesize the sex attractant and developed attract and kill strategy using impure sex attractant as lure and cypermethrin as killing agent. Field tests conducted in Malaysia during 2014⿿2016 indicated that attract-and-kill strategy is superior to and more feasible than the currently applied conventional synthetic pesticide treatment in CPM management. This will result in reduction of conventional pesticide application in cocoa plantation. 02 Discovery of the harlequin bug pheromone biosynthesis intermediate. ARS researchers at Beltsville, Maryland, in cooperation with Virginia Polytechnic Institute and State University, identified a gene, encoding of which produces an enzyme responsible for the formation of a key intermediate in the biosynthesis if the harlequin bug pheromone. Harlequin bug is a widespread key pest in the southern U.S. that heavily damages cabbage and related crops. Application of the aggregation pheromone for monitoring is an important part in the management of this pest. Earlier ARS researchers at Beltsville identified the two aggregation pheromone components of the harlequin bug and now they demonstrate the structural relationship of the pheromone molecules and the newly discovered biosynthesis intermediate. The knowledge of a total pheromone biosynthesis would allow incorporation of pheromone-producing genes into host plants to pursue new attract-and-kill strategies and develop biopesticides via RNAi- based molecular techniques (gene silencing) for controlling this pest.
Impacts
(N/A)
Publications
- Harrison, R.L., Mowery, J.D., Bauchan, G.R., Theilmann, D.A., Erlandson, M. A. 2019. The complete genome sequence of a second alphabaculovirus from the true armyworm, Mythimna unipuncta: Implications for baculovirus phylogeny and host specificity. Virus Genes. 55:104-116.
- Harrison, R.L., Rowley, D.L. 2019. The complete genome sequence of an alphabaculovirus from the southern armyworm, Spodoptera eridania. Microbiology Resource Announcements. 8:e01277-18.
- Escasa, S.R., Harrison, R.L., Mowery, J.D., Bauchan, G.R., Cory, J.S. 2019. The complete genome sequence of an alphabaculovirus from Spodoptera exempta, an agricultural pest of major economic significance in Africa. PLoS One. 14(2):e0209937.
- Feng, Y., Bruton, R.G., Park, A.L., Zhang, A. 2018. Identification of attractive blend for spotted wing drosophila, Drosophila suzukii from apple juice. Journal of Pest Science.
- Zhang, A., Feng, Y., Larson, N., Feldlaufer, M.F. 2018. Eucalyptol detected by aeration from the eggs of the common bed bug (Hemiptera: cimicidae). Journal of Economic Entomology.
- Lancaster, J., Khrimian, A., Young, S., Lehner, B., Luck, K., Wallingford, A., Ghosh, S.B., Zerbe, P., Marek, P., Sparks, M., Tittiger, C., Kollner, T.G., Weber, D.C., Gundersen, D.E., Kuhar, T., Tholl, D. 2018. De novo formation of an aggregation pheromone precursor by an isoprenyl diphosphate synthase-related terpene synthase in the harlequin bug. Proceedings of the National Academy of Sciences.
- Lancaster, J., Lehner, B., Khrimian, A., Muchlinsky, A., Luck, K., Kollner, T., Weber, D.C., Gundersen, D.E., Tholl, D. 2018. An IDS type sesquiterpene synthase produces the pheromone precursor (Z)-a-bisabolene in Nezara viridula. Journal of Chemical Ecology.
- Morrison III, W.R., Blaauw, B.R., Short, B.D., Nielsen, A.L., Bergh, J.C., Krawczyk, G., Park, Y., Butler, B., Khrimian, A., Leskey, T.C. 2018. Successful management of Halyomorpha halys (Hemiptera: Pentatomidae) in commercial apple orchards with an attract-and-kill strategy. Pest Management Science. 75(1):104-114.
- Lawrence, S.D., Novak, N.G. 2018. Over-expression of StZFP2 in Solanum tuberosum L. var. Kennebec (potato) inhibits growth of tobacco hornworm larvae (THW, Manduca sexta L.). Plant Signaling and Behavior.
- Lawrence, S.D., Novak, N.G. 2018. Comparative analysis of the genetic variability within the Q-type C2H2 zinc-finger transcription factors in the economically important cabbage, canola and Chinese cabbage genomes. Hereditas. 155:29.
- Novak, N.G., Perez, F.G., Jones, R.W., Lawrence, S.D. 2019. Detached leaf assays: A simplified approach to study gene expression in potato during infestation by the chewing insect Manduca sexta. Journal of Visualized Experiments. 147:e59153.
- Harrison, R.L., Herniou, E.A., Jehle, J.A., Theilmann, D.A., Burand, J.P., Becnel, J.J., Krell, P.J., Van Oers, M.M., Mowery, J.D., Bauchan, G.R. 2018. ICTV virus taxonomy profile: Baculoviridae. Journal of General Virology. 99:1185-1186.
- Lawrence, S.D., Novak, N.G., Perez, F.G., Jones, R.W. 2019. Over expression of the Q-type ZFP StZFP2 in potato increases resistance to potato late blight (Phytophthora infestans) infection. Journal of Plant Interactions. 14(1):129-136.
- Vanhove, W., Zhang, A., Osman, H.I., Vanhoudt, N., Van Damme, P. 2019. Development of an attract-and-kill strategy for Cocoa Pod Borer (Conopomorpha cramerella Snellen) control. Pest Management Science.
- Sorensen, W., Smith, E.H., Smith, J., Weber, D.C. 2019. Charles Valentine Riley: Founder of modern entomology. Tuscaloosa: University of Alabama Press. 438 p.
- Hall, D.G., Borovsky, D., Chauhan, K.R., Shatters, R.G. 2018. An evaluation of mosquito repellents and essential plant oils as deterrents of Asian citrus psyllid. Crop Protection. 108:87-94.
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Progress 10/01/17 to 09/30/18
Outputs
Progress Report Objectives (from AD-416): Objective 1: Identify, synthesize, and develop semiochemicals (pheromones, kairomones, plant volatiles, and other attractants and repellants) of significant insect crop pests (such as harlequin bug, bagrada bug, spotted wing drosophila, striped cucumber beetle, kudzu bug) and their natural enemies for use in integrated pest management. Subobjective 1a: Develop attractant volatiles and trap designs to monitor and manage the harlequin bug. Subobjective 1b: Assess known stink bug attractants for value in monitoring and/or managing bagrada bug. Subobjective 1c: Identify plant volatiles that attract squash bugs. Subobjective 1d: Identify plant and/or insect volatiles for detection and attraction of kudzu bug. Subobjective 1e: Improve lures for spotted wing drosophila by identification of fruit-based attractants. Subobjective 1f: Synthesize and test aggregation pheromone in combination with baits and traps for striped cucumber beetle management. Objective 2: Develop arthropod biological controls for managing key vegetable pests such as stink bugs, squash bug, and cucumber beetles, including integration of natural enemies with other tactics such as microbial control, semiochemicals, and cultural pest controls. Subobjective 2a: Evaluate the efficacy of the squash bug natural enemy complex for use in pest management and identify the most effective biological control agents. Subobjective 2b: Assess the value of native egg parasitoids to suppress BMSB. Objective 3: Advance effective microbial controls for key crop pests such as stink bugs, noctuid moths, and other seedling pests, including discovery of molecular and ecological mechanisms for sustained field reproduction and persistence, using diverse Bacillus thuringiensis (Bt) strains, Chromobacterium spp., baculoviruses, and other entomopathogens. Subobjective 3a. Discover additional environmental isolates of C. subtsugae and related species, and characterize their insecticidal properties. Subobjective 3b. Determine ability of Bt strains to persist in common vegetable mulches. Subobjective 3c. Develop effective baculoviruses for key pest diamondback moth. Objective 4: Discover naturally occurring biopesticides (such as botanical compounds and/or RNAis) targeting key vegetable pests such as stink bugs and cucumber beetles. Objective 5: Determine and strengthen the genetic basis of plant defense mechanisms (e.g., pest aversion and resistance) for protection of high- value crops such as cole crops on small farms or gardens against destructive insects, and determine the influence of the microflora of pest insects and/or plants on the plant defense response. Subobjective 5a: Determine the effect of knocking out or over-expressing the infestation responsive TF StZFP2 in its native species, and in Arabidopsis. Subobjective 5b: Determine how infestation inducible Q-type C2H2 TFs affect resistance to pest insects in Arabidopsis and B. oleracea. Subobjective 5c: Determine the influence of plant and insect microflora on the infestation response of Arabidopsis to T. ni. Approach (from AD-416): The project brings together a research team with diverse expertise for multiple approaches to insect management. The proposed project will focus on control of key insect pests in small farms and urban gardens, both organic and non-organic. Bio-based integrated pest management approaches to be developed will include: (i) discovery and deployment of natural insect attractants and repellents; (ii) conservation and augmentation of beneficial insects including use of their semiochemicals; (iii) pest-specific microbial controls [bacteria (including Bacillus thuringiensis strains and Chromobacterium spp.) and baculoviruses]; and, (iv) crop genetic resistance using molecular-based gene discovery. Research will target insect pests that cause major damage to key crops such as cucurbits and cole crops, although other important crops such as small fruit, beans, and potatoes may receive attention for specific problems. The combination of semiochemical approaches, biological controls, molecular techniques, and crop resistance, will offer a range of non-chemical tactics useful to integrated pest management strategies for major crop pests in urban small farms and gardens. Objective 1: Identify, synthesize, and develop semiochemicals (pheromones, kairomones, plant volatiles, and other attractants and repellants) of significant insect crop pests and their natural enemies for use in integrated pest management. The aggregation pheromone of the harlequin bug (Murgantia histrionica) is a powerful tool for monitoring and managing this important vegetable pest. The pheromone identified earlier consists of two principal components, both of which are important for the attractiveness. To allow an efficient scale-up of pheromone production, ARS scientists in Beltsville, Maryland, developed a new synthetic procedure that provides both components in improved yields for bioassays. Due to the overlap in chemical composition with brown marmorated stink bug pheromone, this new synthesis is valuable to management of both species. Field attraction of bagrada bug (Bagrada hilaris) to 10,11-epoxy- 1-bisabolen-3-ols which contain pheromones of the brown marmorated stink bug and harlequin bug, was not reproduced by California cooperators, and therefore an APHIS-permitted colony of this western vegetable pest was obtained for further ecological, behavioral, and semiochemical study starting in July 2017. A study of the ovipositional preferences of two squash bug species, Anasa tristis and Anasa armigera, for different cultivars and species of plants in the family Cucurbitaceae was completed. This study provided evidence that A. tristis had an induced preference for the cultivar it was reared on compared with several other cultivars of the same species. Also, A. tristis had a significant preference in paired choice tests for Cucurbita pepo over cucumber and watermelon, but not C. maxima or C. moschata. This study is the first experimental evidence of the ovipositional preferences of the horned squash bug, A. armigera. Horned squash bugs did not show any preference for cucumber over C. pepo, regardless of which species they were reared on. However, they did show a significant preference for cucumber over watermelon and C. moschata, and they preferred C. maxima to cucumber. Ovipositional preference tests provided the groundwork for further research on the responses of squash bugs to host plant volatiles. Methyl benzoate isolated from apple juice, and naturally-occurring in many other plants, was found to possess toxicity against various stages of a variety of insect pests, including the invasive brown marmorated stinkbug (BMSB) and spotted wing drosophila (SWD). Methyl benzoate was at least 5 to 20 times more toxic than a conventional pyrethroid (�-cyfluthrin), sulfur & pyrethrin mixture, and some commercial organic products available on the market against BMSB eggs and SWD larvae and adults. ARS obtained a U.S. Patent (US 9629362 B1) covering this technology. In addition, an attractant blend was developed for spotted wing drosophila (Patent applied for). SWD research is supported by the USDA NIFA CPPM grant, �Managing an Invasive Drosophilid Species in Agriculture using Innovative Behavioral Manipulation.� A pheromone was identified for the peanut burrower bug, the first for this family of true bugs (Cydnidae). Research is supported by the USDA NIFA CPPM grant, �Development of Risk Assessment and Monitoring Tools for Peanut Burrower Bug in Southeastern U.S. Peanut Production Systems.� Objective 2: Develop arthropod biological controls for managing key vegetable pests such as stink bugs, squash bug, and cucumber beetles, including integration of natural enemies with other tactics such as microbial control, semiochemicals, and cultural pest controls. A two-year study of parasitism and predation rates on wild and sentinel eggs of the squash bug, Anasa tristis, in squash fields at BARC was completed and the results were published in 2016. Gryon pennsylvanicum accounted for 99% of the egg parasitism. The egg parasitism rate peaked in late July at 72.8%. In addition, laboratory studies evaluating the efficacy of Gryon pennsylvanicum as a biological control agent have been completed. In the laboratory, the average parasitism rate of a single female without prior exposure to eggs was 59.6%, resulting in a squash bug nymphal survival rate of 22.9%. There were significant differences in the parasitism rate based on egg size of squash bug egg masses, with parasitism rates ranging from < 20% for the largest masses and > 90% for the smallest. A two-year field study of sentinel egg masses of the pest brown marmorated stink bug, and the beneficial stink bug, spined soldier bug (SSB, Podisus maculiventris), deployed in 2015-16 to field sites in Maryland, Virginia, DC, Delaware, and West Virginia, has yielded lesser numbers of the adventive Asian parasitoid, Trissolcus japonicus, in the DC region, but new state records for Delaware and West Virginia. This makes the redistribution of the parasitoid within these states allowable for BMSB suppression. However, T. japonicus only attacks BMSB in certain habitats and is not yet abundant. It successfully attacks the beneficial sentinel egg masses of SSB, however, from laboratory rearings these eggs are too small for optimal parasitoid success. Successful emergence of some native egg parasitoids and higher rates of egg mass consumption by native predators was also notable. Further research is supported by USDA NIFA SCRI CAP grant, �Management of BMSB in US Specialty Crops,� through North Carolina State University in cooperation with several other universities and ARS labs. Objective 3: Advance effective microbial controls for key crop pests such as stink bugs, noctuid moths, and other seedling pests, including discovery of molecular and ecological mechanisms for sustained field reproduction and persistence, using diverse Bacillus thuringiensis (Bt) strains, Chromobacterium spp., baculoviruses, and other entomopathogens. Chromobacterium species were isolated from diverse aquatic habitats and characterized by genomic sequencing, biochemical properties, and their insecticidal properties against diamondback moth, seedcorn maggot and red flour beetle. Two previously unknown species toxic to seedcorn maggot were identified in FY17 by comparing their genomic sequences with known Chromobacterium species. These newly described species can be developed into organic insecticides. To date, four new species of insecticidal Chromobacterium have been identified with lepidopteran and/or dipteran toxicity. The phylogenetic relationship of these species with other species Chromobacterium has been analyzed by analysis of their genomic sequences. A manuscript describing the first of these, C. sphagni, has been accepted for publication. The target pests for Bacillus thuringiensis toxins or modified toxins has been broadened by ARS work with University of Florida with support from a USDA NIFA SCRI grant, �Bt toxin-based strategies for management of Diaphorina citri and citrus greening.� Toxins with promise for control of psyllids, including for control of Asia citrus psyllid, the vector of the devastating citrus greening disease, huanglongbing, will be used to develop resistance in citrus. Determination and analysis of the genomes of two baculoviruses from the true armyworm were completed. A novel virus gene that may be involved in protection of viral DNA during infection was identified. Evidence emerged of extensive exchange of genes among the genomes of different true armyworm and Oriental armyworm baculoviruses. Old World bollworm betabaculovirus isolates from the IIBBL collection were evaluated for pathogenicity in bioassays against bollworm larvae. A group of isolates deposited in 1973, 1978, and 1993 killed larvae with symptoms previously reported for this baculovirus. Objective 4: Discover natural biopesticides (such as botanical compounds and/or RNAi�s) targeting key vegetable pests such as stink bugs and cucumber beetles. ARS scientists in Beltsville, Maryland, made progress in developing RNA interference (RNAi) tools for use with harlequin bug. RNAi deactivates or silences specific genes to disable a target organism. The transcriptome, or the total complement of active genes in the harlequin bug, was determined for different life stages and both sexes of the adult stage. Intraspecific comparison of transcripts was made to identifying a wide variety of differentially expressed transcripts and a reliable source to determine genes involved in key physiological processes. Distinct transcripts associated with particular life stages or sexes, or associated with highly specific biological systems, for example semiochemistry-related genes expressed in adult male during pheromone biosynthesis, were identified. These represent candidate harlequin bug- specific gene targets that may be involved in key synthesis of semiochemicals or may serve as candidates for disruption by molecular biopesticide technologies useful in managing the negative impacts of this insect pest. Specific RNAi- inducing dsRNAs for harlequin bug were developed and used to evaluate RNAi efficacy in vivo. Objective 5: Determine and strengthen the genetic basis of plant defense mechanisms (e.g., pest aversion and resistance) for protection of high- value crops such as cole crops on small farms or gardens against destructive insects. In order to determine the role of the potato gene StZFP2 in resistance to insects or pathogens, transgenic potato lines were produced knocking down or over-expressing StZFP2. The StZFP2 gene in potato is turned on during numerous plant stresses such as insect feeding and late blight infection. StZFP2 encodes a transcription factor that regulates the expression of numerous genes. These transgenic lines have been tested for their level of resistance to potato late blight (Phytophthora infestans) and infestation by Manduca sexta. Several of the over-expressing lines show increased resistance to late blight and several of the knock-down lines are significantly less resistant compared to non-transformed. Accomplishments 01 New species of insecticidal bacteria discovered. Overreliance on a limited number of organic insecticides increases the risk that pests will become resistant to them. Searching for alternatives to existing products, ARS scientists in Beltsville, Maryland, have described a new species of bacteria that produces insecticidal metabolites. The new species, named Chromobacterium sphagni, was discovered growing in Sphagnum bogs in West Virginia and Maine, and is orally toxic to a group of important pests represented by the diamondback moth and gypsy moth. The bacteria produce the toxic factor in artificial culture conditions, and living bacteria are not required to kill moth larvae. Spent cultures of the new species can be processed into an organic insecticide, and will be of interest to manufacturers of organic insecticides. 02 Efficient attractant for spotted wing drosophila (SWD). SWD is an exotic fruit fly from Southeast Asia that was introduced to the temperate regions of North America and Europe in 2008. It attacks a wide variety of fruits and has become a devastating pest of soft- skinned fruit crops. Due to the rapid spread of SWD across the newly- invaded continents, fresh fruit markets have a zero-tolerance policy regarding infestation. Specific and efficient detection tools are needed so that farmers can deliver timely interventions to meet market demands. Since SWD is attracted to damaged and rotting fruits, headspace volatiles from fresh and fermented apple juices were collected and analyzed by advanced scientific instruments to determine the compounds produced and/or enriched during the fermentation process. After performing a series of field tests, ARS scientists in Beltsville, Maryland, identified a five-way blend, which is more efficient and selective for the current standard apple cider vinegar and the commercially available SWD lure. Identification of SWD attractant will help growers accurately detect infestations in orchards, thereby allowing for timely interventions while reducing conventional insecticidal usage to protect our crops, environment, and ecosystem. 03 Economical pheromone for key cucurbit pest. The striped cucumber beetle is a key pest of cucurbit vegetables (squash, cucumbers, and melons) in eastern North America, rapidly colonizing on young plantings, and causing serious feeding damage while transmitting bacterial pathogens. Current control methods are costly. Male beetles produce an aggregation pheromone (attractive to both females and males), that has been identified and synthesized. Until recently, production methods were expensive and did not produce enough pheromone even for field tests. ARS scientists in Beltsville, Maryland, developed a new and less expensive mixed pheromone preparation, and then tested attraction to traps and baits under field conditions in cucurbit vegetable plantings. The results indicate that the new mixed pheromone could enable highly effective, less expensive environmentally-friendly species-specific management of striped cucumber beetle.
Impacts
(N/A)
Publications
- Blackburn, M.B., Farrar, R.R., Sparks, M., Kuhar, D.J., Mitchell, A.D., Gundersen, D.E. 2017. Chromobacterium muskegensis sp. nov., an insecticidal bacterium isolated from Sphagnum bogs. International Journal of Systematic and Evolutionary Microbiology. 67(9):3417-3422.
- Dimeglio, A.S., Kuhar, T.P., Weber, D.C. 2017. Color preference of harlequin bug, Murgantia histrionica (Heteroptera: Pentatomidae). Journal of Economic Entomology. 110:2275-2277.
- Greenstone, M.H., Cornelius, M.L., Olsen, R.T., Payton, M.E. 2017. Test of a natural enemy hypothesis on plant provenance: Spider abundance in native and exotic ornamental landscapes. Journal of Entomological Science. 52(4) :340-351.
- Gundersen, D.E., Adrianos, S.L., Allen, M.L., Becnel, J.J., Chen, Y., Choi, M.Y., Estep, A., Evans, J.D., Garczynski, S.F., Geib, S.M., Ghosh, S.B., Handler, A.M., Hasegawa, D.K., Heerman, M.C., Hull, J.J., Hunter, W.B., Kaur, N., Li, J., Li, W., Ling, K., Nayduch, D., Oppert, B.S., Perera, O.P. , Perkin, L.C., Sanscrainte, N.D., Sim, S.B., Sparks, M., Temeyer, K.B., Vander Meer, R.K., Wintermantel, W.M., James, R.R., Hackett, K.J., Coates, B.S. 2017. Arthropod genomics research in the United States Department of Agriculture-Agricultural Research Service: Applications of RNA interference and CRISPR gene editing technologies in pest control. Trends in Entomology. 13:109-137.
- Hajek, A.E., Solter, L.F., Maddox, J.V., Huang, W., Estep, A.S., Krawcyzk, G., Weber, D.C., Hoelmer, K.A., Sanscrainte, N.D., Becnel, J.J. 2017. Nosema maddoxi sp. nov. (Microsporidia, Nosematidae), a widespread pathogen of the green stink bug Chinavia hilaris (Say) and the brown marmorated stink bug Halyomorpha halys (St�l). Journal of Eukaryotic Microbiology.
- Harrison, R.L., Mowery, J.D., Rowley, D.L., Bauchan, G.R., Theilmann, D.A., Rohrmann, G.F., Erlandson, M.A. 2018. The complete genome sequence of a third distinct baculovirus isolated from the true armyworm, Mythimna unipuncta, contains two copies of the lef-7 gene. Virus Genes. 54(2):297- 310.
- Bakar, S., Latip, S., Awang, A., Zhang, A. 2017. Effects of Zingiber officinale, Curcuma longa and Alpinia galanga essential oils on the morphological characteristic of cocoa pod borer, Conopomorpha cramerella. Journal of Fundamental and Applied Sciences. 9(6s):25-38.
- Thrift, E., Herlihy, M.V., Wallingford, A.K., Weber, D.C. 2018. Fooling the harlequin bug (Hemiptera: Pentatomidae) using synthetic volatiles to alter host plant choice. Environmental Entomology.
- Cornelius, M.L. 2018. Ovipositional preferences of two squash bug species, Anasa tristis, and Anasa armigera (Heteroptera: Coreidae), for different cultivars and species of Cucurbitaceae. Journal of Insect Science.
- Hock, V., Chouinard, G., Lucas, E., Cormier, D., Leskey, T.C., Zhang, A. 2017. Olfactometer responses of plum curculio Conotrachelus nenuphar (Herbst) (Coleoptera: Curculionidae) to host plant volatiles, synthetic grandisoic acid, and live conspecifics. Journal of Insect Behavior.
- Jaffe, B.D., Avanesyan, A., Bal, H.K., Grant, J., Grieshop, M.J., Lee, J.C. , Liburd, O.E., Rhodes, E., Rodriguez-Saona, C., Sial, A.A., Feng, Y., Zhang, A., Guedot, C. 2018. Multistate comparison of attractants and the impact of fruit development stage on trapping Drosophila suzukii (Diptera: Drosophilidae) in raspberry and blueberry. Environmental Entomology.
- Lawrence, S.D., Novak, N.G. 2018. The remarkable plethora of infestation responsive Q-type C2H2 transcription factors in potato. Plant Signaling and Behavior.
- Nixon, L.J., Morrison III, W.R., Rice, K.B., Brockerhoff, E.G., Leskey, T. C., Guzman, F., Khrimian, A., Goldson, S., Rostas, M. 2018. Identification of volatiles released by diapausing brown marmorated stink bugs, Halyomorpha halys (Hemiptera: Pentatomidae). PLoS One. 13(1):e0191223.
- Rice, K.B., Bedoukian, R.H., Hamilton, G.C., Jentsch, P., Khrimian, A., Maclean, P., Morrison III, W.R., Short, B.D., Shrewsbury, P., Weber, D.C., Wiman, N., Leskey, T.C. 2017. Enhanced response of Halyomorpha halys (Hemiptera: Pentatomidae) to its aggregation pheromone with ethyl decatrieonate. Journal of Economic Entomology. 111(1):495-499.
- Feng, Y., Chen, J., Zhang, A. 2018. Commercially available natural benzyl esters and their synthetic analogs exhibit different toxicities against insect pests. Scientific Reports.
- Zeng, F., Liu, H., Sun, S., Zhang, A., Lu, Z., Wang, M. 2018. Three chemosensory proteins from the rice leaf folder Cnaphalocrocis medinalis involved in host volatile and sex pheromone reception. Insect Biochemistry and Molecular Biology.
- Wallingford, A.K., Kuhar, T.P., Weber, D.C. 2018. Avoiding unwanted vicinity effects with attract-and-kill tactics for harlequin bug, Murgantia histrionica (Hahn). Journal of Economic Entomology.
- Weber, D.C., Khrimian, A., Blassioli-Moraes, M., Millar, J.G. 2018. Semiochemistry of Pentatomoidea. In: McPherson, J.E., editor. Invasive Stink Bugs and Related Species (Pentatomoidea): Biology, Higher Systematics, Semiochemistry, and Management. Boca Raton, FL. CRC Press. p. 677-725.
- Welzell, K.F., Lee, S., Chauhan, K.R., Dossey, A.T., Choe, D. 2018. Verification of Argentine ant defensive compounds and their behavioral effects on heterospecific competitors and conspecific nestmates. Scientific Reports. 8:1477.
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Progress 10/01/16 to 09/30/17
Outputs
Progress Report Objectives (from AD-416): Objective 1: Identify, synthesize, and develop semiochemicals (pheromones, kairomones, plant volatiles, and other attractants and repellants) of significant insect crop pests (such as harlequin bug, bagrada bug, spotted wing drosophila, striped cucumber beetle, kudzu bug) and their natural enemies for use in integrated pest management. Subobjective 1a: Develop attractant volatiles and trap designs to monitor and manage the harlequin bug. Subobjective 1b: Assess known stink bug attractants for value in monitoring and/or managing bagrada bug. Subobjective 1c: Identify plant volatiles that attract squash bugs. Subobjective 1d: Identify plant and/or insect volatiles for detection and attraction of kudzu bug. Subobjective 1e: Improve lures for spotted wing drosophila by identification of fruit-based attractants. Subobjective 1f: Synthesize and test aggregation pheromone in combination with baits and traps for striped cucumber beetle management. Objective 2: Develop arthropod biological controls for managing key vegetable pests such as stink bugs, squash bug, and cucumber beetles, including integration of natural enemies with other tactics such as microbial control, semiochemicals, and cultural pest controls. Subobjective 2a: Evaluate the efficacy of the squash bug natural enemy complex for use in pest management and identify the most effective biological control agents. Subobjective 2b: Assess the value of native egg parasitoids to suppress BMSB. Objective 3: Advance effective microbial controls for key crop pests such as stink bugs, noctuid moths, and other seedling pests, including discovery of molecular and ecological mechanisms for sustained field reproduction and persistence, using diverse Bacillus thuringiensis (Bt) strains, Chromobacterium spp., baculoviruses, and other entomopathogens. Subobjective 3a. Discover additional environmental isolates of C. subtsugae and related species, and characterize their insecticidal properties. Subobjective 3b. Determine ability of Bt strains to persist in common vegetable mulches. Subobjective 3c. Develop effective baculoviruses for key pest diamondback moth. Objective 4: Discover naturally occurring biopesticides (such as botanical compounds and/or RNAis) targeting key vegetable pests such as stink bugs and cucumber beetles. Objective 5: Determine and strengthen the genetic basis of plant defense mechanisms (e.g., pest aversion and resistance) for protection of high- value crops such as cole crops on small farms or gardens against destructive insects, and determine the influence of the microflora of pest insects and/or plants on the plant defense response. Subobjective 5a: Determine the effect of knocking out or over-expressing the infestation responsive TF StZFP2 in its native species, and in Arabidopsis. Subobjective 5b: Determine how infestation inducible Q-type C2H2 TFs affect resistance to pest insects in Arabidopsis and B. oleracea. Subobjective 5c: Determine the influence of plant and insect microflora on the infestation response of Arabidopsis to T. ni. Approach (from AD-416): The project brings together a research team with diverse expertise for multiple approaches to insect management. The proposed project will focus on control of key insect pests in small farms and urban gardens, both organic and non-organic. Bio-based integrated pest management approaches to be developed will include: (i) discovery and deployment of natural insect attractants and repellents; (ii) conservation and augmentation of beneficial insects including use of their semiochemicals; (iii) pest-specific microbial controls [bacteria (including Bacillus thuringiensis strains and Chromobacterium spp.) and baculoviruses]; and, (iv) crop genetic resistance using molecular-based gene discovery. Research will target insect pests that cause major damage to key crops such as cucurbits and cole crops, although other important crops such as small fruit, beans, and potatoes may receive attention for specific problems. The combination of semiochemical approaches, biological controls, molecular techniques, and crop resistance, will offer a range of non-chemical tactics useful to integrated pest management strategies for major crop pests in urban small farms and gardens. Objective 1: Identify, synthesize, and develop semiochemicals (pheromones, kairomones, plant volatiles, and other attractants and repellants) of significant insect crop pests and their natural enemies for use in integrated pest management. The aggregation pheromone of the harlequin bug (Murgantia histrionica) is a powerful tool for monitoring and managing this important vegetable pest. The pheromone identified earlier consists of two principal components, both of which are important for the attractiveness. To allow an efficient scale-up of pheromone production, ARS scientists in Beltsville, Maryland, developed a new synthetic procedure that provides both components in improved yields for bioassays. Due to the overlap in chemical composition with brown marmorated stink bug pheromone, this new synthesis is valuable to management of both species. Field attraction of bagrada bug (Bagrada hilaris) to 10,11-epoxy- 1-bisabolen-3-ols which contain pheromones of the brown marmorated stink bug and harlequin bug, was not reproduced by California cooperators, and therefore an APHIS-permitted colony of this western vegetable pest was obtained for further ecological, behavioral, and semiochemical study starting in July 2017. A study of the ovipositional preferences of two squash bug species, Anasa tristis and Anasa armigera, for different cultivars and species of plants in the family Cucurbitaceae was completed. This study provided evidence that A. tristis had an induced preference for the cultivar it was reared on compared with several other cultivars of the same species. Also, A. tristis had a significant preference in paired choice tests for Cucurbita pepo over cucumber and watermelon, but not C. maxima or C. moschata. This study is the first experimental evidence of the ovipositional preferences of the horned squash bug, A. armigera. Horned squash bugs did not show any preference for cucumber over C. pepo, regardless of which species they were reared on. However, they did show a significant preference for cucumber over watermelon and C. moschata, and they preferred C. maxima to cucumber. Ovipositional preference tests provided the groundwork for further research on the responses of squash bugs to host plant volatiles. Methyl benzoate isolated from apple juice, and naturally-occurring in many other plants, was found to possess toxicity against various stages of a variety of insect pests, including the invasive brown marmorated stinkbug (BMSB) and spotted wing drosophila (SWD). Methyl benzoate was at least 5 to 20 times more toxic than a conventional pyrethroid (�-cyfluthrin), sulfur & pyrethrin mixture, and some commercial organic products available on the market against BMSB eggs and SWD larvae and adults. ARS obtained a U.S. Patent (US 9629362 B1) covering this technology. In addition, an attractant blend was developed for spotted wing drosophila (Patent applied for). SWD research is supported by the USDA NIFA CPPM grant, �Managing an Invasive Drosophilid Species in Agriculture using Innovative Behavioral Manipulation.� A pheromone was identified for the peanut burrower bug, the first for this family of true bugs (Cydnidae). Research is supported by the USDA NIFA CPPM grant, �Development of Risk Assessment and Monitoring Tools for Peanut Burrower Bug in Southeastern US Peanut Production Systems.� Objective 2: Develop arthropod biological controls for managing key vegetable pests such as stink bugs, squash bug, and cucumber beetles, including integration of natural enemies with other tactics such as microbial control, semiochemicals, and cultural pest controls. A two year study of parasitism and predation rates on wild and sentinel eggs of the squash bug, Anasa tristis, in squash fields at BARC was completed and the results were published in 2016. Gryon pennsylvanicum accounted for 99% of the egg parasitism. The egg parasitism rate peaked in late July at 72.8%. In addition, laboratory studies evaluating the efficacy of Gryon pennsylvanicum as a biological control agent have been completed. In the laboratory, the average parasitism rate of a single female without prior exposure to eggs was 59.6%, resulting in a squash bug nymphal survival rate of 22.9%. There were significant differences in the parasitism rate based on egg size of squash bug egg masses, with parasitism rates ranging from < 20% for the largest masses and > 90% for the smallest. A two-year field study of sentinel egg masses of the pest brown marmorated stink bug, and the beneficial stink bug, spined soldier bug (SSB, Podisus maculiventris), deployed in 2015-16 to field sites in Maryland, Virginia, DC, Delaware, and West Virginia, has yielded lesser numbers of the adventive Asian parasitoid, Trissolcus japonicus, in the DC region, but new state records for Delaware and West Virginia. This makes the redistribution of the parasitoid within these states allowable for BMSB suppression. However, T. japonicus only attacks BMSB in certain habitats and is not yet abundant. It successfully attacks the beneficial sentinel egg masses of SSB, however, from laboratory rearings these eggs are too small for optimal parasitoid success. Successful emergence of some native egg parasitoids and higher rates of egg mass consumption by native predators was also notable. Further research is supported by USDA NIFA SCRI CAP grant, �Management of BMSB in US Specialty Crops,� through North Carolina State University in cooperation with several other universities and ARS labs. Objective 3: Advance effective microbial controls for key crop pests such as stink bugs, noctuid moths, and other seedling pests, including discovery of molecular and ecological mechanisms for sustained field reproduction and persistence, using diverse Bacillus thuringiensis (Bt) strains, Chromobacterium spp., baculoviruses, and other entomopathogens. Chromobacterium species were isolated from diverse aquatic habitats and characterized by genomic sequencing, biochemical properties, and their insecticidal properties against diamondback moth, seedcorn maggot and red flour beetle. Two previously unknown species toxic to seedcorn maggot were identified in FY17 by comparing their genomic sequences with known Chromobacterium species. These newly described species can be developed into organic insecticides. To date, four new species of insecticidal Chromobacterium have been identified with lepidopteran and/or dipteran toxicity. The phylogenetic relationship of these species with other species Chromobacterium has been analyzed by analysis of their genomic sequences. A manuscript describing the first of these, C. sphagni, has been accepted for publication. The target pests for Bacillus thuringiensis toxins or modified toxins has been broadened by ARS work with University of Florida with support from a USDA NIFA SCRI grant, �Bt toxin-based strategies for management of Diaphorina citri and citrus greening.� Toxins with promise for control of psyllids, including for control of Asia citrus psyllid, the vector of the devastating citrus greening disease, huanglongbing, will be used to develop resistance in citrus. Determination and analysis of the genomes of two baculoviruses from the true armyworm were completed. A novel virus gene that may be involved in protection of viral DNA during infection was identified. Evidence emerged of extensive exchange of genes among the genomes of different true armyworm and Oriental armyworm baculoviruses. Old World bollworm betabaculovirus isolates from the IIBBL collection were evaluated for pathogenicity in bioassays against bollworm larvae. A group of isolates deposited in 1973, 1978, and 1993 killed larvae with symptoms previously reported for this baculovirus. Objective 4: Discover natural biopesticides (such as botanical compounds and/or RNAi�s) targeting key vegetable pests such as stink bugs and cucumber beetles. ARS scientists in Beltsville, Maryland, made progress in developing RNA interference (RNAi) tools for use with harlequin bug. RNAi deactivates or silences specific genes to disable a target organism. The transcriptome, or the total complement of active genes in the harlequin bug, was determined for different life stages and both sexes of the adult stage. Intraspecific comparison of transcripts was made to identifying a wide variety of differentially expressed transcripts and a reliable source to determine genes involved in key physiological processes. Distinct transcripts associated with particular life stages or sexes, or associated with highly specific biological systems, for example semiochemistry-related genes expressed in adult male during pheromone biosynthesis, were identified. These represent candidate harlequin bug- specific gene targets that may be involved in key synthesis of semiochemicals or may serve as candidates for disruption by molecular biopesticide technologies useful in managing the negative impacts of this insect pest. Specific RNAi- inducing dsRNAs for harlequin bug were developed and used to evaluate RNAi efficacy in vivo. Objective 5: Determine and strengthen the genetic basis of plant defense mechanisms (e.g., pest aversion and resistance) for protection of high- value crops such as cole crops on small farms or gardens against destructive insects. In order to determine the role of the potato gene StZFP2 in resistance to insects or pathogens, transgenic potato lines were produced knocking down or over-expressing StZFP2. The StZFP2 gene in potato is turned on during numerous plant stresses such as insect feeding and late blight infection. StZFP2 encodes a transcription factor that regulates the expression of numerous genes. These transgenic lines have been tested for their level of resistance to potato late blight (Phytophthora infestans) and infestation by Manduca sexta. Several of the over-expressing lines show increased resistance to late blight and several of the knock-down lines are significantly less resistant compared to non-transformed. Accomplishments 01 Key genes important for biochemical pathways and pest management identified in harlequin bug by comparative transcriptome analysis. Murgantia histrionica (Hahn), the harlequin bug, is an invasive insect that is a major vegetable insect pest in the United States. Harlequin bug is a piercing/sucking feeder that poses a considerable ecological and economic threat to mustard-family crops (Brassicaceae) especially in the southern U.S. ARS scientists in Beltsville, Maryland, determined the total complement of active genes the transcriptome of different harlequin bug life stages and both sexes of the adult stage. Intraspecific comparison of these transcriptomes identified a wide variety of genetic components involved in key physiological processes. Some genetic components were associated with particular life stages or sexes, or were associated with highly specific biological systems, for example semiochemistry in adult male during pheromone biosynthesis. These represent candidate harlequin bug-specific gene targets that may be candidates for disruption by biopesticide technologies to help manage the negative impacts of this insect pest. This information will be useful to understand genetic components involved in pheromone biosynthesis and pesticide detoxification. 02 Methyl benzoate found toxic to invasive insect pests. ARS scientists in Beltsville, Maryland, discovered that methyl benzoate, isolated from apple juice and naturally-occurring in many other plants, was toxic to various stages of a variety of insect pests, including the invasive brown marmorated stinkbug (BMSB) and spotted wing drosophila (SWD). Against BMSB eggs and SWD larvae and adults, methyl benzoate was found to be 5 to 20 times more toxic than a conventional treatments. Because methyl benzoate is considered environmentally friendly, it has great potential to be used as a safer alternative to synthetic pesticides for sustainable agriculture.
Impacts
(N/A)
Publications
- Harrison, R.L., Rowley, D.L., Funk, C. 2016. The complete genome sequence of Plodia interpunctella granulovirus: Discovery of an unusual inhibitor- of-apoptosis gene. PLoS One. 11(7):e0160389.
- Feng, Y., Zhang, A. 2017. A floral fragrance, methyl benzoate, is an efficient green pesticide. Scientific Reports. 7:42168.
- Harrison, R.L., Rowley, D.L., Mowery, J.D., Bauchan, G.R., Theilman, D.A., Rohrmann, G.F. 2017. The complete genome sequence of a second distinct betabaculovirus from the true armyworm, Mythimna unipuncta. PLoS One. 12(1) :e0170510.
- Weber, D.C., Morrison III, W.R., Khrimian, A., Rice, K.B., Leskey, T.C., Rodriguez-Saona, C., Nielsen, A.L., Blaauw, B.R. 2017. Chemical ecology of Halyomorpha halys: Discoveries and applications. Journal of Pest Science. 90:989-1008.
- Abram, P.K., Hoelmer, K.A., Acebes-Doria, A., Andrews, H., Beers, E., Bergh, C.J., Bessin, R., Biddinger, D., Botch, P., Buffington, M.L., Cornelius, M.L., Costi, E., Delfosse, E., Dieckhoff, C., Dobson, R., Donais, Z., Grieshop, M., Hamilton, G., Haye, T., Hedstrom, C., Herlihy, M. V., Hoddle, M., Hooks, C., Jentsch, P., Neelandra, J., Kuhar, T., Lara, J., Legrand, A., Lee, J.C., Leskey, T.C., Lowenstein, D., Milnes, J., Maistrello, L., Morrison III, W.R., Nielsen, A.L., Ogburn, E., Pickett, C., Poley, K., Pote, J., James, R., Shrewsbury, P., Talamas, E.J., Tavella, L. , Walgenbach, J., Waterworth, R., Weber, D.C., Welty, C., Wiman, N.G. 2017. Integrative review of indigenous arthropod natural enemies of the invasive brown marmorated stink bug in North America and Europe. Journal of Pest Science. 90(4):1009-1020.
- Blackburn, M.B., Sparks, M., Gundersen, D.E. 2016. The genome of the insecticidal chromobacterium subtsugae PRAA4-1 and its comparison with that of chromobacterium violaceum ATC12472. Genomics. doi: 10.1016/j.gdata. 2016.08.013.
- Cornelius, M.L., Dieckhoff, C., Vinyard, B.T., Hoelmer, K.A. 2016. Parasitism and predation on sentinel egg masses of the brown marmorated stink bug (Hemiptera: Pentatomidae) in three vegetable crops: Importance of dissections for evaluating the impact of native parasitoids on an exotic pest. Environmental Entomology. 45(6):1536-1542.
- Dimeglio, A.S., Wallingford, A.K., Weber, D.C., Kuhar, T., Mullins, D. 2016. Supercooling points of Murgantia histrionica (Heteroptera: Pentatomidae) and field mortality in the Mid-Atlantic United States following lethal low temperatures. Environmental Entomology. 45(5): 1294- 1299.
- Meyer, S.L., Chauhan, K.R., Macdonald, M.H. 2016. Evaluation of roselle (Hibiscus sabdariffa) leaf and pomegranate (Punica granatum) fruit rind for activity against Meloidogyne incognita. Nematropica. 46(1):85-96.
- Shirali, S., Guzman, F., Weber, D.C., Khrimian, A. 2017. Expedient synthesis of bisabolenol stink bug pheromones via stereodefined cyclohex-2- enones. Tetrahedron Letters. 58:2066-2068.
- Short, B.D., Khrimian, A., Leskey, T.C. 2016. Pheromone-based decision support tools for management of Halyomorpha halys in apple orchards: development of a trap-based treatment threshold. Journal of Pest Science. doi: 10.1007/s10340-016-0812-1.
- Sparks, M., Rhoades, J.H., Nelson, D.R., Kuhar, D.J., Lancaster, J., Lehner, B., Tholl, D., Weber, D.C., Gundersen, D.E. 2017. A transcriptome survey spanning life stages and sexes of the Harlequin bug, Murgantia histrionica. Insects. doi: 10.3390/insects8020055.
- Ze, S., Zhuang, L., Wen, Z., Huanan, J., Hao, L., Aiming, Z., Zhang, A., Man-Qun, W. 2016. Temporal interactions of plant - insect - predator after infection of bacterial pathogen on rice plant. Scientific Reports. 6:26043.
- Sun, X., Zeng, F., Yan, M., Zhang, A., Wang, M. 2016. Interactions of two odorant-binding proteins from Cnaphalocrocis medinalis G�en�e (Lepidoptera: Pyralidae). Insect Molecular Biology. 25(6):712-723.
- Sun, X., Zhao, Z., Zeng, F., Zhang, A., Lv, Z., Wang, M. 2016. Functional characterization of a pheromone binding protein from rice leaf-folder cnaphalocrocis medinalis in detecting pheromones and host plant volatiles. Bulletin of Entomological Research. 106(6):781-189.
- Thangaiah, S., Webb, M.Z., Ganga, B., Chauhan, K.R. 2016. Synthesis of southern corn root worm pheromone from S-Citronellol and its field evaluation. Journal of Agricultural Chemistry and Environment. 5:223-230.
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Progress 10/01/15 to 09/30/16
Outputs
Progress Report Objectives (from AD-416): Objective 1: Identify, synthesize, and develop semiochemicals (pheromones, kairomones, plant volatiles, and other attractants and repellants) of significant insect crop pests (such as harlequin bug, bagrada bug, spotted wing drosophila, striped cucumber beetle, kudzu bug) and their natural enemies for use in integrated pest management. Subobjective 1a: Develop attractant volatiles and trap designs to monitor and manage the harlequin bug. Subobjective 1b: Assess known stink bug attractants for value in monitoring and/or managing bagrada bug. Subobjective 1c: Identify plant volatiles that attract squash bugs. Subobjective 1d: Identify plant and/or insect volatiles for detection and attraction of kudzu bug. Subobjective 1e: Improve lures for spotted wing drosophila by identification of fruit-based attractants. Subobjective 1f: Synthesize and test aggregation pheromone in combination with baits and traps for striped cucumber beetle management. Objective 2: Develop arthropod biological controls for managing key vegetable pests such as stink bugs, squash bug, and cucumber beetles, including integration of natural enemies with other tactics such as microbial control, semiochemicals, and cultural pest controls. Subobjective 2a: Evaluate the efficacy of the squash bug natural enemy complex for use in pest management and identify the most effective biological control agents. Subobjective 2b: Assess the value of native egg parasitoids to suppress BMSB. Objective 3: Advance effective microbial controls for key crop pests such as stink bugs, noctuid moths, and other seedling pests, including discovery of molecular and ecological mechanisms for sustained field reproduction and persistence, using diverse Bacillus thuringiensis (Bt) strains, Chromobacterium spp., baculoviruses, and other entomopathogens. Subobjective 3a. Discover additional environmental isolates of C. subtsugae and related species, and characterize their insecticidal properties. Subobjective 3b. Determine ability of Bt strains to persist in common vegetable mulches. Subobjective 3c. Develop effective baculoviruses for key pest diamondback moth. Objective 4: Discover naturally occurring biopesticides (such as botanical compounds and/or RNAis) targeting key vegetable pests such as stink bugs and cucumber beetles. Objective 5: Determine and strengthen the genetic basis of plant defense mechanisms (e.g., pest aversion and resistance) for protection of high- value crops such as cole crops on small farms or gardens against destructive insects, and determine the influence of the microflora of pest insects and/or plants on the plant defense response. Subobjective 5a: Determine the effect of knocking out or over-expressing the infestation responsive TF StZFP2 in its native species, and in Arabidopsis. Subobjective 5b: Determine how infestation inducible Q-type C2H2 TFs affect resistance to pest insects in Arabidopsis and B. oleracea. Subobjective 5c: Determine the influence of plant and insect microflora on the infestation response of Arabidopsis to T. ni. Approach (from AD-416): The project brings together a research team with diverse expertise for multiple approaches to insect management. The proposed project will focus on control of key insect pests in small farms and urban gardens, both organic and non-organic. Bio-based integrated pest management approaches to be developed will include: (i) discovery and deployment of natural insect attractants and repellents; (ii) conservation and augmentation of beneficial insects including use of their semiochemicals; (iii) pest-specific microbial controls [bacteria (including Bacillus thuringiensis strains and Chromobacterium spp.) and baculoviruses]; and, (iv) crop genetic resistance using molecular-based gene discovery. Research will target insect pests that cause major damage to key crops such as cucurbits and cole crops, although other important crops such as small fruit, beans, and potatoes may receive attention for specific problems. The combination of semiochemical approaches, biological controls, molecular techniques, and crop resistance, will offer a range of non-chemical tactics useful to integrated pest management strategies for major crop pests in urban small farms and gardens. Objective 1: The harlequin bug aggregation pheromone is a new and important management tool for this key agricultural pest. We identified the pheromone earlier as a mixture of two principal components, both of which are important for attractiveness. To be able to conduct ratio, dose-response, and other field bioassays, we needed sufficient amounts of these pheromone components. Thus, in the current year we developed a new synthetic procedure that provided both components in appreciable yields and allowed scaling-up the pheromone. This new technique was based on an affordable inorganic base used as a catalyst in a key step of the synthesis. An effective tool for detection of Spotted wing drosophila (SWD), an highly damaging invasive fruit fly species which has spread rapidly across the United States, is urgently needed for management of this pest. After performing a series of tests on apple juice volatile aroma, a ternary blend has been identified as the most effective attractant for trapping SWD under laboratory conditions. The activity was confirmed by field tests. In addition, another volatile compound, methyl benzoate (MB), from apple juice was found to exhibit significant acute toxicity against the invasive species, brown marmorated stink bug (BMSB), gypsy moth (GM), SWD, as well as diamondback moth (DBM) and tobacco hornworm (THW). Striped cucumber beetle is a perennial problem with growers of squash, pumpkin, cucumbers, and melons. Its aggregation pheromone was previously discovered but quantities available were not sufficient for field testing. To remedy this need, a facile synthesis was developed producing gram quantities of diastereomeric mixture of the pheromone. We concluded field studies during the 2015 season demonstrating the attractiveness of this synthetic preparation to adult cucumber beetles, and the patent was recently published. Objective 2: Native parasitoid impact on brown marmorated stink bug (BMSB) sentinel egg masses was measured in four crops (bell pepper, tomato, squash, and soybean), in landscaped experimental plots comprised of either native or exotic plant species, and in wooded habitats in Maryland and D.C. The parasitoid species composition and parasitism rate varied in different habitats. Parasitism rates ranged from 12.4% in vegetable crops to 3.7% in landscaped plots. Preliminary bioassays of 65 diamondback moth granulovirus isolates from China, Taiwan, and Japan revealed that 25 of the isolates possessed insecticidal activity against diamondback moth larvae. PCR and DNA sequence analysis of diagnostic genes from these 25 samples revealed that they are all isolates of the betabaculovirus species Plutella xylostella granulovirus. Objective 3: New Chromobacterium isolates were obtained from a variety of aquatic habitats. Bioassays of newly collected isolates that have been identified indicate that within a Chromobacterium species, toxicity to particular insects is relatively constant. Biochemical phenotypes, growth characteristics, and genome level comparisons of two new Chromobacterium species were investigated to determine which features are useful in distinguishing them from other members of the Chromobacterium. A number of genes were investigated for their utility in predicting phylogenetic relationships among Chromobacterium, as 16S rRNA genes are not always reliable. Objective 4: Transcriptome sequence data for Murgantia histrionica, the harlequin bug, was assembled, annotated and quantitatively analyzed. Genes exhibiting differential expression across sexes and developmental stages were identified, and primers have been designed for further inquiry using molecular approaches. Preparation of biological replicates to enable molecular validation efforts is currently in process. These results will assist in the identification of gene targets of potential use in insect biocontrol programs. Objective 5: Arabidopsis transgenics expressing the StZFP2 gene were tested for resistance to diamondback moth using bioassays. Unlike their increased resistance to cabbage looper, evidence indicates that the transgenics do not have increased resistance to diamondback moth. The goal of this project is to determine how StZFP2 affects the plant�s resistance to insects. Genetically modified potato lines were produced in which the level of expression of StZFP2 was increased or decreased. We are currently analyzing genes induced by mechanical wounding, which induces genes associated with defense but is more rapidly performed and reproducible than insect infestation, and is thus a more reliable way to see how changes in StZFP2 levels alter expression of other genes. StZFP2 is produced rapidly after wounding, peaking in expression within 20 minutes. In addition, we are testing whether we can predict the function of StZFP2-like transcription factors by the features of their protein sequence; 27 StZFP2-like genes were identified in potato, and five of these were found to have expression patterns similar to StZFP2 upon wounding. Further studies will determine whether any of these newly discovered transcription factors also play a role in the defense against chewing insect pests. Accomplishments 01 Demonstration of an important native biocontrol for key pest, squash bug. ARS researchers in Beltsville, Maryland, have demonstrated that the egg parasitoid Gryon pennsylvanicum is an important biological control agent of squash bug, a key pest of squash, pumpkins, melon, and watermelon. A two-year study determined that Gryon was able to efficiently track wild squash bug eggs throughout the season. The rate of parasitism on wild egg masses peaked at the end of July at over seventy-two percent. From the end of July until the first week of September, less than 20 percent of squash bug nymphs hatched successfully from wild eggs. These results indicate that Gryon has the potential to be an effective biological control agent and play an important role in the suppression of squash bug populations. 02 Development of spotted winged drosophila attractant. ARS researchers in Beltsville, Maryland, have developed an effective lure for spotted winged drosophila (SWD), a destructive invasive fruit fly native to southeast Asia that is currently spreading across the U.S. Laboratory experiments led to the formulation of four volatile compounds from apple juice that were attractive to the fly under field conditions. Early detection of SWD will allow growers to apply timely pest management interventions and avoid unnecessary applications of insecticides.
Impacts
(N/A)
Publications
- Herlihy, M.V., Talamas, E.J., Weber, D.C. 2016. Attack and success of native and exotic parasitoids on eggs of Halyomorpha halys in three Maryland habitats. PLoS One. 11(3):e0150275.
- Khrimian, A., Shirali, S., Guzman, F. 2015. Absolute configurations of zingiberenols isolated from ginger (Zingiber officinale) rhizomes. Journal of Natural Products. 78(1)3071-3074.
- Mahadwar, G., Chauhan, K.R., Bhagavathy, G., Murphy, C.F., Smith, A.D., Bhagwat, A.A. 2015. Swarm motility of Salmonella enterica serovar Typhimurium is inhibited by compounds from fruit peel extracts. Letters in Applied Microbiology. 60:334-340.
- Wada-Katsumata, A., Zurek, L., Nalyanya, G., Roelofs, W.L., Zhang, A., Schal, C. 2015. Gut bacteria mediate aggregation in the German cockroach. Proceedings of the National Academy of Sciences. 112(51):15678-15683.
- Zeng, F., Zhao, Z., Yan, M., Zhou, W., Zhang, A., Lu, Z., Wang, M. 2015. Identification and comparative expression profiles of chemoreception genes revealed from major chemoreception organs of the rice leaf folder, Cnaphalocrocis medinalis (Lepidoptera: Pyralidae). PLoS One. 10(12) :e0144267.
- Zhang, Q., Zhou, G., Hoover, D.R., Michaelson, N.J., Bryant, P., Margaryan, A., Chauhan, K.R., Aldrich, J., Schneidmiller, R. 2015. (1R,2S,5R,8R)- Iridodial and Z,E-nepetalactol: first long-range 4 chemical attractants for antlions (Neuroptera: Myrmeleontidae). Frontiers in Ecology and the Environment. 2:80.
- Zhang, Z., Gao, Q., Liu, L., Zhang, A., Zhang, B., Luo, J., Chen, L., Wang, M., Lei, C. 2015. Sex pheromone of the mirid bug, Adelphocoris suturalis. Journal of Chemical Ecology. 26(1):25-31.
- Prieto, N., Swift, M., Summerfelt, S., Juarez, M., Zijlstra, R., Aalhus, J. 2015. A feasibility study on the use of near infrared spectroscopy for the authentication of depurated salmon fillets. Journal of Food Analytical Methods. DOI: 10.1007/s12161-015-0168-9.
- Cornelius, M.L., Buffington, M.L., Talamas, E.J., Gates, M.W. 2016. Parasitism and predation rates on sentinel and naturally occurring egg masses of the squash bug (Hemiptera: Coreidae) in Maryland. Environmental Entomology. 45(2) 367-375.
- Harris, C., Abubeker, S.U., Yu, M., Leskey, T.C., Zhang, A. 2015. Semiochemical production and laboratory behavior response of the brown marmorated stink bug, Halyomorpha Halys. Journal of Insect Physiology. 10(1):e0140876.
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