Progress 12/01/07 to 11/30/13
Outputs
Target Audience: Target audiences have been: 1. Cotton producers and cotton researchers and also researchers working on Helicoverpa zea, cotton bollworm. The Proceedings of the Cotton Beltwide Conferences summarized the most important yearly findings of our insecticide resistance monitoring efforts for bollworm. This is also important for corn and sorghum producers and the data and analyzed results are publicly available at http://insecticideresistance.tamu.edu, a web site supported by the Department of Entomology at Texas A&M University. 2. For the more basic aspects of research target audiences have been students and scientists at other Landgrant universities such as the University of Illinois, Iowa State University and Mexico State University. The later is a Hispanic Serving Institution. The PD and her students consistently present their research in International, National and regional and local meetings. Further the PD has developed a micro program through Earth and Sky in Spanish, available through You Tube. The PD maintains a profile in Research Gate and tends to publish in open access journals when possible. The PD has also developed a JoVE publication and has made it available free access; this video and protocol has had more than 10,000 views worldwide and from reputable Universities in other countries. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided? This project provided extraordinary opportunities at a Tier I Research University. Our agency Texas AgriLife Research is the fourth in the Nation in funding for Agriculture. Four Ph.D. students and one M.S. student published their PhD dissertation and Thesis in high quality peer-reviewed journals. One MS student accompanied the PD to Glasgow, Scotland to the SEB (Society of Experimental Biology meeting) in the UK and presented a poster in the Symposium on epithelial physiology honoring Professor Bill Harvey. A PhD student had opportunity to work side-by-side with the PD in learning how to prepare a federal grant to the EPA that was funded. Another PhD student was trained on how to developed State funded proposals, also these were funded. The PI supported students to travel to National meetings (Entomological Society of America) and one of the students obtained the highest honor in research the Comstock Award for Outstanding Achievement as a graduate student (Hsiao-Ling). All of the PD students working under this project received competitive research awards. Through research students have learned bioinformatics, genomics and gene model annotation, statistics, confocal microscopy, molecular biology, they have attended workshops (on tissue imaging), etc. The project has provided opportunities yearly for Research Experiences for Undergraduates. How have the results been disseminated to communities of interest? Activities: The research was additionally disseminated at invited seminars as part of their Teaching and Research mission. These were: 1. Pietrantonio, P.V. 2012. Invited Lecture for Seminar Series. University of Illinois, Urbana-Champain. Department of Entomology. Title “A chamber of secrets: molecular mechanisms involved in reproduction of the red imported fire ant (Solenopsis invicta Buren). February 20, 2012. Invited by Prof. G. Robinson. 2.Pietrantonio, P.V. 2012. Invited Lecture for Seminar Series. New Mexico State University, Department of Biology. Title: Urine of the bloody enemy II: GPCRS involved in regulation of diuresis and excretion in the mosquito Aedes aegypti (L.). Invited by Prof. I. Hansen. April 5, 2012. 3. Pietrantonio, P.V. 2013. Iowa State University, Ames, IA. Department of Entomology. Invited Paul A Dahm Memorial Lecture in Entomology. Dedicated to Insect Toxicologist P.A. Dahm. Spring Semester. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
1. We discovered the renal organ (Malpighian tubule) of mosquito Aedes aegypti presents a variegated pattern of calcitonin (diuretic hormone 31) receptor expression in principal cells as shown by immunohistochemistry. That is, not all principal cells express the receptor. The frequency of receptor-expressing cells decreases from the distal to the proximal end of the M. tubule. The silencing of the receptor decreased in vitro the secretion rate of M. tubules and in vivo the overall volume of urine excreted within 1 h after blood feeding by 30%. This receptor is a critical component of the post-blood feeding diuresis in females. The receptor is glycosylated in M. tubules. The hindgut expression was also confirmed and the receptor is involved in regulation of hindgut contraction; silencing the Cal1 receptor decreases contraction frequency; a video was published as supplementary information. We characterized in 2013 the first tick periviscerokinin receptor from Boophilus microplus. We also clone several neuropeptide receptors from the tick Ixodes scapularis that were submitted to the NCBI and those sequences will be available upon publication of the genome paper of I. scapularis, of which the PD Pietrantonio and members of her laboratory are co-authors. The community effort is led by Dr. C. Hill. Goals were accomplished and results were published in peer reviewed publications and one Ph.D. dissertation in 2013. Other thesis and dissertations are listed in previous years reports. 2. In fire ants we discovered that the receptor for the neuropeptide short NPF (sNPF) is present in brain and ovaries of queens. We developed a highly specific antibody with which the receptor was immunolocalized in tissues and its expression verified by western blots. We found the receptor also in brains of workers and a publication was submitted. We cloned both insulin receptors, and these sequences from fire ant queens allowed us to correct the previously incorrectly predicted sequences from the honey bee genome. We validated RNAi by injecting pupae of female reproductives. Work on the Vitellogenin receptor was published in 2011 and we demonstrated the proof of principle that interfering with this receptor reduces ovary size in queens. 3. In Helicoverpa zea (corn earworm or cotton bollworm), a critical polyphagous pests of many crops including sorghum, we elucidated the mechanisms of pyrethroid resistance present in field-collected adult males: target site insensitivity of the voltage gated sodium channel and associated overexpression of CYP450 enzymes CYP6B8 and CYP6B9.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2013
Citation:
Journal Articles Published
Yang, Y., P. Bajracharya, P. Castillo and Pietrantonio, P.V. 2013. Molecular and functional characterization of the first tick CAP2b (periviscerokinin) receptor from Rhipicephalus (Boophilus) microplus. General and Comparative Endocrinology 194: 142151.
- Type:
Journal Articles
Status:
Published
Year Published:
2013
Citation:
Kwon H, Pietrantonio PV. 2013. Calcitonin receptor 1 (AedaeGPCRCAL1) hindgut expression and direct role in myotropic action in females of the mosquito Aedes aegypti (L.). 2013. Insect Biochem Mol Biol. 43: 588-593.
- Type:
Theses/Dissertations
Status:
Published
Year Published:
2013
Citation:
Ph.D. Dissertation: Cloning, immunolocalization and functional analyses of calcitonin receptor 1 (AedaeGPRCAL1; diuretic hormone 31 receptor) in females of mosquito Aedes aegypti (Diptera: Culicidae).Department of Entomology, TAMU.
- Type:
Journal Articles
Status:
Under Review
Year Published:
2013
Citation:
Journal Article Submitted in review.
Castillo, P. and Pietrantonio PV. 2013.Differences in sNPF receptor-expressing neurons in brains of fire ant (Solenopsis invicta Buren) worker subcastes: indicators for division of labor and nutritional status?- Submitted to PLOS ONE- Reviewed with Minor revisions; was resubmitted and awaiting editorial acceptance.
- Type:
Conference Papers and Presentations
Status:
Awaiting Publication
Year Published:
2013
Citation:
Pietrantonio, P., H. Kwon and R. Nachman. 2013. A diuretic kinin analog elicits food aversion in females of Aedes aegypti mosquitoes, apparently through a sensory function of the kinin receptor in legs. EMBO Conference. July 2013. Kolimbari, Crete. Will be Published in Pathogens and Global Health.
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Progress 01/01/12 to 12/31/12
Outputs
OUTPUTS: OUTPUTS. Activities: The research was disseminated at two invited seminars. These were: 1. University of Illinois, Urbana-Champain. Department of Entomology. Departmental Colloquium. February 20, 2012 Title: A chamber of secrets: molecular mechanisms involved in reproduction of the red imported fire ant Solenopsis invicta Buren. 2. New Mexico State University. Las Cruces, NM. Department of Biology. Departmental Seminar, April 5, 2012. Urine of the bloody enemy II: GPCRs involved in regulation of diuresis and excretion in the mosquito Aedes aegypti (L.). Mentoring: 1. The PI served as Judge for the Entomological Society of America Graduate Student Poster Competition for the President's Prize. ESA Annual Meeting, Knoxville, TN, November 11-15, 2012. 2. The PI is a University mentor for faculty in the NSF-funded ADVANCE Scholar Program for minority faculty in STEM fields at TAMU;several activities within the University. Events: International conferences. Presentations 1. Martinez-Valenzuela, M., G. Damas, M. Iracheta-Cardenas, J. A. Valadez-Lira, K. P. Jalomo Ortiz, P. Pietrantonio, C.A. Blanco, R. Gomez-Flores, P. Tamez-Guerra. 2012. Humoral immune response and enzymatic activity by Heliothis virescens after Bacillus thuringiensis exposure. 2012. March 19-20. Biotechnology Summit, Merida, Yucatan, Mexico. 2. Pietrantonio, P.V. 2012.Title: Molecular and signaling mechanisms controlling diuresis and water balance in the context of the blood feeding adaptation in tick and mosquitoes. Baltimore Vector Encounter. John Hopkins Malaria Institute, Baltimore, MD. June 7-10, 2012. 3. Pietrantonio, P.V. 2012. From getting the keys to an empty lab to full professor, a case study. Presented at the Entomological Society of America Annual Meeting, Symposium How Entomologists Manage: The Global Skills We Often Overlook. November 14, 2012. PARTICIPANTS: One Ph.D. student, Hyeogsun Kwon participated. A female post-doctoral student Prati Bajracharya and a technician Paula Castillo-Bravo participated. A Ph.D. graduate student in rotation, S. Kanameni also participated on fire ant research. TARGET AUDIENCES: Not relevant to this project. PROJECT MODIFICATIONS: Not relevant to this project.
Impacts
We discovered that the renal organ (Malpighian tubule) of mosquito Aedes aegypti presents a variegated pattern of calcitonin (diuretic hormone 31) receptor expression in principal cells as shown by immunohistochemistry. That is, not all principal cells express the receptor. The frequency of receptor-expressing cells decreases from the distal to the proximal end of the M. tubule. The silencing of the receptor decreased in vitro the secretion rate of M. tubules and in vivo the overall volume of urine excreted within 1 h after blood feeding by 30%. This receptor is a critical component of the post-blood feeding diuresis in females. The receptor is glycosylated in M. tubules. Currently the hindgut expression is being studied.
Publications
- PEER-REVIEWED Kwon, H.-S., H.-L. Lu, M. Longnecker, P.V. Pietrantonio. 2012. Role in diuresis of a calcitonin receptor (GPRCAL1) expressed in a distal-proximal gradient in renal organs of the mosquito Aedes aegypti (L.). PLOS One. http://dx.plos.org/10.1371/journal.pone.0050374. October 2012
- PROCEEDINGS Martinez-Valenzuela, M., G. Damas, M. Iracheta-Cardenas, J. A. Valadez-Lira, K. P. Jalomo Ortiz, P. Pietrantonio, C.A. Blanco, R. Gomez-Flores, P. Tamez-Guerra. 2012. Humoral immune response and enzymatic activity by Heliothis virescens after Bacillus thuringiensis exposure. Biotechnology Summit 2012, March 19-20, Merida, Yucatan, Mexico. Eds. F. Fernandez-Luqueno, F. Lopez-Valdez, S. Lozano-Muniz. International Foundation for biotechnology research & early stimulation in the culture of health, nutrition, sport, art, science, technology and society- A.C. Nonprofit Organization. Pages 114-119. ISBN WEB VERSION: 978-607-9023-12-6.
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Progress 01/01/11 to 12/31/11
Outputs
OUTPUTS: We discovered that the sNPF receptor in fire ant queens is also present in the ovary in addition to the brain. We cloned and analyzed the two insulin receptors from fire ant queens, and also discovered there are two insulin receptors in the aphid genome. We quantified the relative receptors' expression in different castes and in different queen tissues. We continued to maintain the web site for insecticide resistance in bollworm Helicoverpa zea : http//: insecticideresistance.tamu.edu. Discovered that overexpression of two highly similar CYP6B8 and CYP6B9 enzymes is associated with pyrethroid-resistant moths. We determined that the kinin receptor in Malpighian tubules of female of the yellow fever mosquito Aedes aegypti is critical for fluid excretion post blood feeding, its knock down reduced fluid output. Partial silencing of this receptor was, however, not deadly for females. We also solved a long standing controversy in the field of renal physiology in mosquitoes. We unequivocally demonstrated that the kinin receptor is expressed in the stellate cells of the Malpighian tubule and not in principal cells as previously reported. We also localized for the first time the kinin receptor in female rectum and midgut endocrine cells. We performed bioassays with deltamethrin coated silver nanobeads applied to glass vials with females of Aedes aegypti. Proved that the silver nanobeads penetrated the cuticle and were recovered from the mosquito hemolymph and are deadly to mosquitoes, although the toxicity is one order of magnitude lower than the pesticide by itself. A peer-reviewed video and protocol on a calcium bioluminescence assay to study structure-activity relationships of neuropeptides on arthropod G protein-coupled receptors received more than 7,000 views in the Journal of visualized experiments, less than a year after publication. PARTICIPANTS: Ph.D. students Hsiao Ling Lu, Hyeogsun Kwon and M.S. student Cymon Kersch benefiting from participating in this project. Also an undergraduate female student, first generation college, from Cedar Crest College, PA, participated in research through a Research Experience for Undergraduates supported by NSF through the EXCITE program in the Entomology Department. C. Kersch, a M.S. master student traveled with the P.I. to Glasgow for the Society of Experimental Biology meeting where both presented a poster. A graduate student Sasha Kay also worked in the laboratory during the summer in research and directed studies. This project was funded by Cotton Incorporated core program. Two Ph.D. students, H.L. Lu and H. Kwon presented their research at the Entomological society of America annual meeting. The Ph.D. student Hsiao Ling Lu was the recipient of the Entomological Society of America Henry Comstock Award for outstanding achievement by a graduate student. Southwestern Branch, November 2011. Only 5 awards are given nationwide. TARGET AUDIENCES: Research results from the Cotton Incorporated funded project on H. zea were presented at the Beltwide Cotton Conferences and results from the fire ant project and mosquito projects were presented at the Entomological Society of America Annual meeting. The PI paid to make the fire ant publication in BMC Neuroscience as open access. The PI maintains a publicly available georeferenced database at http://insecticideresistance.tamu.edu. A female undergraduate, first generation in college participated in research during 10 weeks of summer under the Research Experiences for undergraduates EXCITE program in our Dept., funded by the NSF. PROJECT MODIFICATIONS: Not relevant to this project.
Impacts
The presence of the sNPF receptor in fire ant queen ovaries suggests for the first time for insects that the sNPF neuropeptide may act as a brain-ovary neurohormone, affecting reproduction. The short neuropeptide F localization in the brain is the first for any adult insect. This will allow to test hypothesis of the relationship between nutrition and the expression of the short NPF pathway in the brain. Our sequences for the insulin receptors of fire ants allowed us to correct the incomplete sequences of the insulin receptors in the honey bee genome by third party annotation. Both were incompletely or wrongly predicted. Our finding of sites for differential post translational modifications in both insulin receptor sequences suggests the possibility of differential regulation and therefore that the presence of two insulin receptors in social insects and aphids is not for redundancy in function but for regulation of different functions. In mosquito Aedes aegypti,the demostration that deltamethrin-coated silver nanobeads can be found in the hemolymph and kill mosquitoes is the proof of principle that other compounds could be delivered through the cuticle using nanobeads in a non-invasive manner, without injecting the insects. In Aedes aegypti the finding of the kinin receptors in multiple tissues involved in digestion and excretion indicates the kinins may have been critical for the evolution of the blood feeding adaptation in mosquitoes. One Ph.D. student graduated and one M.S. student graduated under this project.
Publications
- Castillo,L., B.W. Hopkins, H.-L. Lu, S. Singh, and P.V. Pietrantonio. 2011. Transcriptional overexpression of cytochrome P450 gene(s) Cyp6B8/Cyp6B28 is associated with cypermethrin survivorship of field-collected bollworm males (Helicoverpa zea). In Proc. Beltwide Cotton Conferences, January 4-7, Atlanta, Georgia. National Cotton Council, Memphis, TN.
- Lu, H.-L., Pietrantonio P.V. 2011. Insect insulin receptors: insights from sequence and caste expression analyses of two cloned hymenopteran insulin receptor cDNAs from the fire ant. Insect Mol Biol. 20 (5): 637-649.
- Lu, H.-L. and P. V. Pietrantonio. 2011. Immunolocalization of the short neuropeptide F receptor in queen brains and ovaries of the red imported fire ant (Solenopsis invicta Buren). BMC Neuroscience 12: 57.
- Hopkins, B.W., M. Longnecker, and Pietrantonio, P.V. 2011.Transcriptional overexpression of CYP6B8 and CYP6B9 is associated with pyrethroid resistance in Texas Helicoverpa zea. Pest Management Sci. 67(1):21-25.
- Lu, H.-.L., C.N. Kersch and Pietrantonio, P.V. 2011. The mosquito Aedes aegypti kinin receptor is expressed in stellate cells: A new model needed for ion transport Insect Biochemistry and Molecular Biology 41(2):135-40.
- Kersch, C. and Pietrantonio, P.V. 2011. Mosquito Aedes aegypti (L.) leucokinin receptor is critical for in vivo fluid excretion post blood feeding. FEBS Letters 585: 3507-3512.
- Kersch, Cymon N. 2011. M.S. Thesis title: Immunolocalization and in vivo functional analysis by RNAi of the Aedes kinin receptor in female mosquitoes of Aedes aegypti (L.) (Diptera, Culicidae). M.S. Entomology. Supervisor: P.V. Pietrantonio, Chair. Department of Entomology, Texas A&M University.
- Proceedings Pietrantonio, P.V., S. Singh, D. Kim, L. Castillo, L. Nemec, R.D. Parker, B. Reed, K. Siders, M.G. Cattaneo, M. Vandiver and N. Troxclair. 2011. Texas resistance monitoring program reveals increase in cypermethrin susceptibility in Helicoverpa zea (Boddie) populations in 2010. In Proc. Beltwide Cotton Conferences, January 4-7, 2011, Atlanta, Georgia. National Cotton Council, Memphis, TN.
- Published Abstracts International: Kersch, C.N. and Pietrantonio, P.V. 2011. Expression and functional analyses of the kinin receptor in females of the Aedes aegypti (L.) mosquito. Invited poster at the Symposium Molecular Physiology of Epithelial Transport in Insects: a tribute to William Harvey. Society of Experimental Biology Annual Main Meeting. July 1- 4th, 2011. Glasgow, Scotland. Published abstract SEB A1.35: 69.
- Pietrantonio, P.V. 2011. Arthropod vector G protein-coupled receptors involved in fluid excretion: tissue expression, cell specialization revealed in renal organs and RNAi phenotypes. Fifth EMBO Conference "Molecular and Population Biology of Mosquitoes and Other Disease Vectors". 24-30 July, 2011. Kolymbari, Crete, Greece.
- Published abstracts national: Kwon, H. and P.V. Pietrantonio. 2011. Cloning, immunolocalization and functional analysis of calcitonin receptor like receptor 1 (AaegGPRCAL1; Diuretic Hormone 31 (DH31) receptor) in females of mosquito Aedes aegypti (Diptera: Culicidae). Abstract and oral presentation at Graduate student 10 min paper competition IPMIS. Entomological Society of America Annual meeting, Nov. 12-16, 2011, Reno, NV, USA.
- Lu, H.-L. and P.V. Pietrantonio. 2011. Insect insulin receptors: insights from sequence and caste expression analyses of two cloned hymenopteran insulin receptors cDNAs from the fire ant. Abstract and poster presentation by H.L. Lu, IPMIS. Entomological Society of America Annual meeting, Nov. 12-16, 2011, Reno, NV, USA.
- Lu, Hsiao-Ling. 2011. Ph.D. Dissertation title: Vitellogenin receptor and neuropeptide receptors involved in reproduction of the red imported fire ant (Solenopsis invicta Buren). Ph.D. Entomology. Supervisor: P.V. Pietrantonio, Chair. Department of Entomology. Texas A&M University.
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Progress 01/01/10 to 12/31/10
Outputs
OUTPUTS: The immunolocalization of the sNPF receptor in the fire ant queen brain is the first of such in the brain of any adult insect. We reported novel mutations in the bollworm, Helicoverpa zea, voltage gated sodium channel that are associated with cypermethrin survivorship of adult males in the field. Additionally, overexpression of two cytochrome P450 enzyme transcripts, Cyp6B8/Cyp6B28 and Cyp6B9 was found in field collected insects that survived high cypermethrin (pyrethroid) dosages in the adult vial test. PARTICIPANTS: Ph.D. Graduate Students: Bradley W. Hopkins (Ph.D., Graduated May 2010), Hsiao-Ling Lu (fire ant and tick project), Hyeog Sun Kwon (mosquito project), Donghun Kim (tick project and bollworm project). M.S. student: Cymon Kersch (mosquito kinin receptor project). Extension personnel and IPM agents from AgriLife Research collaborate in the statewide pyrethroid resistance monitoring project. An undergraduate Hispanic student, Liliana Castillo from Greenville College in Illinois participated in research through the National Science Foundation Research Experiences for Undergraduates (NSF-REU) EXCITE program from the Dept. of Entomology. She worked on the CYP450 enzyme overexpression detection and the bollworm monitoring program. The monitoring program is supported through the Texas State Support program from Cotton Incorporated and research on mechanisms by the Insecticide Resistance Action Committee (IRAC). Students Hsiao-Ling Lu, Bradley W. Hopkins and Liliana Castillo presented research at meetings and students Donghun Kim and Hyeog Sun Kwon participated in the ESA Annual Meeting in San Diego, CA. TARGET AUDIENCES: Cotton producers and consultants and pesticide industry. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
Identifying the neurons involved expressing the short NPF receptor in fire ant queens will contribute to our understanding of both, neuronal circuits and gene networks that control insect feeding, locomotion and perhaps reproduction. In the bollworm, identification of the sodium channel mutations associated with survival to cypermethrin will allow to implement fast molecular diagnostic tests based on PCR or qPCR that can be applicable to population-level studies of resistance. Further, one mutation identified in the voltage-gated channel cDNA could be due to mRNA editing. The statewide cypermethrin monitoring program provides information for better insecticide selection for bollworm control (see http://insecticideresistance.tamu.edu). Two mechanisms, target site insensitivity and increased metabolism by CYP450 enzymes are associated with cypermethrin survivorship in bollworm field collected adults. This indicates that bollworm populations may already be or could also become resistant to other insecticides through the overexpression of these CYP450s. One Ph.D. student graduated in May 2010 (B.W. Hopkins) and is currently working for Dow AgroSciences Research and Development, Ohio. The bollworm project also allowed training in molecular entomology of a foreign visiting scholar, Dr. Satnam Singh, Assistant Professor, from the Punjab Agricultural University in India.
Publications
- Hopkins, B.W. and P.V. Pietrantonio. 2010. The Helicoverpa zea (Boddie) Lepidoptera: Noctuidae) voltage gated sodium channel and mutations associated with pyrethroid resistance in field collected adult males. Insect Biochem. Mol. Biol. 40 (5):385-93.
- Nachman, R. and P.V. Pietrantonio. 2010. Interaction of mimetic analogs of insect kinin neuropeptides with arthropod receptors. Chapter 3. In: Neuropeptide systems as Targets for Parasite and Pest Control. Eds. Professor Tim Geary and Aaron G. Maule. Landes Bioscience, Austin, TX. Springer series: Advances in Experimental Medicine and Biology vol. 692:27-48. Springer Science+Business Media, LLC. ISBN: 978-1-4419-6901-9.
- Hopkins, B. W. 2010. PhD Dissertation: Monitoring resistance to pyrethroid insecticides in Helicoverpa zea (Boddie) (Lepidoptera, Noctuidae): bioassay validation, voltage-gated sodium channel mutations and Cyp6B overexpression analysis. Department of Entomology. Texas A&M University.
- Hopkins, B.W., L. Bright, P.V. Pietrantonio et. al. 2010. Monitoring for Pyrethroid Resistance in Bollworm (Helicoverpa zea) Populations in Texas and Tamaulipas, Mexico; 2009. In Proceedings 2010 Beltwide Cotton Conferences. January 2010. New Orleans, Louisiana. National Cotton Council, Memphis, TN. Electronic publication and CD.
- Pietrantonio, P.V. and B.W. Hopkins. 2010. Spatial and temporal diversity of polyphagous pests: Pyrethroid resistance in the corn earworm (Helicoverpa zea, Lepidoptera, Noctuidae). Presentation at P-IE section symposium: Understanding and Capitalizing on Agricultural Biodiversity in IPM/IRM. ESA Annual Meeting, San Diego, CA, Dec. 12-15, 2010.
- Lu, H.-L. and P.V. Pietrantonio. 2010. Distribution of short neuropeptide F receptor in virgin and mated queens of the fire ant, Solenopsis invicta Buren (Hymenoptera: Formicidae). Abstract and oral presentation at Graduate student 10 min paper competition IPMIS. Entomological Society of America 58th Annual Meeting. San Diego, CA, Dec. 12-15, 2010.
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Progress 01/01/09 to 12/31/09
Outputs
OUTPUTS: This Hatch project involves numerous projects. 1. Bollworm project: The objective this year was to contribute knowledge on the status of Helicoverpa zea pyrethroid resistance in Texas as in previous years with the addition of the analysis of molecular mechanisms of resistance. We found multiple mechanisms: one involving cytochrome P450 enzymes and the second involving target site insensitivity in the sodium channel. Project Output: a product is a database for public access. The database project was funded by the EPA and it will be released to the public in January 2010. 2. Fire ant reproduction project: we are investigating the reproductive transition from virgin queens to colony foundress in the red imported fire ant Solenopsis invicta. The objective is to understand changes in ovaries at the molecular level and identify signaling receptors involved in reproduction in the fire ant. One goal is to find molecular markers that correlate with behaviors during this transition In fire ants, knowledge of the molecular mechanisms and genes involved in the transition from virgin queens to colony foundress will allow understanding gene networks controlling behavioral outputs (readiness to fly or sexual maturation, mating, etc.) and their correlation with gene expression with the goal of identifying the genes that are involved or ultimately control these behaviors or other physiological processes such as vitellogenesis. This knowledge will allow a more rational management of fire ant populations for a reduction of their reproductive success. 3. Mosquito Aedes aegypti water balance: we are systematically analyzing the function of GPCRs in mosquito excretory system in females pre- and post-blood feeding. The goal is to learn the biological significance of each of these receptors, their activity in critical time periods and their overall contribution to water loss in vitro and in vivo. We focused on various GPCRs involved in diuresis/excretion. 4. An international collaboration on Bemisia tabaci, the whitefly is ongoning with Dr. Vassilis Vassiliou in Cyprus. The goal was to quantify by qPCR the transcript of a cytochrome Cyp6CM1 enzyme as a candidate for the mechanism of resistance to neonicotinoids in Cyprus. We also completed the survey of populations for biotypes (B or Q) in Cyprus. A second international collaboration in Mexico with Dr. Almazan is on Rhipicephalus microplus (ex Boophilus) tick but this is reported under the NRI-USDA grant project. PARTICIPANTS: PhD graduate students: Christopher L. Jagge (graduated 2009), Bradley W. Hopkins (bollworm project), Hsiao Ling Lu (fire ant project), Hyeog Sun Kwon (mosquito project) and Dongun Kim (whitefly project). Master of Science student Cymon Kersch (mosquito project). Collaborators for bollworm project include many extension personnel from this Dept (see publication Beltwide Cotton Conferences). International collaborator whitefly project: Dr. Vassilis Vassiliou, Agricultural Research Institute,Nicosia, Cyprus. Dr. Suparna Taneja-Bageshwar is a PhD Research Associate. TARGET AUDIENCES: Growers, scientists, extension agents, students can be educated through the http://insecticideresistance.tamu.edu web site. The site provides geographical, seasonal and yearly data on the evolution of resistance to pyrethroids in Texas. Efforts include yearly presentations to growers and consultants at the Beltwide cotton conferences. PROJECT MODIFICATIONS: Not relevant to this project.
Impacts
1. The bollworm project involves analysis of bioassays through probit analysis with POLO software.The outcome of the Bollworm project resulted in the completion of the web accessible database on a google-earth type map and can be seen at http://insecticideresitance. tamu.edu. We also published a paper in proceedings with the monitoring results and a second publication in which we proved that the adult vial test for bollworm accurately reflects results with larvae, an assumption that has been finally confirmed experimentally. We also determined that bifenthrin is effective in susceptible insects but may not be successful in the field to control pyrethroid resistant populations giving resistance ratios of up to nine, while cypermethrin or esfenvalerate give lower resistant ratios of about 3-4. For sodium channel analysis we utilize conventional cloning methods and sequencing. For analysis of CYP expression we utilized qPCR. Evolution of resistance to pyrethroids in Helicoverpa zea (bollworm, corn eaworm) threatens economic sustainability in Texas, other southern states and the northeast because this pest migrates even to states where it does not typically overwinter. The availability of knowledge of the molecular mechanisms of resistance present in field insects will allow more precisely estimate frequencies of resistant individuals. This in turn will inform management and pyrethroid use leading towards reduction in use, or a more appropriate timing of application or a change in pesticide altogether. The web site with information will also allow gaining perspective of the geographic distribution of resistance and risk assessment. 2. In fire ants we established the technique of RNAi to investigate ovarian gene expression function by silencing the vitellogenin receptor. This was published. We determined that it is MATING and not FLIGHT what triggers high vitellogenin receptor expression within hours and correct receptor localization at the oocyte membrane appears as a marker for readiness to fly. Silencing of the vitellogenin receptor mRNA reduces ovarian growth pre-mating. 3. In mosquitoes we attempt to identify which signaling receptors contribute to water loss in females after a blood meal. We are quantifying the biological contribution of each receptor to water loss with the goal of being able to manipulate this mechanism in the future through synergists or novel pesticides promoting insect desiccation or death. We determined the expression pattern of DH44 receptor in female mosquitoes and this was the subject of a PhD dissertation. This basic knowledge may contriubute to target validation or discard some putative receptors as targets for pesticides and provides basic knowledge on the functional evolution of family B GPCRs for CRF-like peptides. 4. Whitefly project: we found high levels of Cyp6CM1 enzyme in whiteflies that survived treatment with Imidacloprid in Cyprus. We found overall resistance to different neonicotinoids, acetamiprid being the most effective. A full technical report was presented to the Research Promotion Foundation of Cyprus. One outcome is the graduation of one PhD Entomology student, Christopher Jagge.
Publications
- H.Ling Lu, B. Vinson and P. Pietrantonio. 2009. Oocyte membrane localization of vitellogenin receptor coincides with queen flying age, and receptor silencing by RNAi disrupts egg formation in fire ant vigin queens. FEBS J. 276: 3110-31123.
- Hopkins, B. W. and P. V. Pietrantonio. 2009. Differential efficacy of three commonly used pyrethroids against laboratory and field-collected larvae and adults of Helicoverpa zea (Lepidoptera: Noctuidae) and significance for pyrethroid resistance management. Pest Manag. Sci. 66:147-154.
- Christopher Jagge. 2009. Ph.D. Dissertation: The molecular characterization of a diuretic hormone receptor (GPRDH1) from females of the yellow fever mosquito, Aedes aegypti (L.).
- Pietrantonio, P.V., B. W. Hopkins, J.L. Moore, A. Abrameit, E. Bynum, G. Cronholm, T.X. Liu, R.R. Minzenmayer, G. Moore, R.D. Parker, C.G. Sansone, K. Siders, N. Troxclair, J. Vargas-Camplis. Monitoring for Pyrethroid Resistance in Bollworm (Helicoverpa zea) Populations in Texas and Tamaulipas, Mexico 2008. In Proceedings, 2009 Beltwide Cotton Conferences, January 2009, San Antonio, TX. National Cotton Council, Memphis, TN.
- Vassiliou, V., Emmanoulidou, M., Pietrantonio P., Tsagkarakou, A., J. Vontas and E. Roditakis. 2009. Insecticide resistance in Bemisia tabaci Gennadius (Hemiptera: Aleyrodidae) field populations, in Cyprus. In: Proceedings of the 13th Pan Hellenic Entomological Congress of the Hellenic Entomological Society. Alexandroupolis, Greece, 3-6 November, 2009. pp. 354 - 356. [Abstracts in Greek and English].
- Vassiliou, V., Emmanoulidou, M., Pietrantonio P., Dong Hun Kim, Tsagkarakou, A., J. Vontas and E. Roditakis. 2009. Insecticide resistance in Bemisia tabaci Gennadius (Hemiptera: Aleyrodidae) field populations, in Cyprus. In Proceedings of the 5th International Bemisia Workshop, 9-12 November, 2009. Guangzhou, China. p. 57.
- Hopkins, Bradley W. and P.V. Pietrantonio. 2009. Differential Efficacy of Three Commonly Used Pyrethroids for Laboratory and Field-Collected Larvae and Adults of Helicoverpa zea. Entomological Society of America Annual Meeting. Student competition. Oral presentation by B. Hopkins. Dec. 13-16, Indianapolis, IN. Published abstract.
- Hsiao-Ling Lu, S. B. Vinson and Patricia V. Pietrantonio. 2009. Oocyte membrane localization of vitellogenin receptor coincides with queen flying age, and receptor silencing by RNAi disrupts egg formation in fire ant virgin queens. Entomological Society of America Annual Meeting. Student competition. Oral presentation by H. L. Lu. Dec. 13-16, Indianapolis, IN. Published abstract.
- Lu, H. L., Vinson, B. and P.V. Pietrantonio. 2009. The queen of endocrinology: receptors in ovarian development of fire ant queens analyzed by RNAi and immunohistochemistry. Entomological Society of America Annual Meeting. Indianapolis, Indiana, Dec. 13-16, 2009. Poster presentation by PVP in symposium honoring Professor Judith Willis. Display D0188- IPMIS section.
- Pietrantonio, P.V. 2009. Characterization and in vivo functional analysis of the calcitonin receptor-like receptor 1 (AaegGPRCAL1) in female mosquitoes of Aedes aegypti (Diptera: Culicidae). Invited to the EMBO Conference Series: Molecular and population biology of mosquitoes and other disease vectors. 19-26 July 2009. Published abstract. Kolymbari, Crete, Greece.
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Progress 01/01/08 to 12/31/08
Outputs
OUTPUTS: Invited international presentation. Pietrantonio, P.V., H.-L. Lu and M.-E. Chen. 2008. Reproduction in fire ant (Solenopsis invicta Buren) queens: vitellogenin receptor biology. 2008. Symposium No. 13.07 Current advances in ant physiology and behaviour at the XXIII Intl. Congress of Entomology, Durban, South Africa. July 2008. Published in Congress Proceedings on CD. Volunteer international presentation and published abstract. Pietrantonio, P.V., R. Nachman, S. Taneja-Bageshwar, and C. Jagge. G protein-coupled receptors from tick (Rhipicephalus microplus) and mosquito (Aedes aegypti): biological significance and peptidomimetic design for target validation. Symposium on Signaling (GPCRs) for the XXIII Intl. Congress of Entomology in Durban, S.Africa. July 2008. Published in Congress Proceedings, CD. Pietrantonio, P., Nachman and Taneja-Bageshwar, S. 2008. Functional expression and target validation of tick kinin receptor,a GPCR putatively involved in water balance from Southern Cattle Tick, Rhipicephalus (Boophilus) microplus. VI International Conference on Tick and Tick-Borne Pathogens (TTP-6). Sept. 21-26, 2008. Published in Proceedings, page 88. Bs. As., Argentina. National invited presentation with published abstract. Pietrantonio, P.V. 2008. Can neuropeptide GPCRs be useful acaricide targets? Endocrinology and the road to validate the kinin receptor from tick Rhipicephalus microplus. Invited presentation at the Entomological Society of America Annual Meeting. IPMIS Section Symposium Tick genomics and beyond- New advances in tick-borne disease systems. ESA Annual Meeting web site. Reno, Nevada, Nov. 16-19, 2008. National volunteer presentations with published abstracts. Lu, H.-L., and P.V. Pietrantonio. 2008. Developmental expression pattern and RNA interference of the vitellogenin receptor in fire ant (Solenopsis invicta Buren) queens. Entomological Society of America Annual Meeting. November 16-19, 2008. Reno, Nevada. ESA web site; IPMIS, Lepidoptera and Hymenoptera. Hopkins, B.W. and P.V. Pietrantonio. 2008. Resistance to pyrethroid insecticides in Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae). Entomological Society of America Annual Meeting. November 16-19, 2008. Reno, Nevada. ESA web site, IPMIS, Lepidoptera and Hymenoptera. Kersch, C.N., HyeogSun Kwon and P.V. Pietrantonio. 2008. Cloning and sequencing the calcitonin receptor 3 (AaegGPRCal3) from female mosquitoes of Aedes aegypti (L.) (Diptera: Culicidae). Entomological Society of America Annual Meeting. November 16-19, 2008. Reno, Nevada. Abstract in ESA web site;IPMIS, Diptera. Regional invited presentation. Pietrantonio, P.V. 2008. Biochemical and Polymerase Chain Reaction (PCR) Techniques to Detect Insecticide Resistance and/or Biotypes Associated with Resistance. Annual Meeting of the Entomol. Soc. America, South Western Branch. Ft. Worth, TX. February 24, 2008. Symposium Molecular tools in IPM: research and applications. Regional volunteer presentation. Hopkins, B. W. and P. V. Pietrantonio. 2008. Pyrethroid Insecticide Resistance in Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae). Entomological Society of America Southwestern Branch. Ft. Worth, TX. Feb. 29, 2008. PARTICIPANTS: Post-doc. Dr. Suparna Taneja-Bageshwar. She worked in Pietrantonio's laboratory but collaborated in the laboratory of Dr. Ron Nachman in the ARS-USDA College Station by performing bioluminescence assays. Tick project also in collaboration with student Chris Jagge. Graduate students: Fire ant project: Hsiao-Ling Lu, PhD student. Cotton incorporated project on resistance in bollworm: Terry Junek (MS) and Brad Hopkins (PhD student). EPA project newly awarded: Donghun Kim, PhD student. Student began editing data to develop an interactive web site for bollworm resistance real-time communications to growers and stakeholders. Other arthropod G protein-coupled receptors: PhD students Christopher Jagge and Hyeogsun Kwon. An undergraduate student participated in the project through an REU from NSF. Funding was from IRAC (Insecticide resistance action committee), Cotton Incorporated, Fire Ant Research and Management Plan, NRI-USDA and EPA Region 6. Internationally: projects with Mexico through CONACyT (Collaborator Consuelo Almazan) and with Cyprus through the Research Development Foundation (collaborator Vassilis Vassiliou). A student Diana Carreon-Camacho, from Mexico, came to train in Pietrantonio's laboratory. Student is MS student of collaborator Consuelo Almazan, Univ. Autonoma of Tamaulipas, Tamaulipas, Mexico. TARGET AUDIENCES: The PI yearly publishes reports on the status of resistance in bollworm targeting growers. This information is communicated through Proceedings of Beltwide Cotton Conferences in January. Currently we are developing a web site that will also target IPM agents, growers, agricultural consultants, this project is funded by EPA. The PI regularly publishes in peer reviewed journals the discoveries on physiology and toxicology of arthropods. These publications mainly originate in the PI's laboratory and experiments are mainly conducted by graduate students in Entomology. Additionally the PI presents in multiple scientific forums nationally and internationally (see Outputs). PROJECT MODIFICATIONS: Dr. S. Taneja-Bageshwar has left the laboratory and a replacement must be found shortly to avoid delays in the project.
Impacts
Results from the bollworm monitoring program have helped growers apply pyrethroid at the correct rate even in crops such as sorghum in which lower rate applications of pyrethroids were targeted towards sorghum midge control. Research showed this is likely the reason for increased resistance in bollworm populations in South East Texas. Resistance frequency and resistance ratios have decreased in this area. International collaborative research resulted in no evidence of whitefly biotype Q in Cyprus which indicates that insecticide resistance is developing in the less harmful biotype B. Novel peptidomimetics proved highly active in both tick and mosquito kinin receptors in vitro; these are good small molecule leads to design other synthetic molecules acting on these receptors.
Publications
- Peer reviewed journal articles Taneja-Bageshwar, S., A. Strey, Kaczmarek, K., Zabrocki, J. Pietrantonio, P.V. and Nachman, R. 2008. Comparison of arthropod kinin analogs with cis-peptide bond, type VI-turn motifs identifies optimal stereochemistry for interaction with a recombinant insect kinin receptor from the Southern cattle tick Boophilus microplus. Peptides 29: 295-301.
- Taneja-Bageshwar, S., Zubrzak, P., Strey, A., Williams, H., Reyes-Rangel, G., Juaristi, E., Pietrantonio, P.V. and Nachman, R.J. 2008. Identification of selective and non-selective, biostable β-amino acid agonists of recombinant insect kinin receptors from the Southern cattle tick Boophilus microplus and mosquito Aedes aegypti. Peptides 29: 302-309,
- Jagge, C. and Pietrantonio, PV. 2008. Diuretic hormone 44 receptor in Malpighian tubules of the mosquito Aedes aegypti: evidence for transcriptional regulation paralleling urination. Insect Molecular Biology 17: 413-426.
- Vassiliou V.A., C. Jagge, M. Grispou, P. V. Pietrantonio and A. Tsagkarakou. 2008. Biotype status of Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) from various crops in Cyprus. Phytoparasitica 36, 400-404.
- Paper in proceedings. Junek, T.A., P.V. Pietrantonio, B.W. Hopkins, E. Bynum, M. Cattaneo, G. Cronholm, R.R. Minzenmayer, G. Moore, D. Mott, R. Parker, C.G. Sansone, K. Siders, N. Troxclair, M.R. Vandiver, and J. Vargas-Camplis. 2008. Monitoring for Pyrethroid Resistance in Bollworm (Helicoverpa zea) Populations in Texas and Tamaulipas, Mexico - 2007. In Proceedings, 2008 Beltwide Cotton Conferences, 8-11 January 2008, Nashville, TN. National Cotton Council, Memphis, TN.
- Book Chapter. Vinson, B., P. V. Pietrantonio, C. Coates, H.-L. Lu. 2008. The physiology of reproduction in the red imported fire ant Solenopsis invicta Buren. In: Recent advances in Insect Physiology, Toxicology and Molecular Biology. Editor Nanan Liu. Research Signpost, Kerala, India. 19 pp. ISBN: 978-81-308-0242-8.
- Published abstracts. Pietrantonio, P.V., Hsiao-Ling Lu and Mei-Er Chen. 2008. Reproduction in fire ant (Solenopsis invicta Buren) queens: vitellogenin receptor biology. 2008. Presenter PVP; power point). Oral presentation in the Symposium No. 13.07 Current advances in ant physiology and behaviour at the XXIII International Congress of Entomology, Durban, South Africa. July 2008. Published in the Congress Proceedings on CD.
- Pietrantonio, P.V., R. Nachman, S. Taneja-Bageshwar, and C. Jagge. G protein-coupled receptors from tick (Rhipicephalus microplus) and mosquito (Aedes aegypti): biological significance and peptidomimetic design for target validation. Symposium on Signaling (GPCRs) for the XXIII International Congress of Entomology in Durban, South Africa. July 2008. Published in Congress Proceedings, CD.
- Pietrantonio, P., Nachman and Taneja-Bageshwar, S. 2008. Functional expression and target validation of tick kinin receptor,a GPCR putatively involved in water balance from Southern Cattle Tick, Rhipicephalus (Boophilus) microplus. International Conference on Tick and Tick-Borne Pathogens (TTP-6), Buenos Aires, Argentina. Sept. 21-26, 2008. VI International Conference on Ticks and Tick-borne Pathogens. Book of Proceedings, page 88. Buenos Aires, Argentina.
- Pietrantonio, P.V. 2008. Can neuropeptide GPCRs be useful acaricide targets? Endocrinology and the road to validate the kinin receptor from tick Rhipicephalus microplus. Invited presentation at the Entomological Society of America Annual Meeting. IPMIS Section Symposium Tick genomics and beyond- New advances in tick-borne disease systems. Electronic Abstract in ESA Annual Meeting web site. Reno, Nevada, Nov. 16-19, 2008.
- Lu, Hsiao-Ling, and P.V. Pietrantonio. 2008. Developmental expression pattern and RNA interference of the vitellogenin receptor in fire ant (Solenopsis invicta Buren) queens. Entomological Society of America Annual Meeting. November 16-19, 2008. Reno, Nevada. Electronic Abstract in ESA web site. Student oral competition, IPMIS, Lepidoptera and Hymenoptera.
- Hopkins, B.W. and P.V. Pietrantonio. 2008. Resistance to pyrethroid insecticides in Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae). Entomological Society of America Annual Meeting. November 16-19, 2008. Reno, Nevada. Electronic Abstract in ESA web site. Student oral competition, IPMIS, Lepidoptera and Hymenoptera.
- Kersch, C.N., Hyeogsun Kwon and P.V. Pietrantonio. 2008. Cloning and sequencing the calcitonin receptor 3 (AaegGPRCal3) from female mosquitoes of Aedes aegypti (L.) (Diptera: Culicidae). Entomological Society of America Annual Meeting. November 16-19, 2008. Reno, Nevada. Electronic Abstract in ESA web site. Student oral competition, IPMIS, Diptera. REU NSF Undergraduate student project.
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