Progress 01/01/07 to 12/31/07

Outputs
OUTPUTS: 1- KSU Field Studies in Summer & Fall 2007: Fifty seeds of six hot pepper accessions (PI-438614, PI-370004, PI-438649, PI-435916, Grif-9354 and PI-224448) were planted in the greenhouse. Plants were watered daily and fertilized weekly with Peter's Professional that contains the elements NPK (20:20:20). At KSU field, an area measuring 55 ft X 76 ft was disked and rototilled. Stakes and twine were used to mark off four treatment areas measuring 12ft by 76ft with a foot of buffer between each treatment area. Four soil amendments were applied: a) yard waste compost; b) sewage sludge; c) chicken manure; and d) no mulch treatment used for comparison purposes. Each soil amendment was mixed with the native soil. Irrigation was set out and three rows of hot pepper accessions were planted. The plants were spaced with 2 ft between within rows and 4 feet between rows. Plants were harvested 3 times during September 2007 and the fruits were weighed, counted and graded. 2- KSU Laboratory Studies During July - September, 2007: A- Capsaicinoids Analysis and Current Collaboration with USDA/CRS (January - December, 2007): A total of three hundred and seven (307) hot pepper fruit samples were received from Dr. Robert L. Jarret, USDA/ARS, Plant Genetic Resources Conservation Unit, Griffin, Georgia. Fruit samples were analyzed for capsaicin and dihydrocapsaicin at KSU Land Grant Environmental Toxicology Laboratory. B- Hot Pepper Fruit Analyses & Quality of KSU Field Grown Fruits Pepper fruits were harvested and sorted into marketable and culls. Fruits were weighed, counted, and related to soil management used in the study. Fruit samples were taken for quality parameters (weight, length, width, wall thickness and color). Plants height and width as well as heavy metals accumulated in fruits were also recorded. 3- KSU Undergraduate Students Mentored by Antonious and Trained in Analysis of Natural Products: - Ms. Yoon, Hyeon Hu: KSU Undergrad Student, Spring-07 & Fall-07. - Mr. Lance Lofton: KSU Undergrad Student, Spring-07 & Fall-07. - Ms. Embry Maleka: KSU Undergrad Student, Fall-07. 4- Research results obtained from this hot pepper grant dealing with natural products for pest control were disseminated to Kentucky small farmers at the KSU Research Farm Third-Thursday monthly meetings. Three KSU undergraduate students, working as part-time research assistants in this grant, have gained the technical training and expertise in the area of natural products and analytical chemistry. Three undergraduates have presented their research findings at the 2007 Annual Meeting of the Kentucky Academy of Science, Posters-at-the-Capitol-2007 and coauthored published abstracts and one article published in peer-reviewed journal. PARTICIPANTS: 1- Dr. John C. Snyder, University of Kentucky, Department of Horticulture, Lexington, KY 40546 has collaborated in testing hot pepper extracts against spider mites. 2- Dr. Robert Jarret, USDA/ARS, Plant Genetic Resources (1109 Experiment Street, Griffin, GA 30224) has collaborated in the project and provided 307 new USDA genotypes for capsaicinoids analysis. Dr. Jarret has coauthored articles published in peer-reviwed jounal. 3- Dr. Tejinder S. Kochhar, Kentucky State University, Department of Biology (219A Carver Hall) has collaborated in hot pepper analysis for antioxidants. 4- Mr. William Kerst, Director of Shawnee Research & Development (Box 1659, Jamestown, TN 38556) has collaborated in this project and provided stickers and emulsifiers used in preparation of hot pepper formulations. 5- Dr. Sanjun Gu has collaborated in a new trial for testing the performance of new hot pepper formulation prepared by Antonious against the Japanese Beetles. TARGET AUDIENCES: 1- Two Kentucky limited resource farmers (Mr. Norman Watts & Mr. Morris Roman, Ekron, KY)are currently growing hot peppers in their farms as a cash crop. KSU research results are shared with small farmes involved in the project. 2- The KSU Land Grant Program has presented the hot pepper research findings and updated information regarding new hot pepper geotypes to all Kentucky farmers through the Third-Thursday monthly meetings and farm field days at KSU Research Farm. 3- Many KSU part-time students paid by this grant have become our target audiences in this area of natural products for pest control.

Impacts
There is a concern over the risks of contamination of food and drinking water by residues of synthetic agrochemicals, and the negative impact on the countryside. Farmers are in need of insect pest management strategies that are effective, affordable and environmentally sound. A simplified recipe for preparing natural pesticide formulation from hot pepper fruits for use by Kentucky growers and home gardeners to control insects and spider mites has been described and presented at state and national meetings. Two hot pepper accessions PI-547069 and PI-239703 contained the greatest concentrations of capsaicin (1.34 and 0.93 mg capsaicin/g fruit, respectively) among all accessions analyzed. These two accessions can be explored for mass production of capsaicin. Crude extracts from hot pepper fruits can be explored for developing natural products for use as biodegradable alternatives to synthetic pesticides with both toxic and repellent effects. Using repellent chemicals for crop protection is a unique way to prevent insects and spider mites from laying eggs on target plants and preventing leaf and fruit damage. Some hot pepper species might be incorporated into plant breeding programs to produce fruit with high concentration of pest control agents and provide alternatives to farmers dealing with the sharply-escalating production costs associated with the increasing costs of energy and pesticides. Hot pepper extracts are affordable, environmentally safe, inexpensive, and can be prepared by any grower. This is a significant saving to limited resource farmers.

Publications

• 1- Antonious GF, Meyer J, Rogers Jami, and Hyeon-HU Yoon (2007). Growing hot pepper for cabbage looper and spider mite control. J. Environ. Sci. Health, B42: 559-567.
• 2- Antonious GF, Frank Sikora , and John C. Snyder (2007). Variation in Heavy Metals Accumulation Among Hot Pepper Species. UK Agricultural Experimental Station, Fruit & Vegetable Crops Research Report, Publication No. 555, Coolong T., Smigell C., and Snyder J. (Eds.), University of Kentucky, Lexington, KY, pages 79-81.
• 3-Kochhar TS and Antonious GF (2007). Natural Pesticides in Hot Pepper Species, a poster presentation at the International Congress of Toxicology (7-15-07), Montreal, Canada. The abstract has been published in the International Congress Proceeding in July 2007, page 102.
• 4- Antonious, GF (2007) presented a research poster entitled AOrganic Pesticides: Quantification of Decanoic Acid Methyl Esters in Hot Pepper Accessions@ at the 44th Annual Florida Pesticide Residue Workshop, 7-25-07, St. Pete Beach, FL, page 15.
• 5- Hu Yoon-Hyeon (undergraduate student competition) and Antonious GF (2007). Spectrometric Analysis of Decanoic Acid Methyl Esters in Hop Pepper. Transactions of the Kentucky Academy of Science, Volume 68 (1), Published Abstract, page 105.
• 6- Jami A. Rogers, Janet E. Meyer, and Antonious GF (2007): Yield and Quality of Hot Pepper Grown at Kentucky State University. Transactions of the Kentucky Academy of Science, Volume 68 (1), Published Abstract, page 104.
• 7- Janet E. Meyer and Antonious GF (2007): Lethal Effects of Hot Pepper Extracts on the Cabbage Looper, Trichoplusia ni. Transactions of the Kentucky Academy of Science, Volume 68 (1), Published Abstract, page 104.
• 8- Hu Yoon-Hyeon (KSU undergraduate student), Michael O. Somuah, Antonious GF, and Tejinder S. Kochhar (2007). Organic Pesticides from Hot Pepper Extracts. Posters-at-the-Capitol, February 15, 2007, Governor State of the Commonwealth of Kentucky Building, Frankfort, KY 40601, Published Abstract, page 27.

Progress 09/01/04 to 08/31/07

Outputs
OUTPUTS: crude extracts from hot pepper fruits were explored for developing natural products for use as biodegradable alternatives to synthetic pesticides. A simplified recipe for preparing natural pesticide formulation for use on home garden vegetables to control insects and spider mites is described (see publication record). The impact of soil amendments on quality and capsaicin content of pepper fruits was investigation in 2008. In addition, seeds of 63 accessions of hot pepper, Capsicum chinense were obtained from USDA/ARS Griffin, GA. These accessions of C. chinense represented germplasm originally acquired from a variety of locations including: Belize (n=9), Brazil (n=7), Colombia (n=8), Ecuador (n=6), Mexico (n=10), Peru (n=10), Puerto Rico (n=6), and United States (n=7). KSU Undergraduate Student Lab Workers in 2008: 1-Ms. Yoon, Hu, spring semester-2008 KSU undergraduate student hired by this grant has worked as part-time student on the separation and quantification of capsaicin and pentadecanoic acid methyl ester from 307 hot pepper genotypes. 2-Mr. Michael Somuah, spring semester-2008 has worked on soil amendments and hot pepper yield. 3-Lance Lofton: KSU Undergraduate Student, Spring-2008 4-Lauren Lobel,Summer-2008 undergraduate student. 5-Lauren Lobel, Fall 2008 6-Maleka Embry,Fall Semester-2008 7- Rachel Hayden, Fall-2008 8- Akeem Smith, Spring-08. Fruits of three hundred and seven (307) hot pepper genotypes received from USDA/ARS National hot pepper germplasm collection, Griffin, GA were analyzed for capsaicin and dihydro-capsaicin contents in collaboration with Dr. Robert L. Jarret (USDA project collaborator). Products and results were disseminated via several channels: 1)Presentation of research findings by RA's, students, PI and collaborators at national scientific meetings (Kentucky Academy of Science-2008, Annual Meeting of Florida Pesticide Residue Workshop, Tampa, FL. 2)Research findings were submitted to scientific journals for publication 3)Results have been included in the USDA/ARS Grin databases with collaboration with USDA Agency Cooperator (Dr. Robert Jarret), Dr. John Snyder at the University of Kentucky, and organic and limited resource farmers in three Kentucky Counties (Meade, Adair, and Franklin), and with two chemical companies: Shawnee Research & Development (Shawnee Town, TN), and Organic Solutions Inc. (San Antonio, TX). In addition, since part of the mission of the KSU/Land-Grant Program is to serve the needs of limited resource farmers, results were discussed and disseminated through KSU on-farm monthly meetings and distributed to organic growers and limited resource farmers by the KSU Extension Service through the Third-Thursday monthly meetings at KSU Research Farm. In addition, Four (4)undergraduate students have presented their research findings at the 94th Annual Meeting of the Kentucky Academy of Science, October 31- November 1, 2008, Kentucky Posters-at-the-Capitol-2008, and Presentation at the Annual Biomedical Research Conference for Minority Students (AMRCMS), Orlando, FL, November 5-8, 2008, and coauthored published abstracts. PARTICIPANTS: 1- Antonious, GF has collaborated with Drs. Robert Jarret and Bard Morris and visited their USDA/ARS hot pepper research farms in Byron, GA and visited USDA Labs at the University of Georgia in Griffin, GA. 2- Antonious, GF has collaborated with Dr. John Snyder, Associate Professor of Horticulture, Horticulture Department, University of Kentucky, Lexington, KY on the repellency and mortality of capsaicin, dihydrocapsaicin, and decanoic acid methyl-esters to several arthropods (spider mites and cabbage loopers) of agricultural importance. 3- Antonious, G.F. has collaborated with Dr. TS Kochhar, Professor of Biology, Carver Hall, Division of Mathematics and Sciences, Kentucky State University, Frankfort, KY. This collaboration has provided this project with salary and chemicals for 3 additional KSU undergraduate students in Fall-2008. Several manuscripts and several students' abstracts have been published as a result of this collaboration (See publication record). TARGET AUDIENCES: 1- Conducted research projects with Kentucky small farmers in Meade County and completed the research experiment dealing with hot pepper as a cash crop in Mr. Norman Watts field, Brandenburg Road, Ekron, KY (July-August, 2008). 2- KSU Undergraduate Students: Antonious, GF is currently involved in KSU undergraduate students projects in the area of Environmental Pollution and Bioremediation. All students involved ion the project i.e., Ms. Yoon Hu (KSU Campus, 021 Kentucky Hall); - Mr. Michael Somuah (Michael.somuah@kysu.edu); Mr. Akeem D. Smith (3306 McClain Ct., Louisville, KY 40299); Ms. Lauren Lobel (Lauren.lobel@kysu.edu); Ms. Maleka Embry (maleka.embry@kysu.edu), and other students (see students involvement in environ. Science research projects) were all provided with the state-of-the-art analytical and equipment training they need for the job-market. 3- Antonious, GF has been working with Mr. Louie Rivers, Manager of small farmers outreach program (Mr. Louie Rivers, KSU Extension Bldg., Land Grant Program, phone: (502) 597-6327, e-mail: louie.rivers@kysu.edu) who has demonstrated several on-farm soil management practice, water quality issues, and the safe use of alternative pesticides (hot pepper extracts) to all Kentucky growers during KSU Third-Thursday meetings at KSU Research Farm as well as State and National Meetings. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

Impacts
The literature contains little information on variability in capsaicinoid content among varieties of C. chinense and among their countries of origin. There is a very limited amount of research on capsaicin and other content of hot pepper fruits at land grant universities. This project has highlighted the importance of hot pepper fruits of C. chinense and other hot pepper species in the U.S.Department of Agriculture germplasm collection for variability in fruit that might subsequently utilized to enhance fruit content of capsaicinoids. New soil management practices were used to increase fruit contents of capsaicin and dihydrocapsaicin. New analytical procedures have been developed and validated at KSU research labs for quantifying capsaicin, dihydrocapsacin, and hexadecanoic acid esters in hot pepper fruit extracts of over 300 genotypes. The persistence of capsaicin in soil and plants sprayed with a new formulation of capsaicinoids was determined. The project has provided many Kentucky growers and undergraduate students and growers with the training and expertise in growing hot peppers for capsaicin content and as a cash crop. Impacts- Pungency is key characteristic associated with members of the genus Capsicum and is also an important fruit quality attribute. The USDA pepper (Capsicum spp.) germplasm collection contains several thousand members or accessions. Many of these species and cultivars have not been analyzed for their concentrations of antioxidant compounds such as capsaicinoids that can be used as pest control agents to reduce dependence on synthetic pesticides. Chemical analyses of hot pepper fruits of plants grown from seeds of 63 hot pepper genotypes of C. chinense revealed that accessions PI-152452 (Brazil) and PI-360726 (Ecuador) contained the greatest concentrations of ascorbic acid (1.2 and 1.1 mg g-1 fresh fruit, respectively), while PI-438648 (Mexico) contained the greatest concentration of total phenols content (349 microgram g-1 fresh fruit) among the other 63 accessions tested. PI-640900 (USA) contained the greatest concentration (P< 0.05) of capsaicin (1.52 mg g-1 fresh fruit) and dihydrocapsaicin (1.16 mg g-1 fresh fruit), while total major capsaicinoids in the fruits of PI-438648 (Mexico) averaged 2 mg g-1 fresh fruit. PI-355817 from Ecuador contained the greatest concentrations of B-carotene (8.3 mg g-1 fresh fruit). Among the 63 accessions analyzed, concentrations of total phenols were significantly higher in PI 438648 and PI 159248. These selected accessions were identified as potential candidates for mass production of capsaicin that can be used as alternative to many synthetic pesticides and as potential candidates for mass production of major capsaicinoids that have health-promoting properties. The project has identified hot pepper genotypes for USDA/ARS breeding programs for use as parents in hybridizations to produce high capsaicin, phenol, ascorbic acid, and B-carotene containing varieties.

Publications

• 1- Antonious GF, Berke T, and Jarret RL (2009). Pungency in Capsicum chinense- Varitation among countries of origin. J. Environ. Science and Health B44,1-6.
• 2- Antonious GF and Kochhar TS (2009). Mobility of heavy metals from soil into hot pepper fruits: A field study. Bulletin of Environmental Contamination and Toxicology, in press.
• 3- Antonious GF (2008). Gas chromatographic/ Mass spectrometric analysis of capsaicin. Poster presentation at the 45th Annual Meeting of Florida Pesticide Residue Workshop, July 20-25, 2008, Tampa, FL, published abstract, page 28.
• 4- Lauren M. Lobel (KSU undergraduate student), George F. Antonious and Tejinder S. Kochhar (2008). Antioxidants in hot pepper fruits: Variation among countries of origin. Presentation at the Annual Biomedical Research Conference for Minority Students (AMRCMS), Orlando, FL, November 5-8, 2008, published abstract pp 332-333.
• 5- Hu Yoon-Hyeon (KSU undergraduate student), George Antonious, Terry Berke, and Robert Jarret (2008). Pungency in hot pepper, a poster presented at the Posters-at-the-Capitol, January 28, 2008, Governor=s State of the Commonwealth of Kentucky Building, Frankfort, KY 40601, published abstract, page 53.
• 6- Lauren Lobel (KSU undergraduate student), George Antonious, and Tejinder Kochhar presented a research paper titled- Ascorbic acid and phenol contents of hot pepper fruits from eight countries of hot pepper origin. An oral presentation at the agricultural sciences section at the 94th Annual Meeting of the Kentucky Academy of Science, October 31- November 1, 2008, University of Kentucky, Lexington, KY 40546.
• 7- George Antonious presented a research paper titled- Heavy metal concentration in the fruits of Capsicum chinense. An oral presentation in the agricultural sciences section at the 94th Annual Meeting of the Kentucky Academy of Science, October 31- November 1, 2008 hosted by the University of Kentucky, Lexington, KY 40546.
• 8- Antonious GF, Lauren Lobel (undergraduate student), tejinder Kochhar, Terry Berke, and Robert Jarret (2008). Screening Capsicum chinense for antioxdant contents, UK Agricultural Experimental Station, 2008 Fruit and Vegetable Crops Research Report # PR572, December 2008 issue, University of Kentucky, Lexington, KY.
• 9- Jami A. Rogers and George F. Antonious (2008).Yield and Quality of Hot Pepper Grown with Various Soil Amendments at Kentucky State University Farm. J Ky Acad. Sci., 69, published abstract, page 76.
• 10- Hu Yoon-Hyeon (undergraduate student): George Antonious, Terry Berke, and Robert Jarret (2008). Oral presentation titled Natural capsaicin in Capsicum chinense- Concentration Vs. Origin. J Ky Acad. Sci., 69, published abstract, page 75.
• 11- Yoon-Hyeon Hu and Antonious GF (2008). Poster titled -Quantification of capsaicinoids in hot pepper extracts- presented at the Annual Regional Undergraduate Chemistry Posters, April 18, 2008, University of Kentucky Department of Chemistry, 137 Chemistry-Physics Building, Lexington, KY 40506, published abstract, page 8.

Progress 01/01/06 to 12/31/06

Outputs
-Two Research Assistants: 1-Ms. Janet Meyer (time allocated 100%) & 2-Ms. Jami Rogers (time allocated 100%). As proposed in the project plan of work they are currently working on the project plan of work to achieve the project objectives. -Students Involvement in the Capacity Building Grant in Year 2006: Seven KSU Undergraduate students have been involved in the project during FY'06: 1-Ms. Yoon, Hyeon Hu, KSU Undergraduate Student, Spring Semester-2006, Summer-2006 & Fall Semester-2006. 2- Mr. Daddy N. Boateng, KSU Undergraduate Student, Spring 2006. 3- Ms. Regina Hill, KSU Undergraduate Student, Spring Semester-2006. 4- Mr. Michael Somuah, KSU Undergraduate Student, Summer Session-2006 & Fall Semester-2006. 5- Ms. Shaquita Brown, KSU Undergraduate Student, Summer Session-2006. 6- Ms. Kaisha Hurst, KSU 2006 Summer KSU/REAP Training Program. 7- Ms. Kamille M. Colombel, KSU Undergraduate Student, Spring 2006. -KSU Field Studies in Year 2006: A- One-hundred fifty (150) hot pepper seeds of 5 accessions (PI-438614; PI-435916; PI-370004: PI-438649; and Grif-9354) were selected from 99 accessions (last year study) based on concentration of capsaicin and dihydrocapsaicin in their fruits. Seeds were planted in the greenhouse at KSU Research Farm on March 30, 2006 and transplanted into the field on June 7, 2006. Plants were irrigated using drip-tape irrigation. Fruits were collected and counted for use in preparation of extracts and mass production of capsaicinoids. B- KSU Laboratory Studies in Year 2006:Capsaicinoids from selected hot pepper accessions were extracted, purified, and concentrated. 1- Aliquotes of each extract were used for bioassays using 3rd instar larvae of cabbage looper, Trichoplusuia ni (Hubner) and two-spotted spider mite, Tetranychus urticae Koch. 2- Bioassays were also performed using decanoic acids methyl esters extracted from hot pepper fruits. 3- Each extract was injected into GC/MS for identification of unknown peaks. 4- Capsaicinopids Analysis for USDA/CRS Collaborators: Four-hundred (400) hot pepper samples from USDA/ARS, Plant Genetic Resources Conservation Unit, Griffin, Georgia have been received from Dr. Robert L. Jarret for identification and determination of capsaicin and dihydrocapsaicin content using GC/Mass Spectrometry. This information will be used by USDA to select candidate accessions of hot pepper for pungency and commercial propagation.

Impacts
With the public perception that synthetic pesticides leave residues in the crop produce they eat, there has been increased interest in using natural products for pest control. The potential of using fruit extracts of hot pepper for controlling the cabbage looper, Trichopulsia ni Hubner and spider mite, Tetranychus urticae Koch is explored in this investigation. Crude extracts from hot pepper fruits were prepared and tested for their insecticidal and acaricidal performance. Mortality was greatest (94%) when fruit extract of accession PI-593566 (C. annuum) was spayed on larvae of the cabbage looper, while crude extracts of accessions PI-241675 (C. frutescens) and PI-310488 (C. annuum) were repellent to spider mite. Spectrometric analysis and toxicity to cabbage looper larvae revealed that pentadecanoic acid methylester is likely related to cabbage looper mortality. Fruit extracts of accessions PI-593566 (C. annuum) and PI-241675 (C. frutescens) could be useful for managing populations of cabbage looper and spider mite, which could reduce reliance on synthetic pesticides. Results were disseminated to Kentucky growers through the KSU Third-Thursdays meetings. In addition, this project has provided several training and research opportunities for employment in the area of natural products chemistry and performance of organic pesticides under laboratory and field conditions for minority graduate, undergraduate and high school students (KSU 1890 Research Apprenticeship Program) at KSU and University of Kentucky (UK), a nearby institution.

Publications

• 17- George F. Antonious, Janet E. Meyer, and Jami A. Rogers (2006). Hot Pepper Extracts for Cabbage Looper (Trichopulsia ni Hubner) Control. The 2006 Fruit and Vegetable Crops Research Report, Kentucky Agricultural Station, University of Kentucky, College of Agriculture, Lexington, KY 40546, PR-521, pages 77-78.
• 1- Antonious GF, Meyer J, Snyder JC (2006). Toxicity and Repellency of Hot Pepper Fruit Extracts to the two-spotted spider mite, Tetranychus urticae Koch. J. Environ. Science Health, B41 (8): 1383-1391.
• 2- Antonious GF, Kochhar TS, Jarret R, and Snyder JC (2006). Antioxidants in Hot Pepper: Variation Among Accessions. J. Environ. Science Health, B41, 1237-1243.
• 3- Antonious GF and Snyder JC (2006). Natural products: Repellency and Toxicity of Wild Tomato Leaf Extract to the Two-Spotted Spider Mite, Tetranychus urticae Koch. J. Environ. Science Health, B41 (1): 43-55.
• 4- Antonious GF and Jarret R (2006). Screening Hot Pepper Accessions for Capsaicinoids Content. J. Environ. Science Health, B41: 717-729.
• 5- Hu H. Yoon (KSU undergraduate student), George Antonious, and Robert L. Jarret (2006). Screening hot pepper for capsaicin content, 14th Biennial Research Symposium, Association of Research Directors, Inc., April 1-5, 2006, Hyatt Regency, Atlanta, GA. Published Abstract, page 169-170.
• 6- Daddy N. Boateng (KSU undergraduate student), George Antonious, Tejinder S. Kochhar, and and Robert L. Jarret (2006). Variations in Ascorbic Acid and phenols among hot pepper species, 14th Biennial Research Symposium, Association of Research Directors, Inc., April 1-5, 2006, Hyatt Regency, Atlanta, GA. Published Abstract, page 46.
• 7-Yoon Hyeon Hu (KSU undergraduate student), Daddy Boateng, Antonious GF, and Tejinder Kochhar (2006). Variation in Capsaicinoids content of hot pepper extracts. Posters-at-the-Capitol, February 2, 2006, Governor=s State of the Commonwealth of Kentucky Building, Frankfort, KY 40601, published abstract, page 1.
• 8- Daddy N. Boateng (KSU undergraduate student), George F. Antonious, Yoon H. Hu, Tejinder S. Kochhar, and Robert L. Jarret (2006). Natural Products: Screening Capsicum Accessions for Antioxidants Content. Posters-at-the-Capitol, February 2, 2006, Governor State of the Commonwealth of Kentucky Building, Frankfort, KY 40601, published abstract, page 53.
• 9- Kochhar TS and Antonious GF (2006). Variability in phenolics, ascorbic acid, and capsaicin contents among hot pepper accessions. Oral presentation at the International Conference on Biotechnology Approaches for Alleviating Malnutrition and Human Health, January 9-11, 2006, University of Agricultural Sciences, Gandhi Krushi Vignyan Kendra, Bangalore,Karnataka, India. Published Abstract, page 13.
• 10- Antonious, GF (2006) a research poster entitled Insecticidal and Acaricidal Performance of Hot Pepper Extracts, presented at the 43ndAnnual Florida Pesticide Residue Workshop, July 16-19, 2006, Orlando, FL, page 18.
• 11- Antonious, GF (2006). Capsaicinoids Content of Five Hot Pepper Species. Transactions of the Kentucky Academy of Science 67 (1): Spring 2006, Published Abstract, page 50.
• 12- Janet Meyer, George Antonious, Richard Thacker, and John Snyder (2006). Repellency of Hot Pepper Extracts to the Two-spotted Spider Mite, Tetranychus urticae Koch. Transactions of the Kentucky Academy of Science 67 (1): Spring 2006, Published Abstract, page 51.
• 13- Daddy Boateng (undergraduate student competition), George Antonious and Tejinder Kochhar (2006). Variation in Antioxidant Compounds Among Capsicum Species. Transactions of the Kentucky Academy of Science 67 (1): Spring 2006, Published Abstract, page 50.
• 14- Hu Yoon-Hyeon (undergraduate student competition) and George Antonious (2006). Mass Spectrometric Analysis of Decanoic Acid Methyl Esters in Hop Pepper Accessions. Oral presentation at the Kentucky Academy of Science Annual Meeting, Morehead State University, November 9-10, 2006, Morehead, Kentucky. This presentation has received the First Place Award in Undergraduate Student Competition in Agricultural Sciences.
• 15- Jami A. Rogers, Janet E. Meyer, and George F. Antonious (2006). Yield and Quality of Hot Pepper Grown at Kentucky State University. Oral presentation at the Kentucky Academy of Science Annual Meeting, November 9-10, 2006, Morehead State University, Morehead, Kentucky.
• 16- Janet E. Meyer and George F. Antonious (2006). Lethal Effects of Hot Pepper Extracts on the Cabbage Looper, Trichoplusia ni, Hubner. Oral presentation at the Kentucky Academy of Science Annual Meeting, November 9-10, 2006, Morehead State University, November 9-10, 2006, Morehead, Kentucky.

Progress 01/01/05 to 12/31/05

Outputs
1- Hiring Personnel: Two Research Assistants and six KSU part-time undergraduate students have started the work on the project. 2- Field Experiment at KSU Research Farm: Hot pepper seeds of 24 accessions were planted in the greenhouse at KSU Research Farm on April 20, 2005 and transplanted in the field on June 15, 2005. Plants were transferred to the field, irrigated using drip-tape irrigation system, and the fruits were collected and counted for mass production of capsaicinoids. Yield of hot pepper fruits was obtained for each of the 24 accessions tested. Bulk extraction of capsaicinoids from pepper fruits collected from the field study has been completed. USDA/ARS Plant Genetic Resources Conservation Unit, Griffin, GA, has provided KSU with ninety-nine (99) hot pepper accessions in a trial to select candidate accessions for mass production of the active ingredients (capsaicin, dihydrocapsaicin, and nordihydrocapsaicin). The 99 accessions represent a large array of the different taxa of Capsicum chinense, Capsicum frutescens, Capsicunm baccatum, Capsicum annuum, and Capsicum pubescens. This large collection of hot pepper were extracted, purified, and analyzed at KSU for capsaicinoids using GC/NPD and GC/MS. Capsaicinoids in the fruits were separated, purified, identified, and confirmed using GC/MS and pure standard materials of capsaicinoids. 3- Laboratory Tests: Out of the 99 accessions, twenty-four (24) hot pepper accessions have been selected: Grif-9217, PI-387838, PI-555644, Grif-9324, PI-593925, PI-238057, PI-585253, Grif-9169, PI-224424, PI-497985, Grif-9149, Grif-9317, PI-633758, PI-439522, Grif-9117, PI-257059, Grif-9273, PI-224416, Grif-9320, PI-438622, Grif-9270, PI-596057, PI-159261, PI-195299) for spider mite (Tetranychus urticae Koch) repellency. Mortality tests were also carried out using spider mites. Of the 99 hot pepper accessions tested, satisfactory results were obtained from eight accessions. Significant repellency of spider mite was observed in five accessions (PI-224424, POG-191, POW-47, POW-62, and PRG-25), at various dilutions. Concentrated extracts provided increased repellency in all 8 accessions tested.

Impacts
The pungent compounds of hot pepper, known as capsaicinoids, offer a partial solution as substituents for synthetic pesticides, particularly when two or more active compounds are combined to provide novel modes of action against a wide variety of insects and spider mites. Our results have indicated that crude extracts from accessions PI-596057 (C. baccatum), PI-195299 (C. annuum), GRIF-9270 (C. annuum), and GRIF-9320 (C. frutescens) were repellent. Extracts from Grif-9270, PI-159261 (C. frutescens), and PI-195299 caused the highest mortality. Accordingly, Grif-9270 and PI-195299 are potential candidates for spider mite control by both repellency and kill. These results would be of interest to organic growers who are seeking alternatives for spider mite control on vegetables and other crops. Accession PI-441624 (C. chinense) had the greatest capsaicin content (2.9 mg g-1 fruit) and accession PI-497984 (C. frutescens) had the greatest dihydrocapsaicin content (2.3 mg g-1 fruit), while PI-439522 and PI-497984 (C. frutescens) are the two accessions containing the highest concentration of total capsaicinoids. PI-441624, PI-497984, and PI-439522 were identified as potential candidates for the mass production of capsaicinoids, or for breeding of varieties enhanced for capsaicin content. The project also has provided several training and research opportunities for employment in the area of environmental science for minority graduate, undergraduate and high school students (KSU 1890 Research Apprenticeship Program).

Publications

• 1- Antonious GF and Snyder JC (2005). Repellency of hot pepper extracts to spider mites. Fruit and Vegetable Crops Research Report, PR-521, University of Kentucky, Lexington, KY, pages 76-78.
• 2- Antonious GF, Kochhar TS, Jarret RL and Snyder JC (2005). Antioxidants in hot pepper: Variations among accessions. The 2005 Fruit and Vegetable Crops Research Report, Pest Management, Health, and Sustainable Horticulture, University of Kentucky, Lexington, KY, December, 2005, pages 78-81.
• 3- Daddy N. Boateng (KSU undergraduate student), George F. Antonious and Tejinder S. Kochhar (2005). Quantification of phytochemicals of selected Capsicum accessions. A poster presentation at the Annual Biomedical Research Conference for Minority Students (ABRCMS), Hyatt Regency, Atlanta, Georgia, November, 2005. Published Abstract, page 281.
• 4- Antonious, GF (2005). Capsaicinoids content of five hot pepper species. An oral paper presented at the 91st Kentucky Academy of Science Annual Meeting, Eastern Kentucky University, Richmond, Kentucky, November, 2005.
• 5- Janet E. Meyer, George F. Antonious, Richard R.Thacker, and John C. Snyder (2005). Repellency of hot pepper extracts to the two-spotted spider mite, Tetranychus urticae Koch. An oral paper presented at the 91st Kentucky Academy of Science Annual Meeting, Eastern Kentucky University, Richmond, Kentucky, November, 2005.
• 6- Daddy N. Boateng (KSU undergraduate student), George F. Antonious, and Tejinder S. Kochhar (2005). Variations in antioxidant compounds among Capsicum accessions. An oral paper presented at the 91st Kentucky Academy of Science Annual Meeting, Eastern Kentucky University, Richmond, Kentucky, November, 2005.
• 7- Antonious GF (2005). Insecticides from Hot Pepper, 42nd Annual Florida Pesticide Residue Workshop, July 2005, St. Pete, FL, page 18.

Progress 01/01/04 to 12/31/04

Outputs
A research assistant position has been advertised. Two KSU undergraduate students (Clarence Jordan and Helen Miyasato) have been started in the project as part-time research assistants. Several methods for the separation and quantification of the active ingredients in pepper fruits are currently under study. The selected methods will be used based on the efficiency of the analytical procedure to recover the active ingredients from hot pepper fruit samples at trace levels. Hot peeper seeds have been received from USDA Cooperator and will be planted at Kentucky State University Research Farm under greenhouse conditions. Hot pepper fruits (PI-633754, PI-633755, PI-633756, PI-633757, PI-633834, PI-438622, and PI-585253) were purchased, extracted, and analyzed for two capsaicinoids (capsaicin and dihydrocapsaicin) using GC/MS. Fruit contents were identified, confirmed and compared with standard materials of capsaicinoids.

Impacts
The overall goal of this study is develop an insecticide/acaricide formulation and offer the opportunity for commercial propagation of Capsicum varieties as a cash crop and source of botanical pesticides. The efficiency of using crude extracts of Capsicum species against vegetable pests will be tested in several small-scale organic farmers in Kentucky as an alternative to the declining tobacco production in southeastern U.S. Many analytical procedures will be developed and used to test capsaicinoids dissipation from the treated plant surface into soil and water. Many farmers may be able to grow and produce Capsicum species under contract for industrial uses which will help limited resource farmers in Kentucky to develop new industrial crops. This project will provide a new formulation of pesticides from hot pepper that can be used safely against vegetable pests.

Publications

• No publications reported this period