Progress 10/01/15 to 09/30/16

Outputs
Target Audience:Target audience is graduate students, undergraduate students, high school students, small scale farmers, limited resource farmers, pharmaceutical industries, and natural products weight-loss markets. Changes/Problems:Delays in the construction of suitable greenhouses for the maintenance of Hoodia collections, and for the implementation of the extension training of farmers on the horticultural production of camphor-free P. dulcis plants. All the Hoodia collection died in the inclement conditions in the greenhouse. We have not identified authentic Hoodia germplasm since then. What opportunities for training and professional development has the project provided?a. Eleven undergraduates and two graduate student were trained under all the research subject areas (agronomy, horticulture, analytical chemistry, genomics, biochemistry etc) of the project through the students' hourly work study program of the university in FY'2016-2017. b. Nine undergraduates and a graduate student made oral and/or poster presentations at four learned society (Atlanta ARD Symposium, Pittsburg MANRRS conference, 13th Annual Pathway Symposium, and iRISE symposium) conferences in 2016-2017 and most of them received prizes for excellent research topics in plant systems, and eloquent communication skill. c. Two postdocs were hired to train in the biochemistry, and genomics areas of the research project. d. A research specialist is being trained to mechanize the farm operations in the medicinal plants garden, fruit and vegetable garden, and greenhouse operations. How have the results been disseminated to communities of interest?Through cooperative extension program workshops for high school students from limited resource families, small scale farmers during PVAMU State-wide agriculture field days, on-line on the web site of the College of Agriculture and Human Sciences, and during beginner-farmer training sessions. What do you plan to do during the next reporting period to accomplish the goals?1. Plant protection studies on P. dulcis. 2. Cell and organ tissue culture genomics experimentation on P. dulcis. 3. Further optimization of P. dulcis sweetener and minimization of camphor contents through microbial actions. 4. Purification of monoterpene synthase and sesquiterpene synthase, preparation of polyclonal antibodies, and Western Blot assays of P. dulcis protein extracts.

Impacts
What was accomplished under these goals? a)Established best horticultural and sustainable agronomic practices under goal 1 for P. dulcis production in Texas. b)Synthesized cDNA probes for analyses of the carbon, nitrogen, terpenoid, shikimate etc. metabolic pathways under goal 3. c)Established HPLC, MS-GC protocols for identification and quantitation of primary metabolites, and terpenoids under goals 2 and 3. d) Submitted samples of the leaves, flowers, seeds, and roots of P. dulcis to external laboratories to validate the MS-GC, and metabolomics quantitation of the hernandulcin (zero calorie sweetener) and camphor compositions under goal 2. e) Three original scientific research publications were made under goals 1, 2, and 3 on the medicinal plants, Phyla dulcis: 1. Biochemical research on P. dulcis produced a journal publication co-authored by plant research scientists in conjunction with research specialists, undergraduate and graduate students, and research extension leaders in compliance with the one college infracture vision of the College of Agriculture and Human Sciences: (i) Osuji, G.O., Weerasooriya, A., Ampim, P.A.Y., Carson, L., Johnson, P., Jung, Y., Duffus, E., Woldesenbet, S., South, S., Idan, E., Johnson, D., Clarke, D., Lawton, B., Parks, A., Fares, A., Johnson, A. (2015) Molecular regulation of the metabolic pathways of the medicinal plants: Phyla dulcis. American Journal of Plant Sciences, 6: 1717-1726. 2. Metabolomics research on P. dulcis produced another journal publication co-authored by plant research scientists in collaboration with research specialists, undergraduate and graduate students in compliance with the one college infracture vision of the College of Agriculture and Human Sciences: (i) Osuji, G.O., Gao, M., Carson, L., Weerasooriya, A., Ampim, P., Johnson, P., Duffus, E., Woldesenbet, S., Kirven, J. Williams, E.L., Johnson, D., Clarke, D. (2017) Biotechnological induction of shikimate-based antioxidant accumulation in Phyla dulcis. The Natural Products Journal, 2017: 7, 104-111. 3. The opening publication in the medicinal plants project was in 2014. (i) Adams, R., Weerasooriya, A., Gao, M. (2014) Comparison of volatile leaf terpenoids from Lippia dulcis (Verbenaceae) obtained by steam distillation and pentane liquid extraction. Phytologia 96: 252-259. f) The high water mark of our accomplishment is the provisional application for USA patent: "Production of camphor-free and high hernandulcin Phyla dulcis plants' with the Texas A&M Technology Commercialization. The inventors are Plant Systems Scientists: Osuji, G.O, Carson, L.E., Gao, M., Weerasooriya, A., Ampim, P., Johnson, P., Woldesenbet, S. (2017). This is in recognition of the scientific breakthrough in the unique silencing of camphor biosynthesis in P. dulcis, under goals 1, 2, and 3. g) Under goals 1-5, for strengthening of the capacity of plant systems scientists to conduct relevant pioneering projects on medicinal plants, Dr. A. Weerasooriya received USDA-NIFA Capacity building research funding ($295.132) to establish DNA barcoding identification profiling of male health-related medicinal plant materials (2015). h) Dr. Peter Ampim received in 2017 another NIFA Capacity Building research funding of $596,395 to support small acreage farmers in Texas to produce high value specialty crops.

Publications

• Type: Journal Articles Status: Published Year Published: 2015 Citation: Osuji, G.O., Weerasooriya, A., Ampim, P.A.Y., Carson, L., Johnson, P., Jung, Y., Duffus, E., Woldesenbet, S., South, S., Idan, E., Johnson, D., Clarke, D., Lawton, B., Parks, A., Fares, A., Johnson, A. ((2015) Molecular regulation of the metabolic pathways of the medicinal plants: Phyla dulcis. American Journal of Plant Sciences, 6: 1717-1726.
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Osuji, G.O., Gao, M., Carson, L., Weerasooriya, A., Ampim, P., Johnson, P., Duffus, E., Woldesenbet, S., Kirven, J., Williams, E.L., Johnson, D., Clarke, D. (2016) Biotechnological induction of shikimate-based antioxidant accumulation in Phyla dulcis. The Natural Products Journal, 7, 104-111.
• Type: Journal Articles Status: Published Year Published: 2014 Citation: Adams, R., Weerasooriya, A., Gao, M. (2014) Comparison of volatile leaf terpenoids from Lippia dulcis (Verbenaceae) obtained by steam distillation and pentane liquid extraction. Phytologia 96: 252-259.

Progress 10/01/14 to 09/30/15

Outputs
Target Audience:Target audience isgraduate students, undergraduate students, high school students, small scale farmers, limited resource farmers, pharmaceutical industries, and natural products weight-loss markets. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Eleven undergraduates and two graduate student were trained under all the research subject areas (agronomy, horticulture, analytical chemistry, genomics, biochemistry etc) of the project through the work study program of the university. Two postdocs were hired to train in the biochemistry and genomics areas of the research project. Two research specialists are being trained to mechanize the farm operations in the medicinal plants garden, fruit and vegetable garden, and greenhouse operations; and the second specialist in the HPLC, GC-MS analyses of phytochemicals. How have the results been disseminated to communities of interest?Through cooperative extension program workshops for high school students from limited resource families, small scale farmers during PVAMU campus-wide agriculture field days, and during beginner-farmer training sessions. What do you plan to do during the next reporting period to accomplish the goals? Cell and organ tissue culture experimentation on P. dulcis. mRNA purification and sequencing experiments on P. dulcis. Optimization of P. dulcis sweetener and minimization of camphor contents through biotechnology. Plant protection experimentation on P. dulcis. P. dulcis terpenoid metabolomics panel. Preparation of cDNA probes for the regulatory steps in the steroidal metabolic pathway of Hoodia species.

Impacts
What was accomplished under these goals? a) Established best horticultural and sustainable agronomic practices under goal 1 for P. dulcis production in Texas. b) Synthesized cDNA probes for analyses of the carbon, nitrogen, terpenoid, shikimate etc. metabolic pathways under goal 3. c) Established HPLC, MS-GC protocols for identification and quantitation of primary metabolites, and terpenoids under goals 2 and 3. d) Three original scientific research publications were made under goals 1, 2, and 3 on the medicinal plants: Phyla dulcis: 1. Biochemical research on P. dulcis produced one journal publication co-authored by plant research scientists in conjunction with research specialists, undergraduate and graduate students, and research extension leaders in compliance with the one college infracture vision of the College of Agriculture and Human Sciences: (i) Osuji, G.O., Weerasooriya, A., Ampim, P.A.Y., Carson, L., Johnson, P., Jung, Y., Duffus, E., Woldesenbet, S., South, S., Idan, E., Johnson, D., Clarke, D., Lawton, B., Parks, A., Fares, A., Johnson, A. ((2015) Molecular regulation of the metabolic pathways of the medicinal plants: Phyla dulcis. American Journal of Plant Sciences, 6: 1717-1726. 2. Metabolomics research on P. dulcis produced a journal publication co-authored by plant research scientists in collaboration with research specialists, undergraduate and graduate students in compliance with the one college infracture vision of the College of Agriculture and Human Sciences: (i) Osuji, G.O., Gao, M., Carson, L., Weerasooriya, A., Ampim, P., Johnson, P., Duffus, E., Woldesenbet, S., Kirven, J., Williams, E.L., Johnson, D., Clarke, D. (2016) Optimal induction of shikimate family antioxidant accumulation in Phyla dulcis. The Natural Products Journal (accepted for publication). 3. The opening publication in the medicinal plants project was in 2014. (i) Adams, R., Weerasooriya, A., Gao, M. (2014) Comparison of volatile leaf terpenoids from Lippia dulcis (Verbenaceae) obtained by steam distillation and pentane liquid extraction. Phytologia 96: 252-259. e) Under goals 1-5, for strengthening of the capacity of plant systems scientists to conduct relevant pioneering projects on medicinal plants, Dr. A. Weerasooriya received USDA-NIFA funding ($295.132) to establish DNA barcoding identification profiling of male health-related medicinal plant materials (2015)

Publications

• Type: Journal Articles Status: Published Year Published: 2014 Citation: Adams, R., Weerasooriya, A., Gao, M. (2014) Comparison of volatile leaf terpenoids from Lippia dulcis (Verbenaceae) obtained by steam distillation and pentane liquid extraction. Phytologia 96: 252-259.
• Type: Journal Articles Status: Published Year Published: 2015 Citation: Osuji, G.O., Weerasooriya, A., Ampim, P.A.Y., Carson, L., Johnson, P., Jung, Y., Duffus, E., Woldesenbet, S., South, S., Idan, E., Johnson, D., Clarke, D., Lawton, B., Parks, A., Fares, A., Johnson, A. ((2015) Molecular regulation of the metabolic pathways of the medicinal plants: Phyla dulcis. American Journal of Plant Sciences, 6: 1717-1726.
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Osuji, G.O., Gao, M., Carson, L., Weerasooriya, A., Ampim, P., Johnson, P., Duffus, E., Woldesenbet, S., Kirven, J., Williams, E.L., Johnson, D., Clarke, D. (2016) Optimal induction of shikimate family antioxidant accumulation in Phyla dulcis. The Natural Products Journal (accepted for publication).

Progress 08/14/14 to 09/30/14

Outputs
Target Audience:Limited resource families and minority land owners identified by the Cooperative Extension Program of the University. Changes/Problems:Unpredictable changes in Texas weather including delayed rain fall, flooding of University field plots at crop planting period, delayed assumption of duties by postdocs and specialists, lack of students to be trained on project. What opportunities for training and professional development has the project provided? It has provided opportunities for experiential learning by many students: Sanique South, Edna Idan, Dewisha Johnson, Diadrian Clarke, Feyisayo Akindoju in biochemistry and molecular biology. Some of the students have leveraged the superior training to gain admission to doctoral programs in human nutrition (Sanique South), masters degree program in chemical engineering (Dewisha Johnson). Feyisayo Akindoju's chemistry masters degree research project was on the extraction of camphor from Phyla dulcis leaves. Medicinal plants research project has provided the opportunity to employ a post-doctoral student to undertake deeper enzymology, immunochemical, and toxicological studies on Phyla and Hoodia leaf extracts. How have the results been disseminated to communities of interest?Though research on these plants is still on-going, limited resource farmers and other interest groups are frequently introduced to this research program and the concept during field days and other activities organized by the cooperative extension program of the College of Agriculture and Human Sciences. Our observations show that these farmers are highly interested in pursuing medicinal plants as new crops. In addition, some of the farmers are enthusiastically contacting the Plant Systems Group for information and recommendations on other medicinal plants of interest. To this end, the Plant Systems Group is expanding its research focus to cover specialty crops which also include medicinal vegetables and highly nutritious fruits with a high potential of generating extra cash for the approximately 8.4 million limited resource citizens in the State if the crops are pursued as alternative to traditional crops. What do you plan to do during the next reporting period to accomplish the goals?Research efforts will include: Development of rapid propagation technology for Hoodia Characterization of Hoodia mRNAs encoding steroid metabolic enzymes. Development of cell culture technology for optimized sweetener production by Phyla Identification through molecular biology procedures the metabolic variants of Phyla with diminished compositions of camphor increased compositions of sweetener Dissemination of technology for production of Phyla metabolic variants to limited resources farmers. Preparation of antibodies to Phyla terpene synthases and Western blot applications for characterization of Phyla metabolic variants. Metabolomic and genomic characterization of Phyla metabolic variants.

Impacts
What was accomplished under these goals? Since 2013, the new leadership at the CAHS has re-focused efforts of the Plant Systems research branch to medicinal plants and related initiatives that have the best potential to help a very large number of limited-resource farmers in Texas to increase their income. The medicinal plant research will provide knowledge, knowledge-based solutions and enabling technology for developing high-valued medicinal plants into specialty crops for limited-resource farmers to grow in Texas. It will enable limited-resource farmer in Texas to share the financial benefits of a rapidly developing industry of high-margin dietary supplements and natural products. Towards this goal, researchers at CAHS have made substantial progress in the past two years, as highlighted below. Establishment of living collections of more than 500 medicinal plant species, which lays the foundation for basic and applied research on these plants by researchers of CAHS, and collaborators in many medical research organization in the greater Houston area. Full registration of the living collection, and collaboration with the Botanic Garden Conservation International (BGCI) and the International Plant Exchange Network (IPEN), which enable our Medicinal Plant Research program to access other relevant germplasm world-wide. Initiation of basic and applied research on two high-valued medicinal plant species, Phyla dulcis and Hoodia gordonii, which have a great potential to be developed into special crops in Texas for value-added dietary supplements. Phyla dulcis is native to South America and has a natural zero caloric (non-sugar) sweetener that is a thousand time sweeter than sugar. Hoodia gordonii originates from Namibia and is locally used as an appetite suppressant. Both plants contain compounds that can be used in commercial dietary products such as supplements to manage obesity and diabetes. Two more species, Siraitia grosvenori (formerly called Momordica grosvenori) and Helianthus tuberosus which contain zero caloric sweeteners have been acquired also. They are growing well in the green house and ready to go to outside plots. These species will be expanding our studies. All the medicinal plants contain compounds that can be used in commercial dietary products such as supplements to manage obesity and diabetes. Some of the results of the basic and applied research have already yielded two peer-reviewed publications: Godson O. Osuji, Aruna Weerasooriya, Peter A.Y. Ampim, Laura Carson, Paul Johnson, Yoonsung Jung, Eustace Duffus, Sela Woldesenbet, Sanique South, Edna Idan, Dewisha Johnson, Diadrian Clarke, Billy Lawton, Alfred Parks, Ali Fares, Alton Johnson. (2015). Molecular regulation of the metabolic pathways of the medicinal plants: Phyla dulcis. American Journal of Plant Sciences, 6 (11) in press. R. Adams, A. Weerasooriya, M. Gao (2014). Comparison of leaf terpenoids from Lippia dulcis (Verbenaceae) obtained by steam distillation and pentane liquid extraction. Phytologia. 96 (3): 252-259.

Publications