Progress 09/01/09 to 08/31/11
Outputs
OUTPUTS: A habitat model based on existing locations of black cohosh was developed. From known occurrences, Mahalanobis distance techniques were used to predict the location of additional populations. Conditions most likely to predict populations were determined using GIS layers. Populations of black cohosh (115) were found in Potomac-Garrett, Savage River, and Green Ridge State Forests in western Maryland. Seventy-three percent of observed locations were classified as ideal habitat. Ideal habitat occurred on lower slopes, higher values of topographic convergence index, smaller "distance to stream" values and higher pH than average. Over 128,000 seeds were collected from individual plants in different physiographic regions of western Maryland. Researchers in the chemistry department conducted chemical analysis and functional analysis of key constituents. Results were presented at national meetings of the American Chemical Society, as well as the American Society of Pharmacognosy and the Phytochemical Society of North America. A study on the ecological relations of black cohosh was also conducted. Field data was collected on insect visitors and pollinators of black cohosh, with pollen loads from bees examined under scanning electron microscopy and much of the pollen confirmed to be black cohosh. This data was used to teach and mentor students in several Biology courses and was presented at a summer 2011 ACES symposium. Methods for increasing levels of terpenoid active ingredients in black cohosh and eludication of their role in plant defense were also examined, with results presented at several plant society meetings. Additional outputs achieved by our collaborating partner, University of Maryland Biotechnology Institute, include: 1. Conducted an EST sequencing/gene discovery effort using verified plant specimens that identified roughly 70 genese involved in secondary metabolism in black cohosh. 2. Efficient seed germination schemes were developed for the reproducible growth of certified plants, enabling the establishment of a diverse crop of black cohosh for molecular studies. 3. An invtro cell suspension culture of black cohosh was established and its ability to synthesize potential bioactive compounds was confirmed. 4. DNA barcoding for species authentication of wild and commercial accessions of black cohosh was conducted. 5. A detailed molecular characterization of bioactive tryptamine metabolism in black cohosh to elucidate the mechanisms for the production of N-methyserotonin was undertaken. Outreach achievements included hosting a Mountain Herbalist workshop series in Frostburg from September 2009-May 2010, with over 200 individuals participating. In 2010-2011, a Mountain Roots series was offered in West Virginia with over 600 participants in 13 West Virginia counties attending. An online newsletter was established and circulated to approximately 1600 individuals. Two symposia and one workshop for growers and harvesters were also held. PARTICIPANTS: Dr. Joseph Hoffman, Dean of the College of Liberal Arts and Sciences at Frostburg State University, served as principal investigator on this project, overseeing the research efforts from faculty within the various disciplines at FSU. Other key personnel from FSU were Mimi Hernandez (Outreach Coordinator) who conducted workshops and symposia for growers and harvesters in Maryland and West Virginia, Dr. Peggy Biser from the Chemistry Department who performed the chemical analysis, Dr. Sunshine Brosi from the Biology Department who worked on locating and documenting black cohosh populations in Western Maryland, as well as Dr.Dan Fiscus and Dr. David Puthoff from the Biology Department who conducted the analysis on ecological relations of black cohosh. Dr. James Howell from Allegany College of Maryland partnered with Dr. Brosi on locatintg and documenting black cohosh populations. Joseph Ferrari from the Appalachian Environmental Laboratory partnered with Drs. Brosi and Howell on the habitat modeling. UM faculty participating were Edward Eisenstein, Martin Spiering, and Bhavneet Kaur. In addition to the faculty working on this project, eight undergraduate students and five graduate students from FSU were involved in the research. TARGET AUDIENCES: Outreach activities for this project were targeted toward growers, harvesters, and herbal practitioners in the Appalachian region. Ethnobotany and science students at Frostburg State University were also a target audientce, participating in experiential learning opportunities and receiving classroom instruction related to the project research. Scientists from federal agencies and other academic institutions were also targeted as workshop participants. PROJECT MODIFICATIONS: Not relevant to this project.
Impacts
1) Locating black cohosh populations and conducting habitat modeling - Outcomes include: a) increased understanding of black cohosh habitat requirements which can be used to guide forest management plans for non-time forest products, b) Information generated on the plant's ecological communities, current availability, health, and reproduction. 2) Chemical analysis - New approaches were developed for chemical analysis of key constituents. New protocols for fingerprinting rhizome samples were also developed. 3)Insect visitors and pollinators of black cohosh - Unique bee visitors (568) to black cohosh were captured, with most of them likely pollinators. In addition, new insect exclusion techniques were developed. Added and improved skills and capacities were developed for determination of insect pollinators and for ecological network analysis. 4) Methods for increasing levels of terpenoid active ingredients in black cohosh and eludication of their role in plant defense - Black cohosh plants that have flowered during the current year have a higher level of the active ingredient deoxyactein. Wounding the inactive side of a harvested rhizome leads to increased levels of actein and deoxyactein. This suggestes that these two compounds are associated with defending black cohosh from a predatory attack. Salicylic acid treatment of rhizomes also increases, while treatment with jasmonic acide and ethylene do not increase the levels of actein and deoxyactein, suggesting that these two active ingredients may also play a role in plant defense against microbial pathogens. 5) UMBI outcomes - Results have laid a solide foundation for optimizing plant growth and culture conditions for sustainable propagation of black cohosh, enabling a more comprehensive analysis of the biosynthesis of triterpene glyscosides, phenylpropanoid esters and tryptamines, the key bioactive compounds found in black cohosh. This should facilitate future efforts to develop plant lines with improved phytochemical qualities. 6) Outreach efforts - Establishment of a network of growers and harvesters in western Maryland and the surrounding tri-state area.
Publications
- Brosi, S., 2010. Sustainability of an Appalachian herb, black cohosh, Actaea racemosa L.: determining factors of habitat and abundance in western Maryland. Proc. of the Society for Economic Botany Conference, Xalapa, Mexico.
- Ferrari, J., Brosi, S., and Howell, J. 2010. Habitat suitability for the medicinal plant black cohosh (Actaea racemosa, L.) in western Maryland - a baseline study for management of non-timber forest resources. Proc. of the 2010 International Association of Landscape Ecology, US-IALE Twenty-fifth Anniversary Symposium, University of Georgia, Athens, GA.
- Brosi, S. 2010. People and Plant Communities of the Appalachian Mountains: Ethnobotany in western Maryland. Proc. of the 33rd Annual Appalachian Studies Conference, Engaging Communities, North Georgia College and State University, Dahlonega, GA.
- Martin J. Spiering, Lori A. Urban, Donald L. Nuss, Vivek Gopalan, Arlin Stoltzfus and Edward Eisenstein (2011) "Gene identification in black cohosh (Actaea racemosa L.): Expressed sequence tag (EST) profiling and genetic screening yields candidate genes for production of bioactive secondary metabolites" Plant Cell Reports 30: 613-629.
- Bhavneet Kaur and Edward Eisenstein (2011) "Establishment of black cohosh suspension cell cultures for the production of bioactive secondary metabolites" submitted.
- Bhavneet Kaur and Edward Eisenstein (2011) "High percentage, season-independent seed germination in black cohosh (Actaea racemosa L.)" submitted.
- Chanea Fraser, Akinola Vaughan, Anne Osano, Martin J. Spiering and Edward Eisenstein (2011) "DNA barcoding for species authentication of wild and commercial accessions of black cohosh (Actaea racemosa L.)" in preparation.
- Abstract: A. J. Vickers, S. L.Brosi, J. A. Howell and D. P. Puthoff, Concentrations in triterpene glycoside levels found to differ between reproductive and non-reproductive black cohosh. Plant Biology 2011, American Society for Plant Biologists, Minneapolis, MN, Aug. 6-10, 2011.
- Abstract: Annette DeCapite, Tyler Lancaster, and David Puthoff, Elucidation of terpenoid medicinal compound levels in Black cohosh (Actaea racemosa) rhizomes in response to plant defense signaling molecules Mid-Atlantic Plant Molecular Biology Society Meeting, Laurel, MD Aug. 15-16, 2011.
- Abstract: A. J. Vickers, S. L.Brosi, J. A. Howell and D. P. Puthoff, Concentrations in triterpene glycoside levels found to differ between reproductive and non-reproductive black cohosh. Mid-Atlantic Plant Molecular Biology Society Meeting, Laurel, MD Aug. 15-16, 2011.
- Abstract: Amanda Vickers, Sunshine L. Brosi, James Howell, and David Puthoff, Chromatographic quantification of medicinal compounds in flowering and non-flowering wild-harvested Actaea racemosa L. Botany 2011, Healing the Planet, St. Louis, Missouri, July 9-13, 2011.
- Carter, M. and Biser, P.: Evaluation of solvent affects on anti-oxidant capacity levels of Black Cohosh (Actaea racemosa) General Poster Session: Analytical Chemistry. The 241th ACS National Meeting; 2011, March 27-31; Anaheim, CA.
- Amanda Vickers, Sunshine L. Brosi, James A. Howell, and David P. Puthoff, Assessment of five medicinal compounds in flowerring and non-flowering black cohosh, Appalachian Studies Conference Proceedings, Eastern Kentucky University, Richmond, KY, March 11, 2011.
- Sadeghipour, M., Biser, P., Brosi, S., and Howell, J. 2010. Analysis of solvent and the environmental effect on the Phenloic contents of Black Cohosh. Proc. of the 51st Annual meeting of the American Society of Pharmacognosy and the Phytochemical Society of North Ameerica, St. Petersburg, FL.
- Brosi, S., and Walsh, N., 2010. Stratification protocols and germination rates of black cohosh (Actaea racemosa L.) populations from western Maryland. Proc. of the Society for Economic Botany Conference, Xalapa, Mexico.
- Sadeghipour,M., Biser, P., Brosi, S., and Howell, J.: Analysis of solvent and the environmental effect on the Phenolic contents of Black Cohosh: The 51st Annual meeting of the American Society of Pharmacognosy and the Phytochemical Society of North America; 2010, July 10-14; St. Petersburg, FL.
- Brosi, S. and Walsh, N. 2010. An Herb Grows in Appalachia: Sustainability of Black Cohosh Populations in Western Maryland. Proc. of the 33rd Annual Appalachian Studies Conference, Engaging Communities, North Georgia College and State University, Dahlonega, GA.
- Minnick, A.J. and Biser, P.: Black cohosh: Soxhlet extraction vs. low temperature method. Poster session presented at: General Poster Session: Analytical Chemistry. The 239th ACS National Meeting; 2010, March 21-25; San Francisco, CA.
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