Source: MIDDLE TENNESSEE STATE UNIVERSITY submitted to
EVALUATING DISEASE PREVENTION METHODS FOR ORGANIC AMERICAN GINSENG PRODUCTION IN APPALACHIA, AN INTEGRATED RESEARCH, EDUCATION, AND EXTENSION APPROACH
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
ACTIVE
Funding Source
OTHER GRANTS
Reporting Frequency
Annual
Accession No.
1027241
Grant No.
2021-51106-35583
Cumulative Award Amt.
$455,000.00
Proposal No.
2021-04836
Multistate No.
(N/A)
Project Start Date
Sep 1, 2021
Project End Date
Aug 31, 2025
Grant Year
2021
Program Code
[112.E]- Organic Transitions
Project Director
Gao, Y.
Recipient Organization
MIDDLE TENNESSEE STATE UNIVERSITY
1301 EAST MAIN STREET
MURFREESBORO,TN 37132
Performing Department
Agriculture
Non Technical Summary
American Ginseng (AG, Panax quinquefolius L.) is one of the highest unit value cash crops and one of the most well-known medicinal plants originating from North America. After being harvested for over 300 years from the Appalachian Mountains in North America, wild AG has become scarce and now almost all the AGs sold worldwide is cultivated (Liu et al., 2021). Despite its production only contributing to less than 8% of the total world AG production, the demand for high-quality AG has increased steadily in the past decade both domestically and internationally, especially those produced from wild-simulated and/or organically managed systems. Because ginseng is well recognized for its health-promoting properties such as boosting immunity and preventing respiratory diseases (Yuan et al., 2010; Predy et al., 2005), the demand has increased during the COVID-19 pandemic. Particularly, due to the scarcity of dedicated certified organic land in the major ginseng production countries (USA, Canada, China, and South Korea), American farmers have a unique organic market niche available that can greatly enhance sustainability of rural areas that are not suitable for large-scale agronomic crop production but perfectly suited for ginseng growth (e.g. wooded, slopping, and rocky areas). However, the greatest immediate problem challenging organic American ginseng production is fungal diseases control, which typically requires frequent, sometime even daily use of costly fungicides (Davis, 2002). Persistent fungal pathogen issues can not only jeopardize biomass production but decrease microbial diversity in soils, creating a "replanting issue" against continuous monoculture production (Li et al., 2020; Yang et al., 2015). Presently, organic and transitional farmers cannot find scientifically published data or Extension publications on how to organically manage fungal pathogen issues in ginseng production. Thus, our goals for this proposed project are to (1) evaluate the effectiveness of biological control microbial agents and botanical extracts on fungal disease prevention and treatment for organic AG production; (2) stabilize changes to the soil microbiome that occur under continuous AG production by increasing native bacterial species known to be antagonist to fungal pathogens and toxin accumulation; and (3) promote sustainable organic production and certification to local, regional and national ginseng producers by emphasizing both the consumer's market preference and higher profit margin; (4) develop a comprehensive educational and outreach program to efficiently translate our project findings to stakeholders and the general public. The research components of this multipronged project will increase agroecosystem resilience while attenuating fungal pathogen pressure and lessening the replanting digression issue. Additionally, we will rely on established education and Extension network to disseminate our findings to different stakeholder groups on a regional and national scale. The proposed program would be the first multi-institutional, multidisciplinary, and collaborative project on organic AG production across the Appalachia region; which could cohesively bolster organic AG production, increase market competitiveness and diversity for organic ginseng producers, enhance the economic sustainability in rural areas that mainly supports ginseng production, and maintain values and ethnic traditions in American rural communities as a whole.
Animal Health Component
35%
Research Effort Categories
Basic
35%
Applied
35%
Developmental
30%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
2152220116050%
1230680106050%
Knowledge Area
215 - Biological Control of Pests Affecting Plants; 123 - Management and Sustainability of Forest Resources;

Subject Of Investigation
2220 - Medicinal crops, non-narcotic; 0680 - Other products of the forest;

Field Of Science
1060 - Biology (whole systems); 1160 - Pathology;
Goals / Objectives
The four pillars in the sustainable agricultural system are commonly considered as production efficiency, economic dynamics, ecological impacts, and social responsibility. Our ultimate goal of this project is to develop scientific solutions for and to promote sustainable organic AG production in Appalachia using an integrated research, education, and extension approach (Fig. 1). A four-stage research effort will be conducted sequentially in laboratory, the green house, small-plot organic lands; and producer sites. All four research stages will be integrated with education and extension components. This multipronged system approach will help addressing key managerial issues challenging organic AG production and help increase biodiversity both above and below ground resulting in a more resilient ecosystem while attenuating fungal pathogen pressure and lessening the replant issue. This approach may be easily adapted by regionally divergent farms by adjusting selected bacterial endophyte and cover crop species. Specifically, we have five objectives of this program.Objective 1: Evaluate the effectiveness of biological control microbial antagonists on fungal disease prevention and treatment for organic AG production.The use of biological control agents to both suppress pathogen presence and promote growth in organic agriculture is well documented (O'Callaghan, 2016). However, few studies have demonstrated their effectiveness on ginseng production. Many of these studies were carried out using P. ginseng. Some bacterial antagonists are already on the market for control of Pythium; Companion (B. subtilis), Primastop (Gliocladium caterulatum), Actinovate (Steptomyces lydicus), Mycostop (Steptomyces griseovirdis), PlantShield (Trichderma harzanum), and SoilGard (Trichoderma virens) are all marketed for use against Botrytis, Fusarium, Phytophthora, and Rhizoctonia (Moorman, 2011). These products however, do not specifically mention their use or efficacy on AG (P. quinquefolius). Here we seek to fill in gaps in our collective knowledge of how the use of biological agents will protect AG production.Fungal pathogens tend have a limited range of host species that they harm, often refereed to has host specificity (Li, Cornelissen, & Rep, 2020). Because of this circumstance, the fungal pathogens effecting AG will be collected and identified from wild environments, collaborating ginseng growers and academic researchers. Meanwhile, bacterial endophytes from regional AG genotypes will also be collected, screened against the isolated pathogens, and identified. The effect of the bacterial antagonists on the growth of AG plant and plant resistance to fungal disease will be investigated. In addition, because phytochemical content is one of the key factors determine ginseng's value and it has been reported that bacterial antagonist could increase the phytochemical contents, the phytochemical accumulation after bacterial inoculation will also be investigated.Objective 2: Evaluate the effect of biological control microbial antagonists on soil physiochemical and biological conditions.Soil-borne ginseng fungal pathogens alter the soil microbiome by reducing the species and growth of beneficial bacteria. Our hypothesis is that by introducing or increasing native bacterial species known to be antagonist to fungal pathogens, we can restore the bacterial diversity and population size of antagonistic bacteria that were altered by the fungal pathogen or continuous ginseng production. Our approach will be to assess bacterial diversity and population size in the rhizospheric soil of healthy anddiseased AG. Metabarcode analysis will be used to survey prokaryotic and fungal taxa associated with healthy and diseased AG plants. If specific taxa are found to be associated with healthy AG plants it may be possible to identify those taxa and introduce them into the soils as part of an organic cultivation strategy. These results will allow us to quantify bacterial population and diversity reductions in soil involved in AG production and offer an important baseline number. Bacterial strains shown to be antimicrobial will then be reintroduced into ginseng production fields at levels at or above our baseline finding.Objective 3: Evaluation of the antifungal effect of natural botanical extracts against AG's fungal pathogens.The severity of ginseng replant disease appears linked to the production environment. While replant failure is common in cultivated AG production, it is much less common in forest farmed AG cropping systems. However, fungal disease tends to increase with increasing intensification of forest farming practices. Wild populations of AG contain assemblages of associated plant species, and growers evaluating sites for wild-simulated plantings use these associated plants as 'indicator species' in determining suitability for AG reintroduction. Some of these indicator plant species (i.e. Adiantum pedatum, Arisaema triphyllum, Botrychium virginianum, Caulophyllum thalictroides, and Podophyllum peltatum, Polygonatum biflorum), as well as some woodland companion crops sometimes interplanted with AG (i.e. Allium tricoccum, Cimicifuga racemosa, Hydrastis canadensis), are reported to possess anti-fungal properties. We hypothesize that these plant associates contribute to the reduced disease incidence found in wild or wild-simulated AG populations through antifungal root exudates or compounds in decomposing leaf litter. To test this hypothesis, we will evaluate the antifungal potential of ethanol and aqueous extracts made from select indicator species and companion crops with the goal of developing simple DIY anti-fungal soil treatments that will be shared with growers through our outreach efforts. We will identify these select candidates for study using grower experience coupled with our knowledge of associated flora and possible novel antifungal chemistry associated with a particular species.Objective 4: Promote sustainable organic production and certification to local, regional and national ginseng producers by emphasizing both the consumer's market preference and higher price yield the anticipated research solutions will deliver.The authentication of herbal products through organic certification provides confidence and quality assurances to consumers who are willing to pay for the price premium of certified ginseng products that involve no chemical inputs. Two current examples are the 'Grown in Wisconsin' Seal - simple geographic certification that has increased demand and relative market price (Ginseng Board of Wisconsin 2021); and in Asia, the Chinese "Green Food Certificate" (a partial equivalent to our USDA organic certification) also drives both demand and price for organic products (Liu et al., 2021; China Green Food Development Center). We will develop a market analysis for the cost/return of AG organic transitioning and production and design training models that will help AG producers to become certified. Our outreach will include a training component for ginseng producers by promoting sustainable organic management techniques and certification and on how to niche-market their product at higher prices.Objective 5: Disseminate and transfer the project results to stakeholders and public individuals.Results and best practices identified through this work will be made available through various educational and outreach efforts conducted by project team members and collaborators in various Appalachian states (TN, PA, OH, VA, NC, KY, and NY). An educational component includes teaching activities at MTSU, University of Tennessee, UVA-Wise, and Penn State University and undergraduate research activities in these institutions will be developed.
Project Methods
3.1 Methods for Objective 1.Solicit stakeholder involvement to identify and collect plant materials, bacterial endophytes, and pathogenic fungi The IGI at MTSU will collect both healthy and diseased samples from growers, researchers, and community donors actively solicited by the Tennessee Department of Environment and Conservation (TDEC), the Soil, Plant, and Pest Lab at the University of Tennessee Extension Service, and the North Carolina State University Extension.Collection and Identification of pathogens affecting AG Pathogenic fungi will be isolated or obtained from: 1) symptomatic plants produced from fields and forests of our regional farmer cooperators, and 2) academic plant pathology research laboratories. Infected samples will be triaged in the plant pest diagnostic laboratory at the University of Tennessee Soil, Plant and Pest Center in Nashville, TN.Collection and Identification of bacterial biological control agents (BCAs) and plant growth promoting rhizobacteria (PGPR) from regional AG populations Bacterial BCAs and plant growth promoting rhizobacteria will be isolated from roots of regionally produced AG from our farmer cooperators fields and forests.In vitro & in vivo evaluation of antifungal activity of bacterial BCAs Upon successful isolation, these bacterial endophytes will be screened for their ability to: 1) suppress fungal pathogen growth in vitro by direct (physical contact) inhibition via dual culture assay.2) Suppress fungal pathogen infection in vivo by inoculating AG plants with suspected bacterial antagonist (identified previously) and planting these into soil infested with a pathogen.3) Promote AG growth rate in vivo by inoculating seedlings with liquid cultures of the bacteria and recording growth rate, plant bio-mass, and chlorophyll content using AG seedlings grown in a controlled growth chamber environment. 4) Bacterial endophytes shown to possess efficacy against fungal infection from the growth chamber experiments will advance for further evaluation under greenhouse conditions.Field test of antifungal activity of bacterial BCAs The field trial site is located at the MTSU Experiential Learning and Research Center.Bacterial endophytes shown to possess good efficacy for disease control and/or growth promotion will advance to field trials. These isolates will be cultured in appropriate liquid media and applied to AG by basal drench to established AG plants actively growing in the organic field. Alternatively, when establishing new field sites, AG seedling roots will be dipped liquid suspension of bacterial endophyte and transplanted in the organic field under the artificial shade.3.2 Methods for Objective 2.Soil sample collection Soil samples will be collected from April through October during ginseng's growing season.Assess the changes of soil physicochemical properties and nutrients with the presence of fungal pathogens Soil samples associated with healthy and symptomatic AG plants will be analyzed and compared for soil physiochemical properties and nutrients.Assess the changes of soil biological condition with the presence of fungal pathogens Bacteria and fungi in soil samples associated with healthy and symptomatic AG plants will be surveyed with metagenomic barcoding and compared. For bacterial metabarcoding, the V4 region of the 16S rRNA gene is used to estimate microbial biomass and identify microbial species.Evaluate the potential of bacterial BCAs on restoring the soil microbiota altered by fungal pathogens.3.3 Methods for Objective 3.Solicit stakeholder involvement to identify and collect plant material and pathogenic fungi A meta-analysis will be conducted in conjunction with solicitations of AG growers within our network for historical disease incidence within their AG plots containing differing quantities and densities of indicator species or interplanted companion crops.Identify and collect plant materials Plant material from indicator and companion crop species that have been shown to exhibit antimicrobial activity will be collected from actively growing wild or wild-simulated ginseng populations in our farmer cooperators fields and forests.The aqueous and ethanolic extracts of plant materials from indicator and companion crop species will be prepared using a sonication extraction method (Vinatoru, 2001).In vitro & in vivo evaluation of antifungal activity of plant extracts Plant extracts will be assessed for their antifungal activities in vitro and in vivo.In vivo evaluation will be conducted in the MTSU greenhouses using methods described in the in vivo fungal suppression portion of Objective 1, but substituting plant extracts for the bacterial cultures.Phytochemical analysis of effective plant extracts Plant extracts that are shown to possess efficacy against fungal infection from the in vitro and in vivo experiments will be subjected to phytochemical analysis using ultra high performance liquid chromatography (UHPLC) to determine the main active phytochemicals responsible of the antifungal activity.Field test of botanical extract pathogen suppression Field tests will be conducted in the certified organic land the MTSU Experiential Learning and Research Center in Lascassas, TN. Plant extracts that exhibited antifungal activity in the lab and greenhouse studies will be used as a seed treatment prior to field planting.3.4 Methods for Objective 4.A market analysis for the cost/return of organic transitioning and production This analysis will include budgeting methodology and key component sensitivity tools for the producer to estimate the annual cost of organic production once the transition phase is complete. Taken together with market price data for the organic product, this information will provide a break even estimate for the organic producer.Training events in Tennessee The IGI at MTSU and UT Extension will host training events.Training events in Pennsylvania Team member Dr. Burkhart will organize and lead the development of extension and outreach events through the Shaver's Creek Environmental Center and Forest Resources Extensionat PSU.Training events in Ohio Projector collaborator Dr. Leopold will lead the outreach using existing events hosted by United Plant Savers (UpS), Ohio.Training events in North Carolina Projector collaborator Dr. Hamilton manages a robust program of ginseng and forest-farming outreach in North Carolina offering production workshops and other demonstrations including field days. He will lead the outreach using this existing program at NC Cooperative Extension.3.5 Methods for Objective 5.Dissemination methods Results and best practices identified through this work will be made available through the following dissemination methods by project team members and collaborators: 1) Print media including journal articles, extension publications, brochures, etc.; 2) Existing electronic media including the International Ginseng Institute (IGI)'s website and TDEC's ginseng website, newsletters, and Departmental Twitter and Facebook accounts; 3) Presentations at local, regional, and national academic conferences; and 5) Grower meetings, demonstrations, field visits, and workshops that will be held on multiple sites.Teaching activities at MTSU, UT, UVA-Wise, and PSU Thirteen undergraduate courses taught at MTSU, UVA-Wise, and PSU could greatly benefit from this project.Undergraduate student research activities at MTSU and UVA-Wise The project team members at both institutions will recruit more than ten undergraduate researchers during the project period. These students will be encouraged to disseminate their research associated with organic agriculture production by joint scientific publication and conference presentation.

Progress 09/01/23 to 08/31/24

Outputs
Target Audience:Our target audience included undergraduate and graduate students at the five participating and collaborating institutions, as well as ginseng growers, producers, dealers, and researchers across Tennessee, Virginia, and Pennsylvania. Additionally, our outreach events drew public attendees and engaged diverse groups, including local herbal business owners, state park representatives, and state foresters. Changes/Problems:We requested and received approval for a one-year no-cost extension (NCE), extending the termination date to 8/31/2025. Additionally, we submitted a budget revision to address changes in personnel (a co-PI moved to another institution, the evaluator passed away, and a technician was replaced with a postdoctoral research fellow) as well as adjustments to subawards. Both the extension and budget revision have both been approved. What opportunities for training and professional development has the project provided?The project continues to impact students' professional development through six renovated courses that influenced over 100 students at the principal investigator's institution. Additionally, during the third reporting period, the project provided experimental and professional training to nine students, enhancing their academic development. Internship opportunities created within the project facilitated institutional exchange and furthered student career development. As an extension of the current grant, the project team submitted a proposal to the USDA Organic Transition Grant in 2023 (not funded) and another to the USDA NRCS Conservation Innovation Grant in 2023 (funded). How have the results been disseminated to communities of interest?Results and best practices identified through this work were disseminated through various educational and outreach efforts as shown in Accomplishments of Objective 5. What do you plan to do during the next reporting period to accomplish the goals?We plan to continue our research agenda into the next reporting period as we work toward concluding the project. At the same time, our outreach efforts to promote sustainable organic production and certification among ginseng producers will remain active, along with ongoing dissemination of results to stakeholders.

Impacts
What was accomplished under these goals? Objective 1: Evaluate the effectiveness of biological control microbial antagonists on fungal disease prevention and treatment for organic American Ginseng (AG) production. Accomplishments:We have designated a 10-by-50 ft plot in the experiential learning laboratory at the university farm for our study. Raised beds and a shade structure have been constructed, and the plot is certified organic. However, due to extreme drought in the summer of 2024, we were unable to conduct the field study. With the grant extended by one year, we plan to resume the study in the summer of 2025. Objective 2: Evaluate the effect of biological control microbial antagonists on soil physiochemical and biological conditions. Accomplishments:We are continuing the study comparing the bacterial soil microbiome of healthy and diseased American Ginseng for the potential development of organic fungicides. Our manuscript "The Soil Microbiomes of American Ginseng (Panax quinquefolius) Cultivated in North America Differ by Location But Not Always by Disease Status" was published on PhytoFrontiers in 2024. Objective 3: Evaluation of the antifungal effect of natural botanical extracts against AG's fungal pathogens. Accomplishments:We are continuing our study on antifungal botanical extracts identified during the previous reporting period. This includes examining the phytochemical profile and antifungal properties of goldenseal extract, along with several other promising candidates. Our manuscript "Goldenseal (Hydrastis canadensis L.) extracts inhibit the growth of fungal pathogens in American ginseng (Panax quinquefolius L.)" was published on Molecules Special Issue: Phytochemicals: Extraction,Bioactivities, and Applications in 2024. Additionally, we presented findings on botanical extracts from goldenseal, ginseng, and sweet wormwood (Artemisia annua L.) at four national, regional, and local conferences and meetings. Objective 4: Promote sustainable organic production and certification to local, regional, and national ginseng producers by emphasizing both the consumer's market preference and the higher price yield the anticipated research solutions will deliver. Accomplishments:We have actively promoted sustainable organic management techniques and certification through workshops and presentations at local and regional conferences for ginseng producers. We co-hosted and presented at the Tennessee ginseng diggers and growers workshop in Pikeville, TN in the fall of 2023 to encourage organic certification for ginseng cultivation. Additionally, we participated in the 2024 "Gather to Grow" Forest Farming Conference in Roanoke, Virginia, where Dr. Eric Burkhart (Co-PI) hosted a roundtable discussion on ginseng pest management and highlighted organic approaches for managing ginseng diseases. Objective 5: Disseminate and transfer the project results to stakeholders and public individuals. Accomplishments:Results and best practices identified through this work have been shared through various educational and outreach initiatives. In addition to the workshops and conferences mentioned under Objectives 3&4, we presented on American ginseng conservation at the Conservation Days event at the Barfield Crescent Park in May 2024. We continue to offer six renovated undergraduate and graduate courses to over 100 students at MTSU, which incorporate elements of organic cultivation and sustainable agriculture. Furthermore, nine students (one Ph.D. student, one Master student, and seven undergraduate students) at MTSU, UVA, and PSU have actively participated in research related to this project. Of these, four students presented findings at local, regional, and national conferences, showcasing results and contributing to broader discussions of the project.

Publications

  • Type: Peer Reviewed Journal Articles Status: Published Year Published: 2024 Citation: Maggard, I. J., Deel, K. B., Gao, Y, Cahoon, A. B. (2024) The Soil Microbiomes of American Ginseng (Panax quinquefolius) Cultivated in North America Differ by Location But Not Always by Disease Status. PhytoFrontiers.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2024 Citation: Alnassari, Z., Alnassari, K., Gao, Y., Zhang, M. (2024). Optimization of a Rapid and Efficient Method for Ginsenoside Extraction. 2024 National Conference on Undergraduate Research (NCUR). Long Beach, CA. April 8-10, 2024.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2024 Citation: Isaiah Kam, Gao, Y. (2024). Extraction of sweet wormwood (Artemisia annua L.), a medicinal plant, and evaluation of its terpene content. MTSU Scholar Week. Murfreesboro, TN. March 15, 2024.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2024 Citation: Moore, R., Zhang, M., Gao, Y. (2024). An Untargeted Metabolomic Approach to Studying Anti-Fungal Treatments American Ginseng (Panax quinquefolius). MTSU Scholar Week. Murfreesboro, TN. March 15, 2024.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2024 Citation: Gao, Y. (2024). Inhibitory Effects of Goldenseal (Hydrastis canadensis L.) Extracts on Fungal Pathogens in American Ginseng (Panax quinquefolius L.) 2024 American Phytopathological Society (APS) Southern Division Meeting. February 26-29, 2024, Columbia, SC.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2024 Citation: Isaiah Kam, Gao, Y. (2024). Extraction of sweet wormwood (Artemisia annua L.), a medicinal plant, and evaluation of its terpene content. 2024 Annual Posters at the Capital. Nashville, TN. February 14, 2024.
  • Type: Peer Reviewed Journal Articles Status: Published Year Published: 2024 Citation: Gao, Y.*, Swiggart, E., Wolkiewicz, K., Liyanapathiranage, P., Baysal-Gurel, F., Avin, F. A., ... & Burkhart, E. P. (2024). Goldenseal (Hydrastis canadensis L.) Extracts Inhibit the Growth of Fungal Isolates Associated with American Ginseng (Panax quinquefolius L.). Molecules, 29(3), 556.


Progress 09/01/22 to 08/31/23

Outputs
Target Audience:The target audience we reached includes undergraduate and graduate students at the five participating and collaborating institutions, as well as over 200 ginseng growers, producers, dealers, and researchers in Tennessee, Maryland, North Carolina, and Wisconsin. Additionally, our outreach events attracted public attendees who participated in our outreach events, and we engaged with interested groups such as local herbal business owners, state parks, and state foresters. It's worth noting that ginseng growers come from traditionally underserved communities of all ethnicities. Changes/Problems:We experienced two changes in personnel during the 2022-2023 project period. Unfortunately, our project external evaluator, Mr. Robert "Bob" Beyfuss, passed away in 2023. We have now identified a replacement, Dr. Ying Jin, who will be involved in this project during the 2023-2024 time period.Another change involves the project co-PI, Dr. Seockmo Ku, who transitioned from the project host organization (MTSU) to Texas A&M University in August 2023. Dr. Ku will continue collaborating with the project team on the ongoing research. What opportunities for training and professional development has the project provided?The project significantly impacted students' professional development through six renovated courses that influenced over 100 students at the principal investigator's institution. Additionally, during the second reporting period, the project provided experimental and professional training to 11 students, enhancing their academic development. Internship opportunities created within the project facilitated institutional exchange and furthered student career development. Furthermore, the grant elevated the visibility of the research work leading to invitations to the investigators for seminars, conferences, and academic exchanges. The subsequent section details the major events attended. Beyond these formal gatherings, the increased visibility generated numerous phone calls, emails, and follow-up contacts with individuals who might not have otherwise been aware of the program's work. Furthermore, the work covered in this grant has paved the way for our next grant application. The project team submitted a grant proposal to the USDA Federal-State Marketing Improvement Program (FSMIP) and a grant proposal to the USDA NRCS Conservation Innovation Grant in 2023. How have the results been disseminated to communities of interest?Results and best practices identified through this work were disseminated through various educational and outreach efforts as shown in Accomplishments of Objective 5. What do you plan to do during the next reporting period to accomplish the goals?We intend to carry on our research agenda in the next reporting period. Having identified and tested promising antifungal biocontrol agents in vitro, we will further assess their effectiveness in the greenhouse and field. Meanwhile, our outreach efforts to promote sustainable organic production and certification to ginseng producers will persist, along with the ongoing dissemination of the results to stakeholders.

Impacts
What was accomplished under these goals? Objective 1: Evaluate the effectiveness of biological control microbial antagonists on fungal disease prevention and treatment for organic American Ginseng (AG) production. Accomplishments:We are continuing our work on the isolated and identified ginseng fungal pathogens from the first reporting period. To date, three manuscripts detailing the identification of novel ginseng pathogens have been published in Plant Disease. We also presented the pathogenicity of fungal and bacterial species associated with American ginseng at the Tennessee Academy of Science annual meeting and shared insights into the diversity of the fungal community associated with American ginseng at the 2023 American Phytopathological Society (APS) Annual Meeting - South Division. Additionally, our efforts persist in exploring the antifungal properties of the bacterial endophytes isolated and identified during the first reporting period, utilizing in vitro bioassays. This work is currently in progress. In preparation for the live plant study, we have established two raised beds for planting ginseng stock plants and constructed six plant growth chambers for antifungal assays with live plants. We have acquired approximately 150 three-year-old ginseng plants for use in live assays. Objective 2: Evaluate the effect of biological control microbial antagonists on soil physiochemical and biological conditions. Accomplishments:We are continuing the study comparing the bacterial soil microbiome of healthy and diseased American Ginseng for the potential development of organic fungicides. Soil samples from both healthy and diseased plants in Maryland and Wisconsin were analyzed using environmental DNA metabarcoding to identify predominant bacteria and fungi. Statistical analyses revealed significant differences in alpha- and beta-diversity between locations and inconsistencies in microbial communities associated with diseased plants, advancing our understanding of ginseng soil rhizomes. The findings on the soil microbiomes of healthy vs. blighted cultivated ginseng roots were presented at the 2022 annual meeting of the American Society for Microbiology and the 84th annual meeting of the Association of Southeastern Biologists. Our manuscript on this work was recently accepted by PhytoFrontiers. Objective 3: Evaluation of the antifungal effect of natural botanical extracts against AG's fungal pathogens. Accomplishments:We are continuing the study of antifungal botanical extracts selected during the first reporting period. We have examined the phytochemical profile and antifungal properties of goldenseal extract, along with a few other promising botanical extracts. Our manuscript "Goldenseal (Hydrastis canadensis L.) extracts inhibit the growth of fungal pathogens in American ginseng (Panax quinquefolius L.)" was recently submitted to the Molecules Special Issue: Phytochemicals: Extraction,Bioactivities, and Applications. Additionally, we presented this work at the Tennessee Academy of Science annual meeting. Objective 4: Promote sustainable organic production and certification to local, regional, and national ginseng producers by emphasizing both the consumer's market preference and the higher price yield the anticipated research solutions will deliver. Accomplishments:We conducted an economic study to assess the implications of transitioning from traditional methods of producing ginseng to organic cultivation. The study evaluated the profitability of each method by analyzing costs, expenses, and yields. Our findings suggest that transitioning to organic ginseng reduces the ginseng yield per acre, but the net income for farmers increases substantially. This implies that organic ginseng has significant agribusiness entrepreneurial potential, fostering profitable and sustainable agricultural practices. Our work on the financial implications of transitioning to organic ginseng was presented at the 2023 International Conference of the Institute for Global Business Research (IGBR). Furthermore, we are actively promoting sustainable organic management techniques and certification through workshops, as well as local and regional conferences aimed at ginseng producers. Notably, we hosted a statewide ginseng cultivation workshop in Tennessee with approximately 50 participants and co-hosted a ginseng growers' workshop, where we provided training and spoke about organic ginseng cultivation and certification. Objective 5: Disseminate and transfer the project results to stakeholders and public individuals. Accomplishments: Results and best practices identified through this work have been made available through various educational and outreach efforts. Workshop, growers meeting, field day, demo day: We organized and hosted the Fall 2022 Ginseng workshop at Barfield Crescent Park, Murfreesboro, TN, in October 2022. Four invited speakers presented ginseng harvest data, regulation, site evaluation, planting techniques, intercropping, and growth management techniques. Around 50 participants learned the proper use of dedicated hand tools and planting techniques through expert demonstrations in the Park's natural forest using native seeds. We also distributed 5 pounds of ginseng seeds and goldenseal rootlets to participants for free. Additionally, we participated in and presented American ginseng organic cultivation at the Virtual Field Day at MTSU in March 2023. We co-hosted and presented at Boring Ginseng Roots and Herbs' annual growers' meeting and training workshop in August 2023, engaging in discussions with growers, diggers, buyers, and representatives from herbal companies. Public outreach: We participated in and presented American ginseng conservation at the Barfield Crescent Park Conservation Days event in June 2023. The project team transplanted 50 native ginseng rootlets in the hardwood forest at the Biltmore Estate, Ashville, North Carolina, as part of their Naturalist program. The project team has also conducted over ten meetings (both in-person and virtual) and farm visits with ginseng growers, stakeholders, and collaborators. Digital outreach: Ginseng newsletters (Fall 2022 and Spring 2023 issues) were distributed to 200+ participants via email. Educational component: A component supporting teaching activities at MTSU, University of Tennessee, UVA-Wise, and Penn State University, as well as undergraduate research activities at these institutions, was developed. We continue to offer six renovated undergraduate and graduate courses that include organic cultivation and sustainable agriculture elements, to over 100 students at MTSU. A total of 11 students (one Ph.D. student, three Master students, and seven undergraduate students) at MTSU and UVA have participated in the research of this project, with seven of them presenting at local, regional, or national conferences to report results from the project.

Publications

  • Type: Journal Articles Status: Accepted Year Published: 2023 Citation: Maggard, I. J., Deel, K. B., Gao, Y, Cahoon, A. B. (2023) The Soil Microbiomes of American Ginseng (Panax quinquefolius) Cultivated in North America Differ by Location But Not Always by Disease Status. Accepted by PhytoFrontiers on Nov 19, 2023.
  • Type: Journal Articles Status: Published Year Published: 2023 Citation: Liu, Z., Moore, R., Gao, Y., Chen, P., Yu, L., Zhang, M. and Sun, J. (2023). Comparison of Phytochemical Profiles of Wild and Cultivated American Ginseng Using Metabolomics by Ultra-High Performance Liquid Chromatography-High-Resolution Mass Spectrometry. Molecules, 28(1), p.9.
  • Type: Journal Articles Status: Published Year Published: 2023 Citation: Liyanapathiranage, P., Avin, F.A., Swiggart, E., Lopez, E.F., Parajuli, M., Oksel, C., Gao, Y. , Baysal-Gurel, F. (2023). First Report of Leaf Spot of Panax quinquefolius Caused by Pestalotiopsis nanjingensis in Tennessee and the United States. Plant Disease. ( Co-corresponding authors).
  • Type: Journal Articles Status: Published Year Published: 2023 Citation: Liyanapathiranage, P., Avin, F.A., Oksel, C., Swiggart, E., Gao, Y. , Baysal-Gurel, F. (2023) First Report of Rusty Root of Panax quinquefolius Caused by Pseudomonas marginalis in Tennessee and the United States. Plant Disease. ( Co-corresponding authors).
  • Type: Journal Articles Status: Submitted Year Published: 2024 Citation: Goldenseal (Hydrastis canadensis L.) extracts inhibit the growth of fungal pathogens in American ginseng (Panax quinquefolius L.). Submitted to Molecules Special Issue: Phytochemicals: Extraction, Bioactivities and Applications.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Maggard, I. J., Deel, K. B., Gao, Y., Cahoon, A. B. (2022). Soil Microbiomes of Healthy vs Blighted Cultivated Ginseng (Panax quinquefolius) Roots. 2022 annual meeting of the American Society for Microbiology, Kentucky/Tennessee Brach. Johnson City, TN. October 21-22, 2022.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Wolkiewicz, K., Swiggart, E., Gao, Y. (2022). Extraction efficiency of goldenseal (Hydrastis canadensis L.) using different solvents and plant tissues. The 132nd Tennessee Academy of Science annual meeting, Nashville, TN. November 18, 2022.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Liyanapathiranage, P., Avin, F.A., Oksel, C., Swiggart, E., Gao, Y., Baysal-Gurel, F. (2022). Pathogenicity of fungal and bacterial species associated with American ginseng (Panax quinquefolius). The 132nd Tennessee Academy of Science annual meeting, Nashville, TN. November 18, 2022.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: Oksel, C., Liyanapathiranage, P., Avin, F.A., Swiggart, E., Gao, Y., Baysal-Gurel, F. (2023). Insights into the diversity of the fungal community associated with American ginseng (PanaxquinquefaliusL.). 2023 American Phytopathological Society Annual Meeting - South Division. Durham, NC. February 13-16, 2023.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: Maggard, I. J., Deel, K. B., Gao, Y., Cahoon, A. B. (2023). Soil Microbiomes of Healthy vs Blighted Cultivated Ginseng (Panax quinquefolius) Roots. The 84th annual meeting of the Association of Southeastern Biologists. Winston-Salem, NC. March 23-26, 2023.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Conference Oral Presentation: Bernard, E.C., Lopez, E.F., Gao, Y., Swiggart, E. (2022). Leaf lesions on American Ginseng associated with a foliar nematode, Aphelenchoide ssp., in Tennessee. The 61st Annual Meeting of the Society of Nematologists. Anchorage, Alaska, September 26-29, 2022.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Conference Oral Presentation: Moore, R., Liu, Z., Sun, J., Gao, Y., Zhang, M. (2022). Metabolomic Study of Wild American Ginseng and Cultivated American Ginseng Roots by UHPLC-HRMS and Chemometrics. 2022 SCIX Conference, October 2-7, 2022.
  • Type: Other Status: Published Year Published: 2023 Citation: 2023 Spring Ginseng Newsletter
  • Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: Gao Y. (2023) Evaluating disease prevention methods for organic American ginseng (Panax quinquefolius) production in Appalachia. USDA NIFA: OREI & ORG Project Directors Meeting. Washington D.C. Apr 19-20, 2023.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: Arik, M., Gao, Y., Ho, H. (2023). Financial Implications of Transitioning to Organic Ginseng for Agribusiness. Institute for Global Business Research (IGBR) International Conference. Nashville, TN. April 19-21, 2023.


Progress 09/01/21 to 08/31/22

Outputs
Target Audience:The target audience we reached during the first reporting period includes undergraduate and graduate students at the five participating and collaborating institutions, over 200 ginseng growers, producers, dealers, and researchers in Tennessee, Maryland, North Carolina, and Wisconsin, over two thousand general public attendees who participated in our outreach events (e.g. Conservation Days and Smithsonian's Folklore Festival), and interested groups such as local herbal business owners, state parks, and state foresters. Ginseng growers are traditionally from underserved communities of all ethnicities. No economic demographic data was collected. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?This grant has provided some exceptional and even unexpected opportunities for both training and professional development. It has had an impact beyond the anticipated goals in ways that were not envisioned when the grant was sought. In short, early success brought publicity and the publicity opened new avenues for stakeholders, students, and the principal investigators. As an example, the grant activities were noticed by the Smithsonian Institution where the PD, PIs, and the project's lab assistants were invited to present and disseminate knowledge, techniques, and results to approximately 500,000 visitors over seven days at the Smithsonian's Folklife Festival held on the National Mall in Washington DC. Numerous professional contacts were made that have resulted in a broader mind share of the issues involved. An entirely new stakeholder constituency was created by this exposure and individual student interactions allowed for sharing of techniques that made an immediate impact on their work. The project provided educational opportunities that have impacted the professional development of students. For example, six courses were renovated that impacted more than 150 students at the PI's institution. In the first-year grant cycle, the project also provided 12 students with experimental and professional training for their academic development. Internships were created in the project that facilitated institutional exchange and student career development. This grant raised the profile of the research work creating invitations to the Investigators for seminars, conferences, and academic exchange. The section following dissemination lists the major events that were attended. In addition, there were countless phone calls, emails, and follow-up contacts with parties that would not otherwise have known of the program's work. Also importantly, the grant's attention attracted some donations to the overall program. How have the results been disseminated to communities of interest?Results and best practices identified through this work were made available through various educational and outreach efforts as below. Digital outreach: Regular Ginseng newsletters (Fall 2021, Spring 2022, and Fall 2022) were distributed to 200+ participants via email. A blog was posted on theSoil, Plant and Pest Center, UT Ag extension website to disseminate the information on labeled fungicides for ginseng. An Inaugural Podcast Episode was produced featuring Mr. Bob Beyfuss. A project member was interviewed by SeedBroadcast and WPLN about American ginseng. The story was later picked up by WBUR and aired nationally on NPR's Here & Now program. Workshop, growers meeting, field day, demo day: We organized and hosted Fall 2021 Ginseng workshop at Barfield Crescent Park, Murfreesboro, TN. Four invited speakers presented ginseng relevant harvest data, regulation, pest control, site evaluation, seed planting, and growth management techniques. Thirty participants learned how to use dedicated hand tools and planting techniques through expert demonstrations in the Park's natural forest using native seeds. Project team also participated in the annual Ginseng Field Day at Wausau, Wisconsin, hosted by the Ginseng Board of Wisconsin. We visited six large ginseng farms, visited field test plots, and met with approximately 20 growers in Wausau. Dr. Gao (PD) presented an organic ginseng cultivation workshop at the Organic Growers School 2022 Spring Conference in North Carolina with approximately 40 participants in attendance. Dr. Gao also spoke at Boring Ginseng Roots and Herbs annual growers' meeting and met with a group of growers, diggers, buyers, and representatives from herbal companies. The project team gave six presentations as shown in the following list in the Product section. In addition, PD and PIs have had over 20 meetings (in-person and virtual) with ginseng growers, stakeholders, and collaborators. Public outreach: The project team participated and presented American ginseng conservation in the Conservation Days event at Barfield Crescent Park on Earth Day. The project team members also presented American Ginseng's story on the National Mall in Washington D.C. for a week in June 2022 through an invitation from the Smithsonian Institution to participate in the Smithsonian's Folklife Festival. There we presented talks on forest pharmacy and conservation practices relevant to this endangered plant and engaged Festival attendees in one-on-one discussions. Note, participation was funded by the Smithsonian Institution and no USDA funds were used for this event. An educational component supporting teaching activities at MTSU, University of Tennessee, UVA-Wise, and Penn State University, as well as undergraduate research activities at these institutions, was developed. Six undergraduate and graduate courses with over 150 students at MTSU were renovated to include organic cultivation and sustainable agriculture elements (PLSO 1610 Elements of Plants Science (65 students), PLSO 4670 Herbs, Spices, and Medicinal Plants (34 students), PLSO 3350 Soil Fertility and Fertilizers (23 students), FERM 6450 Industrial Applications of Fermentation (5 graduate students), FERM 3710 Brewing, Distilling, and Fermentation Safety and Sanitation (7 students), FERM 1000 Introduction to Fermentation Science (18 students)). A total of 12 students (one Ph.D. student, five Master students, and six undergraduate students) at MTSU and UVA have been participating in research of this project and ten of them have presented at local, regional, or national conferences to report results from the project. In addition, this project provided internship opportunities for two interns who have been sent to the Soil, Plant, and Pest lab of UT Ag Extension in Nashville for training. What do you plan to do during the next reporting period to accomplish the goals?We plan to continue the research agenda during the next reporting period. As in the first reporting period, promising antifungal biocontrol agents were identified and tested in vitro. We will now further investigate the effectiveness of these agents in the greenhouse and field. Meanwhile, we will continue outreach efforts to promote sustainable organic production and certification to ginseng producers and continue to disseminate the results to stakeholders.

Impacts
What was accomplished under these goals? Objective 1: Evaluate the effectiveness of biological control microbial antagonists on fungal disease prevention and treatment for organic American Ginseng (AG) production. Accomplishments: 1. Collaborating with ginseng growers in Tennessee, Maryland, and Wisconsin, as well as academic researchers in pathology labs at Tennessee State University (TSU) and the University of Tennessee at Knoxville (UTK), we have collected more than 70 diseased AG plants from 5 locations in TN, MD, and WI, and isolated, cultured, and identified 21 fungal pathogens (both morphologically and molecularly) that specifically affect AG. To date, 3 species of Alternaria and 6 species of Fusarium have been isolated and identified from the diseased AG samples collected. A total of 102 healthy ginseng plants were prepared and used for fungal pathogenicity studies. These findings resulted in two manuscripts (one has been published) and were reported at the 2022 Tennessee Academy of Science annual conference. 2. Eight bacterial endophytes were isolated from regional AG genotypes and identified molecularly. The bacterial endophytes were screened against isolated pathogens for their antifungal potentials using dual culture assay, cell-free assay, and split plate assay methods. Two of the bacterial endophytes showed effectiveness against fungal pathogens. 3. We have summarized the fungicides labeled for use on ginseng in TN. A total of 138 labeled fungicides including organics were summarized in an excel spreadsheet. The summary results are available for the first time for the TN ginseng growers. The results were disseminated in the Fall 2021 ginseng workshop and a blog "Fungal Pathogens of American Ginseng, Panax quinquefolius" was posted on the Soil, Plant and Pest Center webpage on the University of Tennessee Ag extension website. Objective 2: Evaluate the effect of biological control microbial antagonists on soil physiochemical and biological conditions. Accomplishments: Collaborating with ginseng growers in Maryland and Wisconsin, we have collected a total of 40 rhizospheric soils of healthy and diseased AG plants. Metabarcode analysis was used to assess bacterial diversity and population size in the rhizospheric soil of healthy and diseased AG. A total of 4,372,803 sequences were generated among which 229,051 high-quality sequences were available for analysis. Soil analysis identified statistically significant differences between soil samples taken from healthy and diseased plants.One predominant bacterium species in the soil was found to be exclusively associated with healthy AG plants. In addition, we found that soil samples collected from two different cultivation methods (field and woods-cultivated) had a statistically significant difference in their microbiomes. These findings were reported at the 2022 American Society for Microbiology regional conference and a manuscript is under preparation. Objective 3: Evaluation of the antifungal effect of natural botanical extracts against AG's fungal pathogens. Accomplishments: We prepared 12 ethanol or aqueous extracts from selected plant species that have shown promising pathogen suppression properties either in growers' practices or reported laboratory research. We also evaluated their antifungal potential against the AG-specific pathogens using agar dilution assay and split plate assay. Among them, 6 botanical extracts showed effectiveness against AG-specific fungal pathogens. The preliminary results of this study were presented at the South Division of the 2022 American Phytopathological Society (APS) Annual Meeting and a manuscript is under preparation. Objective 4: Promote sustainable organic production and certification to local, regional, and national ginseng producers by emphasizing both the consumer's market preference and higher price yield the anticipated research solutions will deliver. Accomplishments: We hosted workshops and presented at local and regional conferences targeting ginseng producers where we promoted sustainable organic management techniques and certification. Specifically, we hosted a state-wide ginseng cultivation workshop in Tennessee with 30 participants, presented an organic ginseng cultivation workshop at the Organic Growers School 2022 Spring Conference in North Carolina, and also spoke at the 2022 New Crops for Southeast Agriculture Conference in Tennessee. Additionally, a market cost/benefit analysis for AG organic transition and production is under development. Objective 5: Disseminate and transfer the project results to stakeholders and public individuals. Accomplishments: Results and best practices identified through this work have been made available through various educational and outreach efforts. Please see the results disemmination section below for detailed activities.

Publications

  • Type: Journal Articles Status: Submitted Year Published: 2022 Citation: Liyanapathiranage, P., Avin, F.A., Oksel, C., Swiggart, E., Gao, Y. , Baysal-Gurel, F. ( Co-corresponding authors). First Report of Rusty Root of Panax quinquefolius Caused by Pseudomonas marginalis in Tennessee and the United States. Submitted to Plant Disease.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2022 Citation: Gao, Y. Lopez, E.F., Baysal-Gurel, F., Eichas R., Swiggart, E. Fungal and Oomycete diseases in American ginseng (Panax quinquefolius L.) and current project evaluating organic disease control. 2022 American Phytopathological Society Annual Meeting - South Division. March 7-11, 2022.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2022 Citation: 2. Lopez, E.F., Bernard, E.C., Gao, Y., Swiggart, E. Preliminary examination of Aphelenchoides sp. detected from American ginseng, Panax quinquefolius, in Tennessee. 2022 American Phytopathological Society Annual Meeting - South Division. March 7-11, 2022.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2022 Citation: Gao, Y., Alnassari, K.A., Alnassari, A.A., Alnassari, F.A., Alnassari, Z.A. Moore, R., Zhang, M. Optimization of Ultrasound Assisted Extraction of Ginsenosides from Roots of Panax quinquefolius. 2022 ASP Annual Meeting. Charleston, South Carolina. July 21-24, 2022.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2022 Citation: Oral presentation - Gao, Y. Growing ginseng in Tennessee. 2022 New Crops for Southeast Agriculture Conference. Nashville, Tennessee (hybrid mode). March 16-18, 2022.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2022 Citation: Oral presentation - Gao, Y. Organic Ginseng Cultivation: Growing Appalachian Roots. Organic Growers School 2022 Spring Conference. Mars Hill University, North Carolina. March 19-20, 2022.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2022 Citation: Oral presentation - 6. Bernard, E.C., Lopez, E.F., Gao, Y., Swiggart, E. Leaf lesions on American Ginseng associated with a foliar nematode, Aphelenchoide ssp., in Tennessee. The 61st Annual Meeting of the Society of Nematologists. Anchorage, Alaska, Sept 26-29, 2022.
  • Type: Websites Status: Published Year Published: 2021 Citation: Lopez, E.F. & Eichas, R. (2021). Fungal Pathogens of American Ginseng, Panax quinquefolius. https://soillab.tennessee.edu/fungal-pathogens-of-american-ginseng-panax-quinquefolius/
  • Type: Websites Status: Published Year Published: 2021 Citation: Fall 2021 Ginseng Newsletter https://mtsu.edu/ginseng/docs/newsletter-2021-fall.pdf
  • Type: Conference Papers and Presentations Status: Published Year Published: 2022 Citation: Spring 2022 Ginseng Newsletter https://mtsu.edu/ginseng/docs/newsletter-2022-spring.pdf
  • Type: Websites Status: Published Year Published: 2022 Citation: Fall 2022 Ginseng Newsletter https://mtsu.edu/ginseng/docs/newsletter-2022-fall.pdf
  • Type: Journal Articles Status: Accepted Year Published: 2022 Citation: Avin, F.A., Liyanapathiranage, P., Shishkoff, N., Swiggart, E., Gao, Y. , Baysal-Gurel, F. ( Co-corresponding authors). First Report of Powdery Mildew of American Ginseng Caused by Erysiphe heraclei in Tennessee and the United States. Plant Disease. https://doi.org/10.1094/PDIS-10-22-2310-PDN