Item No. 1 of 1

INVESTIGATOR: Colova (Tsolova), V. M.; Ananga, A.; Leong, S.; Legaspi, J.; Mobley, R.; Jacono, C.

PERFORMING INSTITUTION:
FLORIDA A&M UNIVERSITY
TALLAHASSEE, FLORIDA 32307

BUILDING CAPACITY FOR CLEAN PLANT DIAGNOSTIC OF GRAPE & SMALL FRUITS AT FAMU TO ENHANCE RESEARCH & EXTENSION IN FLORIDA & SOUTHEASTERN US

NON-TECHNICAL SUMMARY: The rapid expansions of wine industry in Florida and southeastern U.S. have already predisposed the viticulture enterprise to several disease problems such as Eutypa dieback, Crown gall, leafroll and Pierce's disease. Due to the detrimental impact of these diseases, we have identified them as the high priority diseases for research and extension in the state and the region. Our strategic location in the Southeast region coupled with our success in working with the American native grapes and being part of the National Clean Plant Network (NCPN) for Grape mandate us to develop Diagnostic Facility for American Native Grapes. This project is intended to conduct applied research for grape and small fruits disease diagnostic in NIFA's Emphasis Areas of Plants Sciences, Biotechnology and Pest Management; to build human capacity by improving research training and experiential learning of junior faculty and graduate students at Florida A & M University and to develop "Diagnostic Facility for American Native Grapes" serving as a centralized research support unit and student training platform in pathogen screening and prevention and extension program development for grape clean plant material (Program's Need Area: 2b. and 3c.). Our program will impact the state and the southern viticulture industry by providing key pathogen testing and diagnostic of diseases transmitted with planting stock and associated research, methods development and education and outreach for extension personnel and nursery producers. Developing of a "Diagnostic Facility for American Native Grapes" at FAMU will complement the national efforts to enhance the overall supply of clean plant material for grape and other specialty crops.

OBJECTIVES: The primary objective of this capacity building effort is to enhance and strengthen research, teaching, and extension programs at CAFS, FAMU in agriculture science by focusing on a comprehensive screening and diagnostic of grapevine diseases to ensure that our Certified Mother Blocks are maintained disease-free for propagation, and clean plant material is distributed to growers for the sustainability of grape and wine industry in the state of Florida and Southeastern U.S. Maintaining the sanitary status of certified clean plants will offer competitive advantage to nurseries in the southeast region and increased sales of planting materials to growers within and outside Florida leading to maximized profits. To achieve this objective, we will revolve around following specific objectives:  To build institutional capacity and conduct research and service in plant disease testing and diagnostic;  To survey the existing vineyards and nursery producers in the region for occurrence of transmitted through planting stock diseases;  To provide diagnostics of economically important muscadine and American native hybrid varieties by focusing on viruses, crown gall, Eutypa, and Pierce's disease;  To minimize the economic losses by implementing Community of Practice (CoP) extension service for grape clean planting stock and disease best management practices programs. The success of our project will "advance the body of knowledge" in disease diagnostic and sanitary status of the American native grapes and will help the regional grape and wine industry to grow and sustain.

APPROACH: Our strategy is aimed at preventing the spread of grapevine diseases such as PD, Eutypa, crown gall, and leafroll viruses through regular testing and providing growers with disease free planting material, through our work as a regional clean plant center under the USDA/NCPN for Grape. Diagnostic Facility for American Native Grapes at the FAMU Center for Viticulture and Small Fruit Research will be set up. It will utilize the existing Genomic Facility at the Center. In addition a LightCycler system from Roche Molecular Biochemicals, as well as Microplate Absorbance Reader and Immunowash Microplate Washer from Biorad will be acquired under the live of this grant. Therefore, the purchase of these equipments will facilitate the detection of pathogen infections before symptoms are revealed. The equipments will allow us to: 1) precisely quantify DNA, mRNA, and the protein of pathogens, and 2) to complete the experiments in a short time. Other advantages of these methods include highly informative, sensitive and simultaneous analysis of multiple samples. The LightCycler combines fluorescent technologies with ultra-rapid thermal cycling that allows complete amplification and analysis of up to 32 or more samples in less than 20 minutes. We will perform amplification and detection in the same tube, so that results can be monitored online and in real-time. It will also allow us to study multiple sequences concurrently within the same reaction. The sensitivity of this instrument can allow detection of 100 mRNA copies in 10pg of total RNA. This means that we will be able to detect the viral RNA or specific genes from the pathogens even if they are present in very small amount in the sample.

PROGRESS: 2012/09 TO 2016/08
Target Audience:Scientist, grape and wine producers, any research and enterprise involved in production and distribution of grape planting stock. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Our program is the training ground for students and grape growers in phytosanitary evaluation of grape and clean planting stock practices.Dr. James Obuya, post -doctoral Plant Pathologist was recruited from University of Wyoming: UW - Laramie, Wyoming and joined our program in February 2015. How have the results been disseminated to communities of interest?Results of our research were presented as the refereed publications, presentaions at the profesional meetings and comodity groups annual confreces in adition the results were delivered to stakeholders and clienteles via on site visits, field days and workshops: We presented special presentation at the 2015 Annual Florida Grape Growers Conference, January, 15-16, 2015 in Palatka, FL. We conducted Winter Vineyard BMP practices workshop on February 11, 2015. We hosted a visit for the USDA/ARS Administrator Dr. Chavonda Jacobs-Young and team of scientists. We cooperated with UF/IFAS Extension Office at Gadsden County to offer workshop on dormant pruning and sanitation of the vineyard on February13, 2014 for participants from Florida and neighboring states of Alabama and Georgia. We conducted two field days with visiting fresh fruit growers from South Carolina and Florida on best management practices for clean vines . What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

IMPACT: 2012/09 TO 2016/08
What was accomplished under these goals? 1.Buildinstitutional capacity and conducted research and service in plant disease testing and diagnostic for southerngrapes. Provideddiagnostics of economically important muscadine and American native hybrid varieties by focusing on viruses, crown gall, Eutypa, and Pierce's disease.Provided asistance in grapevine disease diagnosticin for the vineyards in Putnam and Orange County, FL. 2.Discoveredthe 3th muscadine variety affected with Grape leafroll virus 1. Initiated survey work for native vectors of the virus in our region. We recruited the assistance and lunch cooperation for pest survey in our Foundation Block from USDA/APHIS PPQ in Gainesville, FL. 3.We worked closely with the industry by vineyard site visits, workshops, field days, lectures and demonstrations at the growers meetings in Florida and neighboring states to implement the clean planting stock and pest and disease "good agricultural practices". 4. We supplied with planting material growers in Florida, North and South Carolina and Texas. 5.We provided protocols to the Dept. of Plant Pathology at NC State University for introduction in vitro and clonal microprpagation of the muscadine grapes.

PUBLICATIONS (not previously reported): 2012/09 TO 2016/08
1. Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Mariah Henry, Anthony Ananga, Vasil Georgiev and Violeta Tsolova: ?Molecular identification and detection of Eutypa lata in muscadine grapes (Vitis rotundifolia Much.)? Proceeding 112th Annual Meeting of the Southern Association of Agricultural Scientists (SAAS): Biochemistry and Biotechnology Division, February 2, 2015, Atlanta, GA.
2. Type: Journal Articles Status: Published Year Published: 2016 Citation: Oglesby, L., Ananga, A., Obuya, J., Ochieng, J., Cebert, E., & Tsolova, V. (2016). Anthocyanin accumulation in muscadine berry skins is influenced by the expression of the MYB transcription factors, MybA1, and MYBCS1. antioxidants-129686 (accepted)
3. Type: Other Status: Published Year Published: 2016 Citation: Obuya, J. O., Ananga, A., & Franc, G. D. (2015). Silent Mutation: Characterization of its Potential as a Mechanism for Sterol 14 [alpha]-Demethylase Resistance in Cercospora beticola Field Isolates from the United States. Journal of Plant Pathology & Microbiology, 6 (6), 1.
4. Type: Other Status: Published Year Published: 2015 Citation: FACT SHEET: Diagnostic and Prevention of of Eutypa Dieback in Muscadine Grapes, Obuya, J., Ananga, A., and V. Tsolova

PROGRESS: 2014/09/01 TO 2015/08/31
Target Audience:Scientist, grape and wine producers, any research and enterprise involved in production and distribution of grape planting stock. Changes/Problems:We would need one year no-cost extension to completae the following objective: 1. To survey the existing vineyards and nursery producers in the region for occurrence of transmitted through planting stock diseases; For collecting samples for Summer and Fall panel respectively for virus and crown gall testing. Eutypa and PD testing will be done in our newly established diagnostic lab. The parameters used to determine the number of 90 vines to be tested from the foundation block: probable infection rate 5% confidence level 99%, total of 210 vines. What opportunities for training and professional development has the project provided?Our program is the training growund for students and grape growers in phytosanitary evaluation of grape and clean plantingstock practices. Dr. James Obuya, post -doctoral Plant Pathologist was recruited from University of Wyoming: UW - Laramie, Wyoming and joined our program in February 2015. He will be overseeing the disease diagnostic and the audit of the foundation vineyard. How have the results been disseminated to communities of interest? We presented special presentation at the 2015 Annual Florida Grape Growers Conference, January, 15-16, 2015 in Palatka, FL. We conducted Winter Vineyard BMP practices workshop on February 11, 2015. We hosted a visit for the USDA/ARS Administrator Dr. Chavonda Jacobs-Young and team of scientists. What do you plan to do during the next reporting period to accomplish the goals?To continue to provide diagnostics of economically important muscadine and American native hybrid varieties by focusing on viruses, crown gall, Eutypa, and Pierce's diseases. To survey the existing vineyards and nursery producers in the region for occurrence of transmitted through planting stock diseases.

IMPACT: 2014/09/01 TO 2015/08/31
What was accomplished under these goals? Field observation and scouting for disease symptoms (presence of Eutypa, Agobacterium vitis and leafroll virus GLRaV -1, was completed in November -December 2014 and March 2015 for all the 23 varieties in the foundation block. The expansion of the G1-nuclear disease free planting block in protective environment was completed and the vines are in the 2nd and 3 growing season respectively. Total of 23 varieties southern grapes in 2 copies each are organized in 2 screen houses as follow: American native hybrid varieties (Florida bunch grapes and rootstocks) and muscadine grape varieties. All the BMP management operations for Fall-Winter 2014 and Spring 2015 were completed. Agreements are under way with Ison's Nursery, Ga for 5 additional fresh fruit muscadine varieties and 3 proprietary selections to be included in the foundation block. 51 clean dormant cutting of our proprietary selection C30-5-1 was received by FPs and planted in the Clean Vine Greenhouse.

PUBLICATIONS: 2014/09/01 TO 2015/08/31
Type: Conference Papers and Presentations Status: Accepted Year Published: 2015 Citation: Mariah Henry, Anthony Ananga, Vasil Georgiev and Violeta Tsolova: ?Molecular identification and detection of Eutypa lata in muscadine grapes (Vitis rotundifolia Much.)? Proceeding 112th Annual Meeting of the Southern Association of Agricultural Scientists (SAAS): Biochemistry and Biotechnology Division, February 2, 2015, Atlanta, GA.

PROGRESS: 2013/09/01 TO 2014/08/31
Target Audience: Scientist, grape and wine producers, any research and enterprise involved in production and distribution of grape planting stock. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Our program is the training growund for students and grape growers in phytosanitary evaluation of grape and clean planting stock practices. How have the results been disseminated to communities of interest? We cooperated with UF/IFAS Extension Office at Gadsden County to offer workshop on dormant pruning and sanitation of the vineyard on February13, 2014 for participants from Florida and neighboring states of Alabama and Georgia. We conducted two field days with visiting fresh fruit growers from South Carolina and Florida on best management practices for clean vines . What do you plan to do during the next reporting period to accomplish the goals? We will continue to work closely with the industry by vineyard site visits, workshops, field days, lectures and demonstrations at the growers meetings in Florida and neighboring states to implement the clean planting stock and pest and disease “good agricultural practices”.

IMPACT: 2013/09/01 TO 2014/08/31
What was accomplished under these goals? We worked closely with the industry by vineyard site visits, workshops, field days, lectures and demonstrations at the growers meetings in Florida and neighboring states to implement the clean planting stock and pest and disease “good agricultural practices”. We supplied with planting material growers in Florida, North and South Carolina and Texas. We continued to survey the insect population in the foundation block for identifying possible native vectors for leafroll virus 1 in our region. We implemented variety trials to introduce new cultivars and expand the foundation block. By the request of the local growers we are working to add 5 more fresh fruit muscadine varieties. We provided protocols to the Dept. of Plant Pathology at NC State University for introduction in vitro and clonal microprpagation of the muscadine grape.

PUBLICATIONS: 2013/09/01 TO 2014/08/31
Type: Websites Status: Submitted Year Published: 2014 Citation: http://www.famu.edu/cesta/main/index.cfm/research/center-for-viticulture-and-sm all-fruit-research/florida-am-university-national-clean-plant-ncpn-center-for-g rape/

PROGRESS: 2012/09/01 TO 2013/08/31
Target Audience: Our project is helping the local grape and wine industry to grow and sustain and assiting entrepreneuship and economic development in rural comunities. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Nothing Reported How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals? Discovery of the 3th muscadine variety affected with Grape leafroll virus 1, make us to look and try to initiate survey work for the possible vectors of the virus in our region. We recruited the assistance and lunch cooperation for pest survey in our Foundation Block from USDA/APHIS PPQ in Gainesville, FL. We are just in the beginning. Traps was installed at the end of the previous growing season and will be installed again at the beginning of the season in March.

IMPACT: 2012/09/01 TO 2013/08/31
What was accomplished under these goals? Building institutional capacity and conduct research and service in plant disease testing and diagnostic for southrn grapes. Providing diagnostics of economically important muscadine and American native hybrid varieties by focusing on viruses, crown gall, Eutypa, and Pierce's disease. Asistance in grapevine disease diagnostic provided in the vineyards in Putnam and Orange County, FL and experimental plots of Auburn University, AL.

PUBLICATIONS: 2012/09/01 TO 2013/08/31
No publications reported this period.