Source: CORNELL UNIVERSITY submitted to
DIAGNOSTIC TECHNOLOGIES AND DISEASE TESTING FOR GRAPEVINE FOUNDATION PLANTING MATERIAL
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
TERMINATED
Funding Source
HATCH
Reporting Frequency
Annual
Accession No.
0227326
Grant No.
(N/A)
Project No.
NYC-153420
Proposal No.
(N/A)
Multistate No.
(N/A)
Program Code
(N/A)
Project Start Date
Oct 1, 2011
Project End Date
Sep 30, 2014
Grant Year
(N/A)
Project Director
Perry, KE, LL.
Recipient Organization
CORNELL UNIVERSITY
(N/A)
ITHACA,NY 14853
Performing Department
Plant Pathology
Non Technical Summary
Planting material free of pathogens is essential for high quality grape production and the sustainable management of vineyards. Viruses and virus-like diseases are graft- and vector-transmissible. Viruses negatively affect fruit quality and yield. If present in the propagation material, these pathogens will be perpetuated in progeny vines. Addressing this problem requires three coordinated efforts. Effective pathogen detecting technologies and services must be in place to support a nursery industry. Foundation planting stocks for grapevine propagation need to be established and monitored. Lastly, a certification program is required to provide quality assurance. This project addresses the first step of pathogen detection technologies to support the nursery industry and will support the second step by facilitating the development of foundation planting stocks. Foundation grape planting blocks in the eastern U.S. are likely to be developed as part of the National Clean Plant Network (NCPN), but commercial growers independently maintain planting blocks and will benefit from support for disease testing. At this stage the NCPN is building the infrastructure for a grapevine clean plant program in the eastern U.S., but has not established a foundation vineyard. The use of public funds to support research related to disease-testing research is quite appropriate; phytosanitary issues are very much in the public domain. The health of a vineyard is a function of the health of neighboring vineyards. The proposed work will improve the competitiveness of the grape and wine industries in New York and the eastern U.S. The expected outcomes are: i) through ongoing interactions with nursery growers, grapevine rootstocks and varieties of greatest interest to the eastern U.S. grape industry will be identified, disease-testing performed and foundation planting stocks propagated, ii) the planting selections above will be comprehensively tested using an array-based diagnostic procedure capable of detecting all known viruses of grapevine, and iii) foundation planting stocks will be re-examined using other established biological and biochemical assays to confirm analyses and detect any additional viral pathogens.
Animal Health Component
(N/A)
Research Effort Categories
Basic
(N/A)
Applied
(N/A)
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
2121139110180%
2161139110120%
Knowledge Area
212 - Pathogens and Nematodes Affecting Plants; 216 - Integrated Pest Management Systems;

Subject Of Investigation
1139 - Grapes, general/other;

Field Of Science
1101 - Virology;
Goals / Objectives
Objective 1) To work with nursery growers and extension personnel to identify grapevine rootstocks and varieties of greatest interest to the eastern U.S. grape industry, and to facilitate the development of a foundation planting blocks and a certification program in NY through the propagation of disease-tested clones. Objective 2) To test grape planting stocks for all known viruses of grapevine, emphasizing the use of array-based nucleic acid hybridization assays to detect a wider range of pathogens than currently tested for. Serological (ELISA) and polymerase chain reaction (PCR) assays will be performed in parallel; array-based testing will enhance rather than replace existing pathogen detection assays. The work will be conducted over the three year period of October 1, 2011 - September 30, 2014. Milestones / Timetable for Objective 1: Meetings with four commercial nursery growers will be held in fall / winter or the spring / summer of the first year (Oct. 2011-Sept. 2012). The most important rootstock varieties will be identified and propagated (Oct. 2011-Sept. 2014). Follow up meetings with the four commercial nursery growers will be held in the last year to summarize the priorities and describe the materials propagated (Oct. 2013-Sept. 2014). Milestones / Timetable for Objective 2: Collected vine materials will be disease tested. (Oct. 2011-Sept. 2014). The presence of newly detected viruses will be confirmed and the viruses characterized. Factors affecting the pathogen spread and transmission will be determined. (Oct. 2012-Sept. 2014). Results from disease testing and the development of material appropriate for foundation planting blocks will be communicated to nursery growers and other stakeholders at the annual grape and wine winter meetings (Oct. 2012-Sept. 2014). Preparation of research reports describing disease testing results and the characterization of viruses observed.(Oct. 2012-Sept. 2014). The expected outputs are: i) through ongoing interactions with nursery growers, grapevine rootstocks and varieties of greatest interest to the eastern U.S. grape industry will be identified, disease-testing performed and foundation planting stocks propagated, ii) the planting selections above will be comprehensively tested using an array-based diagnostic procedure capable of detecting all known viruses of grapevine, and iii) foundation planting stocks will be re-examined using other established biological and biochemical assays to confirm analyses and detect any additional viral pathogens.
Project Methods
The essential working plan will involve three phases: i) discussions and planning with growers, ii) the acquisition and propagation of vines, and iii) the disease testing of vines. The discussions and work with growers will take place at nurseries in western New York. The acquisition, propagation and testing of vines will take place in Geneva and Ithaca, New York. The highest priority in the first year will be discussions with nursery growers to identify grape clones of greatest interest for testing; this will include clones under propagation in NYS and those to be imported. Because rootstocks are critical to most all vinifera plantings, one focus will be on their acquisition, growing and testing. Per discussions with extension personnel (Martinson, Walter-Peterson, Wise), clones of 3309C, MGT 101-14, and Riparia Gloire will be an initial priority. Green cutting and dormant canes will be collected and rooted. Plants will be grown as potted plants to ensure an availability of materials for testing and propagation. An array with oligonucleotide probes for the detection of all 46 described viruses of grapevine developed in the Perry laboratory will be employed to test grape clones. This is based upon the techniques established by Agindotan and Perry (2007, 2008) for solanaceous plant virus detection. Nucleic acids will be extracted from acquired clones and those requested by nursery growers. Methods will follow those developed and published on for the solanaceous plant virus array (Jaubert et al., 2011; Webster et al., 2011). Serological assays (ELISA), PCR, and bioassays (indexing) will be used to complement and validate array findings. Results from disease testing and the development of material appropriate for foundation planting blocks will be communicated to stakeholders, and as appropriate published in peer-reviewed journals; confidentiality will be maintained. Stakeholders will be involved in the design and implementation of the project, through the planned meetings with the major nursery growers in New York. These growers will be involved in prioritizing the materials to be disease tested and propagated.

Progress 10/01/11 to 09/30/14

Outputs
Target Audience: New York grape growers New York nursery growers California and national grape growers Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? A total of eight undergraduate students received training in detecting and analyzing viruses associated with grapevine. These internships ranged from periods of six months to three years. Additionally, one postdotoral associate, one research associate, and three visiting scientists have participated and received training in diagnostic methods. How have the results been disseminated to communities of interest? Results from this project were primarily communicated in regional and national meetings with members of the grape and wine industry. Meetings included: for 2013 Western Grape Conference, Dunkirk, NY, Viticulture 2013, Rochester, NY, and for 2014 Finger Lakes Grape Growers' Conference: Business, Enology and Viticulture NY, Waterloo, NY and the Long Island Agriculture Forum, Riverhead, NY. A national webinar on grapevine health and red blotch disease was held in 2013. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Over the course of this project, we have worked with four of the largest nursery growing operations in the eastern US and with the New York State Department of Agriculture and Markets (NYSDAM). We have developed strategies to facilitate the development of foundation planting blocks and a certification program in NY. This will ensure the greater health status of vines being used to establish new vineyards for eastern grape production. The prerequisite for being able to vegetatively propagate healthy grapevines (and other fruit and vegetable crops) is a disease testing operation to determine the health status of the source or mother plants. We have established a robust testing program for viruses of grapevine that are the most limiting factor in clean plant production. These methods have been used to provide commercial nursery growers with guidance to limit the spread of viruses in their planting stocks. (1) The first project objective has been to work with nursery growers and extension personnel to identify grapevine rootstocks and varieties of greatest interest to the eastern U.S. grape industry, and to facilitate the development of a foundation planting blocks and a certification program in NY through the propagation of disease-tested clones. We held annual meetings with the major nursery growing operations in the eastern US and NYSDAM. From these and additional individual meetings with nursery growers, there was a clear consensus to move forward and formalize a re-establishment of a certification system for grapevines in NY. The process is ongoing; documentation is now being worked on by NYSDAM. The rootstock 3309C is the most commonly employed (upstate region), and of the greatest importance to the nursery industry. (2) The second objective has been to test grape planting stocks for all known viruses of grapevine, emphasizing the use of array-based nucleic acid hybridization assays to detect a wider range of pathogens than currently tested for. Serological (ELISA) and polymerase chain reaction (PCR) assays were performed in parallel; array-based testing enhances rather than replace existing pathogen detection assays. Approximately 8500 nursery and commercial vine samples were tested for viruses serologically. Grapevine leaf roll virus-3 and nepoviruses were the most commonly detected. A smaller subset of samples were tested using the array-based methods. Viruses that would have escaped serological detection were observed, namely a number of members of the family Tymoviridae (Grapevine asteroid mosaic virus, Grapevine syrah virus-1, Grapevine red globe virus) and of the genus Vitivirus (Grapevine virus B, Grapevine virus D, Grapevine virus E). None of these viruses have previously been reported in NY State. Using PCR-based methods, Grapevine red blotch-associated virus was detected in a number of commercial vineyards. As a consequence, this newly recognized virus impacting grape production is now being monitored for in nursery production operations.

Publications

  • Type: Journal Articles Status: Published Year Published: 2014 Citation: Thompson, J., Fuchs, M., McLane, H., Toprak-Celebi, F., Fisher, K., Potter, J., and Perry, K. L., 2014. Multiplex detection of grapevine viruses using a randomly primed RT-PCR/Macroarray platform. Phytopathology 104, 211-219.
  • Type: Journal Articles Status: Published Year Published: 2014 Citation: Krenz, B., Thompson, J. R., McLane, H., Fuchs, M., and Perry, K. L. 2014. Grapevine red blotch-associated virus is widespread in the United States. Phytopathology 102:1232-1240.


Progress 10/15/12 to 09/30/13

Outputs
Target Audience: New York grape growers New York nursery growers California and national grape growers Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? One graduate student received training in plant disease and virus diagnostics, including field observations and laboratory testing. Five undergraduates worked during the academic year training in laboratory analyses of plant viruses, viral nucleic acids and proteins, and plant viral sequence analysis. How have the results been disseminated to communities of interest? Yes, via national meetings, a webinar, a web broadcast, and state grower meetings. What do you plan to do during the next reporting period to accomplish the goals? For Objective 1: Meetings with three commercial nursery growers will be held in the winter/spring and the summer of 2014. The most important rootstock varieties will continue to be propagated. Follow up meetings with commercial nursery growers will be held to summarize the priorities and describe the materials propagated. For Objective 2: Collected vine materials will be disease tested. The presence of newly detected viruses will be confirmed and the viruses characterized. Any issues with regard the pathogen spread and transmission will be determined Results from disease testing and the development of material appropriate for foundation planting blocks will be communicated to nursery growers and other stakeholders at an annual grape and wine winter meeting Research reports will be prepared describing disease testing results and the characterization of viruses observed.

Impacts
What was accomplished under these goals? Two meetings were held jointly between representatives of NYS Dept. Ag. & Mkts, Cornell, and NY nursery growers on 20 March and 2 August 2013 and in Geneva, NY. Cornell researchers Keith Perry, Tim Martinson, and Marc Fuchs were in attendance with representatives from Grafted Grapevine Nursery, LLC, Hermann J. Wiemer Nursery, and Double A Vineyards. Priorties were discussed for the planned grape certification program. Meetings with Double A Vineyards on 30 January 2013 in Freedonia, NY. Dormant wood cuttings from Concord vines were collected from multiple sites to test for the presence of viruses. Macroarray testing of 39 Concord vines revealed a common presence of Grapevine virus E. Issues regarding the propagation of healthy and disease-tested planting stocks were addressed at additional meetings with the following nursery operations: Dr. Frank Vinifera Vineyards on February 11, Double A vineyards on August 28., Grafted Grapevine Nursery, LLC on Sept. 4 and Herman F. Wiemer, Nursery on September 5, 2013. Visual survey of six commercial vineyards was made in September and October 2013 looking for vines infected with grapevine red blotch-associated virus (GRBaV). From this and other times of the year, a total of 2,895 samples were processed and tested, including materials from all of the above nursery operations. Ongoing testing for grapevine red blotch-associated (GRBaV) and other viruses in mother vines from commercial nursery production operations & field collected vines, approximately two hundred samples. This virus has been identified in samples from seven states. Parallel testing for GRBaV along with the virus testing indicated above was done 39 samples received from Long Island vineyards; 73 vines samples received from mother plants in a NY commercial nursery; and 39 plants received from NY extension personnel. Test results were shared with growers and cooperating researchers.

Publications

  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Celebi-Toprak, F., Thompson, J., Perry, K. L., and Fuchs, M., 2013. Arabis mosaic virus in grapevines in New York State. Plant Disease 97:849.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Thompson, J., Fuchs, M., McLane, H., Toprak-Celebi, F., Potter, J., Vargas, J. A., and Perry, K. L., 2013. A routine crop-specific diagnostic macroarray for profiling viral infections in grapevine. Phytopathology 103:145.
  • Type: Journal Articles Status: Accepted Year Published: 2014 Citation: Thompson, J., Fuchs, M., McLane, H., Toprak-Celebi, F., Fisher, K., Potter, J., and Perry, K. L., 2014. Multiplex detection of grapevine viruses using a randomly primed RT-PCR/Macroarray platform. Phytopathology (accepted for publication 20 August, 2013).


Progress 10/15/11 to 10/14/12

Outputs
OUTPUTS: Three of the project participants attended the International Council for the Study of Virus and Virus-like Diseases of the Grapevine in Davis, CA October 7-14, 2012. This meeting is held every three years and provided an excellent forum for the exchange of information. Presentations were made to describe: i) the emergence and detection of a novel DNA virus associated with poor vine vigor and delayed fruit maturity, ii) the development, validation and application of an array-based multiplex detection system for all of the reported viruses of grapevine for which sequences are available, iii) novel strategies for controlling fanleaf virus, and iv) dynamics in the vector transmission of grapevine leafroll viruses. Three site visits for collection and consultation were made to three commercial nursery / vineyard operations. Vine samples for consultation and analyses were received from the three of the primary NY State nursery operations. Two trips were made to the USDA grapevine repository in Geneva, NY to assess symptoms and test for viruses in vines. PARTICIPANTS: Keith L. Perry, PI Marc Fuchs, co-PI Jeremy Thompson, Research Associate Bjoern Krenz, Postdoctoral Associate TARGET AUDIENCES: Commercial grapevine nursery operators, vineyard growers, wineries PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

Impacts
It was determined that dormant canes of all of 16 Concord tested were virus infected with at least one virus, Grapevine leafroll virus 3. This would suggest that most commercial plantings of Concord are virus infected. At least two sources of Concord vines from the eastern US (New York and Massachusetts) were identified as virus free; these were from collections, not from commercial plantings. Two viruses not previously reported in NY State have been detected, Grapevine virus E (GVE) and Arabis mosaic virus. GVE, appears to be well established in NY, as it was observed in vines from multiple sites in multiple counties. Three of four Riesling vines selected as mother plants in a commercial nursery production where shown to be free of regulated viruses. The newly described and emerging DNA virus, named Grapevine cabernet franc-associated virus in NY and Grapevine red blotch-associated virus (GRBaV) in CA, was sequenced and diagnostic primers designed and validated for polymerase chain reaction methodologies. GRBaV was detected in vines from five states (NY, VA, MD, PA and CA).

Publications

  • Krenz, B., Thompson, J., Fuchs, M., and Perry, K. L. (2012). Complete Genome Sequence of a New Circular DNA Virus from Grapevine. Journal of Virology 86:7715
  • Thompson, J., M. Fuchs, K. Fisher, and Perry, K. L. (2012). Macroarray detection of grapevine leafroll associated viruses. Journal of Virological Methods 183:161-169.
  • Thompson, J. R., Fuchs, M. & Perry, K. L. (2011). Genomic analysis of Grapevine leafroll associated virus-5 and related viruses. Virus Research 163:19-27.
  • Fuchs, M., and Oliver, J. E. (2012). A novel approach for engineering resistance to Grapevine fanleaf virus. Page 34 in Proceedings of the 17th Congress of the international council for the study of virus and virus-like diseases of the grapevine (ICGV), Davis, CA.
  • Krenz, B., Thompson, J., Fuchs, M., and Perry, K. L. (2012). Complete Genome Sequence of a New Circular DNA Virus from Grapevine. Page 102 in Proceedings of the 17th Congress of the international council for the study of virus and virus-like diseases of the grapevine (ICGV).
  • Thompson, J., Fuchs, M., Fisher, K., and Perry, K. L. (2012). Multiplex detection of grapevine viruses using a randomly primed RT-PCR/Macroarray platform. Page 132 in Proceedings of the 17th Congress of the international council for the study of virus and virus-like diseases of the grapevine (ICGV), Davis, CA.
  • Celebi-Toprak, F., J. Thompson, K. L. Perry, and M. Fuchs. 2013. Arabis mosaic virus in grapevines in New York State. Plant Disease (accepted).