Brief Summary of Minutes of Annual Meeting

Thursday, May 29, 2014
(Ala Moana Hotel, 410 Atkinson Dr, Honolulu, HI 96814)

Part 1. General session

Opening of meeting by the Chair of the WERA 20 group, Dr. John Hu.

Welcome by Dr. Kenneth J. Grace, Interim Associate Dean of College of Tropical Agriculture and Human Resources at University of Hawaii

Part 2. State Reports

Wyoming (Sadanand Dhekney):
Reported on the identification of promising grape cultivars with increased tolerance to different abiotic stresses, especially cold tolerance; as part of this screening process, sampled cultivars are also tested for the presence of different viruses. An overview on Precision Breeding was presented, that is defined as using sequences from the grape genome rather than from other species (cis-genic rather than trans-genic). In this regard, functionally equivalent segments of the grape genome to replace the left and right borders of the Agrobacterium transformation vector was reported along with using anthocyanin grape sequences as reporter gene.

A group photo was taken during the coffee break time.

Washington (Kenneth Eastwell and Dan Edward Villamor):
Reported on the update of the little cherry disease (LCD) outbreak in Douglas and Chelan counties of the state. Approximately 400 acres of sweet cherry block have been eradicated due to the disease and an estimated 300 acres will be eradicated this year. A recombinase-polymerase amplification assay (RPA) was developed for Little cherry virus 2, the main virus associated with LCD. This assay, which can be used in the field as a rapid and sensitive assay for the virus, is now commercially available as a kit. A total of 242 grapevines from the screenhouse and foundation block of the Clean Plant Center Northwest (CPCNW) were indexed for Grapevine red blotch associated virus (GRBaV) and Grapevine vein clearing associated virus (GVCV). None except one recent accession was tested positive for GVCV. This accession has been removed from the foundation block. Data on deep sequencing of 30 grapevine and 31 stone and pome fruit tree accessions showed the potential of deep sequencing as tool in grape and fruit tree virus certification programs.

Washington (Naidu Rayapati):
The genetic diversity of Grapevine virus A in California (with Maher Rwahnih) and Washington vineyards was analyzed using two functional classes of virus genes, a transcription-related (RNA-dependent RNA polymerase [RdRp]) and a structural (CP) gene. The data showed presence of divergent variants of GVA in both Washington and California vineyards. In many cases, distinct variants of GVA were found as mixtures in individual grapevines. Survey of juice grape (Vitis × labruscana L. ‘Concord’) vineyards indicated the presence of three grapevine leafroll-associated viruses (GLRaVs) and Grapevine fanleaf virus. Among the GLRaVs, GLRaV-2, -3, -4 and its strain -9 were documented so far in ‘Concord’ vineyards. GLRaV-3 was found to be widely distributed in ‘Concord’ vineyards. In collaboration with entomology (D. Walsh), we demonstrated that grape mealybug (Pseudococcus maritimus Ehrhorn) and European fruit lecanium scale (Parthenolecanium corni Bouché) are capable of transmitting GLRaV-3 between wine grapes (V. vinifera) and juice grapes. The results were disseminated through extension and outreach presentations for the benefit of grape growers, certified nurseries, regulatory agencies and other industry stakeholders.

Mississippi (Sead Sabandzovic):
Reported on the identification of six viruses associated with Blackberry yellow vein disease (BYVD), two of which are newly described. Viruses that infect muscadines were also reported, which are the following: Grapevine Syrah virus 1, Grapevine virus B, Grapevine leafroll associated virus 2, a new marafivirus and two other new viruses that are still being characterized. In addition, he shared the results of the screening of Muscadines and other grape cultivars for Rupestris stem pitting-associated virus (RSPaV); RSPaV amplicons that were sent for sequencing corresponded to host sequences, highlighting the need to further investigate PCR results and proceed to downstream sequencing when the PCR data seems questionable. A petition for NCPN support through the creation of a grape center in Mississippi was mentioned.

Maryland (Erich Rudyj):
Reported on the history of the National Clean Plant Network (NCPN); its creation, roles and responsibilities and source of funding. Originally under 2008 Farm Bill (2008-2014), the funding for NCPN was renewed for five years from 2014-2018. The US House of Representatives recognized the value of NCPN and made it permanent by including the network in the Plant Protection Act. A consequence of such change is the surrendering of the no-year funding option for the network but a guaranteed funding of no less than $5M annually. Several critical and emerging issues on NCPN activities were discussed, among which include specialty crop expansion such as roses and sweet potato and the adoption of next generation sequencing for pathogen detection.

Maryland (Margarita Bateman):
Reported on the activities of the USDA-APHIS Plant Germplasm Quarantine Program, particularly on the quarantine program for stone fruits. She mentioned that the Post-entry Quarantine is now known as Controlled Import Permits and reported on the development of a tissue culture system for Prunus as the major accomplishment of the program last year. The review of the French Fruit Certification Program was also reported, citing the similarities and differences by which plant material is accepted in the US and Canada. The results of the visit revealed that the French Fruit Tree Certification Program meets the requirements of both US and Canada.

Maryland (Mark Nakhla):
Reported on the activities and functions of the Center for Plant Health Science and Technology (CPHST). The organization certifies laboratories, develops and validates molecular diagnostic methods, provide training on diagnostics and evaluate new diagnostic technologies. He also reported on Biosensors for rapid detection of regulated plant pathogens called CANARY (Cellular Analysis and Notification of Antigen Risks and Yield). These cells are available for Ralstonia, Potyvirus, Phytophthora, Citrus leprosis virus C (CiLV-C) and citrus variegated disease pathogen, Xyllela fastidiosa.

Indiana (Shulu Zhang):
Reported on the development and availability of AmplifyRP kits for the detection of several pathogens such as Plum pox virus PPV), Candidatus liberibacter asiaticus and recently, Little cherry virus 2 (LChV-2). This technology is an isothermal detection based on recombinase-polymerase assay. The kits provide rapids tests for these pathogens, which take only 15 minutes to achieve the result, and are amenable to crude sap preparations, unlike PCR that require purified nucleic acid material. The kits are available as lyophilized reagents containing buffers, enzyme mix and primers for end-point detection and real time detection. A basic kit called Discovery kit does not contain the primers for the pathogen is also available, allowing flexibility for users to design and use their own primers.

Idaho (Alex Karasev):
The survey of Idaho vineyards revealed the presence of Grapevine leafroll associated viruses 1, 3, 4 and 5, Rupestris stem pitting aassociated virus (RSPaV), Grapevine virus A and B and Grapevine fleck virus (GFkV). The most prevalent among these viruses is GLRaV-3 followed by GLRaV-1. The commercially available antibody against GLRaV-3 has been consistently reliable in detecting the virus but not for GLRaV-1. In his laboratory, antibody was produced against GLRaV-1 using bacterially expressed coat protein of the virus as antigen for immunization. The resulting antibodies were optimized for triple antibody sandwich enzyme-linked immonosorbent assay (TAS-ELISA). This assay is now used in the laboratory for detection of GLRaV-1.

Hawaii (Steve Ferreira):
Presented an overview of the papaya industry in Hawaii, with particular emphasis of the devastating effect of the Papaya ringspot virus (PRSV) and how the genetically modified papaya came to rescue the industry. The transgenic papaya cultivar ‘Rainbow’ was developed by inserting a fragment of the coat protein gene of PRSV. The mechanism of resistance operates through RNA silencing.

Hawaii (Richard Manshardt):
Reported on the transgene flow of genetically modified (GM) papaya to non GM papaya through pollen drift study. This study showed that about 22% of GM papaya plants were observed in feral papaya populations, female papaya plants are the only ones susceptible to transgene flow and transgene flows primarily by seed. Overall, transgene flow into feral papaya populations will be slow and gradual.

Formal meeting adjourned at 5:45pm followed by a group dinner.

Friday 30th May, 8:30am (Oahu, HI)

Field trips were provided. The first stop was at the Waimanalo Research Station, College of Tropical Agriculture and Human Resources (CTAHR), University of Hawaii Manoa (UHM). A field project on the screening of banana germplasm for resistance to Banana bunchy top virus (BBTV), the transgenic papaya block, and the sweet potato germplasm collection were shown. The second stop was the tour of the subtropical fruit tree orchard of the Pearl City Urban Garden Center where collections of citrus, mango, guava, breadfruit, avocado and many others were shown. Group lunch was provided followed by the third stop at Dole Plantation. The group started at the visitor center followed by a visit to the cacao and coffee plantations.

Monday 2nd June, 8:30am (Hilo, HI)

Field trips on the Big Island were provided. The group started at Mauna Loa Visitor Center to check the macadamia nut orchards and the nut and chocolate processing plant. A tour of the facilities as well as the subtropical fruit tree germplasm collection of the National Clonal Germplasm Repository of the United Stated Department of Agriculture-Agricultural Research Service at the Pacific Basin Agricultural Research Center (USDA-ARS-PBARC) and Waiakea Research Station, CTAHR, UHM were shown. After group lunch, the group visited Plant it Hawaii to see production methods of a wholesale nursery of subtropical fruit trees. The final stop was at the Volcano winery. A tour of the winery’s vineyard and tea plantation was provided. Foliar symptoms typical of Grapevine leafroll disease was observed in some wine grape cultivars in the vineyard. This was followed by wine and tea tasting at the winery.

Tuesday 2nd June, 8:30am
(University of Hawaii at Hilo, 200 W. Kawili St, Hilo, HI 96720)

Comments by Administrative Advisor, Dr. Jim Moyer, Director of Agricultural Research Center and Associate Dean for Research for the College of Agricultural, Human and Natural Resource Sciences at Washington State University. He reminded that the level of accountability for each one is increasing and stressed the importance of regional projects. He also reminded to be mindful of the submission of final report within thirty days and underscored the impact statement in the report as well as evidence of collaboration among peers.

Two suggestions were proposed for the location of next year’s meeting, Arkansas and Maryland. The group decided to go to Maryland next year. Margarita Bateman will be the host of the next year’s meeting and the Chair of WERA-20 next year. John Hu will be the Vice-chair.

Continuation of Part 2: State Reports

Hawaii (Mike Melzer and John Hu):
Reported that the economic potential of growing fig in Hawaii is huge. Viruses infecting fig has not yet been reported in Hawaii but virus-like symptoms on fig were observed. Next generation sequencing using 454 platform and dsRNAs from symptomatic fig revealed the presence of virus-like contigs that showed high identities with several closteroviruses, an umbra-, myco-, badna- and emaravirus, and Tobacco necrosis statellite virus. Characterization of these virus-like contigs is currently being pursued.

Canada (Anna-Mary Schmidt):
Reported on the activities of Canadian Food Inspection Agency (CFIA). The agency is the national post quarantine facility for grapevines, tree fruit and small fruits in Canada which provide testing activities (bioassays, ELISA, PCR and next generation sequencing), virus elimination, tissue culture, maintenance of generation 1 virus tested grapevine and tree fruit repository. She also reported on the grapevine virus status in Canadian vineyards. GLRaV -3 is most prevalent in British Columbia (BC) followed by GLRaV-2, mixed infection of GLRaV-2 and -3 and GLRaV-1. GLRaV-1, -2 and -3 is also prevalent in Ontario as well as Grapevine red blotch associated virus (GRBaV).

California (Deborah Golino):
Reported on the progress of Grape Tier II of NCPN. The goals and objectives of NCPN were briefly mentioned including the specialty crop groups that are included. Progress on the grape clean plant centers throughout the country was summarized. For each grape center, this included the number of cultivars that were indexed for viruses, number of cuttings released as well as the number of vines planted in the screenhouses and foundation blocks. The renewal of the NCPN funding for 2014 was also mentioned.

California (Maher Al Rwahnih and Adib Rowhani):
Reported on the development and application of next generation sequencing (NGS) as a new technology for testing new grapevine accessions. The advantages and disadvantages of NGS were mentioned. The cost of NGS has significantly decreased over the last seven years from $4K to $500 per sample, making this technology amenable for large scale use. Several grapevine viruses were characterized through NGS such as Grapevine Syrah virus, GRBaV, Grapevine virus F (GVF), Grapevine vein clearing virus (GVCV). These viruses reacted poorly or none at all with known field woody indicators and would not have otherwise been discovered or would take longer time to be discovered using the current tools if not for NGS. Careful attention, however, must be made as virus/viruses discovered by NGS still require the fulfillment Koch’s postulate.

California (Alan Wei):
Reported on the work funded by a USDA SBIR project. They made a new antibody reagent for GLRaV-3 which showed broader detection spectrum than the mostly commonly used antibody reagent from BioReba. Their antibody reacted positively to several samples which the BioReba antibody did not. After testing these samples by RT-PCR, sequencing revealed that the amplicons were possibly divergent strain of GLRaV-3. This discovery should have potential impact on certification programs because most certification programs Bioreba reagent which may have let some strains of GLRaV-3 escape detection. The preliminary data for GLRaV-3 nanobodies was also presented. Nanobodies consist of a 15K single monomeric variable antibody domain. Like regular antibodies, nanobodies show high affinity to their target antigen but are more thermally stable than regular antibodies.

California (Richard Hoenisch):
Reported on the activities of the National Plant Diagnostic Network (NPDN). The network is a consortium of plant diagnostic laboratories all over the country and is subdivided into five regions. The networks primary purpose is to provide a system that quickly detects and identifies pests and pathogens that are of concern to a particular area. This is mainly achieved by providing laboratory infrastructure and training and network development of first detectors. Pest risk assessment was also discussed in order to mitigate or avoid potential outbreak in a particular area. The importance of this activity was highlighted by providing historical case studies of pest and diseases that have devastated agricultural crops in the country. Present day examples of high risk pests were also enumerated.

Arkansas (Ioannis Tzanetakis):
Reported on the bioinformatics pipelines to follow in analyzing NGS data. The software developed in his laboratory, called Virfind, is already available which can be used by outside users with little training and supervision. The software also allows changing analysis parameters depending on user’s requirement. The value of NGS in small fruit virus detection was underscored by the fact that of the 30 viruses already discovered in blueberries, most do now induce symptoms on known indicators as typified by Blueberry mosaic virus. The overview of the certification programs for blueberry and Rubus was also described and a proposal for the process of certification was presented.

State reports finished at 12:00 pm followed by a group lunch. Group discussion about the issues pertaining to NGS:
(2:00 pm, USDA-ARS-PBARC 64 Nowelo St. Hilo, Hawaii 96720)

Welcome by Dr. Marissa Wall, Acting Director of USDA-ARS-PBARC at Hilo, HI. The group was given a brief overview of the research center. Its main objective is to make sure invasive pest species do not enter the state and, if by any chance they get in, prevent at any circumstance dissemination of the pest out of the state. Another purpose is to help farmers transition from plantation to diversified crops. A tour of the laboratory facilities of the center was provided before the group discussion started.

During the group discussion on the impact of NGS on virus diagnostics, it was suggested that separate working group on NGS that focuses on the (1) technical/scientific aspect of the technology and (2) legal/policies involved with it be created.

• (1) The technical working group will provide workshop regarding the standard methodologies that come with different NGS platforms. This group will be responsible for organizing dry laboratory training on the proper analysis and interpretation NGS sequence data. Case studies regarding the development and use of the technology for different crop specialty groups, especially for fruit trees, grapes and berries, will be helpful so that protocols for each crop can be compiled and validated.
• (2) The legal working group will be responsible for convincing regulators to approve the use of NGS technology in certification programs. This will require substantial amount of data, complied from the technical working group, showing the robustness of the NGS over current technology (PCR, ELISA and biological indexing on indicators) in detecting viruses from different crops. An important component of this endeavor is to show the process and speed by which NGS could assist in releasing a crop accession on a shorter period of time.

The group recognized issues that arose with the current technology. The output of NGS has already revealed new viruses which do not either have strong association with a disease or no association with a disease at all. In scenarios like this, whether these types of virus or virus-like agents will be ignored in certification programs are still not clear. Another issue is the international acceptance of the technology by other countries. Data generated by NGS could be used by regulators from other countries to restrict/prohibit the entry of planting material or produce into their respective countries. This is less of an issue with quarantine programs since new viruses detected by NGS will eventually be eliminated. But careful attention must be made in publishing new viruses in commercial orchards. Outreach and education is necessary to address the implications of NGS results.

The meeting ended with a proposed agenda for next year’s meeting in Maryland, which will be composed of three parts:

• (1) NGS workshop (1 day)
• (2) Field trip and state reports (3 days)
• (3) Policy discussion with some invited regulators on NGS (1 day)

Formal annual meeting of WERA-20 adjourned at 4:00 pm.

Minutes prepared and submitted respectfully by Dan Villamor and John Hu.

Collaborative efforts by the members of the WERA-20 have brought together unique experiences and cutting-edge technologies to advance the detection and characterization of new and emerging viruses and virus-like agents from fruit trees, small fruits, citrus, and grapevines in the US and Canada. Powerful next-generation sequencing (NGS) technology was used extensively in the past year to identify new viruses and virus-like agents from grapevines, blueberries, pome and stone fruit, and edible fig. Comprehensive studies have been conducted in various crop systems to compare the current bioassay systems with NGS for their relative sensitivity and reliability. Progress has been made in the application of NGS technology in quarantine systems that regulate trade between states and countries. Sensitive and reliable diagnostic assays that have been developed by WERA-20 members have been shared with labs of various federal and state agencies as well as centers of the National Clean Plant Network. Novel virus detection systems including recombinase polymerase amplification and nanobody technology have been developed and applied for the rapid detection and reliable diagnosis of viral diseases. The molecular diversity of viruses has been revealed using sensitive and high throughput diagnostic methodologies. Science-based information on the distribution of viruses and transmission of new viruses by insect vectors were obtained and shared with industries to develop practical and environmentally sound strategies for managing the virus and virus-like diseases of fruit trees, small fruits, citrus, and grapevines. Latest information on viruses and vectors was also shared with federal and state regulatory agencies for revising policies to contain spread of viruses.

Abou Ghanem-Sabanadzovic N., Tzanetakis, I.E. and Sabanadzovic, S. 2013. Rubus canadensis virus 1, a novel betaflexivirus identified in blackberry. Archves of Virology 158: 445-449.

Al Rwahnih, M., Daubert, S., Sudarshana, M., and Rowhani, A. 2013. Gene from a novel plant virus satellite from grapevine identifies a viral satellite lineage. Virus Genes 47: 114-118.

Al Rwahnih, M., Dave, A., Anderson, M., Rowhani, A., Uyemoto, J. K., and Sudarshana, M. 2013. Association of a DNA virus with Grapevines affected by Red 1 Blotch disease in California. Phytopathology 103:1069-76.

Al Rwahnih, M., S. Daubert, C. Islas, D.A. Golino, and A. Rowhani. 2014. Description of a fifth Vitivirus in grapevine. Journal of Plant Pathology. 96: 219-222.

Alabi, O. J., Al Rwahnih, M., Mekuria, T. A., Rowhani, A., and Naidu, R. A. 2014. Genetically diverse isolates of Grapevine virus A are present in Washington and California vineyards. Phytopathology. 104: 548-560

Alabi, O.J., Al Rwahnih, M., Mekuria, T., and Naidu, R.A. 2014. Genetic diversity of Grapevine virus A in Washington and California vineyards. Phytopathology 104:548-560.

Bahder, B.W., Naidu, R.A., Daane, K.M., Millar, J.G., and Walsh, D.B. 2013. Pheromone-based monitoring of Pseudococcus maritimus (Hemiptera: Pseudococcidae) populations in Concord grape vineyards. Journal of Economic Entomology 106:482-490.

Bahder, B. W., Poojari, S., Alabi, O.J., Naidu, R.A., and Walsh, D. B. 2013. Pseudococcus maritimus (Hemiptera: Pseudococcidae) and Parthenolecanium corni (Hemiptera: Coccidae) are capable of transmitting Grapevine leafroll-associated virus 3 between Vitis labruscana and Vitis vinifera. Environmental Entomology 42: 1292-1298.

Dhekney S.A., Kandel, R., Garcia y Garcia A. 2014. University of Wyoming-Grape Growers Partnership may Improve Sustainable Grape Production in Wyoming. University of Wyoming, Agricultural Experiment Station, Field Days Bulletin, In Press.

Doan, H.K., Zhang, S. and Davis, M.R. 2014. Development and Evaluation of AmplifyRP Acceler8 Diagnostic Assay for the Detection of Fusarium oxysporum f. sp. vasinfectum Race 4 in Cotton. Plant Health Progress 15(1):48-52.

Ghimire., P., Abou Ghanem-Sabanadzovic, N., Tzanetakis, I.E. and Sabanadzovic, S. 2013. Partial characterization of a novel flexivirus from blackberry. 90th Meeting of the Southern Division of the American Phytopathological Society, February 8-10, 2013, Baton Rouge, LA

Golino, D.A., Rowhani, A. and Uyemoto, J.K. 2013. “Grapevine Virus Diseases”. In : Grape Pest Management, third edition. Pp: 157-173. Ed: Larry J. Bettiga, University of California, Agriculture and Natural Resources publication #3343.

Gray, D.J., Li, Z.T. and Dhekney, 2014. S.A. Precision breeding of grapevine (Vitis vinifera L.) for improved traits. Plant Science, http://dx.doi.org/10.1016/j.plantsci.

Hintz, W.E., Carneiro, J.S., Kassatenko, I., Varga, A., and James, D. 2013. Two novel mitoviruses from a Canadian isolate of the Dutch elm pathogen Ophiostoma novo-ulmi (93-1224). Virology Journal 10:252.

James, D. 2013. Challenges and benefits of standardized diagnostic protocols – International Plant Protection Convention’s perspective. Symposium on “Science and Technology Tools Supporting Phytosanitary Work”, 37th NAPPO Annual Meeting, Guelph, Ontario, Canada, October 31, 2013. Book of Abstracts. (Abstr.).

James, D., Sanderson, D., Varga, A., Greig, N., and Stobbs, L. 2013. Relationships and genetic diversity of selected Canadian isolates of Plum pox virus. 2nd International Symposium on Plum Pox Virus, Olomouc Czech Republic, September 3 – 6, 2013. Book of Abstracts. Pg 15 (Abstr.).

James, D., Sanderson, D., Varga, A., Sheveleva A., and Chirkov, S. 2013. Comparison of Plum pox virus strain W isolates to determine their relationships and further understand their genetic diversity. 2nd International Symposium on Plum Pox Virus, Olomouc Czech Republic, September 3 – 6, 2013. Book of Abstracts. Pg 14. (Abstr.)

James, D., Varga, A., and Sanderson, D. 2013. Genetic diversity of Plum pox virus: strains, disease and related challenges for control. Canadian Journal of Plant Pathology 35(4):431-441.

Leal I., Allen, E., Anema, J., Reisle, C., Foord, B., Uzunovic, A., Varga, A., and James, D. 2013. Development of a reverse transcription loop-mediated isothermal amplification (RT-LAMP) method to detect living pinewood nematode, Bursaphalenchus xylophilus, in wood. In:Schroder, T (ed), Pine Wilt Disease Conference 2013, Brunschweig, Germany, ISSN: 1866-590X, pp 39-40.

Leal I., Allen, E., Reisle, C., Anema, J., Uzunovic, A., Varga, A., and James, D. 2013. Development of a reverse transcription loop-mediated isothermal amplification (RT-LAMP) method to detect living pinewood nematode, Bursaphalenchus xylophilus, in wood. Canadian Journal of Plant Pathology. (Abstr.) 35(1):92.

Lu, Q., Abou Ghanem-Sabanadzovic, N., Ghimire, G., Tzanetakis, I.E., Lawrence, A. and Sabanadzovic, S. 2013. New marafivirus identified ib yellow vein disease-affected blackberries. Abstracts Annual Meeting of the American Phytopathological Society, Austin, TX, USA. Phytopathology 103: S2.87

Mekuria, T.A,, Zhang S. and Eastwell, K.C. (2014) Rapid and sensitive detection of Little cherry virus 2 using isothermal reverse transcription-recombinase polymerase amplification. Journal of Virological Methods 205:24-30.

Melzer M.J., Ayin, C., Sugano, J.S., Uchida, J.Y., Kawate, M.K., Borth, W.B., Sether, D.M. and Hu, J.S. 2013. Differentiation and Distribution of Cordyline Viruses 1–4 in Hawaiian ti Plants (Cordyline fruticosa L.). Viruses 5:1655-1663.

Melzer, M.J., Nelson Simbajon, N., Carillo, J., Borth, W.B., Freitas-Astúa, J., Kitajima, E.W., Neupane, K.R. and Hu, J.S. 2013. A cilevirus infects ornamental hibiscus in Hawaii. Archives of Virology doi:10.1007/s00705-013-1745-0

Osman, F., Al Rwahnih, M. and Rowhani, A. 2013. Improved Detection of Ilarviruses and Nepoviruses Affecting Fruit Trees Using Quantitative PCR. Journal of Plant Pathology (in press).

Osman, F., Hodzic, E., Omanska-Klusek, O., Olineka, T. and Rowhani, A. 2013. Development and validation of a multiplex quantitative PCR assay for the rapid detection of Grapevine virus A, B and D. Journal of Virological Methods 194:138-145.

Poojari, S., Alabi, O.J., and Naidu, R.A. 2013. Molecular characterization and impacts of a strain of Grapevine leafroll-associated virus 2 causing asymptomatic infection in a wine grape cultivar. Virology Journal 10:324.

Poudel, B., Ho, T., Laney, A., Khadgi, A. and Tzanetakis, I.E. 2014. Epidemiology of Blackberry chlorotic ringspot virus. Plant Disease 98:547-550.

Sabanadzovic S. and Abou Ghanem-Sabanadzovic, N. 2013. Cryptic viruses in the flora of the Great Smoky Mountains National Park. Abstracts Annual Meeting of the American Phytopathological Society, Austin, TX, USA. Phytopathology 103: S2.125

Thekke-Veetil, T., Aboughanem Sabanadzovic, N., Keller, K.E., Martin, R.R., Sabanadzovic S. and Tzanetakis, I.E. 2013. Molecular characterization and population structure of Blackberry vein banding associated virus, new ampelovirus associated with yellow vein disease. Virus Research 178: 234-240.

Thekke-Veetil, T., Ho, T., Keller, K.E., Martin, R.R. and Tzanetakis, I.E. 2014. A new ophiovirus associated with blueberry mosaic disease. Virus Research 189: 92-96.

Villamor, D.E.V, Ward, K.F., Collman, S.J. and Eastwell, K.C. 2014. First report of infection of cherry rusty mottle associated virus in Portuguese Laurel (Prunus lusitanica) in Washington State. Plant Disease 98: 699.

Villamor, D.E. and Eastwell. K.C. 2013. Viruses associated with cherry twisted leaf and the rusty mottle group of diseases of sweet cherry represent distinct species within the family Betaflexiviridae. Phytopathology 103:1287-1295.

Zhang S, Russell P, McOwen N, Bohannon S and Vrient A. 2014. AmplifyRP Acceler8 – a rapid isothermal test for plum pox virus using reverse transcription-recombinase polymerase amplification (Abstract). American Phytopathological Society Annual Meeting, August 2014, St. Paul, Minnesota.

Video Presentations

Deborah Golino, UCDavis, “Red Blotch Status of the FPS Classic Foundation and the Russell Ranch Foundation” http://lecture.ucanr.org/Mediasite/Play/486e1db1ad5348eaada05ed570cc8c971d

Marc Fuchs, Cornell UNIVERSITY “Red Blotch: Challenges and Opportunities” http://lecture.ucanr.org/Mediasite/Play/7e6250539e5e4676ad4cd8880c51164c1d