Source: UNIVERSITY OF FLORIDA submitted to
IDENTIFICATION, CHARACTERIZATION AND MANAGEMENT OF PLANT VIRUSES OF CULTIVATED AND WILD PLANT SPECIES
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
HATCH
Reporting Frequency
Annual
Accession No.
0230351
Grant No.
(N/A)
Cumulative Award Amt.
(N/A)
Proposal No.
(N/A)
Multistate No.
(N/A)
Project Start Date
Jul 1, 2012
Project End Date
Sep 30, 2017
Grant Year
(N/A)
Program Code
[(N/A)]- (N/A)
Recipient Organization
UNIVERSITY OF FLORIDA
G022 MCCARTY HALL
GAINESVILLE,FL 32611
Performing Department
Plant Pathology
Non Technical Summary
Plant viruses are emerging pathogens in Florida and elsewhere. Many of the newly emerged viruses have caused significant economic losses in crops. Early detection and rapid responses to new plant viruses can help to mitigate this damage. The goal of this project is to respond to new viruses through rapid identification, development of appropriate assays for their recognition and detection, discovery of information about the biological and molecular characteristics of the virus that can be used to design appropriate disease management tactics to mitigate yield losses that the virus can cause.
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
2121499110150%
2121460110125%
2122199110125%
Knowledge Area
212 - Pathogens and Nematodes Affecting Plants;

Subject Of Investigation
1499 - Vegetables, general/other; 1460 - Tomato; 2199 - Ornamentals and turf, general/other;

Field Of Science
1101 - Virology;
Goals / Objectives
The overall goal of this project is to support Florida agriculture through the timely identification of new viruses and the generation of information to help mitigate damage from new and existing plant viruses. Specific Objectives 1. Identify and characterize viruses occurring in horticultural and wild plants in Florida and the Western Hemisphere. 2. Characterize transmission of viruses by whiteflies. 3. Develop and evaluate new approaches for managment of plant viruses. Expected Outcomes This project will generate information on the identity and diversity of plant viruses in Florida; biological and moleculer characteristics of new viruses; and will use that information to design appropriate management tactics when needed.
Project Methods
Objective 1. This objective focuses on the identification of emerging viruses such as begomoviruses as well as others. New viruses will be identified using appropriate techniques - random,degenerate, or broad-spectrum primers, amplified by rolling circle amplification,RT-PCR or PCR, followed by cloning and sequencing. Information on the symptomatology, experimental and natural host ranges, and modes of transmission will be generated as appropriate. Virus specific reagents (probes, primers, etc.) for detection will be developed. This objective will include elucidating the cause of diseases thought to be caused by a virus or viruses, such as Purple leaf disorder of tomato. This research will generate basic information on new viruses, and reagents for the detection of these viruses. Detection reagents will be used in research on these viruses, and will be made available to university, state and private diagnostic laboratories to aid in the rapid identification of the viruses, an important part of successful virus management. Objective 2. Studies will be conducted to improve our understanding of the relationship that has evolved between plant viruses and the whitefly, Bemisia tabaci. Studies will be conducted to identify the constraints of the relationship and what each of the members derives from this relationship. For example, are virus-infected plants better or worse nutrition sources for whiteflies than non-infected plants Are virus-infected plants more or less attractive than non-infected In addition, studies will be conducted to determine what role the vector plays in evolution of begomoviruses. In addition, basic studies may be conducted if other whitefly-transmitted viruses are found for which little information is known. These studies will develop a base of understanding of the transmission process of plant viruses. This information may be used to develop or modify virus management recommendations (Objective 3). Objective 3. Experiments will be conducted to develop improved management practices for emerging plant viruses. This process will require generating data on relevant aspects of virus biology and vector biology not covered under Objective 2. This could include such things as determining the identity of virus reservoirs, using tissue culture and specific detection reagents to remove viral/viroid pathogens from asexually propagated plant material, etc. Also this research could include evaluating new approaches for interfering with insect transmission of plant viruses through the use of repellents, insecticides, mulches, etc. This may include the development and evaluation of virus resistance in crops. Resistance could be through conventional breeding or through genetic engineering; and would include screening for new resistance genes, finding markers for resistance genes, determining copy number of resistance genes, and mechanisms of resistance. This objective will result in the modification of existing and development of new methods to manage viruses in vegetable and ornamental crops. Recommendations will be distributed to growers through growers meetings and county and state extension agents.

Progress 07/01/12 to 09/30/17

Outputs
Target Audience:Vegetable and ornamental producers and related industries, vegetable and ornamental plant breeders, plant virologists and pathologists, plant diagnostic clinics and diagnosticians Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Trained an undergraduate to conductresearch and taught them some molecular techniques. Trained three graduate students to conduct research and how to analyze results from those studies, as well as trained them how to write a referreed paper. How have the results been disseminated to communities of interest?Through referreed publications. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Identified and sequenced the genome of a virus in passionfruit and a wild plant that was never before reported from Florida (published). Obtained the complete sequence of a virus which was reported forthe first time from tomato in Florida (published). Identified a new species of genomovirus, an emerging genus of viruses,from thrips in Florida (published). Described for the first time a new mastrevirus from sugarcane in Florida (published).

Publications

  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Boukari W, Alcal�-Brise�o RI., Kraberger S, Fernandez E, Filloux D, Daugrois J-H, Comstock JC, Lett J-M, Martin DP, Varsani A, Roumagnac P, Polston JE, Rott PC. 2017. Occurrence of a novel mastrevirus in sugarcane germplasm collections in Florida, Guadeloupe and R�union. Virology J. 14:146.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Kraberger S, JE Polston, HM Capobianco, RI Alcal�-Brise�o, RS Fontenele, A. Varsani. 2017. Genomovirus genomes recovered from Echinothrips americanus sampled in Florida, USA. Genome Announc 5 (21) e00445-17.
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Alcal�-Brise�o RI, Co?kan S, Londo�o MA, Polston JE. 2017. Genome sequence of Southern tomato virus in asymptomatic tomato Sweet Hearts. Genome Announc 5 (7):e01374-16. doi.org/10.1128/genomeA.01374-16
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Polston JE, Londo�o MA, Cohen AL, Padilla-Rodriguez M, Rosario K, Breitbart M. 2017. Euphorbia mosaic virus from passionfruit and Euphorbia heterophylla in Florida, USA. Genome Announc 5 (9): e01714-16.


Progress 10/01/15 to 09/30/16

Outputs
Target Audience:Vegetable and ornamental crop producers, vegetable and ornamental associated industries, vegetable and ornamental plant breeders, plant diagnostic clinics and diagnosticians, virologists, plant pathologists, Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?An undergraduate plant science major received training in how to write a proposal, conduct research, and learned several molecular techniques. A graduate student (MS) received traning in how to conduct research, design experiments and analyze results, and in many appropriate molecular techniques. A postdoctoral associate received training in how to develop and evaluate a new technique, and extensive training in how to write a publishable manuscript. How have the results been disseminated to communities of interest?Results were desseminated primarily through refereed papers. What do you plan to do during the next reporting period to accomplish the goals?Continue to develop assays for virus detection by diagnostic clinics. Continue to characterize new viruses.

Impacts
What was accomplished under these goals? Studies were conducted that evaluated and adapted anew molecular assay for plant virus detection for use in diagnostic clinics (published). We determined that there was low diversity of viruses present in caladiums(unpublished). Studies identified and described he diversity of begomovirus satellites in vegetable crops from a number of locations around the world (published).Studies identified and describedhe diversity of begomovirus satellites in the wild plant, Sida acuta, from Homestead, FL(unpublished). We detected the presence of Southern tomato virus in tomato cultivarsin Florida.

Publications

  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Rosario K, Marr C, Arvind Varsani A, Kraberger S, Stainton D, Moriones E, Polston JE, Breitbart M. 2016. Begomovirus-associated satellite DNA diversity captured through vector-enabled metagenomic (VEM) surveys using whiteflies (Aleyrodidae). Viruses 8 (2), 36; DOI:10.3390/v8020036.
  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Londo�o MA, Harmon CL, Polston JE. 2016. Rapid detection of begomoviruses by recombinase polymerase amplification. Virology Journal 13:48; DOI 10.1186/s12985-016-0504-8.
  • Type: Book Chapters Status: Published Year Published: 2016 Citation: Bowman HM, Polston, JE, McGovern RJ. 2016. Caladium Diseases pp. 1-26 In: Handbook of Florist Crops Diseases. RJ McGovern and WH Elmer, Eds. ISBN: 978-3-319-32374-9 (Print) 978-3-319-32374-9 (Online).


Progress 10/01/14 to 09/30/15

Outputs
Target Audience:Target audience are agricultural producers in Florida and the world, diagnosticians in plant diagnostic labs and centers, plant virologists, plant pathologists, plant breeders. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Training was provided to four graduate students (1 MS, 3 PhD), four undergraduate studentsand a visiting scientist. How have the results been disseminated to communities of interest?Results were dessiminated through refereed publications, a book chapter, a all day meeting with the Florida caladium industry in October, and 3 presentations were made at professional meetings. The newly developed RPA assay for detection of TYLCV was deployed in the UF Plant Diagnostic Center. Protocols were written and then tested successfully in the center. What do you plan to do during the next reporting period to accomplish the goals?Continue with reseach projects that improve diagnostics of viruses, identify new viruses and improve managment of diseases.

Impacts
What was accomplished under these goals? Objective 1. Identify and characterize viruses occurring in horticultural and wild plants in Florida and the Western Hemisphere. We have characterized the diversity of isolates of Dasheen mosaic virus and anotherpotyvirus in Florida caladiums. We conducted studies on the diversity of begomoviruses and their satellites. We conducted studies to identify the causal agent(s) of a systemic necrosis of St. Augustinegrass in Florida. Sugarcane mosaic virus was found in association with the disease. Studies were undertaken to determine if that virus is the sole cause or part of a complex that results in plant death. Objective 2. Characterize transmission of viruses by whiteflies. No significant effort was made under this objective. Objective3. Develop and evaluate new approaches for managment of plant viruses. Work was conducted on a new more rapid assay (RPA) for detection of Tomato yellow leaf curl virus and other begomoviruses. The work was submitted for publication. Specific recommendations were made to caladium growers to reduce the impact and incideance of grassy tuber disease and caladium tuber rots.

Publications

  • Type: Journal Articles Status: Published Year Published: 2015 Citation: "Polston JE, Rosario K, Bretibart M. 2015. Molecular surveillance of plant viruses: identification of new and emerging viruses of tomato before they cause epidemics. Acta Hort. 1069:127-132. "Rosario K, Seah YM, Marr C, Varsani A, Kraberger S, Stainton D, Moriones E, Polston JE, Duffy, S, & Breitbart M. 2015. Vector-Enabled Metagenomic (VEM) Surveys Using Whiteflies (Aleyrodidae) Reveal Novel Begomovirus Species in the New and Old Worlds. Viruses 7(10): 5553-5570. "Harmon, PF, Alcal�-Brise�o, RI, Polston, JE. 2015. Severe symptoms of mosaic and necrosis in cv Floratam St. Augustinegrass associated with Sugarcane mosaic virus in neighborhoods of St. Petersburg, FL. Plant Disease 99:557.


Progress 10/01/13 to 09/30/14

Outputs
Target Audience: Florida vegetable and ornamental growers, extension agents, virologists, diagnosticians Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Training was provided for a postdoctoral student, a graduate student and two research scientists (MS level). How have the results been disseminated to communities of interest? An update on progress in the identification of the causal agents of grassy tuber disease was presented in October to the caladium industry. What do you plan to do during the next reporting period to accomplish the goals? Continue with reseach projects that improve diagnostics of viruses, identify new viruses and improve managment of diseases.

Impacts
What was accomplished under these goals? Objective 1. An new begomovirus was identified in an ornamental plant in Florida. The sequence of this virus was the first full sequence reported for this virus species and helped to resolve the characterization of this isolate and others of this species. The introduction and spread of two newly introducedbegomoviruses to the MIddle East was described. This information will aid in understanding how rapidly begomoviruses can spread and evolve. A virus new to Florida and the US, Cowpea mild mottle virus, was identified using viral metagenomics approaches and whiteflies rather than plants. The presence of the virus was confirmed in weeds and crop plants, and this work provided the first full sequence of the new world Cowpea mild mottle virus species which will aid in the resolution of the confusing taxonomy of this species. Objective 2. A review that clearly connects the dramatic changes in vector taxonomy with changes in viral taxonomy was published. This shouldimprove communications between entomologists and virologists.

Publications

  • Type: Book Chapters Status: Published Year Published: 2014 Citation: Lapidot M, Legg JP, Wintermantel WM, and Polston JE (2014) Management Of Whitefly-Transmitted Viruses In Open-Field Production Systems. Pp 147-206 In: Advances in Virus Research: Control of Plant Virus Diseases: Seed-Propagated Crops, vol. 90. Academic Press. Polston JE (2014) Begomoviruses. Pp. 71-80 In: Compendium of Tomato Diseases and Pests, Second Edition. J B. Jones, T. A. Zitter, T. M. Momol, and S. A. Miller, eds. APS Press, St. Paul, MN.
  • Type: Journal Articles Status: Published Year Published: 2014 Citation: Lapidot M, Gelbart D, Gal-On A, Sela N, Anfoka G, Ahmed FH, Abou-Jawada Y, Sobh H, Mazyad H, Aboul-Ata AE, El-Attar AK, Ali-Shtayeh MS, Jamous RM, Polston JE, Duffy S (2014) Frequent migration of introduced cucurbit-infecting begomoviruses among Middle Eastern countries. Virology Journal 11:181 (10 pages) Polston JE, Londo�o M, Capobianco H (2014) The complete genome sequence of the New World Jatropha mosaic virus. Arch. of Virology 159:3131-3136. Rosario K, Capobianco H, Ng TFF, Breitbart M, Polston JE (2014) Metagenomic analysis of DNA and RNA viruses in whiteflies leads to the discovery and characterization of Cowpea mild mottle virus in Florida. PLoS ONE 9(1): e86748. doi:10.1371/journal.pone.0086748. Polston JE, De Barro PJ, Boykin LM (2014) Transmission specificities of plant viruses with the newly identified species of the Bemisia tabaci species complex. Pest Management Science 70(10):1547-1552.


Progress 10/01/12 to 09/30/13

Outputs
Target Audience: Florida vegetable and ornamental growers, Florida extention agents, virologists Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? A graduate student and a postdoctoral research associate received training in virus identification How have the results been disseminated to communities of interest? Progresson the identification of the causal agents of grassy tuber disease were presented to the caladium industry in October. What do you plan to do during the next reporting period to accomplish the goals? Continue identification of new viruses and publishing of the results.

Impacts
What was accomplished under these goals? Objective 1. We identified the causal agent of afunky spotdiseaseof blueberry as a strain of Blueberry necrotic ring blotch virus. Objective 1. We identified and characterized newly discovered satellites of Sida golden mosaic virus. We demonstrated that the satellites are whitefly-transmitted and that they can replicate with a wide range of helper begomoviruses. Objective 1.We identified a new whitefly-transmitted virus of tomato in Venezuela. Objective 3. We published a video of how to use whiteflies to transmit viruse as a means to screen for resistance to whitefly-transmitted viruses.

Publications

  • Type: Journal Articles Status: Accepted Year Published: 2013 Citation: Polston JE, Capobianco, H. 2013. Transmitting plant viruses using whiteflies. 2013. J. Vis. Exp. (81), e4332, doi:10.3791/4332. Nava A, Londo�o, MA, and Polston, JE. 2013. Characterization and distribution of Tomato yellow margin leaf curl virus, a begomovirus from Venezuela. Archives of Virology 158:399-406. Cantu-Iris M, Harmon P, Londo�o A, Polston JE. 2013. A variant of Blueberry necrotic ring blotch virus associated with red lesions in blueberry. Arch Virol: 158: 21972200.
  • Type: Journal Articles Status: Accepted Year Published: 2013 Citation: Polston JE, Rosario K, Breitbart M. 2013. Molecular surveillance of plant viruses: identification of new and emerging viruses of tomato before they cause epidemics. Acta Hort. (in press).


Progress 07/01/12 to 09/30/12

Outputs
Target Audience: Vegetable and ornamentals producers in Florida, Extension agents, Plant Virologists, Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? An undergraduate was trained in DNA and RNA extraction from plants and in how to detect viruses using PCR. A graduate student received training in how to identify viruses using PCR and RCA followed by cloning. A postdoctoral research associate received training in tospovirus detection. How have the results been disseminated to communities of interest? The results have not yet been distributed to communities of interest. What do you plan to do during the next reporting period to accomplish the goals? We will continue the identification of these viruses by obtaining more sequences and through the analysis of those sequences using appropriate software.

Impacts
What was accomplished under these goals? We made progress on the identification of viruses which caused disease in passionfruit, Jatropha multifida, Celtis occidentalis and Phalaenopsis orchids. We found evidence and obtained partial sequences of begomoviruses in Jatropha, Celtis and passionfruit. We were not able to make any progress on the identification of the causal agent of purpling in tomato. We did find evidence for a tospovirus in Phalaenopsis which is not one of the ones reported to infect Phalaenopsis. Studies are in progress to obtain sequences of these viruses for better identification.

Publications