Development of Resistance to Tomato Spotted Wilt and Similar Viruses inFloral Crops Progress Report — June 1999
Project Progress Report Cover Page & Requirement
11 Glen-Ed Professional Park, Glen Carbon, Illinois 62034 Telephone:618.692.0045 Fax: 618:692.4045 Email: bstoeckl@endowment.org
Annual Report Deadline:
Reports must be received no later than June 1, 1999
Reporting Media:
An original hard copy and computer copy must be submitted.
- Submit 30 copies of the report by mail and
- A MSWord computer copy — two media options exist:
(1) Submit on 3.5″ diskette or (2) attach file to email.
Report Content:
Progress reports must include this cover sheet (fully completed). Do not exceed two typed pages (minimum font size = 11). Reports must (1) review industry needs addressed and project objectives, (2) summarize work conducted since inception outlining results and specific benefits to the industry especially new information identified, (3) outline next steps and future for project and anticipated industry benefit(s).
Purpose of Progress Report:
Progress reports enable the board and industry to keep abreast of the work accomplished in each project receiving AFE funding. They provide a tool to evaluate whether the work is meeting industry needs and original objectives. They are also used as a resource to develop publicity for AFE funded research.
IMPORTANT NOTE:
Information provided will be used as needed for publicity purposes unless the investigator indicates otherwise. If the material is confidential, an appropriate summary must be included that can be publicized, so the status of all projects can be released to industry.
Date: June 1, 1999
Title of Project: TOMATO SPOTTED WILT VIRUS RESISTANCE IN CHRYSANTHEMUM
Institution(s): NORTH CAROLINA STATE UNIVERSITY
AFE Grant Amount: 40,000 Grant Period: 9/1/98 to 8/31/99
Project Completion Date (as indicated on proposal application) 8/31/99
Project Leader: James W. Moyer & Margaret Daub
Title: Professors
Address: Dept of Plant Pathology, Box 7716, NCSU, Raleigh, NC 27695-7616
Telephone: 919 515-7984 Fax: 919 515-7716
Email: james_moyer@ncsu.edu
Additional Researchers (Names):
Tomato Spotted Wilt Virus Resistance in Chrysanthemum
James W. Moyer and Margaret Daub
In order to overcome the obstacles of restrictive patents, a vector plasmid containing the TSWV nucleocapsid (N) gene from TSWV isolate 10 was constructed using standard methods. The selected plasmid pE 1120 contains the OCS “super promoter” and “mas” termination signal in addition to the reporter gene GUS. The cassette can be licensed from Purdue and can be used if GUS is removed. First, tobacco plants of the cv Burley 21 were transformed with this vector using an Agrobacterium system. After three months, a total of 31 transgenic tobacco plants were detected by PCR from more than 100 regenerants that survived numerous exposures to the selection agent. The progeny of the transgenic tobaccos are being challenged by mechanical inoculations of TSWV. Second, Chrysanthemum leaf discs were also inoculated with the same vector. Hundreds of regenerants that survived the selection procedure are under evaluation.
A new approach not only to develop but to study TSWV-derived resistance in plants is being implemented by cloning and further transformation of plants with the non-structural protein in the middle RNA (NSm) and the N genes of two TSWV isolates, D and 10. This strategy is being tested because of the recent discovery that N gene resistance can be overcome. Transformants with the N gene will be used as comparative controls. Also, the combination of both genes will be tested to inhibit the ability of the virus to overcome the resistance derived by the N gene alone. First, the NsM and N genes of both isolates will be cloned into a high copy number plasmid to ensure the fidelity of the engineered constructs. The resulting plasmids will be maintained in Escherichia coli cells. This part is nearly completed. Subsequently, the correspondent genes will be subcloned into a selected transformation vector. Agroinoculation will be used to generate transgenic plants. The plants will be challenge with different TSWV isolates. This system allows more flexibility in using different transformation vectors, an essential component in obtaining a more consistent pathogen derived resistance especially with a highly variable and promiscuous virus such as TSWV. This strategy will also benefit studies in defeating host resistance genes by this virus as well as TSWV population monitoring. Successful strategies will be made available to industry breeding programs.
Publications from this project:
- Sherman, J.M., Moyer, J.W. and Daub, M.E. 1998. A regeneration and Agrobacterium-mediated transformation system for genetically diverse Chrysanthemum cultivars.J. Amer. Soc. Hort. Sci. 123: 189-194.
- Sherman, J.M., Moyer, J.W. and Daub, M.E. 1998. Tomato spotted wilt virus resistance in Chrysanthemum expressing the viral nucleocapsid gene. Plant Disease 82:407-414.
- Daughtrey, M.L., Jones, R.K., Moyer, J.W., Daub, M.E., and Baker, J.R. 1997. Tospovirus strike the greenhouse industry: INSV has become a major pathogen on flower crops. Plant Disease 81:1220-1230.
- James W. Moyer & Margaret E. Daub 1998. Status of Management Options forControl of TSWV/INSV in Floral Crop Production Grower Notes Spring 1998.
- J.W. Moyer and M. E. Daub 1998. Control TSWV/INSV in Floral Crop Production: Gene Transfer Technology may offer solutions. Grower Notes. Society of AmericanFlorists and American Horticulturists 3:5-7
- J.W. Moyer & M. E. Daub 1999. Delving into the Enigmatic World of the Tospovirus. Greenhouse Product News 9 (1):18-19.
