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Development of Resistance to Tomato Spotted Wilt and Similar Viruses inFloral Crops Progress Report — September1994

Date August 30, 1994

Title of Project Development of Resistance to Tomato Spotted Wilt and Similar Viruses in Floral Crops

Institution where work is being conducted North Carolina State University

Amount of Endowment Grant $ 15,000
Covering Period 1/1/94 to 8/30/94

Anticipated Date of Project Completion/Final Report 12/31/95

Individual(s) Conducting Project:

(List Project Leader First)

James W. Moyer - Title Professor

Telephone Number 919/515-5802

Margaret E. Daub - Title Professor

Development of Resistance to Tomato Spotted Wilt and Similar Viruses in Floral Crops

James W. Moyer and Margaret E. Daub

North Carolina State University

Progress Report to the American Floral Endowment, 8/30/94
A. Project Objectives:
To use chrysanthemum as a model system to test the efficacy of geneticallyengineering floral crops for resistance to tomato spotted wilt-like viruses.

B. Summary of Work Conducted:
Over the past five years we have been engaged in researchdesigned to characterize the tomato spotted wilt-like viruses which infect floral crops. This

research led to the discovery that more than one virus is responsible for this disease in floral

crops and to the development of a serological assay for accurate diagnosis which is now the

industry standard. Although accurate diagnosis has helped reduce the viruses from planting

stock, they will probably never be eliminated from the production cycle and thus other measures

are necessary to provide adequate levels of control. During the last two years we have focused

our research on the development of techniques for genetic engineering of resistant plants. We

began our investigation with chrysanthemum, since this crop is economically important, is highly

susceptible to TSWV, and extensive information is available on techniques for growing these

plants in tissue culture. We are thus using chrysanthemum as the experimental model for

transformation of floral crops.

C. Results to Date:
Disease-resistant plants are being developed by ‘genetically-engineered crossprotection”, a process which involves transfer of a virus gene into host plants. Although the

mechanism is not understood, presence of the gene imparts virus resistance to the plant carrying

the gene. In our previous work we successfully developed, for three varieties of

chrysanthemum, protocols for regenerating plants from single cells, a necessary first step for

gene transfer. During the last year we have developed an efficient procedure for moving foreign

genes into these varieties. This procedure has proved effective for gene transfer into both pot

and cut flower mum varieties. We have recently successfully transferred a gene from TSWV

into both a cut flower and pot mum variety. Using both mechanical and thrips inoculation with

a highly virulent chrysanthemum isolate of TSWV, we have identified plants which show a high

level of virus resistance.

D. Future Plans Covered by the Endowment Grant:
Our goals for the coming year are focused in two major areas. First, weare working to make alterations in the virus gene. Information from other systems have

identified gene alterations which can increase the effectiveness of resistance imparted by that

gene. These altered genes are being transferred into the mum varieties and the level of

resistance tested. Second, we are testing our genetically engineered mums for resistance to

diverse isolates of TSWV. As with all disease resistance, virus isolates may exist which are able

to overcome the resistance. One of the factors which we will be investigating is how the

different altered genes affect how the plants respond to diverse isolates. Our inoculation studies

utilize both artificial mechanical inoculation and more natural inoculation using thrips. These

studies will determine the potential usefulness of the resistant varieties for commercial use.

Benefits to the Industry: The overall goal of our work is to define the potential of genetic

engineering for the development of virus resistance in floral crops. Genetic engineering is a

promising strategy for crop improvement, but has yet to be exploited by the floral crops

industry. Through our studies we are producing virus-resistant plants in one crop,

chrysanthemum, and these can be used as breeding stock for the floral industry. The

development of gene transfer protocols developed through this work can also be used for transfer

of other horticulturally relevant genes such as ones affecting flower color or senescence.