Articles
TRANSFORMATION OF GERBERA PLANTS WITH TOMATO SPOTTED WILT VIRUS (TSWV) NUCLEOPROTEIN GENE
Article number
572_17
Pages
149 – 157
Language
English
Abstract
Tomato spotted wilt virus (TSWV) is identified as a causal agent of an important disease in many vegetable and ornamental crops.
In the Research Institute of Pomology and Floriculture, studies on the introduction of resistance-related, TSWV nucleoprotein gene to the gerbera genome (Alaska, Paul’, Prince, Zuzanna) were undertaken.
Gerbera explants (shoots, bases of shoot clumps and leaves with 2-3 mm lamina length) were transformed by Agrobacterium tumefaciens (strain LBA 4404). Construction of the expression vector containing the 35S CaMV promoter, the selectable nptII gene and the N-gene of the virus naturally infecting gerbera was based on pBin19 plasmid.
After 10 months, seventy-four (5.4%) transgenic gerbera plants were selected on medium with the maximal concentration of kanamycin (7 mg l -1). The presence and integration of the N-transgene into the plant genome was confirmed by specific-PCR and Southern-blotting.
The level of nucleoprotein estimated by ELISA was low in transformed tissues, but mechanically inoculated, transgenic plants demonstrated resistance to TSWV.
In the Research Institute of Pomology and Floriculture, studies on the introduction of resistance-related, TSWV nucleoprotein gene to the gerbera genome (Alaska, Paul’, Prince, Zuzanna) were undertaken.
Gerbera explants (shoots, bases of shoot clumps and leaves with 2-3 mm lamina length) were transformed by Agrobacterium tumefaciens (strain LBA 4404). Construction of the expression vector containing the 35S CaMV promoter, the selectable nptII gene and the N-gene of the virus naturally infecting gerbera was based on pBin19 plasmid.
After 10 months, seventy-four (5.4%) transgenic gerbera plants were selected on medium with the maximal concentration of kanamycin (7 mg l -1). The presence and integration of the N-transgene into the plant genome was confirmed by specific-PCR and Southern-blotting.
The level of nucleoprotein estimated by ELISA was low in transformed tissues, but mechanically inoculated, transgenic plants demonstrated resistance to TSWV.
Publication
Authors
M. Korbin, M. Podwyszynska, B. Komorowska, D. Wawrzynczak
Keywords
transformation, gerbera, TSWV nucleoprotein gene
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