ISHS Acta Horticulturae 78: Symposium on Tissue Culture for Horticultural Purposes
PRODUCTION OF BEAN MESOPHYLL PROTOPLASTS AND INOCULATION WITH TOBACCO NECROSIS VIRUS OR TOBACCO MOSAIC VIRUS |
| Author: | D.H. Wieringa-Brants |
| Abstract:
Mesophyll protoplasts have been isolated from leaf tissue of several plant species. So far in most studies tobacco and tomato protoplasts have been used for successful virus infections and cowpea is also reported as being suitable. We used bean mesophyll protoplasts for study of virus behaviour. Protoplasts were isolated from primary leaf tissue of several cultivars of Phaseolus vulgaris L. The protoplast yield differed with the cultivar used. Many factors in the virus inoculation procedure were varied such as incubation time and temperature, buffer, pH, molarity, composition and virusconcentration in the inoculation medium. Despite many trials with different combinations in none of the experiments virusmultiplication of TMV or TNV could be demonstrated at 24, 48 or 72 hr after inoculation of the bean protoplasts. Multiplication of TNV in intact bean leaves at different temperatures was measured until 12 days after inoculation. The first 2 days after inoculation the virus content of the leaves was low. This could be the reason why we did not detect virusmultiplication in bean mesophyll protoplasts. | |
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