ISHS Acta Horticulturae 651: XXI International Eucarpia Symposium on Classical versus Molecular Breeding of Ornamentals - Part II
OPTIMISATION OF RELATIVE QUANTITATIVE RT-PCR FOR EXPRESSION ANALYSIS IN AZALEA FLOWER COLOUR SPORTS |
| Authors: | E. De Keyser, J. De Riek, E. Van Bockstaele |
| Keywords: | real-time RT-PCR, expression analysis, azalea, flower colour, sporting |
| Abstract:
The fastest way to create new azalea (Rhododendron simsii hybrids) cultivars is by making use of flower colour sports, which appear spontaneously on azalea plants. Unfortunately, there is still very little known on how bud sport induction occurs. Therefore, genes coding for two key enzymes of the azalea flavonoid biosynthesis pathway, chalcon synthase (chs) and dihydroflavonol 4-reductase (dfr) that were reported before to be apt for modification by the action of bud sporting, were isolated and characterized. The expression of these two flower colour genes in the petals of azalea flowers will be compared between all ‘Hellmut Vogel’ flower colour sports. To measure the expression levels of both genes, relative quantitative RT-PCR analysis will be worked out on a real-time PCR machine. The expression of housekeeping genes, which is expected to be the same for all sports, will be used to calculate the relative expression level of the two genes of interest. The optimisation of this technique will be discussed. | |
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