ISHS Acta Horticulturae 546: International Symposium on Molecular Markers for Characterizing Genotypes and Identifying Cultivars in Horticulture
AFLP MARKER DEVELOPMENT IN ROSE FOR GENETIC MAPPING: COMPARISON OF THREE RESTRICTION ENZYME PAIRS |
| Authors: | S. Rajapakse, L. Zhang, R.E. Ballard, D.H. Byrne |
| Keywords: | Rosa, DNA markers, linkage maps, marker assisted selection |
| Abstract:
AFLP technology was applied to construct genetic linkage maps in rose for use in marker assisted selection. The mapping progeny was created by crossing a black spot resistant amphidiploid (86-7) with a susceptible tetraploid (82-1134). One F1 hybrid of this cross, 90-69, was open-pollinated to obtain 115 seedlings. AFLP markers were first used to identify and eliminate seedlings produced through cross-fertilization. The remaining progeny set of 52 F2 plants were used in developing AFLP markers with three combinations of restriction enzymes, EcoRI/MseI, KpnI/MseI. and PstI/MseI. With EcoRI/MseI, primers with three selective bases for each restriction enzyme (+3/+3) yielded an optimum number of bands per gel. However, with KpnI/MseI and PstI/MseI, primers with +2/+3 selective bases produced well-separated bands equivalent in number and dispersion to those produced by +3/+3 selective bases of EcoRI/MseI. One linkage map was constructed for each parent using only single-dose markers. The map of 82-1134 consists of 167 markers assigned to 14 linkage groups and covers more than 682 cM of the genome. The map of 86-7 consists of 171 markers assigned to 15 linkage groups and covers more than 902 cM of its genome. A gene controlling the presence of prickles on the petiole was located at the end of linkage group 7 on the map of 86-7. EcoRI/MseI generated nearly twice as many markers per run than PstI/MseI. Markers developed with three restriction enzyme combinations showed a mixed distribution throughout the maps. Compared to EcoRI/MseI and KpnI/MseI AFLP markers, PstI/MseI markers had a slightly greater distribution in both maps. Overall, PstI/MseI AFLP markers provided the highest percentage of mapped markers. | |
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