ISHS Acta Horticulturae 805: VIII International Protea Research Symposium
DEVELOPING DNA MARKERS FOR THE EARLY IDENTIFICATION OF LEUCADENDRON HYBRIDS |
| Authors: | Hui Liu, Guijun Yan, Ralph Sedgley |
| Keywords: | hybrid identification, Leucadendron, PCR-RFLP, RAMP |
| Abstract:
A rapid and reliable method to accurately identify hybrids at an early stage is essential in Leucadendron breeding. Identification based on morphology can be difficult or impossible when the seedlings are small. DNA based PCR-RFLP and RAMP (random amplified microsatellite polymorphisms) markers were developed for this purpose. PCR amplification of the nuclear ITS region followed by restriction digestion with MboІ generated restriction profiles which were effective in the unequivocal identification of hybrids in L. gandogeri × L. procerum, L. eucalyptifolium × L. procerum and L. salignum × L. discolor. Polymorphic bands were also generated by RAMP analysis using the combinations of RAPD and SSR primers. Compared with PCR-RFLP markers, RAMP markers showed more discriminative information on the verification of hybrids from their parents and also on the heterozygosity/homozygosity status of certain loci. The developed DNA markers have been used in the Australian Leucadendron breeding project for the early identification of interspecific hybrids to increase the breeding efficiency. | |
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