ISHS Acta Horticulturae 734: VIII International Symposium on Plum and Prune Genetics, Breeding and Pomology
MOLECULAR GENOTYPING OF SELF-INCOMPATIBLE PLUM CULTIVARS |
| Authors: | B.G. Sutherland, K.R. Tobutt, T.P. Robbins |
| Keywords: | S-RNases, Prunus domestica, Prunus cerasifera, self-incompatibility |
| Abstract:
Consensus primers flanking the first and second intron were designed for amplifying S-alleles in myrobalan (P. cerasifera L.) and domestic plum (P. domestica L.), based on conserved regions of other Prunus S-RNase sequences. Consensus primers spanning the second intron amplified either 1 or 2 bands per diploid myrobalan accession, ranging between 0.7 to 3 kb in size. Sequence information was obtained for 15 of these products. Second intron consensus primers produced 2 to 4 bands when tested in 20 hexaploid domestic plum (P. domestica L.) accessions. Bands in domestic plum ranged between 0.6 to 1.2 kb, from which twelve products were sequenced. First intron consensus primers produced one or two bands per myrobalan accession, with bands ranging from 210 bp to 500 bp in length. Domestic plum cultivars produced between three and six bands, and ranged in size between 228 bp and 493 bp. Sequence diversity was employed to develop allele-specific primers for all 27 sequenced alleles. Data from second intron amplification, first intron amplification, allele-specific PCR and DNA sequencing was then used to provide genotypes for each myrobalan and domestic plum accession. Full S genotypes were obtained for 23 of the 25 myrobalan accessions tested. Partial S genotypes were obtained for 20 domestic plum cultivars, though this does not provide information regarding allele copy number or genomic organisation. | |
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