ISHS Acta Horticulturae 546: International Symposium on Molecular Markers for Characterizing Genotypes and Identifying Cultivars in Horticulture
CHLOROPLAST DNA DIVERSITY AND DISTINGUISHING SWEET CHERRY CULTIVARS: A NEW APPROACH |
| Authors: | A. Mohanty, J.P. Martín, I. Aguinagalde |
| Keywords: | Prunus avium, sweet cherry, universal primers, cpDNA, PCR-RFLP, haplotypes, cultivar identification |
| DOI: | 10.17660/ActaHortic.2001.546.63 |
| Abstract:
The first step before recommending a selected cultivar to an agriculturist is its correct identification for future reference. Techniques that use molecular markers for accurate identification of cultivars must show reproducibility, easy accessibility and facilitate easy manipulation in the laboratory. Chloroplast DNA, because of its conserved nature, is commonly not recommended as a good molecular marker to distinguish cultivars. In a preliminary study we applied a PCR-RFLP technique using universal primers to assess cpDNA diversity in 15 cultivars (10 identified and 5 unidentified) of sweet cherry (Prunus avium L.). We amplified 11.5% of the chloroplast genome. A set of insertion/deletion mutations, ranging between 5 bp and 10 bp, enabled the distinction of 7 haplotypes in the identified cultivars. Three haplotypes in the unidentified cultivars were found, of which 2 were common with that of the 10 identified cultivars and one was a new haplotype. The haplotypes of unidentified cultivars (except the new haplotype) helped in assigning them to their respective groups. No intra-cultivar variation was found. These results demonstrate that this PCR-RFLP technique can detect sufficient cpDNA diversity to group the cultivars of sweet cherry into common sets based on their cpDNA haplotypes. This technique also helped to determine erroneous labelling in samples of one identified cultivar. We think that by increasing the numbers of primers sets used and/or restriction enzymes differences between each and every cultivar may be pin-pointed. This preliminary study opens a new possibility for horticulturists to refer to the existing chlorotypes of P. avium and select accordingly the primer-enzyme combination for rapid screening, thereby resolving doubtful identities of a cultivar. | |
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