Genetic variation of commercial Anemone coronaria cultivars assessed by AFLP

Anemone coronaria is an out-crossing species grown widely as an ornamental. The genetic divergence between and within five widely commercialised cultivars ('Cristrina', 'Monalisa', 'Tetraelite', 'Wicabri' and 'Mistral') has been evaluated using AFLP markers. Within each cultivar three-to-eight sub-cultivars, which. differ mainly in the colour of their flowers, were assayed. Four plants within each sub-cultivar were genotyped. Due to the large size of the A. coronaria genome, an AFLP protocol based on restriction with Sbf I (an 8 bp cutter), and Mse I (a 4 bp cutter) was developed. Clear and reproducible electrophoretic patterns displaying 56 - 79 bands: (average 67) per primer combination were obtained by applying five (Sbf I+2 /Mse I+3) selective nucleotides. A total of 402 bands were observed, of which 152 were polymorphic. Genetic similarities between accessions were calculated according to the Simple Matching Coefficient and used to construct a dendrogram based on the unweighted pair group method using arithmetic averages (UPGMA). The dendrogram resolved the entries into three major branches: A, which included sub-cultivars of 'Cristina'; B, which included two clusters grouping the sub-cultivars of 'Monalisa' and 'Mistral'; and C which included two clusters grouping the sub-cultivars of 'Tetraelite' and 'Wicabri'. The genetic relationships that emerged from our results are consistent with the known origins of these A. coronaria accessions. The majority of plants within each sub-cultivar clustered with one another, but in some cases, no clear genetic differentiation between subcultivars was detectable. Due to the high level of within-sub-cultivar genetic variation detected, future breeding strategies should be based on stabilising cultivars for agronomic and commercial characters, while simultaneously maintaining a high level of heterozygosity, in order to avoid inbreeding depression.