Evaluation of genetic differentiation among healthy and infected Buxus hyrcana with boxwood blight using RAPD and ISSR markers.
Background: Buxus hyrcana (boxwood) is an endangered species in the Hyrcanian forests in the north of Iran. This tree is threatened by habitat loss but faces additional threats from the introduced disease the boxwood blight (caused by the fungus Calonectria pseudonaviculata syn. Cylindrocladium pseudonaviculatum, Cy. buxicola) and the potential effects of climate change. As wide range of genetic polymorphism is necessary to ensure successful adaptation to rapid climatic changes. Methods: Genetic diversity and differentiation between 15 healthy and 15 infected trees of each of two populations were studied using RAPD and ISSR molecular markers. Results: High-band polymorphism was found in pooled samples of B. hyrcana using both ISSR (58%) and RAPD (73%) markers. The ISSR data and the combined data set classified the trees into two groups. However, data from RAPD clustered the trees into three groups. These results indicate different degrees of genetic variation in the sequences of the tested B. hyrcana genomes targeted by the two marker types used. Genetic variation was found to be relatively high, with most of the diversity occurring within populations. Analyses of healthy versus infected pooled samples based on both marker types indicated that genetic diversity parameters were mostly higher in healthy trees. Conclusions: Boxwood blight has had a major effect on B. hyrcana, killing individual stems quickly especially in dense populations and reducing population size (as observed in all populations). Considerable within-population diversity, and higher genetic variability in healthy trees than infected ones, suggested conservation efforts should focus on survivor trees in each population and consider the establishment of tree reservations. Propagation of plants from seeds is preferred, since it would include the widest range of genetic variation.