Genetic analysis of the genus Diospyros ssp. using RAPD and i-PBS methods

Molecular techniques (RAPD and i-PBS) were used to study genetic diversity within persimmon collection at Horticulture Faculty of Mendel University in Lednice. The aim of the work was to distinguish 14 known and 6 of unknown origin persimmon cultivars. The basic screening of 20 OPT primers was applied to 4 cultivars differring in the place of origin. Within the group of screened primers there were chosen those, which gave polymorphic repeatable strong and middle strong bands. Selected primers were used for the RAPD reactions within the whole persimmon collection. Three OPA primers previously described in the literature were also used for the RAPD reactions within the whole persimmon collection. Additional 16 i-PBS primers previously described in the literature were also used for i-PBS analysis of the whole group of cultivars. Amplification was successful with 12 i-PBS primers. The FreeTree software package was used to generate a similarity matrix and then to produce a dendrogram using UPGMA analyses. The similarity dendrograms of all persimmon cultivars were created based on both approaches and also on combination of both analyses by program Tree View. All the dendrograms clearly separated the assessed cultivars into 4 clusters. There are cluster of American persimmons - Meader' (1), 'Garretson' (2) and 'Early Golden' (3). They are representatives of D. virginiana. Further part of dendrogram includes single D. lotus (5), which is also clearly separated from other cultivars of the genus Diospyros. The third cluster includes interspecific hybrids 'Rossiyanka' (10) and 'Nikitskaiya Bordovaiya' (13), which arised from crosses of D. virginiana and D. kaki. The last cluster is formed by cultivars of Japanese persimmon - 'Mikatani Gosho', 'Zenjimaru', 'Tone Wase', 'Hiratanenashi', 'Fuyu', Chinese cultivar - 'Sansi' and two Italian cultivars 'Vaniglia' and 'Tipo'. They are clustered without significant distinction. The similarities and the differences revealed among incorporation of cultivars into groups were compared with the literature findings.