The present study was undertaken to establish a protocol for plant regeneration of I. adriatica, I. illyrica and I. rotschildii. Leaf base segments were used as explants. For callus induction leaf base explants were incubated in the dark on three media with MS mineral solution containing different content of growth regulators: dichlorophenoxyacetic acid (2, 4-D), 1- naphthaleneacetic acid (NAA) and kinetine (KIN). Logistic model with genotypes and treatments as explanatory variables was used to analyse callus response data. Transfer of embryogenic calli on hormone free medium enabled the development of mature somatic embryos. When plant tissue has passed through in vitro culture, many regenerants appear to be no longer clonal copies of their donor genotypes. For this reason phenotypic variability among regenerants was evaluated. Several morphological characteristics were measured and subjected to a principal-component analysis (PCA). The DNA ploidy level was analysed by flow cytometry analysis. Analysis was conducted among 105 regenerants and showed that the DNA ploidy level remained stable in 104 regenerants. |
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