Henbane lattice (Hyoscyamus reticulatus L.), is one of the most important medicinal herbs of the solanaceae family which due to having Tropane alkaloids, frequently used in medicinal industry. Medicinal plant Regeneration, proliferation and In vitro ploidy induction are one of the interesting issues in biotechnology and plant tissue culture. This study was arranged as a completely randomized factorial design with Two Factors, BAP in three levels (0, 4.4, 8.8 µM) and IAA in three levels (0, 1.1, 2.2 µM) with three replications. Then, In vitro polyploidy induction in regenerated plants was surveyed by 5 different concentrations of colchicine at (0.00, 0.05, 0.1and 0.2 %) at various exposure times (24, 48 and 72 h). in order to determining plant ploidy level, microscopic, flow cytometry and chromosome counting were performed. The results showed that after six months; the best results (13.13 plantlet in each explants) were obtained in 8.8 µM BAP combined with 2.2 µM IAA. Flow cytometry study results showed that highest polyploid plants (72%) were obtained in 0.1% colchicine with 48 h exposure times. Polyploidy induction was associated with significant changes in plant characteristics. The chromosome number of diploid plants was 2n = 2x = 34 and in tetraploid plants was 2n= 4x =68. Therefore, can say that colchicine effectively induce polyploidy in the plant.
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