Callogenesis and plantlet regeneration from two Iranian accessions of bitter melon medicinal plant (Citrulus colosynthis L.) in vitro condition

Background and objectives: Bitter melon is one of the anti-cancer medicinal plants that it’s wide level in vitro propagation and anti-cancer material production was necessary. Different factors such as genotype, selected explants’ type and it`s preparation conditions influenced regeneration content. Plant growth regulator`s effect on callogenesis and indirect regeneration from Parangipetai Indian accession of bitter melon assessed and regeneration method by using stem explants was presented. Their results showed that maximum amount of callus induction was obtained in MS medium supplemented with 0.5 mg/liter of IAA and 2,4-D hormones and 1 mg/liter 6-BA. By attention that we have not any report about indirect plantlet regeneration from Iranian accessions of this plant. For this, in this research we assessed callogenesis and plantlet regeneration in two Iranian accessions of bitter melon.
Materials and methods: After surface sterilization and deleted external hard bark, kernel of this accessions were planted on MS medium and after one month, from obtained plantlets stems and leaves explants were transferred to the MS medium supplemented with different hormonal combinations of BAP, NAA, 2,4-D and Kinetin. For indirect regeneration was also examined on different hormonal combinations of BAP, NAA in MS, ½MS and ¼MS mediums. Regenerated plantlets were transferred in small pots containing sterile soil (soil and sand in 3:1 ratio). This plantlets were changed in complete plants and be able to well established and grow.
Results: Callogenesis in both accessions in different hormonal combinations of 2,4-D and Kinetin more little than mediums supplemented with different hormonal combinations of BAP and NAA. In Dehdasht accession in different hormonal combinations of BAP and NAA and in Gachsaran accession in different hormonal combinations of Kinetin and 2,4-D were obtained approximately 100 and 66.6% Callogenesis, respectively. Shooting in both accessions was started at five weeks after callus segments culture. Leaf explants, in hormonal combination of 3 mg/liter BAP in ½MS medium in Dehdasht accession and stem explants, in hormonal combination of 2 and 3 mg/liter BAP in MS medium in both accession showed that the best indirect plantlet regeneration. The best rooting medium was hormonal combination of 0.5 mg/liter BAP with 0.5 mg/liter NAA. Rooting in both accessions was done with this hormonal combination with this difference that in Gachsaran accession rooting percent more little than Dehdasht accession.
Conclusion: The results of this research showed that Dehdasht accession with regard to Gachsaran accession and stem explants with regard to leaf explants have better response to indirect regeneration. All together, both Iranian accessions of this plant have suitable response to callogenesis and indirect regeneration.