Effect of Pollination with Gamma Irradiated Pollen on In Vitro Regeneration of Ovule Culture in Cyclamen

Abstract

The obtention of homozygous lines through in situ parthenogenesis via gamma irradiation method is a comprehensive application in vegetables. However, there are a limited number of studies on in situ parthenogenesis in ornamentals. Therefore, in situ parthenogenetic capacity of more species needs to be examined. For this purpose, the effects of pollination with gamma-irradiated pollen on in vitro ovule cultured in Cyclamen persicum L. were evaluated in this study. Flower buds were collected before anthesis and irradiated at different doses 50, 100, 150, 200, 300, and 450 Gy of gamma-ray using a Co-60 source. The control group was pollinated with non-irradiated pollen. Fruits were harvested 30 days after pollination, and isolated ovule explants were cultured on four different mediums in in vitro. M0 was control group containing half-strength MS basal media; M1 media additionally has 10 g/L maltose, 1.0 g/L proline, 2.0 g/L peptone, 200.0 mg/L spermidine and 0.5 mg/L kinetin; in addition to basal medium, M2 media additionally contains 10 g/L maltose, 1.0 g/L proline, 2.0 g/L peptone, 200.0 mg/L spermidine, 0.4 mg/L gibberellic acid (GA3) and 0.4 mg/L N6-benzyl adenine (BA). M3 media additionally contains 2.0 mg/L 2,4-D and 0.8 mg/L 6-( γ, γ-dimethylallylamino) purine (2iP). Plantlets started to form 10-12 weeks after the beginning of culture. The effects of nutrient media, irradiation dose, and their interactions on plant formation were statistically significant. The lowest plantlet regeneration (0.33%) was obtained from ovule explants exposed to 50 Gy gamma rays and cultured on M3 media while the highest plantlet regeneration (2.66%) was obtained from ovule explants cultured on M1 media 30 days after pollination with non-irradiated pollen grains. According to stomatal observations, there were no statistical differences between donor plant and in vitro regenerated plantlets.

A Review on Flower Bulb Micropropagation: Challenges and Opportunities

“…Techniques such as callus culture, embryo rescue, in vitro pollination, somatic hybridization, induction of somaclonal variation, protoplast culture, synthetic seed production, in vitro ploidy manipulation, genetic transformation, gene mapping, and DNA fingerprinting play essential roles [22,69,71]. In vitro pollination and fertilization, embryo, ovary, ovule, anther, and pollen culture have been reported for some geophytes such as Lilium, Cyclamen, Anemone, Ranunculus, and Amaryllis [72][73][74][75][76][77][78][79][80]. Further scientific inquiry is warranted to evaluate the feasibility and potential applicability of protoplast culture and somatic cell hybridization techniques in the context of geophytes such as Crocus cancellatus as studied by Karamian and Ebrahimzadeh [81].…”