Micropropagation of Red Bell Pepper hybrid cultivar (Capsicum annuum L.) through in vitro axillary buds proliferation

Document Type : scientific research article

Authors

1 Ph.D. Student of Plant Biotechnology, Dept. of Biotechnology, Imam Khomeini International University, Qazvin, Iran

2 Corresponding Author, Associate Prof., Dept. of Biotechnology, Faculty of Agriculture and Natural Resources, Imam Khomeini International University, Qazvin, Iran

3 Assistant Prof., Dept. of Horticultural Sciences, Faculty of Agriculture, University of Guilan, Rasht, Iran

4 Associate Prof., Dept. of Biotechnology, Faculty of Agriculture and Natural Resources, Imam Khomeini International University, Qazvin, Iran.

Abstract

Abstract

Background and objectives: Production of agricultural crops using imported seeds, causes currency to leave the country and increase the cost of production. In Iran, Bell Pepper as an important vegetable crop with high nutritional and medicinal value, is mainly produced through imported hybrid seeds. Therefore, the use of plant tissue culture techniques is one of the most effective methods to remove dependence on seed imports and mass propagation of plants without morphological and genetic variations. The purpose of this research was to investigate and optimize the micropropagation of the Lorca red bell pepper commercial hybrid cultivar (Capsicum annuum L.) through in vitro axillary buds proliferation using nodal segment explant and finally creating a rapid and efficient protocol for the mass propagation of colored bell pepper tissue culture seedlings.

Materials and methods: In this research, the effect of different concentrations of plant growth regulators, benzyl amino purine (BAP) and indole-3-butyric acid (IBA) on in vitro proliferation of axillary buds was investigated using nodal segment explant in the form of completely randomized design (CRD) with three replications. As well as, the effect of activated charcoal as an absorbent in controlling browning and preventing the formation of callus tissue at the end of explants and stimulating rooting in them was tested. In addition, MS medium containing gibberellic acid (GA3) was used to stimulate the elongation of the propagated axillary buds.

Results: The results showed that the highest rate of axillary buds proliferation (7.98 numbers) was created in the treatment of MS medium containing 5 mg L-1 BAP + 0.2 mg L-1 IBA with 78.33 % shoot induction. The best elongation of shoot buds (2.69 cm) was achieved by stimulating the elongation of shoot buds in the treatment of MS medium containing 0.25 mg.L-1 GA3 and then subculture of them in MS medium supplemented with 0.25 mg L-1 BAP + 0.1 mg L-1 IBA along with 1 g L-1 activated charcoal. The shoot explants that were elongated properly, were rooted during subculture on MS medium containing 1 g L-1 activated charcoal. The rooted plantlets, after the acclimatization stage in Phytotron, were transferred to the greenhouse and 80% of them survived and grew well.

Conclusion: This experiment showed that the highest rate of axillary buds proliferation is created in medium containing high concentration of cytokinin (5 mg L-1 BAP) and low concentration of auxin (0.2 mg L-1 IBA) and by stimulating the mitotic divisions in the shoot cambium layer. Also it was observed that adding a suitable concentration of activated charcoal to the MS medium is an effective method to prevent the oxidative browning on the base of explants and no formation of callus tissue and it has play an important role in the optimal growth of plantlets and their rooting.

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