Optimization of Micropropagation of Chelidonium majus L. and Study on Total Phenol Content in Different Organs in In Vitro Conditions

علوم باغبانی

Article 11, Volume 34, Issue 1 - Serial Number 45, June 2020, Pages 145-159   

Document Type: Research Article

DOI: 10.22067/jhorts4.v34i1.81895

Authors

Masud Ghasemi¹; Hossein Arouiee^* ; Pejman Azadi²; Atoosa Ali Ahmadi³

¹Ferdowsi University of Mashhad

², Department of Genetic Engineering, Agricultural Biotechnology Research Institute of Iran, Karaj

³Shahid Beheshti University

Abstract

Introduction: Greater celandine (Chelidonium sp) is one of the plants that its propagation through seed occurs slowly. In addition, Chelidonium majus L. has limited habitats in Iran. For this reason, micropropagation can be considered as an effective method for its rapid and massive propagation and conservation, which can lead to the production of highly uniform plants. Chelidonium majus L. also contains a large amount of secondary metabolites of Isoquinoline alkaloids, including Chelidonine, Sanguinarine, Captesin, Berberrine and Chloritrine, and phenolic compounds. Therefore, the aim of this study was to investigate micropropagation of Chelidonium majus L. and compare the total phenol content in leaf, stem and root in obtained plantlets.
Materials and Methods: To begin the experiment, seeds of Chelidonium majus L. were first washed with distilled water containing a few drops of tween20. Then they were washed with 70% alcohol for 1 min and were finally washed with double-distilled water. Next, they were disinfected with 1% sodium hypochlorite for 5 min, and again were rinsed with distilled water for 3 times of 5, 15, and 180 min under laminar air flow hood. The effects of TDZ at concentrations 0, 0.25, 0.5, 0.75 and 1 mg/L, BAP at concentrations 0, 0.5, 1, 1.5, 2 mg/L considered. Then, the effect of best treatment in combination with NAA and IBA at 0.25, 0.5, 0.75 and 1 mg/L on growth parameters (number of shoot, shoot length and shoot formation capacity index), were studied. The effect of IBA, NAA and IAA at 0, 0.5, 1, 1.5 and 2 mg/L on rooting parameters (number of root and root length) in MS medium supplemented with 3 g/L activated charcoal in in vitro conditions were evaluated. Then different ratios of cocopeat, perlite and peat moss were used for acclimatization of the obtaining plants. Folin method was used to measure total phenol content. The experiment was conducted as factorial in a completely randomized design with four replications.
Results: The results of analysis of variance for proliferation and rooting traits showed that there were significant differences among the treatments at 1% probability level. The results of means comparison showed that the highest numbers of shoots and shoot formation capacity index were obtained  from the treatment of 0.5 mg/L TDZ with the average of 8.12, which did not show a significant difference from the concentration of 0.25 mg/L TDZ, and the lowest shoot number was related to the control treatment. Increasing the amount of TDZ hormone led to the reduction in shoot number, so that at concentration of 1 mg/L TDZ, the average shoot number per explant was four. Combination of 0.5 mg/L TDZ with IBA and NAA had lower effect on Chelidonium majus L. proliferation. Moreover, the greatest shoot length was observed in the treatment of 2 mg/L BAP. Comparison of means values showed no significant difference between the treatments of 2 and 1.5 mg/L BAP at 1% probability level. In this study, MS medium containing 1.5 mg/L IBA was the most appropriate treatment for root formation. The effect of NAA hormone on root number of Chelidonium majus L. showed that the highest number of root was obtained from the treatment of 2 mg/L NAA. Besides, the effect of IAA on root number of Chelidonium majus L. showed that the highest number of root was observed in the treatment of 1 mg/L IAA, and the lowest number of root was related to the control treatment .The results of means comparison for the percentage of acclimatized plants showed that the ratio of 0:2:1 had a significant difference from the rest of the culture media and 85% of the plants were acclimatized, while the ratio of 1:2:1 showed the lowest percentage of acclimatization (20%). Furthermore, the results showed that the culture media had significant effect on acclimatization stage at 1% probability level. The results of the analysis of variance for total phenol content in leaf, stem and root tissues showed that there were significant differences among these three tissues. The results showed that the amount of total phenol in leaf was higher than in the stem, and the amount of phenol in root was insignificant.
Conclusion: Based on the results of this study, micropropagation can be used as a method for commercial production of this species under in vitro conditions.

Keywords

Chelidonium sp; Micropropagation; Proliferation; Total phenol

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