Abstract
In this study, an efficient micropropagation protocol was developed for A. longiloba and the antioxidant properties of field-grown plant, in vitro-derived greenhouse-grown plant and in vitro-derived callus extracts were compared. The A. longiloba seeds tested using tetrazolium chloride salt exhibited 89% viability. Due to poor germination capacity of A. longiloba seeds, the seeds were treated with gibberellic acid (GA3) or sulfuric acid (H2SO4). The maximum seed germination of 87% was observed at 30% H2SO4 treatment after 19.00 d, whereas GA3 treatment showed maximum germination of 53% after 22 d. In vitro shoot multiplication was carried out using various types of cytokinins alone or in combination with auxin. Among them, 6-benzyl amino purine (BAP) single treatment was found to be the best hormone. The highest shoot-length (7.26 cm) and maximum number of shoots per explant (18) were recorded at 3-mg L−1 BAP. For in vitro rooting, indole-3-acetic acid at 0.5-mg L−1 was found to be the optimum concentration. Callus was induced using various types of auxins alone or in combinations with cytokinins. The highest percentage of callus of 91 and fresh weight of 6 g was obtained with 3-mg L−1 IAA. The plantlets produced in the current study were subjected to acclimatization. The combination of topsoil and peat moss at 1:2 ratio was found to be the best soil media. In this study, in vitro-derived callus extract showed the highest phenolic content (538 mg GAE), followed by extracts of field-grown plant parts, i.e., fruit and petiole (504 and 300 mg GAE) while in vitro plant extract showed the lowest (98 mg GAE). Meanwhile, the highest flavonoids was recorded in petiole extract. Comparative antioxidant activity study shows, in vitro-derived callus exhibited better DPPH-radical-scavenging activity (IC50: 0.113-mg mL−1) whereas the extracts of petiole, fruit and in vitro plant showed 0.126-, 0.137- and 0.173-mg mL−1, respectively. At the same time, the fruit extract showed better (IC50: 0.088-mg mL−1) ABTS radical scavenging activity than all extracts tested. In conclusion, the in vitro-derived callus extract could be favored for high TPC and better DPPH scavenging activity. Hence, the present study was conducted to establish an efficient micropropagation protocol and to compare the antioxidant activity of the field-grown plant, in vitro plant and in vitro derived callus extracts of A. longiloba.
Highlights
IntroductionIn vitro culture techniques are an alternative tool for the production of medicinally important plant metabolites, rapid multiplication of endangered or rare species, production of disease free plants and plant–genome transformation [1]
Over the past decades, the use of modern biotechnological approaches such as in vitro micropropagation techniques has vastly increased due to its great potential for the propagation of high-value medicinal plants and manufacturing of high-quality natural products.In vitro culture techniques are an alternative tool for the production of medicinally important plant metabolites, rapid multiplication of endangered or rare species, production of disease free plants and plant–genome transformation [1]
The current study suggested that 3-mg L−1 of benzyl amino purine (BAP) is the optimal concentration that can be used to induce large number shoots from shoots tips of A. longiloba
Summary
In vitro culture techniques are an alternative tool for the production of medicinally important plant metabolites, rapid multiplication of endangered or rare species, production of disease free plants and plant–genome transformation [1]. These techniques assure sustainable production of plants and bioactive compounds (secondary metabolites) as they are independent of environmental factors [2,3]. Further complications include low amounts of the bioactive compounds of interest and slow growth rate of many medicinal plants. Environmental factors such as light, temperature and CO2 can significantly affect the concentration of bioactive compounds in plants
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