In Vitro Culture of Aegle marmelos Against Media Composition Stress: Molecular Identification, Media, and Enzyme Optimization for Higher Growth Yields

Abstract

Aegle marmelos, known for its spiky appearance, is a versatile tree found worldwide. In the Indian medical tradition, this therapeutic tree is utilized to treat various ailments. It is commonly propagated through seeds, although they have a limited lifespan and are susceptible to insect damage. Due to the variability of seed offspring, standardized varieties are not readily available. Molecular identification was performed for the plant species to be as a fingerprint identification based on genomic basic. Hence, this study manipulated the in vitro multiplication for enhancing Aegle marmelos traits through variation in media type and composition. In phase one of the experiment, successful micropropagation has been easily achieved with shoot tip culture on two growth in vitro media: Murashige and Skoog (MS) medium and woody plant medium (WPM) with different concentrations (one-fourth, one-half, three-fourths, and full power media) with two sucrose concentration 20 and 30 g/L. The growth parameters measured indicated a heightened response to both MS and WPM media, each with its distinct composition. The genetic variation via intersimple sequence repeat (ISSR) molecular marker in the first phase was 35.5%. In phase two, the hormonal treatment was applied for the best media choice from Phase 1. During the second phase of multiplication and rooting stages with phytohormones, the optimal treatments were chosen to maximize yields. In the multiplication stage, the most favorable conditions, as determined by morphological parameters, were achieved with full MS medium supplemented with 30 g sucrose, 0.1 mg/L Kin, and 0.75 mg/L BAP. In contrast, for the rooting stage, the optimal treatment consisted of one-fourth MS medium supplemented with 15 g sucrose, 0.5 mg/L Kin, 0.1 g/L activated charcoal, and 15 mg/L IBA. Physiological parameters exhibited variability, with each metabolite displaying distinct optimal conditions. Catalase plays a crucial role in decomposing hydrogen peroxide to protect cells, tissues, and organs. This research effectively enhanced the in vitro micropropagation of Aegle marmelos by determining the most efficacious medium formulations and hormonal treatments for shoot multiplication and roots, while also illustrating the influence of WPM on catalase enzyme activity enhancement.