Nanobiotechnological approach to stimulate the growth of Astragalus chrysochlorus (Leguminosae) using In-Silico and In-Vivo strategies.

Abstract

Astragalus chrysochlorus has been used in herbal therapy to increase endurance to nephritis, as a heart tonic, as a hepatoprotective, as a diabetes treatment, and as a treatment for viral infections. Our research study was aiming to investigate the In-Silico and In-Vitro effect of chitosan nanoparticles on the growth hormones of Astragalus spp during callus and micropropagation of shoots and roots stages. The plant tissue culture technique allows for the laboratory reproduction of complete plants, organs, tissues, or cells under controlled laboratory conditions. The addition of phytohormones (natural or synthetic growth regulators) at appropriate stages of development or maturity can then be utilised to influence plant development. Callus induction methods, as well as shoot and root micropropagation protocols, were established on Astragalus trigonus seeds. Molecular docking illustrated that chitosan NPS showed an energy binding of -7.86 kcal/ mol against ERF-1 target site. Chitosan formed seven hydrogen bonds with Glu5, Gln282, Asn9, Lys16, Glu324 and Glu390 with bond lengths of 2.05, 1.95, 2.05, 2.02, 2.01, 1.90 and 1.84 Å. Among the various ratios and mixes of four growth regulators, a mixture of iso-pentenyl adenosine (IPA) and isopentenyladenine (2ip) with concentrations of 0.5mg/L from each resulted in the highest Mean number of shoots (MNS) (17.5%) and Mean shoots length (MSL) (12.5cm) (1.28%). The best mixture was 0.5 IPA plus 0.5mg/L 2ip, which was evaluated on five plant samples; MNS obtained a maximum value of 22.5 within two subcultures, second and third, then 18.8 with the first subculture, 15.8 with the fourth subculture, and 13.8 with the fifth subculture. Also, MLS improved in all five subcultures, although the highest value was 15cm with the second subculture and 13.8cm with the fourth subculture. Auxins such as 1-Naphthalene-acetic acid (NAA) and In-dole-3-butyric acid (IBA) were applied to plant samples, with the maximum rooting reaching 100% and the root length improving to 14.4cm in comparison to the control, where rooting and root length were 74% and 12.6cm, respectively. Ionic gelatination was used to create chitosan nanoparticles in this study. These nanoparticles had an average particle size of roughly 6010 nm and a qua-si-spherical shape. Furthermore, the colloidal stability and surface characteristics of the as-prepared nanoparticles were studied using FT-IR and dynamic light scattering techniques. Chitosan NPs were used in various concentrations (0.2, 0.5, 1, 2, 3, and 4mg/L), The best concentration was 0.5mg/L stimulating rooting with 83.3%, 4.7 root/shoot number and root length 6.4cm.