Browsing by Author "Greeshma Baby"

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    Development of micropropagation protocols for aquascape plants
    (Department of Floriculture and Landscaping, College of Agriculture, Vellayani, 2025-05-30) Greeshma Baby; Rafeekher, M
    The thesis work entitled “Development of micropropagation protocols for aquascape plants” was carried out at Department of Floriculture and Landscaping, College of Agriculture, Vellayani during 2022-2024. The study was conducted to standardize in vitro propagation protocol for three aquascape plants Hydrocotyle verticillata, Rotala wallichii, and Echinodorus grisebachii using stem node explants. The study comprised of five experiments. The first experiment was to standardize surface sterilization of explants; explants of three plants were separately treated with mercuric chloride in concentrations (0.08 %, 0.1 %) for durations (5 minutes, 7 minutes), mercuric chloride (0.03 %, 0.05 %) for durations (5 minutes) and sodium hypochlorite (1 %, 1.5 %) for durations (10 minutes, 15 minutes). The experiment was laid out in completely randomized design consisting of eleven treatment combinations along with control (distilled water) with three replications and carried out separately for three plants. In Hydrocotyle verticillata 0.08% HgCl₂ for 7 minutes was the best treatment resulted in maximum survival with no contamination. In Rotala wallichii 0.03 % HgCl₂, 5 minutes was most effective with no contamination and maximum survival. In Echinodorus grisebachii 0.05 % HgCl₂, 5 minutes was the best surface sterilization treatment with maximum survival and no contamination. The second experiment was to standardize tissue culture medium for culture establishment and shoot induction. The explants were inoculated in Murashige and Skoog (MS) medium and half-strength Murashige and Skoog (1/2 MS) medium, with different combinations of growth regulators, namely BAP (0, 0.2, 0.5, 1.0, 1.5, 2.0) mg L⁻¹, NAA (0.2, 0.5, 1.0, 1.5) mg L⁻¹, and Kinetin (0, 1, 3, 5) mg L⁻¹. The experiment was laid out in completely randomized design with twenty-six treatment combinations and three replications. In Hydrocotyle verticillata, MS +1.5 mg L-1 BAP was found to be the best in all shoot initiation parameters. MS + 2 mg L-1 BAP was found to be best shoot initiation medium in Rotala wallichii and Echinodorus grisebachii. Third experiment was to standardize the rooting medium. Explants were inoculated in different combinations of IBA (0, 1, 3, 5) mg L-1 and IAA (0, 0.5, 1, 1.5) mg L-1. The experiment was laid out in completely randomized design with fourteen treatment combinations and three replications. The Best rooting medium in Hydrocotyle verticillata and Rotala wallichii was ½ MS + 1.0 mg L-1IBA with maximum root initiation percentage and rapid root emergence. In Echinodorus grisebachii ½ MS + 3.0 mg L-1 IBA was the best. Fourth experiment was to standardize sucrose concentration for both shooting and rooting medium. The experiment was laid out in completely randomized design with eight treatments each for shooting and rooting medium. Explants were inoculated in best shooting and rooting medium along with different sucrose concentrations (10, 20, and 30) g L-1. For Hydrocotyle verticillata, among various sucrose concentrations tested, 1.5 mg L-1 BAP medium along with 30 g L-1sucrose was found to be most effective for enhancing all vegetative attributes, including shoot initiation, elongation, and leaf production. Additionally, the best rooting response was achieved in half strength MS with 1 mg L-1IBA and 30 g L 1sucrose, resulted in maximum root initiation. The combination of 30 g L-1sucrose with MS medium containing 2 mg L-1 BAP was the most effective for promoting shoot initiation, elongation, and leaf formation in Rotala wallichii and maximum root initiation was obtained in half strength MS medium with 1 mg L-1 IBA and 30 g L-1 sucrose. In Echinodorus grisebachii, supplementation of MS medium with 2 mg L-1 BAP and 30 g L-1 sucrose significantly enhanced shoot initiation, elongation, and leaf production and maximum rooting response was observed in half-strength MS medium enriched with 3 mg L-1 IBA and 30 g L 1sucrose. The fifth experiment was to standardize hardening medium. Soil, aqua soil, LECA pebbles, sand, gravel and coco fiber were used in this experiment. These substrates were filled in net pots and kept inside containers filled with water for hardening. The experiment was laid out in completely randomized design consisting of eight treatments with five replications. Among these aqua soil proved to be the most effective medium for hardening in all three plants. The protocols for Hydrocotyle verticillata, Rotala wallichii, and Echinodorus grisebachii were standardized. For Hydrocotyle verticillata, the shoot initiation medium was 1.5 mg L-1 BAP, rooting with ½ MS + 1.0 mg L-1 IBA, 30 g L-1 sucrose, and aqua soil as the hardening substrate. In Rotala wallichii, 2 mg L-1 BAP for shoot initiation, rooting with ½ MS + 1.0 mg L-1 IBA, 30 g L-1 sucrose, and aqua soil for hardening. For Echinodorus grisebachii, shoot initiation medium was 2 mg L-1 BAP, rooting with ½ MS + 3.0 mg L-1 IBA, 30 g L-1 sucrose, and aqua soil as the substrate for hardening.