Development of Explant Sterilization Protocol for In-vitro Propagation of Hydrocera triflora (Marsh Henna)
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Date
2019
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Uva Wellassa University of Sri Lanka
Abstract
Hydrocera triflora is a demanded, commercially valuable, perennial, ornamental aquatic plant, distributed within the Indo-Malaysian region and still not developed some in-vitro sterilization protocol for this species. This study was conducted to analyze the effect of surface sterilization treatments on H. triflora explants for development of in-vitro explant’s surface sterilization protocol. Explants (shoot tip, node, and Internode) were cultured in full strength Murashige and Skooge (MS) medium, supplemented with 3% sucrose level and 3:1 BAP (6-Benzylaminopurine): NAA (Naphthalene Acetic Acid) concentration at 5.70 pH. Effect of sterilization treatments with factorial combinations; 15% to 30% Sodium hypochlorite (NaOCl) (1 to 5 minutes), 70% Ethanol (30 s) on explant surface sterilization with pre-treatment (Topsin fungicide, Teepol detergent and Tween 20) and without pre-treatment for each explants were determined after 10 days. All the explant types’ sterilized without pre-treatment indicated 100% contamination by fungal infection within 2 days. In 20% to 30% NaOCl with 70% Ethanol (pre-treated) indicated that there was no significant difference with the number of contaminants and the treatments for all explant types. All the explant types in this treatment were contaminated by bleaching effects. In 15% to 20% NaOCl with 70% Ethanol (pre-treated) indicated that there was a significant difference between treatments for the average number of contaminants (fungal infection). Lowest average number of contaminants for shoot tips (0.00), nodes (0.16) and internodes (0.16) represented the best treatment for explant sterilization, which was provided by 18%, 19% and 18% NaOCl and 70% Ethanol (pre-treated) respectively. The most effective explant sterilization protocol (pre-treatment, 18 to 19% NaOCl and 70% Ethanol) developed from this study can achieve the highest number of healthy explants for in-vitro propagation of Hydrocera triflora.
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Genetics, Biotechnology