Browsing by Author "De Silva, E. D. U. D"

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Open Access
CSUP technique: a low cost sterilization method using sodium hypochlorite to replace the use of expensive equipment in micropropagation
(National Science Foundation of Sri Lanka, 2012-03-28) Peiris, S; De Silva, E. D. U. D; Edussuriya, M; Attanayake, A. M. U. R. K; Peiris, B. C. N
The ability of in vitro micropropagtion to produce high quality planting material in large quantities is hindered by their high cost of production. Hence the price of planting material is high, making them unaffordable to growers. However, a larger portion of the capital and running cost of micropropagation can be reduced using CSUP (pronounced as seesap) technique, where sodium hypochlorite is used to sterilize glassware and culture media followed by culturing without using a laminar flow cabinet. This study was undertaken with the objective of evaluating the feasibility of the CSUP technique to replace the sterilization carried out using autoclaves and laminar flow cabinets. Sodium hypochlorite in four concentrations 5, 10, 15 and 20% (v/v) were used to rinse glassware and accessories prior to pouring the appropriate culture medium for anthurium cv ‘Tropical Red’ in vitro shoot multiplication. The culturing of axenic shoots was carried out inside a glass-cage and cultures were placed in the growth room. Autoclaved culture medium and a laminar flow cabinet were used in the case of the control cultures. Results revealed that the percentage of contaminationfree cultures, dry weight increase and the shoot multiplication were not significantly different between the respective treatments and the controls, as well as among treatments of different sodium hypochlorite concentrations. Therefore, the CSUP technique using 5% (v/v) sodium hypochlorite solution can be adopted to replace the use of an autoclave and the laminar flow cabinet in the major micropropagation activity of in vitro multiplication.
Open Access
Genetic modification of rosa pinna (Clerodendrum philippinum Schauer) with gai mutant gene for improved plant architecture
(2013) De Silva, E. D. U. D; Herath, M. N. K; Perera, P. A. G. S. K; Fernando, K. K. S; Peiris, S
Rosa pinna (Clerodendrum philippinum Schauer), is a shrub with attractive and fragrant flowers. However, the tall and spread architecture and high leaf to flower ratio hinder its use as an ornamental plant. Hence, changing the level of acceptance of C. philippinum by incorporating a dwarfing gene to the plant is considered useful. A study was conducted to find a suitable regeneration medium and ultimately to modify the plant architecture through transformation of the gibberellic acid inhibitor (gai) mutant dwarfing gene by particle bombardment. Effects of different concentrations of thidiazuron (TDZ:Nphenyl N’ 1,2,3-thidiazol-5-yl urea) on callus initiation and regeneration of shoots and the effects of microcareer flying distance in gene transformation of C. philippinum were investigated. The regenerated shoots were first cultured on hormone free MS medium and transferred to a MS medium with 0.5 mg/L indole butyric acid for rooting after four weeks. The shortest duration for callus initiation, the largest callus volume (28 days after establishment) and the highest number of shoots (75 days after establishment) were observed in 1.5 mg/L TDZ in the MS medium. Results of transferring the gai mutant gene to in vitro leaves of C. philippinum revealed that both the shortest time duration and the highest plant regeneration were at 9 cm micro-carrier flying distance. Leaf tissues of regenerated plants, which carry the gai mutant gene, positively responded for the beta-glucuronidase (GUS) assay. The regenerated shoots were rooted and successfully acclimatized.