Perfecting Explants Sterilization Procedure and Multiple Shoot Induction Medium for In-Vitro Propagation of Lagenandra Species
No Thumbnail Available
Date
2013
Journal Title
Journal ISSN
Volume Title
Publisher
Uva Wellassa University of Sri Lanka
Abstract
Lagenandra species are important aquatic plants in the aquaculture industry of Sri Lanka
(Yapabandara and Ranasinghe, 2006). In the absence of a regular supply due to lack of effective
propagation methods, Lagenandra species are indiscriminately harvested from the wild to
supply to the export market. In order to overcome the problem of species loss and inadequate
supply to the local and foreign markets, an in vitro micro- propagation method was developed
for Lagenandra lancifolia and Lagenandra ovata.
The maintenance of aseptic or sterile conditions is essential for successful tissue culture
procedures. Various sterilization agents are used for surface sterilization the tissues. These
disinfectants are also toxic to the plant tissues, hence proper concentration of disinfectants,
duration of exposing the explants to the various disinfectants, the sequences of using these
disinfectants has to be standardized to minimize explants injury and achieve better survival
(CPRI, 1992). Mercuric chloride and sodium hypochlorite were used for the present study to
standardize the best sterilization protocol for in vitro culture of Lagenandra ovata and
Lagenandra lancifolia.
Culture initiation and multiplication, to a great extent, are dependent on the type and genotype
of explants as well as the type of hormone and their concentration. Many commercial
ornamental plants are being propagated by in-vitro culture on the culture medium containing
auxins and cytokinins (Peril, 2003).BAP (6- Benzylaminopurine) and IAA (Indole acetic acid)
hormone with different concentration were used in the present study to perfect the best
combination of IAA and BAP in shoot multiplication procedure.
Methodology
The research was conducted at the tissue culture laboratory, Floriculture research and
development unit, Royal botanic gardens, Peradeniya. The rhizomes (1 cm - 2 cm pieces) were
used as explants. In the first experiment, explants were soaked in different concentrations of
sodium hypochlorite (15%, 20%, and 25%) and mercuric chloride (0.1%, 0.2%, and 0.4%) to
perfecting explants sterilization procedure. Each explant was soaked for twenty minutes in
Sodium hypochlorite solution and two minutes in Mercuric chloride solutions. Number of
contaminated cultures was observed after two weeks from explants establishment in hormone
free Murashige and Skoog medium.
In second experiment, effect of different hormone combinations (hormone free MS medium, ,
BAP 7 mgl +IAA 0.1 mgl ) in 0.5 MS semi solid medium, full MS semi solid medium and
full MS liquid medium for shoot initiation and multiplication were evaluated. Number of shoot
initiation, shoot length, number of shoots per culture and number of leaves per culture were
recorded after six weeks from explants establishment in MS medium.All the data were analyzed
using ANOVA in SAS and Minitab statistical package.
Description
Keywords
Animal Sciences, Aquaculture and Fisheries, Fish, fish Industry, Aquatic Resources, Aquatic Plant