Abstract
Surface sterilization is a vital step in preparation of healthy and viable explants in tissue culture. Most surface contaminants can be eliminated by surface sterilization with a suitable sterilizing agent. The study aimed to present an effective disinfection method for Clinacanthus nutans shoot regeneration using nodal segments. A total of four different sterilization approaches were conducted by treating nodal explants with various concentrations of sterilizing agent. Sterilizing agents used were Rhizophora apiculata Pyroligneous acid (PA), sodium hypochlorite (Clorox) thiophanate-methyl (fungicide), and Mercuric chloride (HgCl2). Nodal explant then was cultured on plant growth regulator-free Murashige and Skoog (MS) basal medium. This study sterilizing agents revealed that PA showed strong bactericidal activity. However, it led to a high number of fungal contaminations. The pyroligneous acid did not exhibit a strong potential as a disinfectant for C. nutans nodal explant. Overall, HgCl2 exhibits the best reduction in fungal contamination and gives a significant result with thiophanate-methyl fungicide. Surface sterilization with mercuric chloride (0.2%) for 1 hour was the optimum concentration and duration, which resulted in the highest percentage of nodal explant survival and viability. All viable nodal segments developed into shoots. It had been concluded that the best surface sterilization agent was HgCl2.
Highlights
C. nutans belong to the Acanthaceae family and is known as Sabah snake grass
Various sterilization treatments yielded aseptic cultures but three highest aseptic culture percentage were achieved by treating the explants with 0.2%, 0.1% HgCl2 for 1 hour and 30% thiophanate-methyl for 30 minutes
The maximum percentage of explant viability was achieved when explants were surface sterilized with 0.2% mercuric chloride
Summary
C. nutans belong to the Acanthaceae family and is known as Sabah snake grass. It is a medicinal herb used to treat various diseases, such as skin rashes, insects, scorpion or snake. Production of sterile and viable in vitro plantlets is crucial to increasing the clonal propagation efficacy in plant tissue culture [6]. Regardless of its profound health effect, mercuric chloride is frequently utilized for surface sterilization to reduce microbial contamination in a C. nutans tissue culture. The Thio-m fungicide had never been used in plant tissue culture sterilization before its potential as sterilizing agent at different concentrations was studied. Evidence shows that the major groups of PA compounds are hydroxy aldehydes, hydroxy ketones, sugars, carboxylic acids, and phenolics [16] It consists of 5.5% acetic acid, 3.4% methanol, and 6.5% wood tar. This study's objective was to evaluate effective sterilization protocol of C. nutans prior in vitro culture using different types of sterilizing agent
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