Abstract
The present study investigates the effects of different concentrations, as well as type of plant growth regulators (PGRs) and medium (MS, Duchefa) on the growth and development of Centella asiatica in semi-solid culture. In addition, a protocol for successful sterilization of C.asiatica explants prepared from field-grown plants highly exposed to fungal and bacterial contamination was determined. Results for sterilization treatments revealed that applying HgCl2 and Plant Preservative Mixture (PPM) with cetrimide, bavistin and trimethoprim which were included after washing with tap water, followed by the addition of PPM in the medium, produced a very satisfactory result (clean culture 90 ± 1.33%) and TS5 (decon + cetrimide 1% + bavistin 150 mg/L + trimethoprim 50 mg/L + HgCl2 0.1% + PPM 2% soak and 2 mL/L in medium) was hence chosen as the best method of sterilization for C.asiatica. The synergistic combination of 6 benzylaminopurine (BAP) and 1-naphthaleneacetic acid (NAA) in concentrations of 2 mg/L and 0.1 mg/L, respectively, in Duchefa medium compared with MS induced the most optimal percentage of sprouted shoots (93 ± 0.667), number of shoots (5.2 ± 0.079) and nodes (4 ± 0.067) per explant, leaf per explant (14 ± 0.107) and shoot length (4.1 ± 0.67 cm). Furthermore, optimum rooting frequency (95.2 ± 0.81%), the number of roots/shoot (7.5 ± 0.107) and the mean root length (4.5 ± 0.133 cm) occurred for shoots that were cultured on full-strength MS medium containing 0.5 mg/L indole-3-butyric acid (IBA). In this study, the acclimatized plantlets were successfully established with almost 85% survival. The findings of this study have proven an efficient medium and PGR concentration for the mass propagation of C.asiatica. These findings would be useful in micropropagation and ex situ conservation of this plant.
Highlights
Centella asiatica is a valuable medicinal and aromatic herb which is spread throughout the tropical and sub-tropical regions
HgCl2 and Plant Preservative Mixture (PPM) with cetrimide, bavistin and trimethoprim were included after washing with tap water, followed by the addition of PPM in the medium, to give a very satisfactory result and (TS5) was chosen as the best method of sterilization for C.asiatica (Figure 2)
Joshee et al [7] reported that a 60 min soak in a 2% PPM solution before culture establishment and all culture media supplemented with 2 mL/L PPM helped to control fungal and bacterial contaminations
Summary
Centella asiatica is a valuable medicinal and aromatic herb which is spread throughout the tropical and sub-tropical regions. It has its origin in the wetlands of Asia, such as in China, India, and Malaysia. The most common method employed for the micropropagation of C.asiatica involves the propagation of shoots via a solid or semi-solid system. Such a semi-solid system has successfully contributed to improved multiplication yields, and it has become gradually more important in improving productivity and reducing the time taken to multiply commercially important material
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