Drug Invention Today Vol.2.Issue.1.January 2010 17-21 N. Sarada Mani et al. / Drug Invention Today 2010,2(1),17-21 Research Article Available online through www.ditonline.info *Corresponding author. Prof. N. Sarada Mani Department of Botany, Andhra University, Visakhapatnam, INDIA. 530 003 Tel.: + 91-9949129539 E-mail: varaprasadphd@rediffmail.com INTRODUCTION Centella asiatica is a weekly aromatic smelling herb of the family Apiaceae. In India it has been used widely in Ayurvedic medi- cine for hundreds of years to treat a wide range of illness and anxi- ety 1 . This medicinal plant is widely spread in many Asian countries. It is used in different continents by diverse ancient cultures and tribal groups. In India, it is described under the name of Mandukaparni and has many common names including Gotu Kota and Indian Penny- wort 2 . It is also listed as one of the Traditional Chinese Medicine (TCM) in China and it has also been used as a support for faster healing of small wounds 3 . According to the report prepared by the Export and Import bank of India C. asiatica is one of the important medicinal plants in the International market of medicinal plant trade 4 . However, the wild stock of this species has been markedly depleted Because of large scale and unrestricted exploitation of this natural resource coupled with limited cultivation and insufficient attempts for its replacement and now it is listed as threatened species by the International Union for Conservation of Nature and National Resources (IUCN) 5 and an endangered species 6, 7 . As per our knowledge, there are few reports are available on Conservation of an endangered medicinal plant Centella asiatica through Plant Tissue culture T. Bangaru Naidu 1 , S. Nageswara Rao 1 , N. Sarada Mani 1* , Y.S.Y.V. Jagan Mohan 2 , and Sudhakar Pola 2 1 Department of Botany, Andhra University, Visakhapatnam, INDIA. 530 003 2 Department of Biotechnology, Andhra University, Visakhapatnam, INDIA. 530 003 Received on: 07-08-2009; Revised on: 21-09- 2009; Accepted on:19-11-2009 ABSTRACT Centella asiatica is an endemic herbaceous plant with great medicinal value. Medicinal properties of the plant has led to it’s over exploitation coupled with inadequate cultivation and unsatisfactory attempts for its replacement, the wild stock of this species has been markedly exhausted and now it is listed as threatened species. Hence, there is an urgent necessitate to safeguard this precious germplasm. Plant tissue culture has proved to be helpful in conserving threatened plant species. In this direction, in the present study, an efficient protocol was developed for callus induction and plantlet regeneration from nodal explants of Centella asiatica. Well-organized plant regeneration was achieved on MS medium containing different concentrations and combinations of growth regulators. MS medium supplemented with 4 mg/ l and NAA in combination with 2 mg/l 2, 4-D proved effective for callus induction, with this combination we achieved a frequency of 92% and BAP 1.5 mg/l and KN 1.5 mg/l was effective for regeneration response. Keywords: Endangered plant, medicinal , callus induction, shoots, Plantlet regeneration. conservation of Centella asiatica using plant tissue culture 8 . Plant tissue culture studies 9, 10, 11, and recent studies 12, 13 reported callus development, somatic embryogenesis and regeneration in Centella asiatica. However, the rate of regeneration is not to be practical. Consequently, a reproducible protocol for efficient plant regenera- tion from established callus or cell cultures is needed in Centella asiatica to conserve the species. The objective in the present study is to develop an efficient regeneration system through plant tissue culture and to promote cal- lus formation from nodal explants with high regenerative capacity, and to analyze the effect of different auxin’s on rooting of regenerated shoots. The data collected from this study will be used for further investigations into improved production of Centella asiatica for potential reintroduction into its natural habitat and for commercializa- tion purposes. MATERIALS AND METHODS Plant material and explant source: Healthy explants of nodal segments, leaf petiole were col- lected from Centella asiatica (Figure.1) plants grown in Botanical gardens, Andhra University , Visakhapatnam, were washed under running tap water for at least 30 min, followed by soaking in 5% (v/v) Teepol for 5 min. After thorough washing in sterilized distilled water, the explants were surface sterilized with freshly prepared 0.1% (w/v) HgCl 2 for 3 min. Following repeated washing with sterile distilled wa- ter, the explants were inoculated onto culture media. ISSN: 0975-7619