International Journal of Pharmaceutical Science Invention ISSN (Online): 2319 – 6718, ISSN (Print): 2319 – 670X www.ijpsi.org Volume 3 Issue 1‖ January 2014 ‖ PP.41-45 www.ijpsi.org 41 | Page In Vitro Culture of Highly Valuable Medicinal Plant Bacopa Monnieri (L.) Penn. for Rapid and Mass Multiplication. Yugal Kishore Mohanta 1 *& Souvagyalaxmi Sahoo 2 1 (P.G.department of Zoology,North Orissa University,Baripada,757003,India) 2 (Tectona Biotech resource Centre (TBRC),Shishupalgarh, Bhubaneswar,India) ABSTRACT : A protocol has been developed for micropropagation of Bacopa monnieri (L.) Penn., a medicinal plant of high commercial potential with high reputation as a memory vitalizer or enhancer. Nodal segments containing axillary buds were surface sterilized with 0.1% solution of mercuric chloride for 5 min and were inoculated aseptically on culture medium. In vitro clonal multiplication methods and the elite clones were observed that, MS basal+0.5 mg/l IAA and MS basal+ 0.5 mg/l NAA shown the best results for culture initiation and axillary shoot proliferation. For rooting MS+ Agar 7 g/l, Sugar 20g/l and MS+Agar 8g/l were found to be the best mediumin terms of shoot/root ratio and numberof shoots.Compact globular callus was best initiated and proliferated on MS+0.5 mg/l BAP+1 mg/l 2,4 D and medium with best regeneration in MS+1.0 mg/l BAP+1 mg/l IAA. The experimentation was made successful with 71% survival plantlets producing regeneration of the Bacoppa monnieri L. Penn. for mass cultivation. KEYWORDS: Bacopa monnieri MS medium, Rooting, Shoot multiplication I. INTRODUCTION Bacopa monnieri L. (Family Scrophulariaceae) also referred to as B. monniera, Herpestis monniera, water hyssop, and “Brahmi,” has been used in the Ayurvedic system of medicine for centuries [1]. It is a genus of spreading herbs, commonly growing in damp and marshy places throughout India, ascending up to an altitude of 1320 m, and it is a small creeping, glabrous, succulent, herb rooting at nodes. It is an ancient and renowned medicinal plant with legendary reputation as memory vitalizer [2]. Traditionally, it was used as a brain tonic to enhance memory development, learning, concentration and to provide relief to patients with anxiety or epileptic disorders. The plant has also been used in India and Pakistan as a cardiac tonic, digestive aid, and to improve respiratory function in cases of bronchoconstriction. Bacopa’s antioxidant properties may offer protection from free radical damage in cardiovascular disease and certain types of cancer [3]. It possesses anti-inflammatory, analgesic and antipyretic activity [4]. In very recent study, B. monnieri was placed second position in a priority list of most important Indian medicinal plants evaluated on the basis of medicinal importance, commercial value and potential candiadte for further research and development [5,6]. It is also well known to contain steroidal saponins Bacoside A and steroidal saponins Bacoside B. Some other constituents present in Brahmi are alkaloids brahmine, herpestine etc. Compounds responsible for the pharmacological effects of bacopa include alkaloids, saponins and steroids.The effects of B. monnieri have already been approved by clinical research trials. There is thus an immediate need for assessing the natural populations, developing protocols for micro propagation, regeneration and agronomical practices. The present study deals with the rapid mass scale multiplication of B. monnieri using nodal explants containing axillary bud. II. MATERIALS AND METHODS 2.1 Collection of explants Nodal segments were collected from the juvenile shoots of growing at Experimental Nursery, Tectona Biotech Resource Centre (TBRC). Juvenile segments containing single axillary bud shoots tips were used as source material for micro propagation. 2.2 Sterilization of explants B. monnieri twigs with 3 - 4 nodes were collected, excised from plants maintained in the nursery, Tectona Biotech Resource Centre (TBRC) were washed thoroughly in running tap water to remove the superficial dust and mud, then explants were washed with dilute detergent (1 - 2% Labolene) solution for 15 min and then washed well in running tap water. Again these explants were dipped in to the Bavistene (3-4%) fungiside. Explants were surface sterilized with 5 min treatment with 0.1% (w/v) HgCl 2 . B. monnieri was extremely sensitive to surface sterilizing agent, therefore, the surface sterilization procedure was optimized and this helped in preventing blackening of tissues and establishment of clean cultures. During surface sterilization treatment, it was found that treatment with 0.1% mercuric chloride leads to blackening of the explant [7,8].