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Volume: 3, Issue: 3, May-June, 2015
DOI: 10.7324/JABB.2015.3304

Research Article

An efficient micropropagation protocol of Bacopa monnieri (L.) Pennell through two-stage culture of nodal segments and ex vitro acclimatization

Shashikanta Behera, Nibedita Nayak, Shasmita, Durga P. Barik, Soumendra K. Naik

Abstract

An efficient plant regeneration protocol through two-stage culture of nodal segment has been reported here for a valuable medicinal plant, Bacopa monnieri. Multiple shoots with large number of shoot buds were induced from nodal explant on Murashige and Skoog’s (1962) (MS) medium fortified with (1.0-5.0 mg L-1) N6- benzyladenine (BA). Of the different concentrations of BA tested, 3.0 mg L-1 BA offered the best result, where the nodes swelled and an average of 6.5 shoots with numerous buds/node were recorded. The swollen nodes, cultured on MS + 3.0 mg L-1 BA, were sub-cultured either on MS or MS supplemented with 1.0 mg L-1 GA3 medium. The best result (114.2 shoots/ node) with an average shoot length of 6.4 cm was observed on MS media supplemented with 1.0 mg L-1 GA3. Cent per cent acclimatization of healthy in vitro regenerated shoots was obtained by ex vitro rooting in plastic pots containing garden soil, which saves time and tissue culture cost by abolishing the in vitro rooting step.

Keywords: Axillary shoot proliferation, Bacopa monnieri, Medicinal plant, Two-stage culture, Ex vitro rooting.

References

1. Shrivastava N, Rajani M. Multiple shoot regeneration and tissue culture studies on Bacopa monnieri (L.) Pennell. Plant Cell Rep. 1999; 18(11):919-923.

2. Majumdar S, Garai S, Jha S. Genetic transformation of Bacopa monnieri by wild type strains of Agrobacterium rhizogenes stimulates production of Bacopa saponins in transformed calli and plants. Plant Cell Rep. 2011; 30:941-954.

3. Ahire ML, Walunj PR, Kishor PBK, Nikam TD. Effect of sodium chloride-induced stress on growth, proline, glycine betaine accumulation, antioxidative defence and bacoside A content in in vitro regenerated shoots of Bacopa monnieri (L.) Pennell. Acta Physiol Plant. 2013; 35:1943-1953.

4. Rastogi S, Pal R, Kulshreshtha DK. Bacoside A3 - a triterpenoid saponin from Bacopa monnieri. Phytochemistry. 1994; 36(1):133-137.

5. Banerjee M, Shrivastava S. An improved protocol for in vitro multiplication of Bacopa monnieri (L). World J Microbiol Biotechnol. 2008; 24(8):1355-1359.

6. Elangovan V, Govindasamy S, Ramamoorthy N, Balasubramanian K. In vitro studies on the anticancer activity of Bacopa monnieri. Fitoterapia. 1995; 66(3):211-215.

7. Tripathi YB, Chaurasia S, Tripathi E, Upadhyay A, Dubey GP. Bacopa monnieri L. as an antioxidant mechanism of action. Indian J Exp Biol. 1996; 4(6):523-526.

8. Rahman LU, Verma PC, Singh D, Gupta MM, Banerjee S. Bacoside production by suspension cultures of Bacopa monnieri (L.) Pennell. Biotechnol Lett. 2002; 24: 1427-1429.

9. Kishore K, Singh M. Effect of bacosides, alcoholic extract of Bacopa monnieri (L.) (brahmi), on experimental amnesia in mice. Indian J Exp Biol. 2005; 43:640\—645.

10. Banerjee M, Modi P. Micropropagation of Bacopa monnieri using cyanobacterial liquid medium. Plant Tiss Cult Biotechnol. 2010; 20(2):225-231.

11. Ali G, Shrivastav PS, Iabal M. Responses of Bacopa monniera cultures to cadmium toxicity. Bull Environ Contam Toxicol. 2001; 66:342-349.

12. Shukla OP, Dubey S, Rai UN. Preferential accumulation of cadmium and chromium: toxicity in Bacopa monnieri (L.) under mixed metal treatments. Bull Environ Contam Tox. 2007; 78:252-257.

13. Sharma S, Kamal B, Rathi N, Chauhan S, Jadon V, Vats N, Gehlot A, Arya S. In vitro rapid and mass multiplication of highly valuable medicinal plant Bacopa monnieri (L.) Wettst. Afr J Biotechnol. 2010; 9(49):8318-8322.

14. Ved DK, Goraya GS. Demand and Supply of Medicinal Plants in India. NMPB, New Delhi and FRLHT, Bangalore, India; 2007.

15. Tiwari V, Tiwari KN, Singh BD. Comparative studies of cytokinins on in vitro propagation of Bacopa monniera. Plant Cell Tiss Org Cult. 2001; 66:9\—16.

16. Rathore S, Singh N. In vitro conservation of Bacopa monnieri- an endangered medicinal plant. Global J Bio-Sci Biotechnol. 2013; 2(2):187-192.

17. Shah JD. Studies in growth and ecology of Bacopa monnieri (L.) Pennell.-a medicinal herb. Ph.D. Thesis, Banaras Hindu University, Varanasi, India; 1965.

18. Sharma N. Micropropagation of Bacopa monneiri L. Penn.- an important medicinal plant. M.Phil. Thesis. Thapar Institute of Engineering and Technology Patiala, India; 2005.

19. Tiwari V, Singh BD, Tiwari KN. Shoot regeneration and somatic embryogenesis from different explants of Brahmi [Bacopa monniera (L.) Wettst]. Plant Cell Rep. 1998; 17:538-543.

20. Ceasar SA, Maxwell SL, Prasad KB, Karthigan M, Ignacimuthu S. Highly efficient shoot regeneration of Bacopa monnieri (L.) using a

two-stage culture procedure and assessment of genetic integrity of micropropagated plants by RAPD. Acta Physiol Plant. 2010; 32:443-452.

21. Murashige T, Skoog F. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant. 1962; 15:473-497.

22. Gomez KA, Gomez AA. Statistical procedure for agricultural research, Wiley, New York, USA; 1984.

23. Binita BC, Ashok MD, Yogesh TJ. Bacopa monnieri (L.) Pennell: a rapid, efficient and cost effective micropropagation. Plant Tiss Cult Biotechnol. 2005; 15(2):167-175.

24. Ramesh M, Saravanakumar RM, Pandian SK. Benzyl amino purine and adenine sulphate induced multiple shoot and root induction from nodal explants of Brahmi, Bacopa monnieri (Linn.) Penn. Green Page. 2006; 5(1):44-51.

25. Mohanta YK, Sahoo S. In vitro culture of highly valuable medicinal plant Bacopa monnieri (L.) Penn. for rapid and mass multiplication. Int J Pharma Sci Invent. 2014; 3(1):41-45.

26. Jain A, Pandey K, Benjamin D, Meena AK, Singh RK. In vitro approach of medicinal herb: Bacopa monnieri. Int J Innov Res Sci Eng Technol. 2014; 3(5):12088-12093.

27. Kaur J, Nautiyal K, Pant M. In vitro propagation of Bacopa monnieri (L.) Wettst- A medicinally priced herb. Int J Curr Microbiol Appl Sci. 2013; 2(8):131-138.

28. Iram S, Anis M. In vitro shoot multiplication and plantlet regeneration from nodal explants of Cassia angustifolia Vahl.-a medicinal plant. Acta Physiol Plant. 2007; 29:233-238.

29. Konan NK, Sangwan RS, Sangwan BS. Somatic embryogenesis from cultured mature cotyledons of Cassava (Manihot esculenta Kranz). Identification of parameters influencing the frequency of embryogenesis. Plant Cell Tiss Org Cult. 1994; 37(2):91-102.

30. Kumari S, Starlin NM, Huxley AJ. In vitro propagation of Bacopa monnieri (L.) - a wetland medicinal plant. J Basic Appl Biol. 2010; 4(3):138-142.

31. Showkat P, Zaidi Y, Asghar S, Jamaluddin S. In vitro propagation and callus formation of Bacopa monnieri (L.) Penn. Plant Tiss Cult Biotechnol. 2010; 20(2):119-125.

32. Yusuf A, Rajesh KT, Nikhilesh S, Rao PS. Effects of antioxidants and gelling agents on regeneration, in vitro conservation and genetic stability of Bacopa monnieri (L.) Pennell. Int J Ayur Herb Med. 2011; 1(3):51-67.

33. Mehta J, Ansari R, Syedy M, Khan S, Sharma S, Gupta N, Rathore R, Vaishnav K. An effective method for high frequency multiple shoots regeneration and callus induction of Bacopa monnieri (L.) Pennell: an important medicinal plant. Asian J Plant Sci Res. 2012; 2(5):620-626.

34. Pandiyan P, Selvaraj T. In vitro multiplication of Bacopa monnieri (L.) Pennell from shoot tip and nodal explants. J Agric Technol. 2012; 8(3):1099-1108.

35. Jain R, Prasad B, Jain M. In vitro regeneration of Bacopa monnieri (L.): a highly valuable medicinal plant. Int J Curr Microbiol Appl Sci. 2013; 2(12):198-205.

36. Asha KI, Devi AI, Dwivedi NK, Nair RA. In vitro regeneration of Brahmi (Bacopa monnieri (Linn.) Pennell - an important medicinal herb through nodal segment culture. Res Plant Biol. 2013; 3(1):01-07.

37. Letham DS, Palni LMS. The biosynthesis and metabolism of cytokinins. Annu Rev Plant Physiol. 1983; 34:163\—197.

38. Kumari R, Priyadarshni M, Anjali K, Shukla LN. In vitro mass multiplication of Bacopa monnieri (L.) an endangered and valuable medicinal herb. Indian J Sci Res. 2014; 7(1):1248-1253.

39. Kumar HNK, Chandana E, Chauhan JB. In vitro propagation of Calamus nagbettai: an endangered plant. J Microbiol Biotechnol Res. 2012; 2 (2):270-275.

40. Srivastava J, Raghav PK. Effect of different gibberellic acid concentration on pigeon pea (Cajanus cajan [L.] Millsp.) cv. Manak (H77216) via cotyledonary node explants. Int J Adv Biotechnol Res. 2014; 5(2):151-156.

41. Taiz L, Zeiger E. Plant physiology. 5th ed. Sinauer Associates Inc., Publishers, Sunderland, Massachusetts, USA; 2010.

42. Phulwaria M, Rai MK, Harish, Gupta AK, Ram K, Shekhawat NS. An improved micropropagation of Terminalia bellirica from nodal explants of mature tree. Acta Physiol Plant. 2012; 34:299\—305.

43. Rathore JS, Rai MK, Phulwaria M, Shekhawat NS. A liquid culture system for improved micropropagation of mature Acacia nilotica (L.) Del. ssp. indica and ex vitro rooting. Proc Natl Acad Sci Sect B: Biol Sci. 2014; 84(1):193-200.

44. Sathyanarayana BN, Dalia BV. Plant tissue culture, Practices and new experimental protocols, IK International Publishing House, Pvt. Ltd. New Delhi, India; 2007.

How to cite this article:
Shashikanta Behera, Nibedita Nayak, Shasmita, Durga P. Barik, Soumendra K. Naik. An efficient micropropagation protocol of Bacopa monnieri (L.) Pennell through two-stage culture of nodal segments and ex vitro acclimatization. J App Biol Biotech. 2015; 3 (03): 016-021.

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