Ethnobotanical Leaflets 12: 419-424. 2008.
Conservation Efforts through In Vitro Seed Germination of a Hepatoprotective Plant - Phyllanthus beddomie (Gamble) Mohanan
Maridass, M1 and K. Thangavel2
1Animal Health Research Unit, St. Xaviers College,
2Depart of Biotechnology,
Thirunelveli (D.t), Tamil
Author for correspondence: E-mail: email@example.com
Whole Plants and plant parts used in ethnomedicine contain curative agents that are used in many modern medicines. The Phyllanthus genus contains over 600 species of shrubs, trees and annual or biennial herbs distributed throughout the tropical and subtropical regions of both hemispheres (Figueira et al., 2006), and have long been used in traditional medicine to treat chronic liver diseases (Dhiman et al., 2005). Phyllanthus appears to be promising in patients with chronic hepatitis B virus (HBV) infection (Thyagarajan et al.1988; 1990;1999). Bioactive principles like alkaloids, tannins, flavonoids, lignans, phenols and terpenes have been isolated from various species of Phyllanthus and their compounds showed antinociceptive activity (Cechinel Filho et al.1996). Liu et al. (2001) published a meta-analysis of the effect on and safety of genus Phyllanthus for chronic HBV infection.
In - vitro propagation of endangered plants can offer considerable benefits for the rapid cultivation of species that are at risk, that have limited reproductive capacity and exist in threatened habitats (Fay, 1992). In vitro propagation methods are essential components of plant genetic resources management and they are becoming increasingly important for conservation of rare and endangered plant species (Sudha et al. 1998; Benson et al. 2000; Iankova et al. 2001; Bhatia et al. 2002). The successful use of plant tissue culture techniques for the micropropagation of members of the genus Phyllanthus has been reported for species such as P. emblica, P. urinaria, P. amarus, P. abnormis, P. caroliniensis, P. tenellus, and P. niruri (Unander, 1991; Ishimaru et al., 1992; Santos et al., 1994). Phyllanthus beddomie (Gamble) Mohanan is a sub-shrub found in evergreen forest and the flowering and fruiting season is April - May. Viswanathan et al., (2002) reported the existence of this species (rediscovered) after a lapse of about 73 years in 1998 (collected from Kalakad Mundanthurai Tiger Reserve, Tirunelveli District), and they have treated this species as critically endemic one. Thus, there is an urgent need for the conservation of this species. There are no reports on the in-vitro micropropagation of P. beddomie. The present study represents a preliminary approach for the development of a rapid and reproducible protocol for in vitro multiplication of P. beddomie, which may be helpful in the conservation of this medicinal plant. Moreover, this type of approach may be a vital tool to have a continuous supply of raw materials for the pharmaceutical industry throughout the year to formulate antihepatitic medicines.
Materials and Methods
In vitro culture of Phyllanthus beddomie
Phyllanthus beddomie seeds were collected from Kalakad Mundanthurai Tiger Reserve forest area (Kodhaiyar) and sterilized by immersion in 70% ethanol for 5sec, followed by immersion in 0.5% aqueous solution of sodium hypochlorite with one drop of Tween 20 per 500ml for 20min, and rinsed three times in sterile double distilled water. Surface sterilized seeds were sowed for germination in MS medium. Ten duplicate sets were maintained (one in light and one in complete dark condition). Prior to this, an attempt was made to test the viability of the seeds by sowing them in fertile soil. But there was no appreciable rate of germination.
Sowed seeds in MS medium (Murashige and Skoog, 1962) were
incubated for 21 days in darkness in a growth cabinet at 24 o C. Varying levels cytokinins
Results and Conclusion
Among the tested
concentrations and combinations of PGRs, auxins were able to facilitate the rooting phenomenon
than that of shooting (Benson, 2000). Even in case of the duplicates
supplemented with cytokinins there was no notable
percentage of shooting. This may be
due to the improper placement of plumule during the
initial stage of germination. Among the auxins
Regarding the nature of
emerged roots, sturdy nature of the roots was proportional to the increasing
concentration of auxins like
No responses in root and shoot formation of MS medium containing BA, and Kinetin in low concentrations (0.10 2.00 ΅M). The findings of the present study may be helpful for the development of micropropagation of P. beddomie. Further studies are ongoing for the further establishment and ex situ conservation of P. beddomie and such type of medicinal plants are in need to fulfill the present need of the pharmaceutical industry (Fay, 2002).
Table 1. The frequency (%) of seed germination responses of P. beddomie after 21 days on MS medium supplemented with PGRs in different concentrations.
The authors wish to thank
the Department of Science and Technology, Govt. of India,
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