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. Xavier’s College, 2Depart of Biotechnology, Thirunelveli (D.t), Tamil Author
for correspondence: E-mail: orchideyadass@yahoo.com Issued Introduction 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
like 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
tested, 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 µM). 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, References Benson, E.E., Danaher
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