Journal of Medicinal Plants Research Vol. 4(17), pp. 1822-1824, 4 September, 2010
Available online at http://www.academicjournals.org/JMPR
DOI: 10.5897/JMPR10.060
ISSN 1996-0875 ©2010 Academic Journals
Short Communication
Detection of somaclonal variations using RAPD
fingerprinting in Silybum marianum (L.)
Tariq Mahmood
1
*, Nazia Nazar
1
, Bilal Haider Abbasi
2
, Mir Ajab Khan
1
, Mushtaq Ahmad
1
,
Muhammad Zafar
1
1
Department of Plant Sciences, Quaid-i-Azam University, Islamabad-45320, Pakistan.
2
Department of Biotechnology, Quaid-i-Azam University, Islamabad-45320, Pakistan.
Accepted 27 April, 2010
Random amplification of polymorphic DNA (RAPD) markers was used to detect somaclonal genetic
variations between in vitro and in vivo grown tissues of Silybum “S. marianum”. Ten primers (OPC1-
OPC10) of RAPD OPC were used. All the primers gave reproducible banding pattern except OPC3.
Monomorphic bands were observed in case of all primers, whereas only OPC10 generated different
banding pattern among the samples of S. marianum. OPC 10 produced ten unique bands in each
sample ranging in size from 200 - 1000 bp. On the basis of the results obtained, it was observed that
genetic variation is present in different samples of S. marianum and RAPD technique can be used to
detect genetic similarities and dissimilarities between in vitro- and in vivo-grown tissues of S. (S.)
marianum.
Key words: RAPD, regeneration, Silybum, somaclonal variation, thistle.
INTRODUCTION
Silybum (S.) marianum (L.) Gaertn is used for oral
treatment of toxic liver damage and for therapy of chronic
inflammatory liver diseases and liver cirrhosis (Morazzoni
and Bombardelli, 1995). This activity of S. marianum is
due to a mixture of flavonolignans, known as silymarin
(Kurkin et al., 2001). At present, all of the commercially
available Silybum is obtained from wild. The increasing
worldwide demand for Silybum is endangering the sparse
populations. Thus, to fulfill the increasing demand,
developing an efficient in vitro regeneration protocol of
genetically similar S. marianum plants with desirable
agronomic and chemical traits is needed. In this respect
different studies related to the standardization of its tissue
culture conditions and antioxidants activity has been
reported (Abbasi et al., 2010). Nonetheless, somaclonal
variation frequently occurs as a consequence of the
propagation process in different plant species (Chuang et
al., 2009). Somaclonal variation reduces the commercial
*Corresponding author. E-mail: taariq.mahmood@yahoo.com,
tmahmood@qau.edu.pk. Tel: +92-51-9064 3144. Fax: +92-51-
9064 3004.
value of plants (Oropeza et al., 1995). Thus it is very
important to detect somaclonal variation at earlier stage
of plant growth to avoid economic loss (Chuang et al.,
2009).
Genetic variation analyses can be observed from
morphological characters and other markers as protein or
deoxyribonucleic acid (DNA)-based markers. However,
the morphological characters evaluation requires the
plants to grow to full maturity prior to identification.
Subsequently genetic resources characterization also
needed to establish development of genetic markers for
valuable traits (Paterson et al., 1991).
RAPD is the simplest, sensitive and useful technique
for the analysis of genetic fidelity of in vitro propagated
plants (Chuang et al., 2009; Williams et al., 1993).
Because RAPD polymorphism results from either a
nucleotide base change that alter the primer binding site,
or from an insertion or deletion within amplified region.
Polymorphism usually results in the presence or absence
of amplification products from a single locus (Tingey and
del Tufo, 1993).
In this study, RAPD was employed to evaluate the
genetic reliability of S. marianum plantlets regenerated by
leaf organogenesis. To the best of our knowledge, this is