Assessment of somaclonal variation in Plantago major using molecular markers

Paper Details

Research Paper 01/10/2014
Views (168) Download (2)

Assessment of somaclonal variation in Plantago major using molecular markers

Fatemeh Esmaeili, Lia Shooshtari , Mansour Ghorbanpour, Alireza Etminan
J. Bio. Env. Sci.5( 4), 402-408, October 2014.
Certificate: JBES 2014 [Generate Certificate]


Plantago major is a short stemmed annual herb belongs to Plantaginaceae family which is important because of its wide use in pharmaceutical, medical, healthcare and industrial purposes. The present study was conducted to evaluate the the somaclonal variations within Plantago major callus culture using inter simple sequence repeats markers. Callus induction was optimized using leaf and root as explants and MS medium containing various concentrations of 2,4-D and Kin. Moreover the effect of number of subcultures was considered in assessing genetic variability. Eighteen callus samples derived from different explants, media and subcultures were selected and their genomic DNA were extracted using CTAB method with some modifications. Six ISSR primers were used to estimate genetic diversity among 18 callus samples. The comparison of Banding patterns derived from 6 ISSR primers showed that the calli exhibited conspicuous somaclonal variation that could be eventually exploited for in vitro selection systems.


Amzad H, Kunihiko K, Mineo M, Kazuhiro N. 2003. Somaclonal variation of regenerated plants in chilli pepper (Capsicum annuum L.). Euphytica 130(2), 233-239.

Gaafar RM, Saker MM. 2006. Monitoring of cultivars identity and genetic stability in strawberry varieties grown in Egypt. World Journal of Agricultural Sciences 2(1), 29-36.

Jin S, Mushke R, Zhu H, Tu L, Lin L, Zhang Y, Zhang X. 2008. Detection of somaclonalvariation of cotton (Gossypium hirsutum) using cytogenetics, flow cytometry andmolecular markers. Plant Cell Reports. 27(8), 1303-1316. 10.1007/ s00299-008-0557-2

Kaeppler SM,  Kaeppler  HF,  Rhee  Y.  2000. Epigenetic aspects of somaclonal variation in plants. Plant Mol Biology 43(2-3), 179-188.

Kuiper PJC, Bos M. 1992. Plantago: a Multidisciplinary study. Springer, Berlin.

Kuksova VB, Pieven NM, Gleba YY. 1997. Somaclonal variation and in vitro induced mutagenesis in grapevine. Plant Cell, Tissue and Organ Culture. 49(1), 17-27.

Kumar Sharma S, Bryan GJ, Winfield MO, Millam S. 2007. Stability of potato (Solanum tuberosum L.) plants regenerated via somatic embryos, axillary bud proliferated shoots, microtu-bers and true potato seeds: a comparative phenotypic, cytogenetic and molecular assessment. Planta 226(6), 1449-1458. 10.1007/ s00425-007-0583-2

Labra M, Vannini C, Grassi F, Bracale M, Balsemin M, Basso B, Sala F. 2004. Genomic stability in Arabidopsis thaliana transgenic plants obtained by floral dip. Theoretical and Applied Genetics 109(7), 1512-1518. 10.1007/s00122-004-1773-y

Lambe P, Mutambel H, Fouche J, Deltour R, Foidart J, Gaspar T. 1997. DNA methylation as a key process in regulation of or-ganogenic totipotency and plant neoplastic progression. In Vitro Cellular & Developmental Biology – Plant 33(3), 155–162.

Li P, Chen H, Li YX. 2005. Establishment and optimization of in vitro regeneration system for Plantago major L. Chinese Journal of Biotechnology 21(6), 916-922

Lei CP, Jiun KS, Choo CS, Singh R. 2006. Analysis of tissue culture – derived regenerants using methylation sensitive AFLP. Asia Pacific Journal of Molecular Biology and Biothechnology 14(2), 47-55.

Morcillo F, Gagneur C, Adam H, Richaud F, Singh R, Cheah S.C, Rival A , Duval Y, Tregear J.W. 2006. Somaclonal variation in micropropagated oil palm. Characterization of two novel genes with enhanced expression in epigenetically abnormal cell lines and in response to auxin. Tree Physiology 26(5), 585-94.

Murashige T, Skoog F. 1962. A revised medium for rapid growth and bioassays with tobacoo tissue cultures. Physiologia Plantarum 15(3), 473-496.

Nei M. 1978. Estimation of average heterozygosiy and genetic distance from a small number of individuals. Genetics 89(3), 583-590.

Perrier X, Flori A, Bonnot F. 2003. Methods for data analysis. In: Hamon P, Seguin M, Perrier X, Glazmann JC (eds) Genetic diversity of cultivated tropic al plants. Science Publishers, Inc and Cirad, Montpellier, 31–63.

Pourjabar A, Mohammadi SA, Khosroshahli M, Motalebi AR, Ziai SA. 2009. Optimization of in vitro culture of Milk Thistle (Silybum marianum) and analysis of somaclonal variation by RAPD markers. JOURNAL OF AGRICULTURAL SCIENCE 19(2), 95-106.

Sagar GR, Harper JL. 1964. Plantago major L., P. media L. and P. lanceolata L. Journal of Ecology 52(1), 189–221.

Saghai-Maroof MA, Soliman KM, Jorgensen RA, Allard RW. 1984. Ribosomal Spacer Length Polymorphism in Barley: Mendelian Inheritance, Chromosomal Location and Population Dynamics. Proceedings of National Academy of Science of theU.S.A 81(24), 8014–8018.

Samuelsen AB. 2000. The traditional uses, chemical constituents and biological activities of Plantago major L. A review. Journal of Ethnopharmacology 71(1-2), 1–21.

Skirvin RM, Norton M, Mcpheeters KD. 1993. Somaclonal variation: has it proved useful for plant improvement? . II International Symposium on In Vitro Culture and Horticultural Breeding.

Stavri M, Mathew KT, Bucar F, Gibbons S. 2003. Pangelin, an antimycobacterial coumarin from Ducrosia anethifolia. Planta Medica. 69(10), 856-859.

Tatikonda L, Wani SP, Kannan S, Beerelli N, Sreedevi TK, Hoisington DA, Devi P, Varshney RA .2009. AFLP-based molecular characterization of an elite germplasm collection of Jatrophacurcas L., a biofuel plant. Plant Science 176(4), 505-513.

Wakhlu  AK,  Barna  KS.  1989.Callus  initiation  growth and plant regeneration in Plantago ovate Forssk. Cv.Gl-2 Plant Cell Tissue and Organ Culture 17(2), 235-241.

Zubair M, Rumpunen K, Lindholm C, Nybom H. 2010. Genetic and phytochemical variations in Plantago major. 28th international horticultural congress. Lisboa, Portugal, August 22-27, 2010. Poster presentation.

Zietkiewicz E, Rafalski A, Labuda D. 1994. Genome fingerprinting by simple sequence repeat (SSR)-anchored polymerase chain reaction amplification. Genomics 20(2), 176-183.