*Corresponding Author ________________________________________________ Journal of Plant Development Sciences Vol. 9 (3) : 191-200. 2017 ASSESSMENT OF GENETIC FIDELITY OF MOTHER PLANT AND IN VITRO RAISED MEDICINAL PLANT EPHEDRA GERARDIANA THROUGH MOLECULAR MARKERS Indra Rautela 1, 2 , Manjul Dhiman 2 , Manish Dev Sharma 3 , Naveen Kumar 4 and Pragati Misra* 1 1 Department of Molecular and Cellular Engineering, Jacob Institute of Biotechnology and Bioengineering, Sam Higginbottom University of Agriculture, Technology and Sciences, Allahabad, India 2 Department of Botany K.L. DAV (PG) College, Roorkee, Uttrakhand, India 3 Shri Guru Ram Rai Institute of Technology and Science,Patel Nagar, Dehradun, Uttrakhand,India 4 Molecular Cytogenetic Lab, MBGE department, G.B.Pant University of Agriculture and Technology. Email: pragatimisra3@rediffmail.com Received-25.02.2017, Revised-14.03.2017 Abstract: Ephedra gerardiana is an important medicinal gymnosperm shrub. It has been traditionally use for an assortment of medicinal purpose. Molecular markers analysis was conducted to screen genetic fidelity among in vitro raised plantlets compare with mother plant of Ephedra gerardiana. Genetic fidelity of regenerated plants was assessed using Random Amplified Polymorphic DNA (RAPD) and Simple Sequence Repeat (SSR) Primers. A total of 50 RAPD primers and 30 SSR primers were utilized in the present study to analyze genetic fidelity of mother plant and among tissue culture raised plants of Ephedra gerardiana. Out of 50 RAPD primers, 19 primers exhibited DNA amplification in all the DNA samples and out of 30 SSR primers, 18 were show amplification. The amplified products of the regenerated plants showed similar banding patterns to that of the mother plant thus demonstrated the homogeneity of the micropropagated plants. The banding pattern ruled out presence of any kind of somaclonal variation. Thus, the results revealed that genetic fidelity between the micropropagated and mother plant in Ephedra gerardiana and supports the suitability of tissue culture technique for generation of genetically similar plants. Hence, the results obtained confirmed genetic stability of regenerated plants. Keywords: Ephedra gerardiana, Micropropagation, Genetic fidelity, RAPD, SSR INTRODUCTION phedra is a gymnospermic genus of Gnetales and family Ephedraceae. Ephedra belongs to class Genetopsida having 3 different genera Ephedrales, Gnetales and Welwitschiales respectively (sharma et al., 2012). Ephedra, also known as ma huang, contains the ephedrine alkaloid that stimulates the central nervous system (Tod and Stewart 1997). Ephedra have significant medicinal properties due to the presence of various keenly secondary metabolites. Ephedrine alkaloids have an adrenaline like effect on the body it excites the nervous system, opens blood vessels, and stimulates the heart (www.ephedrafacts.com). The Ephedra sinica plant contains ephedrine alkaloids that are used in the herbal supplements once they have been cultivated from the dried stems of the plant. Some of the main alkaloids in Ephedra are ephedrine and pseudoephedrine, which can be found in over the counter (OTC) drugs, but an important distinction is that the alkaloids in OTC drugs are produced synthetically (Paul 2001). Synthetic ephedrine is more potent than the ephedrine alkaloids found in ephedra used mainly as a bronchial decongestant, ephedrine is found in bronchodialators such as Primatene, while pseudoephedrine is commonly utilized in decongestants such as Sudafed. Physiologically, it acts to expand breathing passages, constrict blood vessels, and increase arterial blood pressure. It is the increase in arterial blood pressure that causes severe hypertension, stroke, or heart attack by Jody (1997). Plant tissue culture appears to be a reliable method for the production of Ephedra within a short time span. Among the various methods of in vitro propagation, shoot proliferation is considered to be the least susceptible to genetic modification (Shenoy et al., 1992). Somaclonal variation has been reported to some extent at different levels i.e. morphological, cytological, cytochemical and molecular in micropropagated plants (Rani and Raina 2000). The genetic analysis of micropropagated plants using a multidisciplinary approach, especially at the DNA sequence level initially and at various cultural stages, it is essential to maintain genetic integrity of micropropagated plants. Several cytological, isozyme and molecular markers have been used to detect the variation and/or confirm the genetic fidelity in micropropagated plants (Gupta et al., 2009; Kumar et al., 2011). The sensitivity, reproducibility and strong discriminatory power of microsatellite and RAPD markers (Parida et al., 2009; Sharma et al., 2013) make them suitable for detecting somaclonal variation. However, their application in the study of somaclonal variation has been quite limited. In the present study, Ephedra gerardiana were propagated using shoot buds as the explants. The E RESEARCH ARTICLE