RESEARCH COMMUNICATIONS CURRENT SCIENCE, VOL. 114, NO. 7, 10 APRIL 2018 1520 *For correspondence. (e-mail: hemantyadav0011@gmail.com) $ Contributed equally. † Deceased. Diversity in a widely distributed dioecious medicinal plant, Tinospora cordifolia (Willd.) Miers. ex. Hook F. and Thomas Suchita Lade 1,$ , Pooja Singh Sikarwar 1,$ , Md. Akram Ansari 1 , Sayyada Khatoon 2 , Nikhil Kumar 3 , Hemant Kumar Yadav 1, * and Shirish A. Ranade 1,† 1 Molecular Biology and Genetics Laboratory, and 2 Pharmacognosy and Ethnopharmacology Division, CSIR-National Botanical Research Institute, Rana Pratap Marg, Lucknow 226 001, India 3 B2/M91 SBI Colony, Sector B, Janakipuram, Lucknow 226 021. India The nature and extent of diversity in Tinospora cordi- folia, a dioecious climber, widely distributed in India, and one of the most important medicinal plants has remained underexplored. The present study reveals wide range of tinosporaside content (0.016–4.523 mg/g), berberine content (0.27–76.9 mg/g) and jaccard dis- tances (0.0341–0.559). The neighbour joining tree, structure simulation and principal coordinate analysis resolved all the accessions into six sub-clusters, four of which were congruent in the analyses. Sub-cluster I uniquely included all male accessions, with above av- erage leaf areas and below average tinosporaside con- tents with hairy and fibrous leaves. Further, analysis of molecular variance considering three populations showed that maximum variance (87%) was within the population. The result of this preliminary study re- vealed genetic diversity, population structure in T. cordifolia. Keywords: AMOVA, berberine, ISSR, NJ tree, RAPD, Tinospora, tinosporaside. MEDICINAL plants in the Indian system of medicine and health include several widely distributed taxa in India; yet, very few studies have reported on the nature and extent of diversity in these plants. Considering their use- fulness, it is essential that detailed studies are undertaken to determine the nature and extent of genetic, phytochem- ical and morphological diversity so as to account for their wide and robust distribution, and also to enable their best utilization for the benefit of humankind. The plant of Tinospora cordifolia (Willd.) Miers. ex. Hook F. and Thomas (family Menispermaceae), is a large spreading, deciduous, succulent climber, widely distributed in almost the entire country 1 from the Himalayas to the southern part of peninsular India. The plant is also found in South East Asian countries such as Borneo, the Philip- pines, Malaysia, Indonesia, Thailand, Vietnam and China; as well as in North, West and South Africa. Its robust and wide distribution in the country has reportedly not been linked to any anthropogenic activities, despite its eco- nomic importance. T. cordifolia is an important medicinal plant commonly called ‘Guduchi’/‘Giloy’ in Hindi and ‘Amrita’ in Sanskrit indicating immortality and signifies its uses for revitalization. In Ayurveda it is designated as a ‘Rasayana’, and is recommended to enhance general body resistance, promote longevity and as an antistress and adaptogen 1,2 . Almost all plant parts have medicinal properties. Leaves, stem, bark, fruits and even the whole plant have been used differently for treatment of a diverse range of disorders including allergies, diabetes, immune system, fever, gout, ulcer, skin inflammation, antidote to venoms, respiratory tract infections, diabetes, rheumatoid arthritis and even cancer 2–5 . Several reports on its chemi- cal constituents, medicinal properties and validation of therapeutic claims have already been published 5–10 . The medicinal properties of the plant have been attributed to a variety of constituents, such as alkaloids, diterpenoids, lactones, glycosides, steroids, sequiterpenoids, phenolics, aliphatic compounds and polysaccharides that have been extracted from T. cordifolia. Of the various components, tinosporaside and berberine are reported as biomarkers for this plant and are dominant compounds 11,12 . Although a number of studies are available on various medicinal or therapeutic constituents isolated from the whole plant or its parts 11–16 , the plant-to-plant variation in the phyto- chemical constituents of T. cordifolia has not been re- ported. An important question arises here on the extent of diversity and the dioecious nature of the plants – whether or not the diversity (morphological, chemical and mo- lecular) is similar in both genders of the plants. Considering the distribution and dioecious nature of the plants, it is expected that there will be extensive vari- ation in genetic as well as phytochemical constituents in male and female plants. Thus, keeping in view the im- portance of T. cordifolia it is important to analyse genetic as well as phyto-chemical diversity in this plant. In this communication we report the extent of diversity in morphology, tinosporaside and berberine content and molecular marker based genetic diversity in a number of T. cordifolia plants. In all, 114 plants were collected from various phyto- geographical zones of the country as stem cuttings, and maintained in the field at CSIR-NBRI garden under natu- ral conditions. Eighty six accessions were selected for the present study which included 29 accessions from Uttar Pradesh, 25 from Madhya Pradesh, 19 from Jammu and Kashmir, 8 from Maharashtra, 3 from Kerala and 2 from Andhra Pradesh ( Supplementary Table 1). Fifty Inter Simple Sequence Repeats (ISSR) primers and hundred Random Amplification of Polymorphic DNA (RAPD) primers were first tested. Ten of each primer (Table 1) that revealed good polymorphism and resolution were