Industrial Crops and Products 76 (2015) 1123–1132 Contents lists available at ScienceDirect Industrial Crops and Products journal homepage: www.elsevier.com/locate/indcrop Micropropagation, molecular profiling and RP-HPLC determination of mangiferin across various regeneration stages of Saptarangi (Salacia chinensis L.) Jaykumar J. Chavan a,∗ , Dhanaji M. Ghadage a , Ashok S. Bhoite b , Suraj D. Umdale c a Department of Botany, Yashavantrao Chavan Institute of Science, Satara 415001, India b Vice Chancellor, Shivaji University, Kolhapur 416004, India c Department of Botany, Shivaji University, Kolhapur 416 004, India a r t i c l e i n f o Article history: Received 7 April 2015 Received in revised form 10 August 2015 Accepted 11 August 2015 Keywords: Antidiabetic Anticancer Callus RP-HPLC ISSR Mangiferin Mature node Micropropagation RAPD Salacia chinensis a b s t r a c t Salacia chinensis L. (family – Hippocrateaceae) is a convenient source for industrial scale isolation of antidiabetic and anticancerous drug ‘mangiferin’ which is extensively used in both traditional and mod- ern medicines. High demands from pharmaceutical industries lead the indiscriminate harvesting from the wild which created the pressure on natural populations. Micropropagation system was developed using different explants for both commercialization and conservation of S. chinensis. Murashige and Skoog’s (MS) medium with different concentrations and combinations of plant growth regulators (PGRs) viz. 6-benzylaminopurine (BAP), kinetin (KN), indole-3-butyric acid (IBA), indole-3-acetic acid (IAA), - naphthalene acetic acid (NAA) and 2,4-dichlorophenoxy acetic acid (2,4-D) were used during various in vitro regeneration stages. Maximum rate of shoot multiplication (75%) and maximum number of shoots (6.7 ± 1.0) were obtained with MS medium fortified with BAP (2.0 mg/l), NAA (0.8 mg/l) and ascorbic acid (100 mg/l). The antioxidants and other additives significantly altered the rate of shoot multiplication, number of shoots as well as survival rate of shoots. The plantlets rooted on ½MS + IBA 1.5 mg/l, showed 80% establishment in soil without any morphological variation. The genetic integrity of the regener- ated plants was further confirmed using random amplified polymorphic DNA (RAPD) and inter simple sequence repeats (ISSR) analysis. A total of 3871 distinct bands were produced by 12 RAPD and 17 ISSR primers in mother and 20 randomly selected micropropagated clones wherein 3858 (99.66%) bands were monomorphic and rest of 13 (0.34%) were polymorphic. The high level of monomorphism confirms the genetic uniformity of the micropropagated clones. Additionally, reverse phase-high performance liq- uid chromatographic (RP-HPLC) analysis at various in vitro regeneration stages showed uniformity in terms of presence of mangiferin however, the amount of mangiferin varied across regeneration stages. Phytochemical and molecular fingerprinting of micropropagated clones revealed that the in vitro regen- eration protocol developed could be successfully used for industrial-scale propagation of S. chinensis with enhanced production of mangiferin. © 2015 Elsevier B.V. All rights reserved. Abbreviations: AA, ascorbic acid; AM, additive mixture; BAP, 6- benzylaminopurine; IAA, indole-3-acetic acid; IBA, indole-3-butyric acid; ISSR, inter simple sequence repeats; MA, mangiferin; MS, Murashige and Skoog’s medium; NAA, -naphthalene acetic acid; PGR(s), plant growth regulator(s); PVP, polyvinylpyrrolidone; RAPD, randomly amplified polymorphic DNA; RP-HPLC, reversed-phase high performance liquid chromatography. ∗ Corresponding author. Fax: +91 2162 234392. E-mail addresses: jaychavansu@gmail.com, jayjchavan@yahoo.com (J.J. Chavan). 1. Introduction Saptarangi (Salacia chinensis L.), a vital member of family Hip- pocrateaceae is distributed in the tropical and subtropical areas of the world, especially in Indian subcontinent (Li et al., 2008). It is commonly known as ‘Saptarangi’ and ‘Saptachakra’ in Ayurvedic medicine, a traditional medicinal system of India (Govindaraj et al., 2009; Singh et al., 2010). Various parts of S. chinensis have been used to treat variety of ailments such as arthritis, rheumatism, leucor- rhoea, inflammation, fever, bronchitis skin, venereal diseases etc. (Li et al., 2008). The extensive role of this species in treating dis- eases like diabetes, obesity, liver disorder, inflammation, useful as http://dx.doi.org/10.1016/j.indcrop.2015.08.028 0926-6690/© 2015 Elsevier B.V. All rights reserved.