RESEARCH ARTICLE Chemical Profiling of Tylophora indica (Burm. F.) Merrill. Shoot Cultures Established Through Leaf Explant Swati R. Patel 1 • Ashutosh R. Pathak 1 • Aruna G. Joshi 1 • Neeta Shrivastava 2 • Sonal Sharma 2 Received: 10 December 2019 / Revised: 26 March 2020 / Accepted: 30 September 2020 Ó The National Academy of Sciences, India 2020 Abstract Tylophora indica (Burm. F.) Merrill. is an important medicinal plant of Asclepiadaceae family. Leaf explants were utilized for establishment of shoot cultures in Murashige and Skoog’s medium fortified with sucrose (3%) and different cytokinins individually as well as in combinations. Optimum 28.90 ± 0.41 shoots regenerated in medium supplemented with 6-benzyladenine (BA, 4 lM) and kinetin (Kn, 8 lM), when leaves of second node were placed adaxially with upright position for 8 weeks. Further transferring it to same medium for another 4 weeks formed 63.80 ± 1.45 shoots. The shoots were successfully rooted using earlier published media for the same plant and healthy rooted shoots were hardened and acclimatized to field. High performance thin layer chromatography fin- gerprint revealed that the in vitro shoots were able to synthesize metabolites at par with in vivo shoots. Keywords Developmental age Á HPTLC fingerprint Á Indirect organogenesis Á Leaf explants Á Medicinal plant Á Plant growth regulator Introduction Tylophora indica (Burm. F.) Merrill. (Asclepiadaceae), commonly known as Indian ipecac or Antamul, is tradi- tionally known to treat asthma and hence is also known as ‘asthma herb’ [1]. The plant is routinely used in whooping cough, dysentery, diarrhea and rheumatic gouty pains [2]. It contains many alkaloids of which tylophorine is major one which have anti-angiogenic [3], anticancerous [4] and antitumor [5] activities. Increasing demand of this plant has led to its depletion which causes a threat to this species in wild as the seed viability and its germination rate is low [6]. This has created a gap in fulfilling the market demand of the plant and it can be overcome by in vitro techniques which have been reliable and continuous source of metabolites [7]. Leaf is suitable for regeneration of large number of shoots in medicinal plants of Asclepiadaceae family [8] and regeneration depends on many factors like explant age and its orientation on culture medium [9, 10]. These in vitro shoots can be used as an alternative to wild shoots and hence they should be assessed for their biosynthetic potential using chromatography techniques like high performance thin layer chromatography (HPTLC) [11, 12]. In the present study, large numbers of shoots were regenerated from leaf explants as compared to earlier reports [6, 13–15]. In the present study, the effect of explant orientation on shoot regeneration as well as HPTLC fingerprint of in vitro shoots is being reported for the first time which will help in mass production and to Significance Statement Maximum 28.90 ± 0.41 shoots were regenerated in BA (4 lM) and Kn (8 lM) within eight weeks and multiplied up to twelve weeks formed 63.80 ± 1.45 shoots. HPTLC profiling proved that in vitro shoots were able to synthesize metabolites as in vivo shoots. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s40011-020-01194-y) contains sup- plementary material, which is available to authorized users. & Aruna G. Joshi aruna.joshi-botany@msubaroda.ac.in 1 Department of Botany, Faculty of Science, The Maharaja Sayajirao University of Baroda, Vadodara, Gujarat 390002, India 2 B.V. Patel Pharmaceutical Education and Research Development (PERD) Centre, Ahmedabad, Gujarat 380054, India 123 Proc. Natl. Acad. Sci., India, Sect. B Biol. Sci. https://doi.org/10.1007/s40011-020-01194-y