Contents lists available at ScienceDirect Industrial Crops & Products journal homepage: www.elsevier.com/locate/indcrop Assessment of genetic and biochemical fidelity of field-established Hedychium coronarium J. Koenig regenerated from axenic cotyledonary node on meta-topolin supplemented medium Shashikanta Behera a , Subrat K. Kar b , Kedar K. Rout c , Durga P. Barik a , Pratap C. Panda b , Soumendra K. Naik a, ⁎ a Department of Botany, Ravenshaw University, Cuttack, 753003, Odisha, India b Plant Taxonomy and Conservation Division, Regional Plant Resource Centre, Bhubaneswar, 751015, Odisha, India c Gonasika Science College, Keonjhar, 758001, Odisha, India ARTICLEINFO Keywords: Cotyledonary node Hedychium coronarium HPTLC ISSR Micropropagation ABSTRACT Hedychium coronarium is an important medicinal plant of Zingiberaceae family and is widely used in cosmetics industry and perfumery. H. coronarium is also the source of a number of valuable secondary metabolites in- cluding coronarin D, a compound with immense potential to treat cancer. Therefore, there is an urgent need to ensure sustainable use of this plant. The current study reports an one-step in vitro plant regeneration (simulta- neous production of shoot and root) protocol for H. coronarium using axenic cotyledonary nodes by optimizing the type and concentration of plant growth regulators. In this study, meta-topolin (mT) alone was found to be optimum, as compared with other individual cytokinins and combinations of growth regulators. The production of shoot/ plantlets was highest on MS medium supplemented with 3.0 mg/L mT. Axenic shoot segments isolated from these primary shoots/ plantlets were cultured in the aforesaid medium to produce about 4590 plantlets. Around 200 plantlets were used for acclimatization; approximately 93% plantlets were successfully acclimatized and established in field. The genetic fidelity of the micropropagated plants with that of the mother plant was ascertained using Inter Simple Sequence Repeats (ISSR) analysis. Quantitative biochemical and High Performance Thin Layer Chromatography (HPTLC) analysis of the micropropagated plant vis-à-vis the mother plant confirmed biochemical similarities. In summary, an efficient plant propagation system was developed for H. coronarium to enable sustainable use of the plant for commercial and conservation purposes. 1. Introduction Medicinal plants are under stress in their natural habitat due to urbanization, defragmentation, deforestation, pollution, climate change, natural calamities and indiscriminate harvesting (Chen et al., 2016). In India, for example, a number of medicinal plants with high trade value are collected from wild habitat unsustainably and nothing is being done to cultivate the plants (Ved and Goraya, 2007). As a result, these plants are at risk of becoming endangered. Such a threat can be addressed through biotechnological intervention such as plant tissue culture to produce, throughout the year, a large number of plants, which are both genetically and biochemically similar to the mother plant (Amoo et al., 2012). Hedychium coronarium J. Koenig (Family: Zingiberaceae) is an erect, rhizomatous herb. It is widely used in traditional systems of medicine, particularly in India and China (Behera et al., 2018a, b). Its attractive butterfly-shaped flower has a sweet smell and hence the plant is pop- ular for ornamental use in tropical region (Yue et al., 2015). While the flower has volatile compounds, the rhizome is also known to have vo- latile oil; thus both the flowers and rhizome are used for perfumery (Chadha, 2005; Chan and Wong, 2015; Yue et al., 2015). In addition, different parts of H. coronarium including rhizomes, flowers, and tender shoots are consumed as vegetables in various parts of the world (Behera et al., 2018a). H. coronarium has already been overexploited in different parts of India. Loss of the plant’s natural habitat has pushed it into the list of threatened plants in various states of India (Chadha, 2005; Ved et al., 2008). The plant is usually propagated through rhizome or seed. However, propagation through its geophytic unit, rhizome, although time consuming, is most common as seed germination is very poor https://doi.org/10.1016/j.indcrop.2019.03.051 Received 5 December 2018; Received in revised form 20 March 2019; Accepted 21 March 2019 ⁎ Corresponding author. E-mail address: sknuu@yahoo.com (S.K. Naik). Industrial Crops & Products 134 (2019) 206–215 0926-6690/ © 2019 Elsevier B.V. All rights reserved. T