Contents lists available at ScienceDirect Industrial Crops & Products journal homepage: www.elsevier.com/locate/indcrop Increased biomass and pigment production from Cassia alata L. callus cultures and their potential as a textile dye Mansi R. Shah, Indu Anna George ⁎ Department of Life Sciences, University of Mumbai, India ARTICLE INFO Keywords: Textile dye Callus Cassia alata Anthraquinones Salicylic acid Ethrel ABSTRACT Cassia alata L. is a medicinally and economically important plant. Various plant parts were analyzed for callus production in this study using diferent plant growth regulators such as IAA, BAP, TDZ, 2-iP, NAA, Kinetin in the concentration range 1 μM–10 μM and their combinations. The hormone combination which gave the best callus weight was selected for callus growth analysis and pigment production. Elicitation with Salicylic acid and Ethrel increased pigment production 12–13 fold. Callus extract was further assessed as a textile dye. The dye obtained had an excellent fastness to washing and perspiration. 1. Introduction Cassia alata is an ethnomedical plant belonging to family Caesalpiniaceae. It is also known as antiringworm plant as it is efective against ringworm. It has been afectively used against a myriad of skin diseases such as fungal infections, insect bites, scabies, herpes, blotch, eczema and mycosis. Traditional remedies for constipation and asthma included Cassia alata as a component. It has been reported to have antiinfammatory, antimutagenic, analgesic, and broad spectrum anti- microbial activities (Ahmed et al., 2013; Singh et al., 2012). Phyto- chemical analysis of Cassia alata reveals that the anthraquinones are one of its major active compounds. However low concentrations of the anthraquinone in the leaf afects the quality of herbal preparation (Dave and Ledwani, 2012; Sakunpak et al., 2009). Anthraquinones have also been employed as natural dyes. There is a surge in the demand for natural dyes in the market due its environment- friendly qualities. This demand strains the natural plant resources whereas its cultivation occupies land that could be used for agriculture. Furthermore the proceeds are obtained when plants are mature. Callus culture on the other hand promises increased pigment production in a short period of time with minimal nonagricultural land requirements. Many species of the Cassia genus such as Cassia fistula and Cassia tora have been recognized as a source of natural dye (Rajesh et al., 2014). This piqued an interest in similar attributes of Cassia alata. The present study traces the growth index of Cassia alata callus cultures with concomitant pigment production, elicitation of pigments from the cultures and its efcacy as a textile dye. 2. Materials and methods 2.1. Plant material Mature brown pods of C. alata were collected from the University of Mumbai Vidyanagari Campus (Fig. 1a). The plant was identifed by Botanical Survey of India (BSI, Pune) where a herbarium sample of the same has been submitted. Pods of C. alata were opened manually. Mature seeds were surfaced sterilized sequentially with detergent (few drops in water, 10 min), 10% Savlon (5 min) and 0.05% mercuric chloride (5 min). The surface sterilized seeds were rinsed thrice with sterile distilled water allowing 5 min for each wash. The seeds were scarifed in a microwave (800 W) for 1 min and cultured in half strength Murashig and Skoog (MS) medium. 2.2. Callus induction Callus was induced from the roots, hypocotyl and cotyledon ex- plants of 7 days old in-vitro germinated seedlings. MS full strength nutrient media supplemented with 6 diferent hormones namely TDZ, BAP, 2-iP; Kinetin (kn), IAA and NAA (concentration range of 1–10 μM) were analyzed for callus induction. The root and hypocotyl explants were of 1 cm in length and cotyledon explants were 1 cm 2 in size. The explants were inoculated such that 10 ml of medium was available for https://doi.org/10.1016/j.indcrop.2018.11.018 Received 16 August 2018; Received in revised form 11 October 2018; Accepted 6 November 2018 Abbreviations: IAA, indole acetic acid; NAA, 1-naphthaleneacetic acid; TDZ, thidiazurone; BAP, 6’benzyl amino purine; 2-iP, 6-(γ, γ-dimethylallylamino) purine; TLC, thin layer chromatography ⁎ Corresponding author. E-mail address: indu.george@lifesciences.mu.ac.in (I.A. George). Industrial Crops & Products 128 (2019) 346–353 0926-6690/ © 2018 Elsevier B.V. All rights reserved. T