Lakshmi et al., IJPSR, 2021; Vol. 12(4): 2211-2224. E-ISSN: 0975-8232; P-ISSN: 2320-5148 International Journal of Pharmaceutical Sciences and Research 2211 IJPSR (2021), Volume 12, Issue 4 (Research Article) Received on 03 April 2020; received in revised form, 10 August 2020; accepted, 21 September 2020; published 01 April 2021 IN-VITRO SEED GERMINATION AND EFFECT OF TDZ AND A g NO 3 ON HIGH FRE-QUENCY SHOOT REGENERATION FROM RUELLIA TUBEROSA L., USING COTYLE-DANARY NODE EXPLANTS P. V. Chaithanya Lakshmi 1 , C. M. Narendra Reddy 1 , P. Rathinapriya 2 , M. Ramesh 2 and B. Srinivas * 1 Department of Biotechnology 1 , Dravidian University, Kuppam - 517426, Andhra Pradesh, India. Department of Biotechnology 2 , Alagappa University, Science Campus, Karaikudi - 630004, Tamil Nadu, India. ABSTRACT: Ruellia tuberosa, commonly called Minnie root, has plenty of applications in herbal medicine due to the presence of several bioactive compounds. A simple, efficient, and reproducible in-vitro plant regeneration protocol was developed for Ruellia tuberosa. The cotyledonary nodal explants were excised from seedlings were inoculated on Murashige and Skoog (MS) medium supplemented with various concentrations of BAP and NAA, respectively. The highest numbers of 9.86 ± 0.15 shoots/explant were obtained in MS medium augmented with BAP (2.0 mg L -1 ) and NAA (1.0 mg L -1 ). The micro shoots induced per explant significantly increased with the supplementation of BAP to auxin-containing medium. The results showed that maximum percentage of multiple micro shoots 89%, with a number (13.80 ± 0.26) shoots / explant was found on MS medium supplemented with BAP (2.0 mg L -1 ) and NAA (1.5 mg L -1 ). Further, the highest frequency of micro shoots was produced on MS medium supplemented with TDZ (0.3 mg L -1 ) and AgNO 3 (0.4 mg L -1 ) for large-scale production. The highest frequency of root induction (96%) was recorded on a half-strength MS medium fortified with 1.5 mg L -1 IBA. The well-rooted plantlets were transferred into the plastic cups containing peat and soil (2:1) and successively established in the greenhouse. In conclusion, our study demonstrates efficient in-vitro regeneration, and in the future, this protocol will be a valuable tool to enhance secondary metabolite production in Ruellia tuberosa. INTRODUCTION: The recent researches on Ruellia tuberosa have shown the presence of rich secondary metabolites, which are in demand commercially in the pharmacology field. These include traces of Luteolin and appreciable amounts of apigenin and Malvidin 5 Glucoside in the aerial parts. QUICK RESPONSE CODE DOI: 10.13040/IJPSR.0975-8232.12(4).2211-24 This article can be accessed online on www.ijpsr.com DOI link: http://dx.doi.org/10.13040/IJPSR.0975-8232.12(4).2211-24 Apigenein and Malvidin 5 glucoside are the anthocyanins that are present in flower buds of Ruellia tuberosa 1 . Anthocyanins are the most studied class of natural polyphenols that provides color for fruits and flowers. Piyaporn et al., 2010 2 conducted research regar- ding dyeing property of flowers of Ruellia tuberosa on cotton fabrics by applying chitosan as a mordant obtained best result. Several studies have also shown that foods rich in anthocyanins are healthy, and they can act against cancer, inflammation, cardiovascular and chronic disorders. Recent advancements in biotechnological techniques such as tissue culture, enzyme and fermentation Keywords: In-vitro propagation, Micro shoots, Ruellia tuberosa, Thiadiazuron and silver nitrate Correspondence to Author: B. Srinivas Department of Biotechnology, Dravidian University, Kuppam - 517426, Andhra Pradesh, India. E-mail: bathulasrinivas71@gmail.com