Research Article Effects of NAA and BAP, Double-Layered Media, and Light Distance on In Vitro Regeneration of Nelumbo nucifera Gaertn. (Lotus), an Aquatic Edible Plant Noraini Mahmad, 1 Rosna Mat Taha, 1 Rashidi Othman, 2 Azani Saleh, 1,3 Nor Azlina Hasbullah, 4 and Hashimah Elias 1 1 Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603 Kuala Lumpur, Malaysia 2 International Institute for Halal Research and Training (INHART), Herbarium Unit, Department of Landscape Architecture (KAED), International Islamic University Malaysia, 53100 Kuala Lumpur, Malaysia 3 Faculty of Applied Science, MARA University of Technology, 40450 Shah Alam, Selangor, Malaysia 4 Department of Agriculture Sciences, Faculty of Technical and Vocational Education, Sultan Idris Education University, 35900 Tanjung Malim, Perak, Malaysia Correspondence should be addressed to Noraini Mahmad; fara aid@siswa.um.edu.my Received 10 February 2014; Revised 25 March 2014; Accepted 28 March 2014; Published 7 May 2014 Academic Editor: Sami Aifa Copyright © 2014 Noraini Mahmad et al. his is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. In vitro direct regeneration of Nelumbo nucifera Gaertn. was successfully achieved from immature explants (yellow plumule) cultured on a solid MS media supplemented with combinations of 0.5 mg/L BAP and 1.5 mg/L NAA which resulted in 16.00 ± 0.30 number of shoots per explant and exhibited a new characteristic of layered multiple shoots, while normal roots formed on the solid MS basal media. he double-layered media gave the highest number of shoots per explant with a ratio of 2 : 1 (liquid to solid) with a mean number of 16.67 ± 0.23 shoots per explant with the formation of primary and secondary roots from immature explants. In the study involving light distance, the tallest shoot (16.67 ± 0.23 mm) obtained from the immature explants was at a light distance of 200 mm from the source of inlorescent light (1000 lux). he plantlets were successfully acclimatized in clay loam soil ater 8 months being maintained under in vitro conditions. 1. Introduction Lotus is in the genus of Nelumbo and belongs to the family of Nelumbonaceae. he Nelumbonaceae family consists of a perennial aquatic and emergent angiosperm plant which consists of two species: Nelumbo nucifera Gaertn. (the Asian or sacred lotus) and Nelumbo lutea (Willd.) Pers. (the Amer- ican lotus or water chinquapin). he former is distributed in Asia and North Australia and the latter is found in North and South America [1, 2]. Lotus is an important economic aquatic plant, not only as a dainty and ornamental lower but also as a source of herbal medicine, with strong bioactive ingredients, including alkaloids and lavonoids, and antioxi- dant, antisteroidal, antipyretic, anticancerous, antiviral, and antiobesity properties [3–6]. Lotus is usually propagated vegetatively through rhizome division or tuber production, but the normal propagation rate is very low [7]. It can also be multiplied through seeds but, for quick and more eicient germination, the seeds need to be scariied by rubbing the outer hard seed coat gently on sandpaper at both ends and inally immersing them in water to initiate germination. Scariied seeds were germinated ater 3-4 days while normal seeds took 10–15 days to germinate. If the hard coating remains intact, the seeds will remain viable for centuries and it may take a few years for the seed to sprout if placed in water [8]. he present research is aimed at studying in vitro regen- eration of immature (yellow plumule) explants on solid Murashige and Skoog (MS) media supplemented with difer- ent combinations and concentrations of -napthaleneacetic Hindawi Publishing Corporation e Scientific World Journal Volume 2014, Article ID 745148, 8 pages http://dx.doi.org/10.1155/2014/745148