ORIGINAL ARTICLE Multiple shoot proliferation, bulblet induction and evaluation of genetic stability in Asiatic hybrid lily (Lilium sp.) Rakesh Yadav • Neha Yadav • Minakshi Pal • Umesh Goutam Received: 15 June 2013 / Accepted: 14 December 2013 / Published online: 29 January 2014 Ó Indian Society for Plant Physiology 2014 Abstract An efficient multiple shoot cluster regeneration and micro-bulb induction protocol was developed for Asiatic hybrid lily (Lilium sp.). Highest multiple shoot cluster formation of 78.4 % was observed on MS medium supplemented with 0.25 mg l -1 BAP and 1.0 mg l -1 GA 3 . The growth regulators, i.e., abscisic acid, daminozide and chlorocholine chloride, significantly affected bulblet quality parameters such as average size, number, fresh mass, and scale count. A dose of 0.1 mg l -1 abscisic acid, 50 mg l -1 daminozide, and 10 mg l -1 chlorocholine chloride exhibited highest micro-bulb regeneration rate per explant of 6.0 ± 0.28, 5.6 ± 0.17, and 6.0 ± 0.34, respectively. A high sucrose concentration of 40 g l -1 also produced highest number of bulblet per explant (6.1 ± 0.05). Amongst the 10 arbitrary decamer primers used to analyze the samples, only 6 primers amplified clear reproducible bands. The 6 primers produced a total of 36 (an average of 6 bands per primer) scorable bands. Phenotypic observations and ran- domly amplified polymorphic DNA (RAPD) analysis established that the regenerated micro-bulbs did not develop any detectable genetic aberration. Thus, the results indicate that in vitro produced bulblets of Lilium spp. (Asiatic hybrid) were clonally identical to their donor plant and were genetically stable. Keywords Bulblet Á Lilium Á Genetic stability Á RAPD Introduction Lilies are well known perennial bulbous plants belonging to the monocotyledonous family Liliaceae. Worldwide, over 100 species reside in Northern Hemisphere and thousands of genotypes are cultivated at commercial scale as floricultural crops, mainly because of their large, attractive flowers (Wang et al. 2009). Bulbs are cultivated commercially for use in the cut flower and potted-plant industries. Additionally, Lilies have also been described for their medicinal values as a stomachic, stimulant and aro- matic drug (Malla 1994). The premier commercial lilies principally belong to three hybrid groups: the Asiatic hybrids (A), the Oriental hybrids (O), and the Longiflorum group (L). The establishment of a reproducible plant regeneration system is necessary to supply true-to-type lily plants for commercial purposes and also for unconven- tional improvement approaches such as genetic transfor- mation and the fusion procedures. Some reports are available for tissue culture for mem- bers of the genus Lilium for various species and hybrids such as L. longiflorum (Nhut 1998), L. rubellum (Niimii et al. 1997), L. lancifolium (Marinengeli and Curvetto 1997), L. auratum (Takayama and Misawa 1979), L. tes- taceum (Wozniewski et al. 1991), L. formosanum (Nakano et al. 2000), L. davidii (LingFei et al. 2009), L. oxypetalum (Joshi and Dhar 2009) L. ledebourii (Bakhshaie et al. 2010) and Lilium hybrids (Nova ´k and Petru ˚ 1981; Marinengeli and Curvetto 1997). However, there is no reliable R. Yadav (&) Á N. Yadav Department of Bio and Nano Technology, Guru Jambheshwar University of Science and Technology, Hisar 125001, Haryana, India e-mail: ry.yadav01@gmail.com M. Pal Centre for Plant Biotechnology, Haryana State Council for Science and Technology, CCSHAU New Campus, Hisar 125001, Haryana, India U. Goutam Department of Biotechnology, Lovely Professional University, Phagwara 144402, Punjab, India 123 Ind J Plant Physiol. (October–December 2013) 18(4):354–359 DOI 10.1007/s40502-014-0060-4