Multiple shoot proliferation, bulblet induction and evaluation of genetic stability in Asiatic hybrid lily (Lilium sp.)

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 GA3. 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.