An efficient protocol for in vitro regeneration and conservation of Shirui lily (Lilium mackliniae Sealy): a lab-to-land approach to save the rare endangered Asiatic lily species
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An efficient protocol for direct and indirect shoot regeneration and proliferation from bulb scales of Shirui lily (Lilium mackliniae Sealy), an endangered Asiatic lily species endemic to the Shirui hill peak, Manipur, India, has been developed. Bulb scales were isolated from mature bulbs and cultured on Murashige and Skoog (MS) basal medium supplemented with different concentrations of 6-benzylaminopurine (BAP), kinetin (KIN), or thidiazuron (TDZ). For direct shoot regeneration from bulb scale explants, 0.5 mg L−1 BAP yielded the highest shoot induction (3.5 shoots per scale; a 96.7% response). For indirect de novo organogenesis, optimum callus induction was achieved with 2.0 mg L−1 2,4-dichlorophenoxyacetic acid (2,4-D), and shoot organogenesis was higher (16.2) when subcultured onto 0.5 mg L−1 BAP medium. Multiple shoot regeneration and pseudo-bulb formation protocols were assessed; the highest shoot proliferation (10.1) occurred with 0.5 mg L−1 BAP and 1.0 mg L−1 gibberellic acid (GA3). Rooting response was 96% with 0.5 mg L−1 1-naphthalene acetic acid (NAA), with multiple roots per shootlet. Plantlet survival was increased to 92.5% during the hardening-off process by using hydroponics with Hoagland’s solution in a mist chamber. Clonal fidelity was assessed through random amplified polymorphic DNA (RAPD) analysis comparing the mother plant and regenerated plantlets. After confirming genetic uniformity, the pseudo-bulblets with four to six leaves and three to four roots were successfully established at the Shirui hills peak. This in vitro regeneration and ex vitro conservation approach could be helpful to save this rare endangered species in a sustainable way.
KeywordsBulb scale Callus Endangered Lilium mackliniae Sealy Micropropagation RAPD
The authors are thankful to the Director and Joint Director of ICAR Research Complex for NEH Region for the infrastructure facilities. The “Shirui village authority” is gratefully recognized for providing the necessary plant materials for the present study. The authors also express gratitude for the support of “Shirui youth club,” Mr. Ajit Kumar Singh, SMS (Plant Protection), and Dr. Y. Ramakrishna, Head, Farm Science Centre, Ukhrul, for the necessary cooperation in awareness and transplantation of the in vitro plantlets at the Shirui hill peak.
MRS and MD participated in designing the experiment, analyzing the data, and drafting the manuscript. MPD and MD executed the experiments. All authors read and approved the final manuscript.
The authors wish to acknowledge the financial support of the Indian Council of Agricultural Research (ICAR), Government of India, under institutional program, and the Department of Science and Technology (DST), Government of India, for internship to MPD for carrying out this work.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
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