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In vitro Propagation of Curcuma caesia Roxb. via Bud Culture Technique and ISSR Profiling of the Plantlets for Genetic Homogeneity

An in vitro propagation protocol has been developed for Curcuma caesia Roxb., an endangered medicinal plant by Foundation for Revitalisation of Local Health Traditions and Central Forest Department of India. The plant bears poorly germinated seeds and produces two-storage organs: rhizomes and multiple root tubers. Only rhizomes have medicinal-economic values. They serve as propagules too, which results in a shortage of planting material. Therefore, a complete one-year production cycle of C. caesia has been standardized through in vitro propagation including explants establishment (one month), subculture cycles (seven months), rooting (one month) followed by primary hardening (one month) and secondary hardening (two months). Dormant shoot buds on rhizome served as explants for culture initiation on Murashige and Skoog (MS) medium supplemented with different concentrations of 6-benzyladenine (BA) and kinetin (KIN) in combination with citric acid (CA), adenine sulfate (AdS) and indole- 3-acetic acid (IAA). Maximum bud break (70%) was obtained on MS with 8 mg L-1 BA, 8 mg L-1 KIN, 100 mg L-1 CA, 200 mg L-1 AdS and 2 mg L-1 IAA (standard medium). Shoot production potential continued on this medium during the subsequent seven-month-long subculture cycle. The in vitro raised shoots rooted best on ½-strength MS containing 1 mg L-1 indole-3-butyric acid. Plantlet survival rate was >95% after acclimatization. The genetic homogeneity of plantlets with the mother plant was analyzed using Inter Simple Sequence Repeats which generated a monomorphic banding pattern to confirm the uniformity of in vitro raised plantlets of C. caesia.