Somatic Embryogenesis and Genetic Fidelity Study of the Micropropagated Medicinal Species, Canna indica

Abstract

Canna indica Linn. (Cannaceae) is used both as medicine and food. Traditionally, various parts of C. indica are exploited to treat blood pressure, dropsy, fever, inflammatory diseases etc. However, to date there is no reliable micropropagation protocol for C. indica. We present here a regeneration technique C. indica with banana micropropagation medium (BM). BM supplemented with 3% sucrose, 0.7% agar, and 0.17% NH4NO3 and different plant growth regulators like BAP (2 mg•L −1) and NAA (0.5 mg•L −1) was found to be effective in inducing callus. BM with BAP (2 mg•L −1) was ideal for somatic embryogenesis and plantlet regeneration. After a period of 3 months, regenerated plantlets were successfully transferred to the field conditions. Appearance of somaclonal variation among the regenerated plants is a common problem which was assessed by DNA fingerprinting. To detect genetic fidelity in C. indica, RAPD and ISSR markers were employed. Ten RAPD primers produced 60 amplicons, while 7 ISSR primers generated 45 bands in both in vitro plantlets and mother plants. RAPD and ISSR analyses showed no evidence of polymorphism between parent plants and the regenerated plants as all the amplified products were found to be monomorphic.

Organogenesis and Ultrastructural Features ofIn VitroGrownCanna indicaL.

“…According to the table, the percentage of shoot formation increased with the increase of BAP up to the optimum concentration of 5.0 mg/L and later gradually decreased. The pattern was in agreement with previous researchers [ 14 , 23 ] which observed a less effective formation of shoots with higher concentrations of BAP. The variants in optimum concentration of BAP for regeneration could be attributed to the types of explants used.…”

“…The variants in optimum concentration of BAP for regeneration could be attributed to the types of explants used. Regeneration of plantlets from somatic embryos of C. indica only required low concentration of BAP (2.0 mg/L) [ 14 ] while, in the present study, rhizome explants required a high concentration of BAP (5.0 mg/L) to induce optimum shoot and root formation. Similarly, rhizome buds of Curcuma manga give the best response of shoot formation in MS medium containing a high concentration of BAP (9.0 mg/L BAP) [ 24 ].…”

“…Despite its many medicinal values and uses, traditional propagation of C. indica is hindered by its extremely hard seed coat [ 9 ] and slow vegetative propagation of rhizome which is also susceptible to viral infection during multiplication [ 10 – 13 ]. Furthermore, common practice of asexual propagation through rhizome produces genetic stagnancy and genetic variation limitations in Canna [ 14 ]. Plant tissue culture provides a solution whereby the method rapidly micropropagates plants of superior qualities and free from microorganisms in a relatively short time and minimal space using few starting materials.…”

“…Later, the same author tried to stimulate shoot development or bud formation from meristem tips with little success [ 16 ]. Recently, a study managed to obtain complete plantlets from leaves explant of C. indica via indirect regeneration through in vitro callus and somatic embryogenesis [ 14 ].…”

Micropropagation, genetic fidelity assessment and phytochemical studies of Clerodendrum thomsoniae Balf. f. with special reference to its anti-stress properties

“…Controlled cultures with no hormone were also prepared. Sub-culturing was done at 2 weeks interval in the same media having the same hormonal composition [15]. BAP was applied at the rate of 1, 2, 3 and 4 mg/l and NAA at the rate of 0.5mg/l to observe various stages of in-vitro callusing and subsequent shoot formation.…”