An improved and efficient micropropagation of Eclipta alba through transverse thin cell layer culture and assessment of clonal fidelity using RAPD analysis

An improved and efficient in vitro regeneration system has been developed for Eclipta alba, a medicinally important plant, through transverse thin cell layer culture (tTCL). The transverse section of the nodal segment of field grown plants was used as tTCL explants for plant regeneration. Shoot multiplication from tTCL nodal explants was influenced by BAP and their interaction with Kin or NAA. MS medium containing 13.2 μM BAP and 4.6 μM Kin was most effective for shoot multiplication from tTCL nodal explants. Upon this medium, percent response for shoot proliferation was 100% with an average of 32.6 shoot buds per tTCL nodal explant. Regenerated shoots from tTCL nodal explants were rooted on the growth regulator free MS medium. The rooted plantlets were successfully acclimatized and established in soil with a survival frequency of 90–100%. Random amplified polymorphic DNA (RAPD) markers were used to evaluate the genetic fidelity of the micropropagated plants. RAPD profile analysis indicated that micropropagated plants were genetically similar to mother plant.

► The present paper, being the first report, describes a regeneration method for Eclipta alba using tTCL (transverse thin cell layer) explants derived from field grown plants.
► MS medium containing 13.2 μM BAP and 4.6 μM Kin was most effective for shoot multiplication.
► In comparison to intact nodal segments (1.0–1.2 cm), tTCL (0.1–0.3 cm) explants had higher shoot multiplication ability.
► The maintenance of genetic integrity of in vitro regenerated plants with the mother plant was evaluated by RAPD markers.