Rapid multiplication and in vitro production of leaf biomass in Kaempferia galanga through tissue culture

An efficient protocol has been established for rapid multiplication and in vitro production of leaf biomass in Kaempferia galanga L, a rare medicinal plant. Different plant growth regulators like Benzyladenine (BA), Indoleacetic acid (IAA), Indolebutyric acid (IBA), Napthaleneacetic acid (NAA) and adenine sulphates (Ads) have been tried for induction of multiple shoots using lateral bud of rhizome as explants. The highest rate of shoot multiplication (11.5 +/- 0.6) shoot/explant as well as leaf biomass production (7.4 +/- 0.3) gram/explant was observed on Murashige and Skoog medium supplemented with Benzyladenine (1 mg/l) and Indoleacetic acid (0.5 mg/l). Data of shoot multiplication and leaf biomass production were statistically analysed. Upon excission of leaves after 2 months of culture under sterile condition, the base of each plantlet was transferred to fresh media which could produce the same leaf biomass within another 2 months in a 50 ml culture tube containing 20 ml and 250 ml conical flasks containing 30 ml Murashige and Skoog medium. The rate of multiplication and leaf biomass production remained unchanged in subsequent subcultures. The regenerated plantlets were acclimatized in greenhouse and subsequently transferred to the field. Survival rate of the plantlets under ex vitro condition was 95 percent. Genetic fidelity of the regenerants was confirmed using random amplified polymorphic DNA (RAPD) marker. The protocol could be commercially utilized for large scale production of true-to-type plantlets and as an alternative method of leaf biomass production in Kaempferia galanga.

High frequency plantlet regeneration from rhizomatous buds in Mantisia spathulata Schult. and Mantisia wengeri Fischer and analysis of genetic uniformity using RAPD markers.

A protocol has been devised for enhanced in vitro regeneration of critically endangered Mantisia spathulata Schult. and Mantisia wengeri Fischer. Highest Bud Forming Capacity (BFC) of 6.10 +/- 0.55 with an average of 19.93 +/- 3.19 roots was obtained for M. spathulata within 5-6 weeks in Murashige and Skoogs (MS) medium supplemented with a combination of 10.0 microM of N6-benzyladenine (BA) and 2.5 microM of alpha-naphtalene acetic acid (NAA). For M. wengeri, BFC of 7.82 +/- 0.73 and 20.86 +/- 1.65 roots was achieved in MS media supplemented with a combination of 5.0 microM BA and 2.5 microM of NAA RAPD markers were used to evaluate the genetic stability of in vitro raised hardened plantlets. Similarity coefficient among the regenerated plants ranged between 0.85-0.98 for M. spathulata and 0.83-0.98 for M. wengeri. Maximum of 88 and 90% genetic similarity were obtained between in vitro raised hardened plantlets and mother stock of M. spathulata and M. wengeri, respectively through RAPD analysis. The hardened plantlets after RAPD analysis on being transferred to soil of experimental garden showed no marked phenotypic variations in vegetative or floral characteristics.