Different protocols are usually used for extracting total deoxyribonucleic acid (DNA) from different plant species of same order and DNA of the associated viruses. Here, we describe a rapid, efficient and universal protocol for isolating total DNA from the members of Zingiberales which harbor a high amount of polysaccharides and secondary metabolites. DNA isolated with this protocol was successfully used for PCR based downstream applications viz. random amplified polymorphic DNA (RAPD), Inter-simple sequence repeats (ISSR), DNA barcoding gene (Internal transcribed spacer and trnl-f) amplification and detection of the viruses.
Nodal segments of Costus speciosus (Koen.) Sm. containing single axillary buds were cultured on Murashige and Skoog medium (MS medium) supplemented with plant growth regulators for inducing plantlets. For breaking of axillary bud dormancy, nodal segments were cultured on 40-70gl -1 sucrose or 1-13 µM adenine sulphate (AdS) supplemented MS basal medium containing 5 µM 6-benzylaminopurine (BAP) and 1µM α-naphthalene acetic acid (NAA). The nodal segments cultured on 1-13 µM AdS, 5 µM BAP, 1 µM NAA and 50gl -1 sucrose showed simultaneous production of shoots and roots while those cultured on 5 µM BAP, 1 µM NAA and 40-70gl -1 sucrose produced shoots only. The most effective media for breaking axillary bud dormancy was 5 µM BAP, 1 µM NAA, 50 gl -1 sucrose and 10 µM AdS supplemented medium. The propagules from 40-70gl -1 sucrose produced roots in shoot multiplication medium, i.e.,10 µM AdS, 1 µM NAA, 50gl-1 sucrose and 3-11 µM BAP supplemented medium. The best response for shoot multiplication was on 10 µM AdS, 1 µM NAA, 50gl-1 sucrose and 7 µM BAP. The well-rooted shoots were hardened and transferred to the soil where they showed 95% survival rate. Results show that axillary bud can be used for micropropagation of Costus speciosus.
Eleven cultivars of C. longa var. Lakadong were collected from Manipur having different topography. Curcumin content in different cultivars has been analyzed by using UV-Visible Spectrophotometer (100 Bio-Carry Spectrophotometer). The curcuminoids content were analyzed and quantified for identification of best quality cultivar. Thoubal Cultivar with highest curcumin content (9.44%) was subjected for tissue culture technique using different culture vessels and silver nitrate for rapid multiplication and scaling up of microrhizome production. High multiplication rate of 27.40±0.47 were obtained in Murashige and Skoog's medium supplemented with 3% sucrose + 1 mg L -1 α-napthalene acetic acid, 4 mg L -1 6-benzyl-amino-purine and 11 µM silver nitrate. Effect of different culture vessels and silver nitrate were studied for microrhizome and multiple shoots formation. Relatively higher rate of shoots along with microrhizome (17.5±0.32) can be seen in Growtek which was grown without any plant growth regulator. Growtek was used for scaling up of microrhizome production in vitro and utmost microrhizome was produced in liquid Murashige and Skoog's medium supplemented with 8% sucrose, 1 mg L -1 α-napthalene acetic acid, 4 mg L -1 6-benzyl-amino-purine and 11 µM silver nitrate (36.25±0.27). Addition of silver nitrate in the medium resulted in improvement of microrhizome induction in vitro. Higher concentration of silver nitrate (33, 44, 66, 88 µM) negatively affected the microrhizome and shoot multiplication and shows inhibition of tissue response completely. Analysis of in vitro derived plantlets during acclimatization shows that the exogenous applied of silver nitrate shows superior growth as compared to control. 90-95% of plantlets with and 75-80% plantlets without silver nitrate treatment were successfully established under ex vitro acclimatization. The protocol could be utilized for large scale production of true-to-type plantlets and as alternative method to step forward towards an improved commercial propagation system for more efficient and productivity.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.