Sterilization procedure and callus regeneration in black turmeric (Curcuma caesia)

Abstract

Sterilization procedure, media composition, explants selection and control of physical environment are critical for successful cultures and callus induction with surface sterilization being very challenging in most plants. Five different sterilization methods were evaluated to come up with the best for subsequent use to establish an in vitro regeneration method for the induction of callus in Curcuma caesia using excised leaf and rhizome explants. Murashige and Skoog (MS) media supplemented with various concentration of 2,4-Dichlorophenoxy acetic acid (2,4-D)/Indole-3-acetic acid (IAA) (0.5- 5.0mg/L), singly or in combination with Benzyl aminopurine (BAP)/Kinetin (KIN) (0.1-5.0mg/L), 0.3% sucrose and 0.08% agar were used. The result of the sterilization procedures showed 15% NaHClO3 (5min) + 70% Ethanol (30s) + 0.1% HgCl2 (5min) to be the most effective in controlling contamination in C. caesia among all the treatments tested. The response to callus induction was found to depend on the type of explants used and growth regulators combination. Leaf explants gave the highest percentage of callus induction. Highest percentage of callus induction (66.70%) was obtained in the growth regulator combination of 2, 4-D (0.5mg/L) + BAP (0.1mg/L) and least (14.29%) in IAA (2.0mg/L) + BAP (0.5mg/L). Equal and higher concentration of 2, 4-D + BAP of 5.0mg/L each also provided better result (40.00%). No callus was obtained in all the single concentration of 2, 4-D used.

A Protocol for Rapid Clonal Propagation and Microrhizome Production of Curcuma caesia Roxb. (Zingiberaceae): A Critically Endangerd Medicinal Plant of North East India

“…The shoot proliferation capacity of C. caesia depended on the concentration of cytokine like BAP. BAP alone or in combination with other growth regulators induced shoot multiplication (Abubakar and Pudake, 2019;Parthasarathy and Sasikumar, 2006).The positive effect of BAP on multiple shoot regeneration was also observed in ginger (Zingiber officinale Rosc.) (Balachandran et al, 1990).…”

Phytochemical Composition, Antioxidant, Antibacterial, and Enzyme Inhibitory Activities of Various Organic Extracts from Apocynum hendersonii (Hook.f.) Woodson

“…The higher antibacterial effect of ethanol extract may have been due to the additional effect of the solvent, as its aqueous form is often used as sterilizing agent [ 60 ] and as seen in the negative control. Tannin, which has strong bactericidal activity [ 61 ], might have strengthened the antibacterial effect of the extract.…”

Callus Production Protocol for Aegle marmelos (L.) Corr.: As a Tool for Extraction of Secondary Metabolites

“…Current study has proved that MS medium supplemented with combinations BAP ranging from of 0.5 mgL -1 to 1.5 mgL -1 and 2,4 D ranging from 1.0 mgL -1 to 2.0 mgL -1 could also be used to induce superior callus from seed explants of A. marmelos as a new addition to the literature. Not only that, a combination of 0.5 mgL -1 2, 4 D and 1.0 mgL -1 BAP resulted in highest percentage of callus production in black turmeric (Curcuma caesia) (Abubakar and Pudake, 2019) Once initiated, callus should be multiplied to obtain a sufficient amount for the secondary metabolite extraction. Auxins alone or together with a cytokinin can be used for the process according to the preference of the plant species.…”