Agricultural Research Management Information System

Home
Research Summary
Search
About Us
- ARMIS
- Brochure
Contact Us
Report
User Request
Data Input
Help
- Operation Manual
  - PDF
  - Video
- Program Area & Commodity
We have reached 37600 number of research entries at this moment.

- Logout

Research Detail

Home
Research
Detail

Efficient Procedure for in Vitro Microrhizome Induction in Curcuma Longa L. (zingiberaceae) – a Medicinal Plant of Tropical Asia

M. A. Islam
Institut für Botanik, University Hannover, Herrenhäuser Str. 2, 30419 Hannover, Germany

K. Kloppstech
Institut für Botanik, University Hannover, Herrenhäuser Str. 2, 30419 Hannover, Germany

H. -J. Jacobsen
Institut für Botanik, University Hannover, Herrenhäuser Str. 2, 30419 Hannover, Germany

Abstract/ Executive Summary:

An improved in vitro microrhizome induction system in Curcuma longa L. has been developed. Freshly sprouted axillary buds from underground rhizomes were used as initial explants and multiplied through established in vitro systems. Multiplied shoots were excised and subcultured on hormone free medium for four weeks to induce microrhizome. Effects of light, sucrose and growth regulators on in vitro microrhizome production have been studied. Nine per cent sucrose was found to be the most suitable for microrhizome production, when incubated in the dark. Various concentrations of BA and Kn, and NAA were tested. BA (12.0 μM) and NAA (0.3 μM) were found suitable for the induction of microrhizomes. Larger microrhizomes performed better under in vivo conditions and developed shoots readily, when they were transferred directly from in vitro to soil without acclimatization. Microrhizomes produced under in vitro conditions can be stored and transported easily which are advantageous for in vitro shoot multiplication.

Keywords: Curcuma longa, Microrhizome induction, Medicinal plant

Location: Bangladesh National Herbarium (DACB), Mirpur, Bangladesh and The Institute of Botany, University of Hannover, Germany

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Turmeric

Objectives:

To see the effects of in vitro culture conditions such as sucrose concentration, strength of MS medium and light illumination on large scale microrhizome induction in C. longa L.

Methodology:

Establishment of contamination free cultures: Underground rhizomes of C. longa L. collected from Bangladesh National Herbarium (DACB), Mirpur, were cultivated in the glasshouse of the Institute of Botany, University of Hannover, Germany to allow sprouting of buds. Sprouted immature shoots (ca. 1 cm long) were collected and used as the source of explants. Immature buds were cleaned with running tap water and then washed with detergent (Tween-20) for 5 min, subsequently rinsed thoroughly under running tap water for 5 min. Explants were then immersed in 70% ethanol for 30 - 40 sec before incubation in disinfectant (0.1% HgCl₂ to which two - three drops of Tween-20/100 ml were added). Under sterile conditions, HgCl₂ solution was decanted and the explants were rinsed five - six times with sterile distilled water. Sterilized sprouts were then dissected to remove the outer few layers of leaf sheaths under aseptic conditions. Excised buds were initially cultured on MS basal medium supplemented with 6 μM BA and 0.3 μM NAA, and 3% sucrose. The medium was solidified with 0.8% agar (Duchefa, NL) after adjusting the pH to 5.8 and sterilization by autoclaving at 121⁰C (1.06 Kg^-1 m^-2) for 20 min. About 25 ml of the medium (M1) were dispensed into the sterile plastic, `De Wit’ culture tubes (Duchefa, NL). Initial explants were cultured on this shoot induction medium in 16 h light (white fluorescent light with 50 μM m^-2 s^-1 ) and 8 h darkness in order to obtain contamination free cultures since contamination of the explants taken from underground rhizomes of C. longa had been a major problem. BA, Kn, NAA and MS salts - An experiment was carried out to determine the effect of the concentration of sucrose using M3 medium (M1 medium minus of agar and 0.75∇ MS salts) with 1, 3, 5, 7, 9 and 11% sucrose treatments. All treatments were investigated under both complete darkness and 16 h photoperiod (50 μM m^-2 s^-1). Sixteen combinations with different concentrations of BA and Kn alone or in combination with 0.3 μM NAA were investigated under complete darkness using M4 medium (M3 medium modified with 9% sucrose). Results presented in the tables and in the figures are the pooled means ± standard errors (SE) of two repeated experiments each with 15 replications with an exception in which ten replicates were used without any repetition. The test of statistical significance was done by applying Tukey’s test at 5% level using SAS statistical software, Release 8 (SAS Institute Inc., Cary, NC).

Source of Information: Plant Tissue Cult. 14 (2) : 123-134, 2004 (December)

Article Link (Online Journal):

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

In the present investigation the in vitro shoot multiplication system for C. longa has been optimised. Microrhizome induction in Curcuma needs further improvement. However, our developed protocol can be used to produce a higher amount of large microrhizomes compared to previously reported protocols. Production of in vitro microrhizomes would be a suitable source of disease free seed rhizomes that could be stored and transported easily. In addition to that, in vitro microrhizomes does not require acclimatization in the field. The present protocol is a step forward towards an improved commercial propagation system for Curcuma longa more efficient and productive than having a crop of plantlets. Various concentrations of BA and Kn, and NAA were tested. BA (12.0 μM) and NAA (0.3 μM) were found suitable for the induction of microrhizomes. Larger microrhizomes performed better under in vivo conditions and developed shoots readily, when they were transferred directly from in vitro to soil without acclimatization. Microrhizomes produced under in vitro conditions can be stored and transported easily which are advantageous for in vitro shoot multiplication.

Knowledge Management: Journal