Micropropagation of Etlingera coccinea (Zingiberaceae)

• Main Author: Nurul Humaira Abdullah Thaddeus
• Format: Thesis
• Language: English; English
• Published: 2013
• Subjects: QK495 Angiosperms
• Online Access: https://eprints.ums.edu.my/id/eprint/41819/1/24%20PAGES.pdf; https://eprints.ums.edu.my/id/eprint/41819/2/FULLTEXT.pdf; https://eprints.ums.edu.my/id/eprint/41819/

An effective protocol was developed for in vitro regeneration of valuable food additive plant Etlingera coccinea (zingiberaceae) through different methods of plant tissue culture. Micropropagation was possible via shoot regeneration from rhizome bud explants, callus induction and proliferation derived from leaf explants, plantlets regeneration from callus culture and shoot multiplication through pseudo stem explants. This study was involved the effect of Plant Growth Regulators (PGRs), types and strengths of basal media, and types and concentrations of carbon sources. The results obtained showed that plant growth factors manipulation as stated markedly influence shoot regeneration, callus induction and proliferation, plantlets regeneration from callus culture, and shoot multiplication of Etlingera coccinea. Rhizome buds were sterilized and cultured on Murashige and Skoog (MS) basal medium containing 0.1-2.0 μM of thidiazuron (TDZ), benzylaminopurine (BAP) or kinetin (KIN) supplied with 3% (w/v) of sucrose. High percentage of shoot regeneration (100%) was achieved by (~1±0.5 cm) of the whole bud cultured on MS medium containing 1.0 μM TDZ after 20 weeks of culture. Shoots were transferred to medium containing 5.0 μM BAP for shoot elongation and multiplication. High frequency of rooting (90%) was observed on medium supplemented with 1.0 μM IBA. The maximum percentage (100%) of callus formation was obtained after 10 weeks of culture from in vitro leaf explants cultured on half strength MS basal medium supplemented with 5.0 μM 2,4- dichlorophenoxyacetic acid (2,4-D) and 10 μM Benzylaminopurine (BAP). In media selection, half strength MS medium was most suitable for callus induction while 3% (w/v) of sucrose was the best carbon source among other sugar types tested. Callus cell proliferation was rapidly increased on MS basal medium supplemented with 2.5 μM 2,4-D within 5 weeks of culture. Combination of 2.5 μM 2,4-D and 5.0 μM BAP improved callus proliferation. 3% (w/v) of sucrose was most effective for callus proliferation and long term maintenance of callus culture. Plantlets regeneration with highest frequency of shoot formation (88%) from callus culture were obtained on MS medium containing 2.5 μM BAP and 0.1 μM IAA with a mean of 5.5 and 7.0 shoots and roots per explant after 6 weeks of culture. The most responsive explants were observed on full strength MS basal medium while 3% (w/v) of sucrose was the best source of carbon among other carbon sources tested. Roots regeneration were observed along with the shoot formation. 2.5 μM BAP was superior to KIN and ZEA on shoot multiplication from pseudo stem explants. Combination of 2.5 μM BAP and 0.7 μM NAA enhance shoots regeneration percentage up to 100% over a 6 weeks period of culture and produced 12.0 shoots per explant with 100% of rooting percentage. Full strength of MS basal medium and 3% (w/v) of sucrose were the best supplements for shoot multiplication via pseudo stem explants. Micropropagated plantlets were grown healthy after transplanted to the pots containing a mixture of 3:1 (w/w) soil: sand with 85% of survival explants after 6 month of transplantation. The procedure reported here offers a potential system for micropropagation of Etlingera coccinea and expected to be useful in improvement and conservation of this species in the future.