Establishment of contamination-free culture and induction of shoot regeneration in vitro in Kapur Bukit (Dryobalanops beccarii Dyer)

Kwan, Pok Li (2008) Establishment of contamination-free culture and induction of shoot regeneration in vitro in Kapur Bukit (Dryobalanops beccarii Dyer). [Final Year Project Report] (Unpublished)

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Abstract

Kapur Bukit (Dryobalanops beccarii Dyer) has been identified as one of the timber species to be used in forest plantation and reforestation in Sarawak. To meet the high demand of planting stock, there is a need to mass produce this species. Since clonal propagation of this species is difficult and slow using conventional methods, micropropagation has been explored as an alternative to mass produce this species. The objective of this study is to establish a protocol to obtain axenic explants from fieldgrown materials and proceed to induce multiple shoot proliferation. Various factors such as serious contamination and browning influence the establishment of axenic explants of this species. These factors have successfully been addressed after a fast and effective protocol has been developed. The present study showed that 20 % Clorox surface sterilization for 15 min, supplementing 2 ml/L PPM and 10 mg/L TET to the culture medium were able to reduce contamination problem. Incorporation of 0.5 % PVP10 into the media, handling of explants under cool condition (approximately 15°C) during surface sterilization steps, culturing explants in dark condition and frequent transfer were found to be helpful in obtaining maximum survival of explants. No sign of multiple shoot formation was observed so far. However, vigorous cell growth, callus formation and growth of leaf primordia observed in medium containing cytokinin. The results showed that high concentration of BAP (3 mg/L), TDZ (3 mg/L) and combination of BAP and NAA (at 3 mg/L and 0.3 mg/L respectively) induced more response in the shoot tips in culture. The combination of BAP and NAA was considered as the most effective treatment in inducing the explants to respond.

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