Establishment of Contamination-free Culture and Induction of In Vitro Plant Regeneration in Dabai (Canarium odontophyllum Miq.)

Chong, Mui Sia (2009) Establishment of Contamination-free Culture and Induction of In Vitro Plant Regeneration in Dabai (Canarium odontophyllum Miq.). [Final Year Project Report] (Unpublished)

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Abstract

Dabai, Canarium odontophyllum Miq. an indigenous fruit tree of Sarawak is dioecious, having male and hermaphrodite trees. Vegetative propagation of by conventional methods though not highly successful is currently the only mean to produce clones of selected genotypes. Micropropagation has been considered as an alternative to multiply the selected clones of Dabai. In this approach, an effective protocol for establishment of contamination-free and viable explants is a prerequisite for regeneration of plant in vitro. This study aimed primarily to improve a protocol developed earlier on establishment of contamination-free and viable culture and then to attempt plant regeneration through proliferation of axillary shoots, organogenesis or somatic embryogenesis. A protocol which can achieve two fold increases in the percentage of contamination-free culture has been developed. The protocol involved spraying of 75% (v/v) Ethanol to stock plants prior to collection of explants which were then surface sterilized for 20 minutes in 20% (v/v) Clorox and then agitated in 0.3% (w/v) Benomyl for one hour before cultured in solid half-strength MS media supplemented with 3 mg/L BAP, 2 ml/L PPM, and 10 mg/L TET. Induction of direct organogenesis has been initiated recently. To date only roots were observed developing from young cotyledon explants. Callus had been induced from the cotyledons in 3.0-9.0 mg/L 2,4-D in the attempt on induction of somatic embryogenesis. In conclusion, 80% contamination-free culture could be achieved using that protocol. Extension of axillary shoot growth and root formation on cotyledon explants could be observed.

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