Establishment of Axenic Culture and In Vitro Shoot Regeneration of Pepper (Piper nigrum L. )

Chen, Teck You (2008) Establishment of Axenic Culture and In Vitro Shoot Regeneration of Pepper (Piper nigrum L. ). [Final Year Project Report] (Unpublished)

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Abstract

Black Pepper (Piper nigrum L. ), the King of Spices, is an important and widely used spice around the world. The objectives of this study are to develop an effective protocol for establishment of contamination-free (axenic) culture from field-grown mature P. nigrum plant as the first step towards micropropagation of selected elite pepper plant, and to establish an effective protocol for in vitro regeneration of P. nigrum for large scale propagation. Shoot-tip explants were surface sterilized with 10% Clorox and 10 minutes exposure time after this was found to be the most effective surface sterilization regime. The nested analysis of variance indicated that there was significant difference in number of axenic explants against number of rounds of pre-sampling treatment. However, analysis of variance signified that there was no significant difference in the number of axenic explants against all treatments in pre-sampling treatment and the number of rounds of pre-sampling treatment when newly emerged shoots were used as the explants. The treatment of 2 ml/L PPM and 10 mg/L tetracycline supplemented in the media ensure the emergence of contaminants was fully prohibited for a period of time. Exudation of phenolics was much reduced if young shoot tips emerged at three weeks after pruning were used as the explants. In the induction of direct organogenesis, MS medium was found to be the most effective medium to form multiple shoots of P. nigrum. Even though axenic explants were used to induce multiple shoot formation but bacterial contamination was recurring. Medium incorporated with 2 mg/L BAP only or with 2.5 mg/L BAP and 0.2 mg/L 2,4-D recorded the highest percentage of axenic explants.

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