The evaluation of sterilization protocol for sprout explants in oil palm (Elaeis guineensis Jacq.) tissue culture

Abstract

The objective of this study was to develop an efficient protocol for sterilization of sprout explants in tissue culture of oil palm. Plant materials used were plumulae and radicle from sprouts of the D x P Sriwijaya variety obtained from Seed Processing Unit PT Binasawit Makmur, Palembang. The medium used was MS composition was supplemented with vitamins, myo-inositol and sucrose, and the pH was set at 5.8 ± 0.02 before being solidified with agar. Cultures were maintained under a light intensity of 1,500 lux and 16-hour photoperiod and temperature of 24 to 26°C. Five protocols for the eradication of explant contamination were tested. i.e. There were 5 (five) different methods of explant sterilization employed, ie: A) explants were washed with sterile aquadest followed by soaking in 70% alcohol for 1 minute; B) explants were washed with sterile aquadest followed by soaking in 1% Benlox 50WP solution for 30 minutes, 1% Agrept 20WP for 30 minutes, and soaking in 70% alcohol for 5 minutes; C) explants were washed with sterile water plus detergent, followed by soaking in 1% Benlox 50WP plus few drops of Tween-80 for 30 minutes, soaking in 1% Agrept 20WP plus few drops of Tween-80 for 30 minutes, soaking in 70% alcohol for 5 minutes; D) explants were washed with sterile water plus detergent, followed by soaking in 0.1% HgCl2 solution for 30 minutes; and E) explants were washed with sterile water plus detergent, followed by soaking in 0.2% Dithane M-45 plus few drops of Tween-80 for 30 minutes, soaking in 1% NaOCl for 5 minutes, soaking in 0.1% HgCl2 plus few drops of Tween-80 for 30 minutes. The results showed that treating the explants with 0.1% HgCl2 for 30 minutes following 0.2% Dithane M-45 and 1% NaOCl applications was proven to be effective for the eradication of contamination.