In vitro Propagation of Polyscias fruticosa Plant

Abstract

The aim of this work was to develop a well defined protocol for in vitro propagation of Polyscias fruticosa plant. A factorial experiment in a complete randomized design was employed in all experiments. Analysis of variance as described to compare statistical differences between treatments using L.S.D at 5% probability level by Snedecor and Cochran. This study was carried out during the period from 2008 to 2011 aiming to achieved the most suitable protocol for micro propagation of Polyscias fruticosa Harms in Laboratory of Tissue Culture, Zohria Botanical Garden, Horticulture Research Institute, Agriculture Research Center. The young shoots were sterilized by immersion in a clorox (commercial bleach 5.25% sodium hypochlorite) solution at the rates of 20, 25 and 30 % for 10, 15, 20 and 25 min. Shoot tips were cultured on (Murashige and Skoog) MS medium supplemented with (Benzylaminopurine) BA at 0.0, 1.0, 3.0, or 5.0 mg/l and (Kinetin) kin at 0.0, 0.5, 1.0, 2.0 or 4.0 mg/l, and 3 g/l (activated charcoal) AC. The shoots obtained from establishment stage were cultured on multiplication media. Three sets of experiments with combination of different phytohormones for shoot regeneration were performed. The first experiment multiplication media consisted of MS medium supplemented with BA at 0.0, 1.0, 3.0 or 5.0 mg/l and kin at 0.0, 0.5, 1.0, 2.0 or 4.0 mg/l, and their combinations. In the second experiment multiplication media consisted of (Woody Plant medium) WPM basal nutrient medium supplemented with BA at 0.0, 1.0, 3.0 or 5.0 mg/l and kin at 0.0, 0.5, 1.0, 2.0 or 4.0 mg/l, and their combinations. In the third experiment multiplication media consisted of (Gamborg medium) B5 basal nutrient medium supplemented with BA at 0.0, 1.0, 3.0 or 5.0 mg/l and kin at 0.0, 0.5, 1.0, 2.0 or 4.0 mg/l, and their combinations. Shoots were cultured on MS basal rooting medium supplemented with (Naphthalene acetic acid) NAA at 0.0, 0.1, 0.5, 1.0, 2.0 and 4.0 mg/l or (Indole butyric acid) IBA at 0.0, 0.1, 0.5, 1.0, 2.0 and 4.0 mg/l for 45 days. Rooted plantlets were singly picked out into 5 cm plastic pots filled with 1:0, 1:1, 1:2 or 1:3 (v:v) peatmoss and sand, respectively. The interaction between 25% clorox for 15 minutes resulted in the highest value for survival. For the establishment stage, 3.0 mg/l BA and 2.0 mg/l kin showed the tallest shoots. For multiplication stage, the highest shoot length, number of shoots, number of leaves and callus formation was obtained at B5 medium supplemented with 5.0 mg/l BA and 2.0 mg/l kin. For NAA on rooting, the highest number of roots and root length was obtained on medium supplemented with 1.0 mg/l NAA. The highest percentage of plant survival was achieved by transplanting of the plantlets to pots containing sand and peatmoss at the ratio of 1:1(v/v). This study was carried to develop the most suitable protocol for micropropagation of Polyscias fruticosa Harms, while ex vitro acclimatization need further work to increase establishment at greenhouse.