Effect of Antioxidants, Culture Conditions and Frequency of Subculture to Control Phenols during in vitro Culture of Phalaenopsis Hybrid Cultivar ORIANA

G. Madhuri, Siddharth Kumar Palai, P. Tripathy, S. Beura
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Abstract

The present study was conducted to control phenols in the media during in vitro cultures using flower stalk explants of Phalaenopsis hybrid cultivar Oriana. MS medium + BAP (1.00 ppm) + NAA (0.50 ppm) + 3% sucrose (w/v) was used commonly for all the treatment. Results revealed that inclusion of 500 ppm citric acid in the media, rinsing the explants with citric acid 500 ppm for 20 minutes before inoculation, keeping culture in dark for 20 days and sub-culturing at 4, 6 and 8 days after inoculation were found to be significantly reducing the browning intensity (very low browning) in medium. The explants remained green even after 4th week of culture with significantly minimum percentage of blackening of cultures (10.00%), minimum percentage of dead cultures (20.00%) and maximum percentage of cultures survived (80.00%).Inclusion of 500 ppm citric acid in the media, rinsing of explant with citric acid 500 ppm for 20 minutes before inoculation, incubating the culture in dark for 20 days and subculturing at an interval of 4, 6, and 8 day after inoculation in case of flower stalk explants were observed to control phenols in the media during in vitro cultures of Phalaenopsis hybrid cultivar Oriana.
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抗氧化剂、培养条件和传代频率对蝴蝶兰杂交品种ORIANA离体培养中酚类控制的影响
本研究以蝴蝶兰杂交品种Oriana的花茎外植体为材料,对离体培养过程中培养基中酚类物质的控制进行了研究。所有处理均采用MS培养基+ BAP (1.00 ppm) + NAA (0.50 ppm) + 3%蔗糖(w/v)。结果表明,在培养基中加入500 ppm柠檬酸,接种前用500 ppm柠檬酸冲洗外植体20分钟,黑暗培养20 d,接种后4、6和8 d继代培养,可显著降低培养基中的褐变强度(极低褐变)。培养4周后,外植体仍保持绿色,培养体发黑率最低(10.00%),死亡率最低(20.00%),存活率最高(80.00%)。在蝴蝶兰杂交品种Oriana的离体培养过程中,观察到在培养基中加入500 ppm的柠檬酸,接种前用500 ppm的柠檬酸冲洗外植体20分钟,在黑暗中培养20天,在接种后4天、6天和8天进行继代培养。
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