Investigation on Protocol of Cryopreservation of Aquilaria agallocha Roxb. and Virus Elimination method of Phalaenopsis
| Autor: | Yu-Wen Lu, 呂玉雯 |
|---|---|
| Rok vydání: | 2015 |
| Druh dokumentu: | 學位論文 ; thesis |
| Popis: | 103 In vitro grown of Aquilaria agallocha Roxb., Phalaenopsis Sogo Yukidian ‘V3’ and Phalaenopsis Unimax Moonlight ‘Taida Green Apple’ were used in this study for cryopreservation, rooting, acclimatization and the virus elimination. Aquilaria agallocha Roxb. were cultured on proliferation medium 3 - 4 months for experiments. Explants were cryopreserved by vitrification. 1 - 2 mm shoot tips were excised from seedling after preculturing on different sucrose concentration medium for 7 day, treated with LS for 60 min, subjected to PVS2 for 90 min then immerged in liquid nitrogen directly for cryostorage, rewarmed after one week and recovery with different medium. The result demonstrated the explants precultured on 0.2 M sucrose medium for 7 days, treated with LS for 60 min and PVS2 for 90 min, recovery with medium containing activated charcoal, reached a survival rate of 27.41% after cryostorage. In addition, an micropropagation protocol for rooting and accimatizating was developed. Plantlets were rooted (92%) on 1/2 WPM medium after treated with 1.0 mg/L IBA for 10 days and acclimatized successfully (94%). CyMV infected Phalaenopsis Sogo Yukidian ‘V3’ and CyMV, ORSV doubly infected Phalaenopsis Unimax Moonlight ‘Taida Green Apple’ cultured on growth medium for 3 months were applied for investigating the effects of meristem tissue culture, cryotherapy, and chemotherapy on virus elimination. 52.17 % of virus-free Phalaenopsis Sogo Yukidian ‘V3’ were obtained when excised shoot tips were cultured on medium containing Ribavirin for 3 months. Up to 100 % of virus-free Phalaenopsis Sogo Yukidian ‘V3’ were obtained when shoot tips excised from previous treated explants were cultured on Ribavirin medium again. On virtrification-cryotherapy, Phalaenopsis Sogo Yukidian ‘V3’ precultured and recoverd with medium containing Ribavirin, reached a survival rate of 30.4%. However, it failed to eliminate virus. For Phalaenopsis Unimax Moonlight ‘Taida Green Apple’, plantlets were treated with Ribavirin for one month then subjected to virtrification-cryotherapy, reached a survival rate of 92.5%. Unfortunately, neither cryotherapy nor meristem tissue culture combined with chemotherapy could achive virus elimination. |
| Databáze: | Networked Digital Library of Theses & Dissertations |
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