Optimization of Transformation Efficiency of Suspension Cultured Vitis vinifera cv. Chardonnay Embryogenic Cells
Vitis vinifera cv. Chardonnay suspension cultures were established from proembryogenic mass and employed for optimizing Agrobacterium-mediated transformation system. One-factor-at-a-time experiment revealed that OD600 of Agrobacterium, time of inoculation, co-cultivation, and cell-drying before inoc...
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| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
KeAi Communications Co., Ltd.
2012-03-01
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| Series: | Journal of Integrative Agriculture |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2095311912600239 |
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| Summary: | Vitis vinifera cv. Chardonnay suspension cultures were established from proembryogenic mass and employed for optimizing Agrobacterium-mediated transformation system. One-factor-at-a-time experiment revealed that OD600 of Agrobacterium, time of inoculation, co-cultivation, and cell-drying before inoculation significantly affected the transformation efficiency which reached maximum 21.5% at the following conditions: 0.8 of OD600, 25 min of inoculation, 2 d of co-cultivation, and 10 min of cell drying. Response surface methodology experiments based on a five-level, four-factor central-composite rotatable design were then used to optimize these selected factors. The optimized conditions for Chardonnay grape transformation were: 0.8711 of OD600, 28.9 min of inoculation, 2.25 d of co-cultivation and 11.76 min of cell drying. After optimization, transformation efficiency was 26.2% and there were no interactions among different factors. |
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| ISSN: | 2095-3119 |