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| Optimization of Transformation Efficiency of Suspension Cultured Vitis vinifera cv. Chardonnay Embryogenic Cells |
| WU Jiao, HE Rong-rong, WANG Chao-xia |
1.College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, P.R.China
2.Center for Viticulture and Small Fruit Research, Florida A&M University, Tallahassee 32317, USA |
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摘要 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 efficiencywhich reached maximum 21.5% at the following conditions: 0.8 of OD600, 25 min of inoculation, 2 d of co-cultivation, and 10min of cell drying. Response surface methodology experiments based on a five-level, four-factor central-compositerotatable design were then used to optimize these selected factors. The optimized conditions for Chardonnay grapetransformation were: 0.8711 of OD600, 28.9 min of inoculation, 2.25 d of co-cultivation and 11.76 min of cell drying. Afteroptimization, transformation efficiency was 26.2% and there were no interactions among different factors.
Abstract 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 efficiencywhich reached maximum 21.5% at the following conditions: 0.8 of OD600, 25 min of inoculation, 2 d of co-cultivation, and 10min of cell drying. Response surface methodology experiments based on a five-level, four-factor central-compositerotatable design were then used to optimize these selected factors. The optimized conditions for Chardonnay grapetransformation were: 0.8711 of OD600, 28.9 min of inoculation, 2.25 d of co-cultivation and 11.76 min of cell drying. Afteroptimization, transformation efficiency was 26.2% and there were no interactions among different factors.
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Received: 12 October 2010
Accepted:
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| Fund: This research was supported by the 948 Program, Ministry of Agriculture, China (2006-G26) and the National Grape Industry Technology System, China (nycytx-30-zy-05). |
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Corresponding Authors:
Correspondence LU Jiang, Tel: +1-850-412-7393, Fax: +1-850-412-7464, E-mail: j.lu.cau@gmail.com
E-mail: j.lu.cau@gmail.com
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| About author: WU Jiao, Tel: +86-10-62737465, E-mail: jiaolong722@gmail.com; ZHANG Ya-li, Tel: +86-10-62737465, E-mail: olivia.yl.zhang@gmail.com |
Cite this article:
WU Jiao, HE Rong-rong, WANG Chao-xia.
2012.
Optimization of Transformation Efficiency of Suspension Cultured Vitis vinifera cv. Chardonnay Embryogenic Cells. Journal of Integrative Agriculture, 12(3): 387-396.
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