Optimization of Transformation Efficiency of Suspension Cultured Vitis vinifera cv. Chardonnay Embryogenic Cells

doi:10.1016/S1671-2927(00)8556

Journal of Integrative Agriculture ›› 2012, Vol. 12 ›› Issue (3): 387-396.DOI: 10.1016/S1671-2927(00)8556

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

收稿日期:2010-10-12 出版日期:2012-03-01 发布日期:2012-03-11
通讯作者: Correspondence LU Jiang, Tel: +1-850-412-7393, Fax: +1-850-412-7464, E-mail: j.lu.cau@gmail.com
作者简介: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
基金资助:
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).

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

Received:2010-10-12 Online:2012-03-01 Published:2012-03-11
Contact: Correspondence LU Jiang, Tel: +1-850-412-7393, Fax: +1-850-412-7464, E-mail: j.lu.cau@gmail.com
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
Supported by:
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).

摘要/Abstract

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.

关键词: transformation, Chardonnay, suspension cultures, response surface methodology

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.

Key words: transformation, Chardonnay, suspension cultures, response surface methodology

WU Jiao, HE Rong-rong, WANG Chao-xia. Optimization of Transformation Efficiency of Suspension Cultured Vitis vinifera cv. Chardonnay Embryogenic Cells[J]. Journal of Integrative Agriculture, 2012, 12(3): 387-396.

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Optimization of Transformation Efficiency of Suspension Cultured Vitis vinifera cv. Chardonnay Embryogenic Cells

Optimization of Transformation Efficiency of Suspension Cultured Vitis vinifera cv. Chardonnay Embryogenic Cells

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