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

• Published in: 农业科学学报（英文版） 2012, Vol.11 (3), p.387-396
• Main Author: WU Jiao ZHANG Ya-li HE Rong-rong ZHU Lei WANG Chao-xia XU Xia LU Jiang
• Format: Article
• Language: English
• Subjects: 悬浮培养; 接种时间; 效率优化; 根癌农杆菌介导; 胚性细胞; 葡萄品种; 转化系统; 霞多丽葡萄
• ISSN: 2095-3119; 2352-3425

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.