GFP-FABD2和GFP-MBD标记蛋白对拟南芥悬浮细胞培养及应激响应能力的影响

Influence of GFP-FABD2,GFP-MBD Fusion Protein on Culture and Stress Response Ability of Arabidopsis Suspension Cells

以转GFP-FABD2和GFP-MBD基因的拟南芥为材料,研究了GFP-FABD2和GFP-MBD这两种细胞骨架标记蛋白对拟南芥愈伤组织诱导、悬浮细胞培养及应激响应能力的影响.结果表明:(1)GFP-MBD标记蛋白延长愈伤的出愈时间,改变愈伤形态,使转基因拟南芥种子的出愈量减少为野生型的59%、悬浮细胞的长短轴比缩小为1.20±0.21、第7天细胞活力下降为0.66±0.09,影响细胞的生长曲线.(2)GFP-FABD标记蛋白虽对愈伤生长影响不大,但却使悬浮细胞的长短轴比显著增加为2.49±1.18、第7天细胞的活力下降为0.87±0.06,造成悬浮细胞生长曲线的改变.(3)通过调整培养条件的激素水平,以上两种细胞骨架标记蛋白对悬浮细胞生长的影响可以得到修复.(4)检测优化条件下培养的GFP-FABD2或GFP-MBD悬浮细胞对温度、渗透压、机械应力等环境改变的应激响应能力,结果未发现与野生型有明显区别.

The effects of the plant cytoskeleton labels, GFP-FABD2 and GFP-MBD were investigated on Arabidopsis callus induction, suspension cells cultivation and capacity of response to stress. The results showed that the expression of these two fusion proteins diversely effected the callus induction and suspen- sion cell growth. （1）GFP-MBD expression extended the time of callus induction, altered the callus mor- phology,reduced the quantity of callus to 59% of wild type,decreased the major/minor axial ratio of sus- pension cells to 1.20±0.21,reduced the relative viability of 7-day-old suspension cultures to 0.66±0.09 and altered cell growth curve. （2）GFP-FABD2 expression had no apparent effects on callus, but it signifi- cantly increased the major/minor axial ratio of suspension cells to 2.49±1.18, reduced the relative viability of 7-day-old suspension cultures to 0.87!0.06 and effected cell growth curve. （3）These effects can be restored by adjusting the hormone level. （4）In the optimized condition,we did not score any significant devia- tion in temperature,osmotic pressure,mechanical stress and other stress from those found in wild type Ar- abidopsis suspension cells.