采用中心复合设计和响应曲面分析法,在实验室条件下研究了温度(16~40℃)、盐度(10~50)对马氏珠母贝鳃热休克蛋白Hsp70基因表达量的联合效应.设定温度范围为16~40℃,盐度范围为10~50.结果表明:温度的一次效应、二次效应对马氏珠母贝鳃Hsp70基因表达量影响显著;盐度的一次效应对马氏珠母贝鳃Hsp70基因表达量无显著影响、二次效应对马氏珠母贝鳃Hsp70基因表达量影响显著,温度、盐度之间的互作效应对马氏珠母贝鳃Hsp70基因表达量无显著影响,且温度的效应大于盐度.建立了马氏珠母贝鳃Hsp70基因表达量模型方程,决定系数98.7%、矫正决定系数97.4%,预测决定系数89.2%,模型的拟合度极高,可用于预测马氏珠母贝鳃Hsp70基因的表达量.通过对模型方程优化,得到在温度26.78℃、盐度29.33时,马氏珠母贝鳃Hsp70基因表达量达到最小值0.5276,满意度达到98%.试验结果可为马氏珠母贝鳃Hsp70基因的上调表达、维持细胞内环境稳定以及提高马氏珠母贝抗逆性提供理论指导.
By using central composite experimental design and response surface method, the joint effects of water temperature ( 16-40 ℃ ) and salinity (10-50) on the expression of gill Hsp70 gene in Pinctada martensii (Dunker) were studied under laboratory conditions. The results showed that the linear and quadratic effects of temperature on the expression of gill HspTO gene were significant, the linear effect of salinity was not significant, while the quadratic effect of salinity was significant. The interactive effect of temperature and salinity was not significant, and the effect of temperature was greater than that of salinity. The model equation of the gill Hsp70 gene expression was established, with the R2, Adj. R2, and Pred. R2 as high as 98.7%, 97.4%, and 89.2%, respectively, suggesting that the overarching predictive capability of the model was very satisfactory, and could be practicably applied for prediction. Through the optimization of the model, the expression of the gill Hsp70 gene reached its minimum (0. 5276) when the temperature was 26.78 ℃ and the salinity was 29.33, with the desirability value being 98%. These experimental results could offer theoretical reference for the high expression of gill Hsp70 gene in P. martensii, the maintenance of cell internal environment stability, and the enhancement of P. martensii stress resistance.