温度、盐度对马氏珠母贝外套膜Hsp70基因表达量的联合影响

Effects of Simultaneous Variation in Temperature and Salinity on the Expression of Mantle Hsp70 Gene in Pinctada fucata

设定温度范围16～40°C,盐度范围10～50,采用复合设计和响应曲面分析法在实验室条件下研究不同温度、盐度对马氏珠母贝(P.fucata)外套膜Hsp70基因表达量的综合效应并建立模型。采用方差分析(ANOVA)、决定系数等对实验所得回归方程模型进行显著性及拟合度检验。结果表明,温度的一次、二次效应对马氏珠母贝外套膜Hsp70基因表达量影响显著(p值<0.05);盐度的一次效应对马氏珠母贝外套膜Hsp70基因表达量无显著影响(p值>0.05),二次效应对马氏珠母贝外套膜Hsp70基因表达量影响极显著(p值<0.05);温度、盐度之间的互作效应对马氏珠母贝外套膜Hsp70基因表达量无显著影响(p值>0.05)且温度的效应大于盐度的效应。经响应曲面分析法和优化,得到马氏珠母贝外套膜Hsp70基因表达量在适宜的温度、盐度范围内呈峰值变化,在温度26.85°C、盐度29.39时,马氏珠母贝外套膜Hsp70基因表达量达到最小值0.95,满意度达到99.36%。建立了马氏珠母贝外套膜Hsp70基因表达量模型方程,其决定系数98.69%,矫正决定系数97.38%,预测决定系数91.42%,模型的拟合度极高,可用于预测马氏珠母贝外套膜Hsp70基因表达量。

The optimal conditions on the expression of mantle Hsp70 gene in Pinctada fucata were determined by response surface methodology and complex design. Model of marked and fit was tested by ANOVA and Determine coefficient. The composition design was applied to evaluating the effects of two independent variables （Temperaturel6℃-40℃, Salinityl0-50）. The results showed that linear effects of temperature and quadratic effects of temperature on the expression of mantle HspTO gene were extremely significant （p〈0.05）; linear effects of salinity on the expression of mantle Hsp70 gene was not significant （P〉0.05）, quadratic effects of salinity on the expression of mantle HspTO gene had significant effect （P〈0.05）; the interactive effects of temperature and salinity on the expression of mantle HspTO gene were not significant （P〉0.05）, effects of temperature was greater than effects of salinity. Building the mathematical-regression model for Hsp70 gene expression in mantle of Pinctada martensii, R-Squared was 98.69%, Adj R-Squared was 97.38% and Pred R-Squared was 89.21%, evincing that the overarching predictive capability of the model was very satisfactory, and could bepracticably applied. Optimizing the model with response surface, the two factors had peaking while temperature and salinity were simultaneously between appropriate range. The optimal conditions to obtain the lowest expression of mantle Hsp70 gene （0.5276） were when temperature was 26.78℃, and salinity was 29.33. When expression of mantle Hsp70 gene got the lowest, desirability was 98%.The experiment result offers theory basis for high expression of mantle Hsp70 gene in Pinctada martensii protecting cell＇s internal environment stability and enhancing the resistance.