Optimization of culture medium for two mixed green algae by response surface methodology
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摘要: 【目的】优化普通小球藻（Chlorella vulgaris）和斜生栅藻（Scenedesmus obliquus） 2种混合绿藻的培养基，为提高混合绿藻的生长速度及推广应用淡水有益藻类混合培养技术提供参考依据。【方法】采用单因素试验筛选BG11培养基中适宜普通小球藻和斜生栅藻2种混合绿藻增殖的营养盐浓度范围，通过Plackett-Burman试验筛选显著影响混合绿藻生长的营养因素，采用响应面法优化主要营养盐的最佳浓度，并进行混合绿藻培养验证试验。【结果】通过单因素试验筛选出相对适宜混合绿藻生长的7种营养盐浓度范围，通过Plackett-Burman试验筛选出显著影响混合绿藻生长的3个营养因素（分别为Na2CO3、K2HPO4和NaNO3）。响应面法分析结果表明，营养因素中NaNO3与K2HPO4交互作用强，对混合绿藻细胞密度具有显著影响（P＜0.05，下同），得到优化后的培养基配方为NaNO3 1600.00 mg/L、K2HPO4 50.85 mg/L和Na2CO3 32.39 mg/L。使用优化后的培养基培养混合绿藻，可显著提高其细胞光合系统PSII最大光能转换效率，混合藻细胞密度最高达36.75×106个/mL，较对照组显著提高24.28%，在整个培养周期内，普通小球藻和斜生栅藻的细胞密度比基本为1.00∶1.00。【结论】通过响应面法优化的普通小球藻和斜生栅藻混合绿藻培养基，能实现混合绿藻的高密度培养，可供大规模推广应用淡水有益藻类混合培养技术时参考；培养2种混合绿藻培养基中的Na2CO3、K2HPO4和NaNO3最佳浓度分别为32.39、50.85和1600.00 mg/L。Abstract: 【Objective】This study optimized the culture medium for Chlorella vulgaris and Scenedesmus obliquus， which provided reference for improving the growth rate of mixed greenalgae and promoting the application of mixed culture technology of freshwater beneficial algae.【Method】The concentration ranges of nutrient salt that suitable for the proliferation of C. vulgaris and S. obliquus in BG11 medium were screened by single factor test. And the nutrient factors that significantly affected the growth of the mixed green algae were determined by Plackett-Burman test. By using response surface methodology，the optimal concentrations of the main nutrient salts in BG11 medium were optimized. Then，confirmatory experiment of the mixed green algae culture was performed.【Result】Relatively suitable concentrations of 7 nutrient salts for the mixed green algae growth were determined by single factor test. Using the Plackett-Burman test， 3 nutrient factors，Na2CO3， K2HPO4 and NaNO3， which significantly impacted the growth of mixed green algae most were determined via Plackett-Burman test. The results of response surface methodology analysis showed that the strong interaction between NaNO3 and K2HPO4 had a significant effect on the cell density of mixed green algae（P＜0.05，the same below）. In the optimized medium， NaNO3， K2HPO4 and Na2CO3 were 1600.00， 50.85 and 32.39 mg/L，respectively. Maximum light conversion efficiency in the photosynthesis PSII system of the mixed green algae in the optimized medium was significantly improved. The cell density of mixed green algae in the optimized medium group peaked at 36.75×106 cells/mL， which was significantly increased by 24.28% than the control group. Furthermore，the cell density rate of C. vulgaris and S. obliquus was basically 1.00∶1.00 throughout the culture cycle.【Conclusion】The mixed green algae culture medium of C. vulgaris and S. obliquus optimized by response surface methodology can realize the high density culture of mixed green algae，which can be used as a reference for the large scale application of mixed culture technology of freshwater beneficial algae；the optimal concentrations of Na2CO3，K2HPO4 and NaNO3 in the culture medium of two mixed green algae are 32.39， 50.85 and 1600.00 mg/L respectively.
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