应用响应曲面法优化萱藻丝状体扩增条件  

Optimization of amplification conditions of filaments of Scytosiphon lomentaria by response surface methodology(RSM)

作　　者：王玉萌 刘德举 张玉敏 张敏[1] 宫相忠[1] WANG Yumeng;LIU Deju;ZHANG Yumin;ZHANG Min;GONG Xiangzhong(College of Marine Life Science,Ocean University of China,Qingdao 266003,China)

机构地区：[1]中国海洋大学海洋生命学院,山东青岛266003

出　　处：《水产学报》2022年第9期1646-1655,共10页Journal of Fisheries of China

基　　金：山东省重点研发计划(2016GSF115042)。

摘　　要：为优化萱藻丝状体扩增条件,提高萱藻丝状体的扩增速率,以实验室萱藻种质库保存的丝状体为实验材料,在单因素实验的基础上,应用Box-Behnken设计对萱藻丝状体扩增条件进行了响应曲面优化探索。结果显示,以增重倍比为指标,确定了扩增萱藻丝状体的最佳条件为温度20.45℃,光照强度78.13μmol/(m^(2)·s),外加氮磷比16.19。在该条件下扩增10 d,萱藻丝状体增重倍比为(299.21%±13.58%),显著高于单因素实验和Box-Behnken设计实验条件下获得的萱藻丝状体增重倍比。实验结果与理论预测值的相对误差小于5%,模型可靠。为评价优化培养后萱藻丝状体的发育情况,对丝状体进行了20 d的诱导,结果显示,优化培养后的萱藻丝状体呈深褐色,细胞质充盈,孢子囊比例(26.37%±5.22%)比对照组的孢子囊比例(18.10%±3.51%)提高了45.69%;优化条件下的孢子囊直径平均为(19.75±0.21)μm,显著高于对照组的孢子囊直径(16.91±0.36)μm。研究表明,在萱藻丝状体扩增过程中,对扩增环境条件进行调控,设置合适的营养盐浓度配比,可以实现较高的扩增速率,并促进丝状体的孢子囊发育。Filament amplification is a key step in artificial seedling cultivation of Scytosiphon lomentaria.In order to optimize the growth conditions of the filament of S.lomentaria,a Box-Behnken design(BBD)was employed for experimental design and analysis of the results to obtain the best possible combination of temperature(X1:17-23℃),light intensity[X_(2):43.2-100μmol/(m^(2)·s)],and added nitrogen:phosphorus(N∶P)ratio(X_(3):7-17).The results show that the mathematical model has a good fitting degree and good regression property.The model predicted and optimized reaction parameters as follows:temperature 20.45℃,light intensity 78.13μmol/(m^(2)·s),added N∶P ratios 16.19.Under the condition of the amplification for 10 d,the increasing ratio of filament biomass was 299.21%±13.58%,which was significantly higher than that measured in the single factor experiment and Box-Behnken design experimental conditions.In order to evaluate the development of filaments of S.lomentaria after optimized culture,the filaments were induced for 20 days.The results showed that the sporangium ratio after optimized culture(26.37%±5.22%)was higher than that of unoptimized culture(18.10%±3.51%).Meanwhile,the average sporangium diameter under optimized conditions was(19.75±0.21)μm,which was also higher than that before optimized culture[(16.91±0.36)μm].The results showed that the high amplification rate and the development of sporangia of filaments could be achieved by purposeful regulation of environmental conditions and appropriate nutrient concentration.

关 键 词：萱藻 丝状体 孢子囊 响应曲面法 扩增 

分 类 号：S968.4[农业科学—水产养殖]