组合处理对新型斜生栅藻(Scenedesmus obliqast)积累虾青素的研究  

Effect of combined treatment on astaxanthin accumulation in the Scenedesmus obliqast

作　　者：凌善锋[1,2,3,4] 韩佳祥 项智锋 蔡刘新 陆冬平 张赢 LING Shan-feng;HAN Jia-xiang;XIANG Zhi-feng;CAI Liu-xin;LU Dong-ping;ZHANG Ying(Jingchu University of Technology,Jingmen 448000,China;Hangzhou Xitai Biotechnology Co.,Ltd,Tonglu 311500,China;Hangzhou Meipeptide Biotechnology Co.,Ltd,Hangzhou 310012,China;USDA-ERRC,Philadelphia 19038,USA)

机构地区：[1]荆楚理工学院生物工程学院,湖北荆门448000 [2]杭州硒泰生物科技有限公司,浙江桐庐311500 [3]杭州美肽生物技术有限公司,浙江杭州310012 [4]美国农业部东部研究中心,费城美国19038

出　　处：《生物技术》2020年第3期285-289,294,共6页Biotechnology

基　　金：浙江桐庐县2019年第二批高层次人才创业创新资助项目C类(100万经费);荆楚理工学院教授博士工作室建设项目(661997);引进博士科研启动金项目计划(QDB201706,2020-2021)。

摘　　要：[目的]研究20μmol/L褪黑素和0.2 mg/L水杨酸(SA)组合3.0 mg/L茉莉酸甲酯等组合处理诱导新型斜生栅藻协同促进积累虾青素的效应。[方法]在优化和改良的BG11培养基中培养,营养细胞培养阶段用一系列平行的40.0W红光灯管和1.2 m长28.0 W蓝光灯管照射藻液(总光照强度为252μmol·m^-2 s^-1),连续培养至对数生长期,每天震荡摇动培养物2次,防止黏壁。对数生长期1 d后,通过1.2 m长蓝光灯管+高光强+氮饥饿+51.8 g/L葡萄糖和20.0 g/L木糖,进行生物量发酵和诱导部分虾青素,继续培养3 d(方案1),然后在广泛诱导阶段,分步提高光照强度高约555μmol·m^-2 s^-1后,培养3 d(方案2),继续光照,再通过2μmol/L褪黑素组合3.0 mg/L茉莉酸甲酯加0.2 mg/L水杨酸诱导获得虾青素,继续培养5 d(方案3)。[结果]3个处理方案均能轻微促进斜生栅藻细胞密度的增加,最大生物量达到0.91×10^6个/mL强度。方案3处理组藻细胞内SA含量适度提升为1.0 mmol/L,藻体内虾青素含量为9.6%,显著高于对照组(P<0.05)。运用荧光定量PCR分析表明:经过方案3处理后,斜生栅藻新藻种(Scenedesmus obliqast)体内ipi、psy、crtR-b基因的表达量分别为对照的15.60、16.20、19.68倍,均有显著差异(P<0.05)。尤其是lyc基因的表达量为对照的31.20倍,达到差异极显著(P<0.01)。从方案1到方案3,bkt、zds和pds基因的表达量逐渐增加,但不明显(P>0.05)。[结论]方案3的组合处理能使藻细胞内积累适度提升的水杨酸信号分子,适度提升的胞内SA含量就显著促进了虾青素的增长,而且组合处理协同调控7种关键酶基因表达。因此,2μmol/L褪黑素组合0.2 mg/L水杨酸加3.0 mg/L茉莉酸甲酯等诱导是非常有效的。[Objective]To study the effects of combined 20μmol/L melatonin and 0.2 mg/L salicylic acid with 3.0 mg/L methyl jasmonateon on astaxanthin accumulation in Scenedesmus obliqast.[Method]Scenedesmus obliqast grew in the optimized and improved BG11 medium.The vegetative cells was under A series of parallel 40.0W red lights and 28.0W blue lights with 1.2 meters long(total light 252μmol·m^-2·s^-1)and cultivated to the logarithmic phase continuously.Shake the culture twice a day to prevent wall sticking.One day after logarithmic growth,biomass fermentation and astaxanthin induction of Scenedesmus obliqast were done through blue light with 1.2 meters plus high light plus N starvation plus 51.8 g/L glucose plus 20.0 g/L xylose.They were cultivated for three days continuously(Project 1).Then,light intensity was increased to 555μmol·m^-2·s^-1 and they were cultivated for three days continuously(Project 2).Finally,combined 20μmol/L melatonin and 0.2 mg/L salicylic acid with 3.0 mg/L methyl jasmonateon on astaxanthin accumulation were performed.They were cultivated for five days continuously(Project 3).[Result]The biomass of Scenedesmus obliqast increased slightly through three projects and the maximum biomass was 0.91×10^6个/mL.After Project 3 treatment,SA content in Scenedesmus obliqast cells was increased to 1.0 mmol/L moderately.And the astaxanthin contents of Scenedesmus obliqast cells increased to 9.6%with the significant difference(P<0.05).qRT-PCR showed that after Project 3 treatment,three key enzyme genes,such as ipi,psy and crtR-b,were 15.60,16.20,19.68 times of the control with the significant difference(P<0.05),respectively.Especially,lyc were 31.20 times of the control with the significant difference at P<0.01.Bkt,zds and pds genes expressions were gradually increased at P>0.05.[Conclusion]Combined treatments through Project 3 could increase SA signal molecules in the cells moderately.Moderate increased SA promoted astaxanthin accumulation significantly.Moreover,combined treatment could regulate seven k

关 键 词：斜生栅藻新藻种 组合处理 协同效应 虾青素 分子机制 

分 类 号：Q945.[生物学—植物学]