诱导子对三角褐指藻岩藻黄素含量和三角褐指藻ξ-胡萝卜素脱氢酶基因表达的影响  

Effects of Elicitors on Fucoxanthin Production and ZDS Gene Expression in Phaeodactylum tricornutum

作　　者：张燕[1] 龚一富[1] 景丹丹[1] 朱帅旗 王何瑜[1] 

机构地区：[1]宁波大学海洋学院,浙江宁波315211

出　　处：《中国药学杂志》2017年第12期1061-1068,共8页Chinese Pharmaceutical Journal

基　　金：浙江省科技厅重点科技创新团队项目资助(2012R10029-07;2010R50029);宁波科技攻关项目资助(2013C10018);浙江省大学生科技创新活动计划暨新苗人才计划项目资助(2014R405050)

摘　　要：目的研究甲基茉莉酸(MeJA)、花生四烯酸(AA)、乙酰水杨酸(ASA)对三角褐指藻岩藻黄素含量及三角褐指藻ξ-胡萝卜素脱氢酶基因(PtZDS)表达的影响。方法采用Illumina HiSeq^(TM)2000高通量测序方法获得PtZDS的cDNA全长序列,并对其进行生物信息学分析。利用HPLC测定MeJA、AA和ASA处理下,三角褐指藻岩藻黄素含量,RT-PCR检测PtZDS基因的表达。结果该基因全长1 905 bp,具有一个1 776 bp的开放阅读框,编码591个氨基酸。推导的氨基酸序列分析结果表明,该蛋白具有一个典型的氨基氧化酶结构域(amine oxidase domain)、NAD(P)-binding Rossmann-like结构域和叶绿体转运肽序列。系统进化树结果表明,PtZDS蛋白与假微型海链藻(Thalassiosira pseudonana CCMP1335)的同源性达76%,亲缘关系较近,共处同一进化支上。0.1 mg·L^(-1)AA、25 mg·L^(-1)ASA和100μmol·L^(-1)MeJA处理下三角褐指藻单位细胞内岩藻黄素的含量达到最高。RT-PCR分析结果表明,0.1 mg·L^(-1)AA、50μmol·L^(-1)MeJA和10 mg·L^(-1)ASA处理下,PtZDS的表达水平达到最高。结论 PtZDS基因的表达可能与三角褐指藻中岩藻黄素的积累存在一定关系。100μmol·L^(-1)MeJA处理下,三角褐指藻合成岩藻黄素的能力最强。OBJECTIVE To explore the effects of methyl jasmonic acid（ MeJA）, arachidonic acid（AA） and acetylsalicylic acid （ASA） on fucoxanthin production and PtZDS gene expression in Phaeodactylum tricornutum. METHODS The full length cDNA of PtZDS gene in P. tricornutum was obtained by HiSeqTM Illumina 2000 and bioinformatics analysis. HPLC was used to determine the production of fucoxanthin in Phaeodactylum tricornutum, and RT-PCR was used to determine the expression level of PtZDS. RESULTS The total length of PtZDS gene was 1 905 bp, containing an open reading frame of 1 776 bp encoding 591 amino acids. The deduced amino acid sequence analysis showed that PtZDS protein contained a typical amino oxidase domain, NAD（ P） -binding Rossmann- like domain and a chloroplastic transit peptide sequence. The phylogenetic analysis demonstrated that PtZDS was homologous with Thalas- siosirapseudonana CCMP1335（76% ）. Under the treatment of 0. 1 mg·L^-1 AA, 25 mg·L^-1 ASA and 100 μmol·L^-1 MeJA, the high- est yield of unit cell fucoxanthin content was achieved. Upon the observation of PtZDS regulation expression, the expression level of PtZDS reached a peak value under the treatment of 0. 1 mg·L^-1 AA, 50 μmol·L^-1 MeJA and 10 mg ·L^-1 ASA. CONCLUSION The expression of PtZDS gene has certain relationship with the fucoxanthin synthesis of P. tricornutum. Under the treatment of 100 μmol·L^-1 MeJA, the ability of synthetic fucoxanthinis the strongest in P. tricornutum.

关 键 词：三角褐指藻 岩藻黄素 甲基茉莉酸 乙酰水杨酸 花生四烯酸 ξ-胡萝卜素脱氢酶 

分 类 号：R284[医药卫生—中药学]