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

发布时间：2018-08-23 20:11

【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。
[Abstract]:Objective to study the effect of (MeJA), arachidonic acid (AA), acetylsalicylic acid (ASA) on the content of algin and the (PtZDS) expression of 尉-carotene dehydrogenase gene in Phaeopsis triangularis. Methods the full-length cDNA sequence of PtZDS was obtained by Illumina HiSeq~ (TM) 2000 high throughput sequencing and bioinformatics analysis. MeJA,AA and ASA were used to detect the expression of PtZDS gene by HPLC, and the content of algin was detected by reverse transcription-polymerase chain reaction (RT-PCR). Results the total length of the gene was 1905 bp, with an open reading frame of 1 776 bp, encoding 591 amino acids. The deduced amino acid sequence analysis showed that the protein had a typical amino oxidase domain (amine oxidase domain) nad (P) binding Rossmann-like domain and chloroplast transport peptide sequence. Phylogenetic tree results showed that the homology of PtZDS protein and (Thalassiosira pseudonana CCMP1335 was 76, and the phylogenetic relationship was close. Under the treatment of 0.1 mg L ~ (-1) AA,25 mg L ~ (-1) ASA and 100 渭 mol L ~ (-1) MeJA at the same evolutionary branch, the content of fucoflavin in the cells of Phaeophyllum triangularis reached the highest. The results of RT-PCR analysis showed that the expression level of PtZDS reached the highest level under the treatment of 0.1 mg L ~ (-1) AA,50 渭 mol L ~ (-1) MeJA and 10 mg L ~ (-1) ASA. Conclusion the expression of PtZDS gene may be related to the accumulation of alginin in Phaeodactylum triangularis. Under the treatment of .100 渭 mol L ~ (-1) MeJA, the ability of algin synthesis is the strongest.
【作者单位】：宁波大学海洋学院;
【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)
【分类号】：Q943.2

【参考文献】