Understanding the functions of endogenous DOF transcript factor in Chlamydomonas reinhardtii
Understanding the functions of endogenous DOF transcript factor in Chlamydomonas reinhardtii
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了解莱茵衣藻内源 DOF 转录因子的功能
DOI：
10.1186/s13068-019-1403-1
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发表时间：
2019-03-27
影响因子：
6.3
通讯作者：
Hu, Zhangli
中科院分区：
文献类型：
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作者：
Jia, Bin;Xie, Xinfeng;Hu, Zhangli
背景：利用基因工程手段调控微藻脂类代谢相关基因是提高微藻脂类积累的重要途径。单指DNA结合因子（DNA binding with one finger，DOF）是高等植物中的一种植物特异性转录因子，它通过调控碳氮代谢途径中的关键基因来调控这些途径。DOF的过表达可以增加植物的脂质产量，但是DOF的过表达是否可以增加微藻的脂质产量还不清楚。该转录因子全长1875 bp，编码624个氨基酸，具有保守的DOF结构域。crDOF在C. reinhardti显著增加细胞内脂质含量。转基因藻株Tranc-crDOF-12的总脂肪酸含量为126.01 μg/mg（干重），比野生型高23.24%。此外，转基因Tranc-crDOF-12品系中的不饱和脂肪酸含量显著增加。荧光定量PCR分析结果表明，在转基因株系Tranc-crDOF-12中，BCC 1、FAT 1、SQD 1、MGD 1、DGD 1和PGP 1基因的表达水平显著上调，而ACP 1、ACS 1、CIS 1和SQD 2基因的表达水平显著下调。reinhardtii通过调节参与脂质代谢的关键基因。根据这些发现，我们建议，通过过表达DOF来提高微藻中的脂质含量可以在其他工业微藻菌株中实现，并用于生物柴油的工业生产。
Background:The regulation of genes related to lipid metabolism by genetic engineering is an important way to increase the accumulation of lipids in microalgae. DNA binding with one finger (DOF) is a plant-specific transcription factor in higher plants, where it regulates carbon and nitrogen metabolic pathways by regulating key genes involved in these pathways. Overexpression of DOF can increase lipid production in plants; however, it is not clear whether overexpression of DOF can increase lipids in microalgae.Results:In this study, we cloned a DOF transcription factor, crDOF, fromChlamydomonas reinhardtii. The sequence of this transcription factor is 1875 bp and encodes a peptide of 624 amino acids with a conserved DOF domain. Overexpression of crDOF inC. reinhardtiisignificantly increased the intracellular lipid content. The content of total fatty acids in the transgenic algae line Tranc-crDOF-12 was 126.01 μg/mg (dry weight), which was 23.24% higher than that of the wild type. Additionally, the content of unsaturated fatty acids in the transgenic Tranc-crDOF-12 line increased significantly. Fluorescence quantitative PCR analysis showed that in the transgenic line Tranc-crDOF-12, the expression levels of BCC1, FAT1, SQD1, MGD1, DGD1 and PGP1 genes were significantly upregulated, while the expression levels of ACP1, ACS1, CIS1 and SQD2 were downregulated.Conclusions:Our results confirm that crDOF increases intracellular lipids inC. reinhardtiiby regulating key genes involved in lipid metabolism. According to these findings, we propose that enhancing the lipid content in microalgae by overexpressing DOF may be achieved in other industrial strains of microalgae and be employed for the industrial production of biodiesel.