HUANG Ying,LI Xiaolian,YIN Jianbo,et al.Transcriptome analysis of Chlamydomonas reinhardtii under the co-regulation of three genes[J].Journal of Shenzhen University Science and Engineering,2021,38(2):194-200.[doi:10.3724/SP.J.1249.2021.02194]





Transcriptome analysis of Chlamydomonas reinhardtii under the co-regulation of three genes
HUANG Ying LI Xiaolian YIN Jianbo LIN Yongqi PENG Xiaobo and JIA Bin
College of Life Sciences and Oceanography, Shenzhen Key Laboratory of Marine Bioresources and Ecology, Guangdong Technology Research Center for Marine Algal Bioengineering, Shenzhen University, Shenzhen 518060, Guangdong Province, P.R.China
基因工程 生物信息学 转录组acs2基因 cis1基因 DOF转录因子 脂质生物合成 莱茵衣藻
genetic engineering bioinformatics transcriptome acs2 gene cis1 gene DOF transcription regulator lipid biosynthesis Chlamydomonas reinhardtii
为研究多基因调控下微藻油脂合成调控的分子机制,对acs2-cis1-dof基因共调控的莱茵衣藻藻株DLC进行转录组测序及分析,获得749个差异表达基因.通过京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes, KEGG)聚类分析发现,有51个差异表达基因参与碳水化合物代谢,15个差异表达基因参与脂质代谢,差异基因主要富集在甘油酯代谢、脂肪酸降解、糖酵解及糖异生、淀粉和蔗糖代谢等通路中.采用BODIPY 505/515荧光染色法检测发现,热激后莱茵衣藻藻株DLC细胞内的甘油三酯质量较野生型提高了193%,通过检测吸光度发现藻细胞的淀粉质量降低了45%,说明基因共调控不仅增强了脂质合成代谢,还使得糖代谢中的碳流重新定向.研究工作为探究基因共调控下脂质合成代谢的分子机理以及微藻产油基因工程提供理论参考.
In order to study the regulatory roles of multiple genes in microalgal lipid metabolism, a Chlamydomonas reinhardtii strain DLC is constructed with overexpression of DOF transcription regulator and silencing of acs2 and cis1. Transcriptome sequencing and analysis are conducted and 749 differentially expressed genes (DEGs) are identified. According to the result of KEGG (Kyoto encyclopedia of genes and genomes) classification, 51 and 15 DEGs are involved in carbohydrate metabolism and lipid metabolism respectively. These DEGs mainly play key roles in glyceride metabolism, fatty acid degradation, glycolysis/gluconeogenesis and starch and sucrose metabolism. In addition, triacylglycerol (TAG) and starch content are analyzed by BODIPY505/515 fluorescence analysis and absorbance detection, which shows that intracellular TAG increases by 193% as compared with wild-type, while starch content decreases by 45%. These results further indicate that the co-regulation of the three genes not only enhances the lipid anabolism, but also reorients the carbon flux in the saccharide metabolism. The research reveals the molecular mechanism of lipid overproduction under co-regulation of three genes and provides a theoretical reference for the genetic engineering of microalgae lipid production.


[1] GIBELLINI F, SMITH T K. The Kennedy pathway: de novo synthesis of phosphatidylethanolamine and phosphatidylcholine[J]. IUBMB Life,2010,62(6):414-428.
[2] JIA Bin, SONG Yanzi, WU Min,et al.Characterization of long-chain acyl-CoA synthetases which stimulate secretion of fatty acids in green algae Chlamydomonas reinhardtii[J].Biotechnology for Biofuels,2016,9(1):184-195.
[3] DENG Xiaodong, CAI Jiajia, FEI Xiaowen.Effect of the expression and knockdown of citrate synthase gene on carbon flux during triacylglycerol biosynthesis by green algae Chlamydomonas reinhardtii[J].BMC Biochemistry,2013(14):38-49.
[4] JIA Bin, XIE Xinfeng, WU Min,et al.Understanding the functions of endogenous DOF transcript factor in Chlamydomonas reinhardtii[J].Biotechnology for Biofuels,2019(12):67-80.
[5] 胡浪,刘烨蓉,许伟钊,等.集胞藻缺氮驯化株的代谢组学分析[J].深圳大学学报理工版,2019,36(5):564-569.
HU Lang, LIU Yerong, XU Weizhao,et al.Metabonomics analysis of nitrogen deficiency-generated strains of Synechocystis sp.[J].Journal of Shenzhen University Science and Engineering,2019,36(5):564-569.(in Chinese)
[6] 娄素琳,朱秀兰,曾志勇,等.盐藻microRNAs高通量测序和生物信息学分析[J].深圳大学学报理工版,2018,35(4):331-333.
LOU Sulin, ZHU Xiulan, ZENG Zhiyong,et al.Bioinformatics analysis of Dunaliella microRNAs by high-throughput sequencing[J].Journal of Shenzhen University Science and Engineering,2018,35(4):331-333.(in Chinese)
[7] 黄瑛,殷剑波,李小连,等.莱茵衣藻三基因共调控油脂积累研究[J].中南民族大学学报自然科学版,2020, 39(5):465-470.
HUANG Ying, YAN Jianbo, LI Xiaolian,et al.Study on the regulation of oil accumulation by three genes of Chlamydomonas reinhardtii[J].Journal of South-Central University for Nationalities Natural Science,2020,39(5):465-470.(in Chinese)

[8] KIM D, LANGMEAD B, SALZBERG S L.HISAT: a fast spliced aligner with low memory requirements[J].Nature Methods,2015,12(4):357-360.
[9] LANGMEAD B, SALZBERG S L.Fast gapped-read alignment with Bowtie 2[J].Nature Methods,2012,9(4):357-359.
[10] LI Bo, DEWEY C N.RSEM: accurate transcript quantification from RNA-seq data with or without a reference genome[J].BMC Bioinformatics,2011,12(1):323-339.
[11] PAN Hong, ZHANG Lihua, LI Xun,et al.Biosynthesis of the fatty acid isopropyl esters by engineered Escherichia coli[J].Enzyme and Microbial Technology,2017(102):49-52.
[12] WEPY J A, GALLIGAN J J, KINGSLEY P J, et al. Lysophospholipases cooperate to mediate lipid homeostasis and lysophospholipid signaling[J].Journal of Lipid Research,2019,60(2):360-374.
[13] PAVLOVIC Z, BAKOVIC M.Regulation of phosphatidylethanolamine homeostasis; the critical role of CTP:phosphoethanolamine cytidylyltransferase (Pcyt2)[J].International Journal of Molecular Sciences,2013,14(2):2529-2550.
[14] 徐靖,李辉亮,朱家红,等.植物脂肪酸β-氧化的研究进展[J].生物技术通讯,2008,19(1):141-144.
XU Jing, LI Huiliang, ZHU Jiahong,et al.Insights into fatty acids β-oxidation in plant[J].Letters in Biotechnology,2008,19(1):141-144.(in Chinese)
[15] MORAIS S, KNOLL-GELLIDA A, ANDR M,et al.Conserved expression of alternative splicing variants of peroxisomal acyl-CoA oxidase 1 in vertebrates and developmental and nutritional regulation in fish[J].Physiological Genomics,2007,28(3):239-252.
[16] JENKINS L S, NUNN W D.Genetic and molecular characterization of the genes involved in short-chain fatty acid degradation in Escherichia coli: the ato system[J].Journal of Bacteriology,1987,169(1):42-52.
[17] HALLER J F, SMITH C, LIU Dalan,et al.Isolation of novel animal cell lines defective in glycerolipid biosynthesis reveals mutations in glucose-6-phosphate isomerase[J].The Journal of Biological Chemistry,2010,285(2):866-877.
[18] RENGEL R, SMITH R T, HASLAM R P,et al.Overexpression of acetyl-CoA synthetase (ACS) enhances the biosynthesis of neutral lipids and starch in the green microalga Chlamydomonas reinhardtii[J].Algal Research,2018(31):183-193.


 GONG Ying-xue,TAI Yan,XIAO Wen-juan,et al.Construction of Saccharomyces cerevisiae integrated expression vector and its application in cellulose bioconvesion[J].Journal of Shenzhen University Science and Engineering,2010,27(2):82.
 MO Bei-xin.RNA localization in plant cells and advances of the research[J].Journal of Shenzhen University Science and Engineering,2011,28(2):230.
 LIU Shi-de,CHEN Han-ying,ZHANG Jian-hua,et al.A novel STI1-like protein from Physarum polycephalum can enhance E.coli response to stress[J].Journal of Shenzhen University Science and Engineering,2011,28(2):347.
 ZHANG Yi-zhe,LIU Qiong,TIAN Jing,et al.Investigation on the interactive protein of iodothyronine deiodinase 3 mutant in human brain[J].Journal of Shenzhen University Science and Engineering,2011,28(2):423.
 LI Jian-cheng,PENG Shi-qing,et al.Expression and immunological activity of enterotoxin C2 from Staphylococcus aureus in Chlamydomonas reinhardti[J].Journal of Shenzhen University Science and Engineering,2012,29(2):159.[doi:10.3724/SP.J.1249.2012.02159]
 WANG Chao-gang,HUANG Hui-zhu,SUN Hai-shan,et al.Construction of Chlamydomonas reinhardtii system for analyzing the function of algal promoter[J].Journal of Shenzhen University Science and Engineering,2012,29(2):534.[doi:10.3724/SP.J.1249.2012.06534]
 LIU Ze-huan,ZHENG Yang-yang,GONG Ying-xue,et al.Direct conversion from cassava to bioethanol with a recombinant strain of yeast[J].Journal of Shenzhen University Science and Engineering,2012,29(2):548.[doi:10.3724/SP.J.1249.2012.06548]
 Hu Zhangli,Li Jiancheng,and Cheng Xuewei.The mitochondrial genetic system and exogenous gene expression of Chlamydomonas reinhardtii[J].Journal of Shenzhen University Science and Engineering,2013,30(2):603.[doi:10.3724/SP.J.1249.2013.06603]
 LI Yuan - le and TAO Lan.Protein secondary structure prediction based on WSVM[J].Journal of Shenzhen University Science and Engineering,2006,23(2):117.
 Wang Chaogang,Li Yi,and Hu Zhangli.Rapid screening of the transgenic algae haboring lpaat and gpd1[J].Journal of Shenzhen University Science and Engineering,2016,33(2):33.[doi:10.3724/SP.J.1249.2016.01033]


Foundation:National Natural Science Foundation of China(31870343);Basic Research and Applied Basic Research Foundation of Guangdong Province(2020A1515010352);Shenzhen Basic Research Foundation(JCYJ20170818095134434,JCYJ20180305125327627)
Corresponding author:Associate professor HUANG Ying.E-mail: huangy@szu.edu.cn
Citation:HUANG Ying, LI Xiaolian, YIN Jianbo, et al. Transcriptome analysis of Chlamydomonas reinhardtii under the co-regulation of three genes[J]. Journal of Shenzhen University Science and Engineering, 2021, 38(2): 194-200.(in Chinese)
基金项目:国家自然科学基金资助项目(31870343);广东省基础与应用基础研究基金资助项目(2020A1515010352);深圳市基础研究计划资助项目(JCYJ20170818095134434; JCYJ20180305125327627)
作者简介:黄瑛(1976—),深圳大学副教授、博士.研究方向:微藻生物质利用.E-mail: huangy@szu.edu.cn
引文:黄瑛,李小连,殷剑波,等.3个基因共调控下莱茵衣藻的转录组分析[J]. 深圳大学学报理工版,2021,38(2):194-200.
更新日期/Last Update: 2021-03-30