光诱导雨生红球藻虾青素积累的信号通路转录组分析  

Transcriptome analysis of signal transduction pathway involved in light inducing astaxanthin accumulation in Haematococcus pluvialis

作　　者：崔红利 许文鑫 崔玉琳[3] 季春丽 张春辉[1,2] 秦松 李润植[1,2] Hongli Cui;Wenxin Xu;Yulin Cui;Chunli Ji;Chunhui Zhang;Song Qin;Runzhi Li(College of Agriculture,Shanxi Agricultural University,Taigu 030801,Shanxi,China;Institute of Molecular Agriculture and Bioenergy,Shanxi Agricultural University,Taigu 030801,Shanxi,China;Yantai Institute of Coastal Zone Research,Chinese Academy of Sciences,Yantai 264003,Shandong,China)

机构地区：[1]山西农业大学农学院,山西太谷030801 [2]山西农业大学分子农业与生物能源研究所,山西太谷030801 [3]中国科学院烟台海岸带研究所,山东烟台264003

出　　处：《生物工程学报》2021年第4期1260-1276,共17页Chinese Journal of Biotechnology

基　　金：国家自然科学基金(No.31902394);山西省重点研发计划(No.201803D31063);山西省应用基础研究项目(No.201801D22125);山西农业大学科技创新基金项目(No.2018YJ16);国家海洋局海洋生物活性物质与现代分析技术重点实验室项目(No.MBSMAT-2016-03)资助。

摘　　要：雨生红球藻Haematococcuspluvialis在逆境胁迫条件下可大量积累虾青素,被认为自然界最理想的虾青素生产者。高光能有效诱导其虾青素的合成与积累,但藻细胞感知和转导光信号进而调控虾青素积累的机制尚不清楚。文中采用Illumina Hiseq 2000高通量测序技术分别获得正常、高白光及高蓝光处理组4.0 G、3.8 G及3.6 G的原始数据量,经质控与拼接之后获得51954条长度至少为200 bp的unigenes基因,经过比对分析,共有20537个unigenes在NR、NT、KO、SwissProt、PfamGO及KOG等数据库中的至少1个数据库中注释成功,达到39.52%。差异表达基因分析显示,高白光vs正常组共获得1255个DEGs;高蓝光vs正常组共获得1494个DEGs;高白光与高蓝光vs正常组共同的DEGs有1008个。KEGG富集分析显示,与对照组相比高白光与高蓝光共同的显著富集通路包括光合作用、类胡萝卜素合成、脂肪酸合成、氧化磷酸化、DNA复制、碳代谢及氮代谢等过程。通过对转录组数据进一步分析,挖掘鉴定了大量光受体及其信号转导通路中的互作蛋白。随机筛选DEGs基因15条,采用荧光定量PCR技术检测其转录水平,结果表明与转录组差异表达数据高度一致。光受体及其信号转导通路中的互作蛋白基因差异表达分析,推测"光信号→光受体→互作蛋白(互作蛋白→转录因子/转录调节子)→功能基因表达→虾青素积累"的信号转导通路可能参与上述调控过程,为深入解析光诱导虾青素合成的转录调控机制奠定了基础。The unicellular green alga Haematococcus pluvialis is the best source of natural astaxanthin(AST)in the world due to its high content under stress conditions.Although high light(HL)can effectively induce AST biosynthesis,the specific mechanisms of light signal perception and transduction are unclear.In the current study,we used transcriptomic data of normal(N),high white light(W),and high blue light(B)to study the mechanisms of light inducing AST accumulation from the point of photoreceptors.The original data of 4.0 G,3.8 G,and 3.6 G for N,W,and B were obtained,respectively,by the Illumina Hi-seq 2000 sequencing technology.Totally,51954 unigenes(at least 200 bp in length)were generated,of which,20537 unigenes were annotated into at least one database(NR,NT,KO,SwissProt,Pfam,GO,or KOG).There were 1255 DEGs in the W vs N,1494 DEGs in the B vs N,and 1008 DEGs in the both W vs N and B vs N.KEGG enrichment analysis revealed that photosynthesis,oxidative phosphorylation,carotenoid biosynthesis,fatty acids biosynthesis,DNA replication,nitrogen metabolism,and carbon metabolism were the significantly enriched pathways.Moreover,a large number of genes encoding photoreceptors and predicted interacting proteins were predicted in Haematococcus transcriptome data.These genes showed significant differences at transcriptional expression levels.In addition,15 related DEGs were selected and tested by qRT-PCR and the results were significantly correlated with the transcriptome data.The above results indicate that the signal transduction pathway of"light signal-photoreceptors-interaction proteins-(interaction proteins-transcription factor/transcriptional regulator)-gene expression-AST accumulation"might play important roles in the regulation process,and provide reference for further understanding the transcriptional regulation mechanisms of AST accumulation under HL stress.

关 键 词：雨生红球藻 高光 虾青素 转录组测序 光信号通路 

分 类 号：Q78[生物学—分子生物学] TQ28[化学工程—有机化工]