莱茵衣藻丝/苏氨酸蛋白激酶突变株蓝光响应的表征

Characterization the response of Chlamydomonas reinhardtii serine/threonine protein kinase mutant to blue light

摘要:

eee) 为研究莱茵衣藻丝/苏氨酸蛋白激酶(silk/threonine protein kinase, STK)介导藻细胞蓝光响应的分子机制，本文对蓝光胁迫下莱茵衣藻STK突变株系crstk11 (AphvIII盒反向插入stk11基因编码区)进行表型鉴定及转录组分析。表型鉴定显示，正常光(白光)下，野生型株CC5325与突变株crstk11的生长和色素含量差异较小；蓝光抑制了crstk11藻细胞生长和叶绿素合成，但显著促进类胡萝卜素积累。转录组分析显示，蓝光处理4 d，突变株(STK4) vs.野生型(wild type, WT4)共检测到差异表达基因(differential expression genes, DEGs) 860条(559个上调，301个下调)。高蓝光处理8 d，STK8 vs. WT8共获得1 088个DEGs (468个上调，620个下调)。KEGG富集分析发现，与CC5325相比，crstk11蓝光响应基因主要参与胞内光合作用催化活性、碳代谢和色素合成等。其中，上调基因包括psaA、psaB和psaC，psbA、psbB、psbC、psbD、psbH和psbL，petA、petB和petD，以及编码ATP合成酶α、β和c亚基的基因。下调基因有petF和petJ等。研究揭示了莱茵衣藻蛋白激酶CrSTK11可能通过介导光合作用、色素合成和碳代谢参与藻细胞蓝光响应，这为深入解析藻类光胁迫抗性机制提供了新知识。

Abstract:

In order to investigate the molecular mechanism of silk/threonine protein kinase (STK)-mediated blue light response in the algal Chlamydomonas reinhardtii, phenotype identification and transcriptome analysis were conducted for C. reinhardtii STK mutant strain crstk11 (with an AphvIII box reverse insertion in stk11 gene coding region) under blue light stress. Phenotypic examination showed that under normal light (white light), there was a slight difference in growth and pigment contents between the wild-type strain CC5325 and the mutant strain crstk11. Blue light inhibited the growth and chlorophyll synthesis in crstk11 cells, but significantly promoted the accumulation of carotenoids in crstk11. Transcriptome analysis showed that 860 differential expression genes (DEG) (559 up-regulated and 301 down-regulated) were detected in mutant (STK4) vs. wild type (WT4) upon treatment under high intensity blue light for 4 days. After being treated under high intensity blue light for 8 days, a total of 1 088 DEGs (468 upregulated and 620 downregulated) were obtained in STK8 vs. WT8. KEGG enrichment analysis revealed that compared to CC5325, the crstk11 blue light responsive genes were mainly involved in catalytic activity of intracellular photosynthesis, carbon metabolism, and pigment synthesis. Among them, upregulated genes included psaA, psaB, and psaC, psbA, psbB, psbC, psbD, psbH, and L, petA, petB, and petD, as well as genes encoding ATP synthase α, β and c subunits. Downregulated genes included petF and petJ. The present study uncovered that the protein kinase CrSTK11 of C. reinhardtii may participate in the blue light response of algal cells by mediating photosynthesis as well as pigment and carbon metabolism, providing new knowledge for in-depth analysis of the mechanism of light stress resistance in the algae.