Hwangbo Kwon, Lim Jong-Min, Jeong Seok-Won, Vikramathithan Jayaraman, Park Youn-Il, Jeong Won-Joong
Plant Systems Engineering Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon, South Korea.
Department of Biological Sciences, Chungnam National University, Daejeon, South Korea.
Front Plant Sci. 2018 May 7;9:590. doi: 10.3389/fpls.2018.00590. eCollection 2018.
Microalgae and higher plants employ an inorganic carbon (Ci) concentrating mechanism (CCM) to increase CO availability to Rubisco. Operation of the CCM should enhance the activity of the Calvin cycle, which could act as an electron sink for electrons generated by photosynthesis, and lower the redox status of photosynthetic electron transport chains. In this study, a hypothesis that microalgal cells with fully operating CCM are less likely to be photodamaged was tested by comparing a mutant with its wild type (WT). The mutant acquired by screening gamma-ray-induced mutant libraries of sp. ArM0029B exhibited constitutively active CCM (CAC) even in the presence of additional Ci sources under mixotrophic growth conditions. In comparison to the WT alga, the mutant named to constitutively active CCM1 (CAC1) showed more transcript levels for genes coding proteins related to CCM such as Ci transporters and carbonic anhydrases (CA), and greater levels of intracellular Ci content and CA activity regardless of whether growth is limited by light or not. Under photoinhibitory conditions, CAC1 mutant showed faster growth than WT cells with more PSII reaction center core component D1 protein (encoded by ), higher photochemical efficiency as estimated by the chlorophyll fluorescence parameter (Fv/Fm), and fewer reactive oxygen species (ROS). Interestingly, high light (HL)-induced increase in ROS contents in WT cells was significantly inhibited by bicarbonate supplementation. It is concluded that constitutive operation of CCM endows cells with resistance to HL partly by reducing the endogenous generation of ROS. These results will provide useful information on the interaction between CCM expression, ROS production, and photodamage in and related microalgae.
微藻和高等植物利用无机碳(Ci)浓缩机制(CCM)来提高核酮糖-1,5-二磷酸羧化酶/加氧酶(Rubisco)可利用的CO。CCM的运作应能增强卡尔文循环的活性,卡尔文循环可作为光合作用产生的电子的电子阱,并降低光合电子传递链的氧化还原状态。在本研究中,通过比较一个突变体与其野生型(WT),对一个假设进行了测试,该假设为具有完全运作的CCM的微藻细胞不太可能受到光损伤。通过筛选嗜盐杜氏藻(Dunaliella salina)sp. ArM0029B的γ射线诱导突变体文库获得的突变体,即使在混合营养生长条件下存在额外的Ci来源时,也表现出组成型活性CCM(CAC)。与野生型藻类相比,命名为组成型活性CCM1(CAC1)的突变体在编码与CCM相关蛋白质(如Ci转运蛋白和碳酸酐酶(CA))的基因上显示出更多的转录水平,并且无论生长是否受光限制,其细胞内Ci含量和CA活性水平都更高。在光抑制条件下,CAC1突变体比野生型细胞生长得更快,具有更多的光系统II反应中心核心组分D1蛋白(由psbA编码),通过叶绿素荧光参数(Fv/Fm)估计具有更高的光化学效率,以及更少的活性氧(ROS)。有趣的是,碳酸氢盐补充显著抑制了高光(HL)诱导的野生型细胞中ROS含量的增加。得出的结论是,CCM的组成型运作部分地通过减少ROS的内源性产生赋予盐藻细胞对高光的抗性。这些结果将为盐藻及相关微藻中CCM表达、ROS产生和光损伤之间的相互作用提供有用信息。