Lozoya-Gloria Edmundo, Cornejo-Corona Ivette, Thapa Hem R, Browne Daniel R, Devarenne Timothy P
Department of Genetic Engineering, CINVESTAV-IPN, Irapuato, Guanajuato, México.
Department of Biochemistry and Biophysics, Texas A&M University, College Station, TX, USA.
Bio Protoc. 2017 Aug 20;7(16):e2508. doi: 10.21769/BioProtoc.2508.
We analyzed the reactive oxygen species (ROS) accumulation in the colony-forming green microalga in response to several stress inducers such as NaCl, NaHCO, salicylic acid (SA), methyl jasmonate, and acetic acid. A staining assay using the fluorescent dye CellROX Green was used. CellROX Green is a fluorogenic probe used for measuring oxidative stress in live cells. The dye is weakly fluorescent inside cells in a reduced state but exhibits bright green photostable fluorescence upon oxidation by ROS and subsequent binding to DNA. The large amount of liquid hydrocarbons produced and excreted by , creates a highly hydrophobic extracellular environment that makes difficult to study short times defense responses on this microalga. The procedure developed here allowed us to detect ROS in this microalga even within a short period of time (in minutes) after treatment of cells with different stress inducers.
我们分析了集落形成的绿色微藻中活性氧（ROS）的积累情况，以响应几种胁迫诱导剂，如氯化钠、碳酸氢钠、水杨酸（SA）、茉莉酸甲酯和乙酸。使用了一种基于荧光染料CellROX Green的染色测定法。CellROX Green是一种用于测量活细胞氧化应激的荧光探针。该染料在细胞内处于还原状态时荧光较弱，但在被ROS氧化并随后与DNA结合后会发出明亮的绿色光稳定荧光。[此处原文缺失具体藻类名称]产生并分泌的大量液态碳氢化合物创造了一个高度疏水的细胞外环境，这使得在短时间内研究这种微藻的防御反应变得困难。此处开发的方法使我们能够在使用不同胁迫诱导剂处理细胞后的短时间内（几分钟内）检测这种微藻中的ROS。
