ROS Detection in Botryococcus braunii Colonies with CellROX Green Reagent

Autor: Edmundo Lozoya-Gloria, Ivette Cornejo-Corona, Hem Thapa, Daniel Browne, Timothy Devarenne
Jazyk: angličtina
Rok vydání: 2017
Předmět:
Zdroj: Bio-Protocol, Vol 7, Iss 16 (2017)
Druh dokumentu: article
ISSN: 2331-8325
DOI: 10.21769/BioProtoc.2508
Popis: We analyzed the reactive oxygen species (ROS) accumulation in the colony-forming green microalga Botryococcus braunii in response to several stress inducers such as NaCl, NaHCO3, 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 B. braunii, 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.
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