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Heat Shock Treatment of Chlamydomonas reinhardtii and Chlorella Cells
Published: Vol 3, Iss 15, Aug 5, 2013 DOI: 10.21769/BioProtoc.849 Views: 9454
Edited by: Ru Zhang
Original research article
The authors used this protocol in:
Mar 2013
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Abstract
The protocol is very reliable and simple for inducing heat shock in unicellular green algae cells. The main purpose was to compare cellular response of three Chlorella species, isolated from different habitats: Chlorella vulgaris 8/1- thermophilic, Chlorella kesslery- mesophilic and C. vulgaris- extremophilic. Species were isolated from different habitats and differ in their temperature preferences and tolerance. Temperature induced stress response was measured as cell survival, induction of chloroplast HSP70B and DSBs induction and rejoining.
Materials and Reagents
- Species:
Three Chlorella species were used: C. vulgaris, isolated from soil samples of Livingston Island, the South Shetland Archipelago, Antarctic; C. vulgaris strain 8/1, isolated in 1968 from thermal springs in the region of Rupite, Bulgaria, and cultivated in our laboratory since 1975 and Chlorella kesslery a mesophile, from the Trebon collection.
Cultivation: Chlorella species were cultivated on Tris Acetate Phosphate (TAP) medium under continuous light of 60 μmol/m2/s and a temperature of 23 °C ± 0.1 °C in a Phytotron GC 400 growth chamber. The species were cultivated at this temperature because it is well known, that eurythermal algae, could be grown at a wide range of temperatures. - TAP medium (see Recipes)
- Sager–Granick medium (see Recipes)
Equipment
- Phytotron GC 400 growth chamber (NUVE Ankara/Turkey 2009)
- Microscope
- Bürker chamber
- Circulating water bath
Category
Plant Science > Phycology > Cell analysis
Cell Biology > Cell signaling > Stress response
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