Chlamydomonas reinhardtii, a model system for functional validation of abiotic stress responsive genes.
Stress tolerance is a multigenic character and there are many stress responsive genes, which are stress specific. Although many of these have been cloned, their functional significance remains fragmentary. Hence it is important to identify the relevant stress genes involved in altering the metabolism for adaptation. Overexpression is one of the several approaches and Chlamydomonas is a suitable system to study the functional relevance of stress genes. Stress responses can only be assessed on prior exposure to sublethal induction stress. In this study the acclimation response of Chlamydomonas was assessed for different abiotic stresses using physiological screens like chlorophyll stability, membrane damage, cell viability, accumulation of free radicals, survival and recovery growth. We demonstrate that Chlamydomonas responds to diverse stresses and is a potential system to study the relevance of stress genes. The relevance of choline oxidase A (codA), a key enzyme in glycinebetaine biosynthesis, was examined by developing transformants expressing codA gene from Arthrobacter globiformis. Southern positive transformants showed enhanced accumulation of glycinebetaine. The transformants also showed enhanced growth under salinity, high light coupled with methylviologen-induced oxidative stress, high temperature and cold stress. However the transgenics were not tolerant to PEG-mediated simulated osmotic stress, LiCl, menadione and UV stress. Increased cell survival and decreased chlorophyll degradation in transformants under acclimated conditions further confirmed the relevance of codA in imparting stress tolerance. Our results indicated that the relevance of stress responsive genes can be efficiently validated for diverse abiotic stresses using Chlamydomonas system.
Hema
R
R
Department of Crop Physiology, University of Agricultural Sciences, GKVK, Bangalore 560 065, India.
Senthil-Kumar
M
M
Shivakumar
S
S
Chandrasekhara Reddy
P
P
Udayakumar
M
M
D000222:Adaptation, Physiological / Q000187:drug effects / Q000235:genetics* / Q000528:radiation effects; D020418:Algal Proteins / Q000235:genetics*; D000818:Animals; D001622:Betaine / Q000378:metabolism; D016825:Chlamydomonas reinhardtii / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development / Q000528:radiation effects; D002934:Cinnamates / Q000494:pharmacology; D003080:Cold Temperature; D005966:Glucuronidase / Q000378:metabolism; D006921:Hygromycin B / Q000031:analogs & derivatives / Q000494:pharmacology; D008027:Light; D018021:Lithium Chloride / Q000494:pharmacology; D008954:Models, Biological; D009997:Osmotic Pressure; D018384:Oxidative Stress / Q000187:drug effects / Q000528:radiation effects; D010269:Paraquat / Q000494:pharmacology; D015203:Reproducibility of Results; D012965:Sodium Chloride / Q000494:pharmacology; D014466:Ultraviolet Rays; D024483:Vitamin K 3 / Q000494:pharmacology
D020418:Algal Proteins; D002934:Cinnamates; D001622:Betaine; D006921:Hygromycin B; C026273:hygromycin A; D012965:Sodium Chloride; D024483:Vitamin K 3; D005966:Glucuronidase; D018021:Lithium Chloride; D010269:Paraquat