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Chlamydomonas reinhardtii, a model system for functional validation of abiotic stress responsive genes

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Abstract

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.

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Abbreviations

APX:

Ascorbate peroxidase

codA:

Choline oxidase A

HSP:

Heat shock protein

LEA:

Late embryogenesis abundant

MDA:

Malondialdehyde

ROS:

Reactive oxygen species

SSC:

Saline sodium citrate

TAP:

Tris–acetate–phosphate

SOD:

Superoxide dismutase

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Acknowledgments

Hema acknowledges the financial assistance from Department of Crop Physiology towards her Ph.D. programme. Senthil-Kumar thanks the Council of Scientific and Industrial Research, New Delhi, for the Senior Research Fellowship award (No. 9/271(86)/2004/EMR-1) for his Ph.D. programme and also American Society for Plant Biologists for award to ‘Plant Biology 2005’. Dr. B. J. Rao BARC Bombay, Dr. M. V. Rajam, South Campus (DU), New Delhi, and Dr. E. H. Harris, Duke University, Durham, NC, USA are acknowledged for the Chlamydomonas strains. The authors are thankful to Dr. Shivaprakash, MRC Bangalore, Dr. Gopalakrishna, Invitrogen Bangalore and Dr. Lalitha Anand, IIHR Bangalore, for the useful discussion during manuscript preparation.

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  1. Department of Crop Physiology, University of Agricultural Sciences, GKVK, Bangalore, 560 065, India

    R. Hema, M. Senthil-Kumar, S. Shivakumar, P. Chandrasekhara Reddy & M. Udayakumar

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  1. R. Hema
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  2. M. Senthil-Kumar
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  3. S. Shivakumar
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  4. P. Chandrasekhara Reddy
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  5. M. Udayakumar
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Correspondence to M. Udayakumar.

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R. Hema and M. Senthil-Kumar contributed equally.

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Hema, R., Senthil-Kumar, M., Shivakumar, S. et al. Chlamydomonas reinhardtii, a model system for functional validation of abiotic stress responsive genes. Planta 226, 655–670 (2007). https://doi.org/10.1007/s00425-007-0514-2

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