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First published online January 14, 2005; 10.1104/pp.104.055244

Plant Physiology 137:545-556 (2005)
© 2005 American Society of Plant Biologists

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Functional Genomics of Eukaryotic Photosynthesis Using Insertional Mutagenesis of Chlamydomonas reinhardtii1

Rachel M. Dent, Cat M. Haglund, Brian L. Chin2, Marilyn C. Kobayashi and Krishna K. Niyogi*

Department of Plant and Microbial Biology, University of California, Berkeley, California 94720–3102

The unicellular green alga Chlamydomonas reinhardtii is a widely used model organism for studies of oxygenic photosynthesis in eukaryotes. Here we describe the development of a resource for functional genomics of photosynthesis using insertional mutagenesis of the Chlamydomonas nuclear genome. Chlamydomonas cells were transformed with either of two plasmids conferring zeocin resistance, and insertional mutants were selected in the dark on acetate-containing medium to recover light-sensitive and nonphotosynthetic mutants. The population of insertional mutants was subjected to a battery of primary and secondary phenotypic screens to identify photosynthesis-related mutants that were pigment deficient, light sensitive, nonphotosynthetic, or hypersensitive to reactive oxygen species. Approximately 9% of the insertional mutants exhibited 1 or more of these phenotypes. Molecular analysis showed that each mutant line contains an average of 1.4 insertions, and genetic analysis indicated that approximately 50% of the mutations are tagged by the transforming DNA. Flanking DNA was isolated from the mutants, and sequence data for the insertion sites in 50 mutants are presented and discussed.

1 This work was supported by the National Science Foundation (grant no. MCB–0235878).

2 Present address: Department of Biology, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA 02139.

Article, publication date, and citation information can be found at www.plantphysiol.org/cgi/doi/10.1104/pp.104.055244.

* Corresponding author; e-mail niyogi{at}nature.berkeley.edu; fax 510–642–4995.

Received October 19, 2004; returned for revision November 11, 2004; accepted November 12, 2004.




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