Functional study of LIL3 through the use of an inducible microRNA system
Jonathan Neilson and Dion Durnford
Department of Biology, University of New Brunswick, Fredericton, NB, CA
During acclimation to high-intensity light, a number of short-term photoprotective mechanisms are induced to protect the photosynthetic machinery from photooxidative damage. The Light Harvesting-Like (LIL) protein family is a diverse group of thylakoid membrane proteins, many of which are thought to be involved in photoprotection. LIL3 is a thylakoid membrane protein, found in all green algae and land plants, that contains two predicted membrane-spanning regions, and a single chlorophyll-binding domain. In land plants, LIL3 is hypothesized to be a pigment carrier protein for newly synthesized chlorophyll molecules; however, its function in algae has not been determined. To examine the function of LIL3 in Chlamydomonas we have created LIL3 RNAi knockdown mutants using an inducible artificial microRNA (amiRNA) based on the system developed by Molnar et al (2009 Plant J, 58, p165-174). This system takes advantage of the reversible inducibility of the mitochondrial alternative oxidase (AOX) promoter when Chlamydomonas reinhardtii cells are grown in the presence of nitrate as their sole nitrogen source. Details of the amiRNA system are presented, as well as preliminary physiological data concerning the LIL3 knockdown phenotype.
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