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BIOENERGETICS AND PHOTOSYNTHESIS

A Phosphofructokinase B-Type Carbohydrate Kinase Family Protein, NARA5, for Massive Expressions of Plastid-Encoded Photosynthetic Genes in Arabidopsis^1,[W],[OA]

Graduate School of Biological Sciences, Nara Institute of Science and Technology, Ikoma, Nara 630–0101, Japan (T.O., K.N., T.A., H.A., A.Y.); and Research Institute of Innovative Technology for the Earth, Kizugawa, Kyoto 619–0292, Japan (H.T., K.-I.T.)

To date, there have been no reports on screening for mutants defective in the massive accumulation of Rubisco in higher plants. Here, we describe a screening method based on the toxic accumulation of ammonia in the presence of methionine sulfoximine, a specific inhibitor of glutamine synthetase, during photorespiration initiated by the oxygenase reaction of Rubisco in Arabidopsis (Arabidopsis thaliana). Five recessive mutants with decreased amounts of Rubisco were identified and designated as nara mutants, as they contained a mutation in genes necessary for the achievement of Rubisco accumulation. The nara5-1 mutant showed markedly lower levels of plastid-encoded photosynthetic proteins, including Rubisco. Map-based cloning revealed that NARA5 encoded a chloroplast phosphofructokinase B-type carbohydrate kinase family protein of unknown function. The NARA5 protein fused to green fluorescent protein localized in chloroplasts. We conducted expression analyses of photosynthetic genes during light-induced greening of etiolated seedlings of nara5-1 and the T-DNA insertion mutant, nara5-2. Our results strongly suggest that NARA5 is indispensable for hyperexpression of photosynthetic genes encoded in the plastid genome, particularly rbcL.

² Present address: Faculty of Bioenvironmental Science, Kyoto Gakuen University, 1–1 Nanjyo, Sogabe, Kameoka, Kyoto 621–8555, Japan.

³ Present address: Plant High Technology Institute, Ltd., Takayama Science Plaza, 8916–12 Takayama, Ikoma, Nara 630–0101, Japan.

The author responsible for distribution of materials integral to the findings presented in this article in accordance with the policy described in the Instructions for Authors (www.plantphysiol.org) is: Akiho Yokota (yokota{at}bs.naist.jp).

^[W] The online version of this article contains Web-only data.

^[OA] Open Access articles can be viewed online without a subscription.

www.plantphysiol.org/cgi/doi/10.1104/pp.109.139683

^* Corresponding author; e-mail yokota{at}bs.naist.jp.

Received April 8, 2009; accepted July 2, 2009; published July 8, 2009.