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JXB Advance Access originally published online on July 30, 2004
Journal of Experimental Botany 2004 55(405):1989-1996; doi:10.1093/jxb/erh218
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Journal of Experimental Botany, Vol. 55, No. 405, © Society for Experimental Biology 2004; all rights reserved

RESEARCH PAPER

Accumulation of menaquinones with incompletely reduced side chains and loss of {alpha}-tocopherol in rice mutants with alternations in the chlorophyll moiety

Masaru Shibata1,2, Michito Tsuyama1,3, Tsuneaki Takami1, Hideyuki Shimizu1 and Yoshichika Kobayashi1,*

1Department of Plant Resources, Graduate School of Agriculture, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka 812-8581, Japan
2Department of Materials Engineering, Nagaoka National College of Technology, 888 Nishikatakai, Nagaoka, Niigata 940-8532, Japan
3Forestry Research Institute, Oji Paper Company Ltd., 24-9 Nobono-cho, Kameyama, Mie 519-0212, Japan

* To whom correspondence should be addressed. Fax: +81 92 642 2880. E-mail: ykoba{at}agr.kyushu-u.ac.jp

The rice mutants M249 and M134 accumulate chlorophyllides a and b which are esterified with incompletely reduced alcohols such as geranylgeraniol, dihydrogeranylgeraniol, and tetrahydrogeranylgeraniol. Quantities of {alpha}-tocopherol, phylloquinone, and menaquinones in leaves of these mutants were determined by high performance liquid chromatography (HPLC) with a fluorescence detector after post-column chemical reduction to convert quinones to fluorescent quinols. Methylnaphthoquinones, varying in the reduction state of the side chain (menaquinones), were detected in leaf segments of the rice mutants on HPLC analyses with both high selectivity and sensitivity to plant quinones. Mutant M249 preferentially accumulated menaquinone, which contains tetrahydrogeranylgeraniol as its side chain. However, mutant M134 exhibited preferential accumulation of menaquinone with a geranylgeraniol side chain. In both mutants, the accumulation patterns of menaquinones with different prenyl side chains were similar to those of chlorophyll with the corresponding prenyl side chains. The content of P700, the photosystem I primary electron donor, in the wild type was greater than that of either mutant, on both a chlorophyll and a fresh weight basis. However, the ratios of total methylnaphthoquinones to P700 were similar in both the wild type and the mutants. Since no comparative large differences in photosynthetic activity exist between the wild type and the mutants, these results suggest that the hydrogenation of the methylnaphthoquinone side chain to phytol is not an essential requirement for it to function as an electron acceptor in photosystem I. On the other hand, {alpha}-tocopherol was detected in fully developed leaves of the wild type, but not in those of the mutants. Accumulation of menaquinones and the loss of {alpha}-tocopherol in mutant leaves suggest that the reduction of chlorophyll-geranylgeraniol to phytol and that of geranylgeranyl pyrophosphate to phytyl pyrophosphate are catalysed by the same enzyme.

Key words: Chlorophyll, geranylgeraniol, menaquinone, phylloquinone, phytol, {alpha}-tocopherol


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