Directed Evolution of Yeast Ferric Reductase to Produce Plants with Tolerance to Iron Deficiency in Alkaline Soils
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概要
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One of the most serious problems in agriculture worldwide is low iron (Fe) availability, due to high soil pH. About 30% of arable land is too alkaline for optimal crop production. Non-graminaceous and dicot plants, which use a reduction strategy to uptake Fe, suffer from Fe deficiency under these conditions, because the ferric chelate reductase in the root plasma membrane functions inefficiently at high pH. The refre1 (reconstructed yeast ferric reductase) gene was subjected to random mutagenesis to obtain variants with high activity under high pH conditions. A mutant library was screened using a yeast in vivo assay system, and screens at pH 8.0 and 8.5 produced 10 candidates. In vivo ferric reductase activity was analyzed quantitatively. Yeast cells carrying the variant with the highest ferric reductase activity showed 6.0, 8.7, and 38 times greater activity at pH 8.0, 8.5, and 9.0, respectively, than did cells containing the original refre1 gene. An amino acid substitution at position 312 was common to most of the high-activity variants. This substitution is believed to play an important role in the increased reductase activity at high pH. Interestingly, this mutation is near a heme-coordinating histidine codon, and the corresponding residue is probably located in the intramembranous region close to the cytoplasm. The variant gene with the highest reductase activity was introduced into tobacco, and transgenic tobacco carrying the gene showed enhanced tolerance to low Fe availability. This result should be useful in the engineering of non-graminaceous and dicot plants tolerant to Fe deficiency in alkaline soils.
- 社団法人日本土壌肥料学会の論文
著者
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MORI Satoshi
Department of Applied Biological Chemistry, Division of Agriculture and Agricultural Life Sciences,
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Nishizawa Naoko
Department of Agricultural Chemistry, Faculty of Agriculture, The University of Tokyo
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Nishizawa Naoko
Laboratory Of Plant Biotechnology Graduate School Of Agricultural And Life Sciences The University O
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Mori S
Laboratory Of Plant Biotechnology Graduate School Of Agricultural And Life Sciences The University O
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Kawasaki S
Kyokugen Research Center For Materials Science At Extreme Conditions Osaka University
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TAKAHASHI Michiko
Department of Pediatrics, Sapporo Kousei Hospital
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NAKANISHI Hiromi
Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, Unive
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Oki Hiroyuki
Department of Applied Biological Chemistyr, The University of Tokyo
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Kim Suyeon
Department of Global Agricultural Sciences, The University of Tokyo
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Yamaguchi Hirotaka
Department of Applied Biological Chemistry, The University of Tokyo
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Kawasaki Shinji
Japex Geosciences Institute Tokyo Japan
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Matsuhashi S
Takasaki Advanced Radiation Research Institute Japan Atomic Energy Agency
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Nishizawa Naoko
The University Of Tokyo
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Nishizawa Naoko-kishi
Lab Plant. Mol. Physiol. Tokyo Univ. Fac. Pharmaceutical. Science. Nagoya City Univ.
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Nishizawa Naoko
Department Of Agricultural Chemistry Faculty Of Agriculture The University Of Tokyo
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Mori S
Dept. Of Appl. Biol. Chem. Univ. Tokyo
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Mori Satoshi
Department Of Agricultural Chemistry Faculty Of Agriculture The University Of Tokyo
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Yamaguchi Hirotaka
Dept. Agriculture And Agricultural Chemistry Tokyo Univ.:crest
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Takahashi Michiko
Department Of Cardiology Kitasato Institute Hospital
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Takahashi Michiko
Department Of Global Agricultural Sciences Graduate School Of Agricultural And Life Science The Univ
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Mori Satoshi
Lab Plant. Mol. Physiol. Tokyo Univ. Fac. Pharmaceutical. Science. Nagoya City Univ.
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Nishizawa N‐k
Univ. Tokyo Tokyo Jpn
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Mori S
Department Of Applied Biological Chemistry The University Of Tokyo
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Kim S
Univ. Tokyo Tokyo
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Yamaguchi H
Akita Prefectual University
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Oki Hiroyuki
Department Of Applied Biological Chemistyr The University Of Tokyo:(present Address)the Graduate Sch
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Nishizawa Naoko
Environmental Science Tohoku University
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Mori S
Meijo Univ. Aichi Jpn
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Nishizawa Naoko-kishi
Laboratory Of Plant Nutrition And Fertilizers Department Of Agricultural Chemistry The University Of
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Takahashi Michiko
Lab Plant. Mol. Physiol. Tokyo Univ. Fac. Pharmaceutical. Science. Nagoya City Univ.
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Nishizawa Naoko-kishi
Laboratory Of Plant Molecular Physiology The University Of Tokyo
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Oki Hiroyuki
Department of Applied Biological Chemistry, The University of Tokyo
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