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Plant and Cell Physiology, 2003, Vol. 44, No. 5 541-548
© 2003 Oxford University Press

Toxicity of Free Proline Revealed in an Arabidopsis T-DNA-Tagged Mutant Deficient in Proline Dehydrogenase

Tokihiko Nanjo1,2,5, Miki Fujita3, Motoaki Seki3, Tomohiko Kato4, Satoshi Tabata4 and Kazuo Shinozaki1,6

1 Laboratory of Plant Molecular Biology, RIKEN Tsukuba Institute, 3-1-1, Koyadai, Tsukuba, Ibaraki, 305-0074 Japan
2 Genesis Research Institute, Inc., 4-1-35, Noritake-Shinmachi Nishi-Ku, Nagoya, Aichi, 451-0051 Japan
3 Plant Functional Genomics Research Group, RIKEN Genomic Sciences Center, 1-7-22, Suehiro-cho, Tsurumi-Ku, Yokohama, Kanagawa, 230-0045 Japan
4 Kazusa DNA Research Institute, 1532-3, Yana, Kisarazu, Chiba, 292-0812 Japan

The toxicity of proline (Pro) to plant growth has raised questions despite its protective functions in response to environmental stresses. To evaluate Pro toxicity, we isolated an Arabidopsis T-DNA-tagged mutant, pdh, that had a defect in Pro dehydrogenase (AtProDH), which catalyzes the first step of Pro catabolism. The pdh mutant showed hypersensitivity to exogenous application of <=10 mM L-Pro, at which wild-type plants grew normally. A dose-dependent increase in internal free Pro accumulation was observed in pdh plants during external Pro supply. These results do not just prove the toxicity of Pro, but also suggest that AtProDH is the only enzyme acting as a functional ProDH in Arabidopsis. To further analyze the targets of Pro toxicity, we compared the expression of thousands of genes by pdh plants with that by wild-type plants by cDNA microarray analysis. Most genes were unaffected. Here we demonstrate Pro toxicity by using the pdh mutant and discuss a cause-and-effect action between an excess of free Pro and growth inhibition in Arabidopsis.

5 Current address: Department of Molecular and Cell Biology, Forestry and Forest Products Research Institute, 1 Matsunosato, Tsukuba, Ibaraki, 305-8687 Japan.

6 Corresponding author: E-mail, sinozaki{at}rtc.riken.go.jp; Fax, +81-29-836-9060.


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