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Plant Physiol, April 2000, Vol. 122, pp. 1239-1248

Ectoine, the Compatible Solute of Halomonas elongata, Confers Hyperosmotic Tolerance in Cultured Tobacco Cells1

Hideki Nakayama, Kazuya Yoshida,* Hisayo Ono, Yoshikatsu Murooka, and Atsuhiko Shinmyo

Graduate School of Biological Sciences, Nara Institute of Science and Technology, 8916-5 Takayama-cho, Ikoma-shi, Nara 630-0101, Japan (H.N., K.Y., A.S.); and Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamada-oka, Suita-shi, Osaka 565-0871, Japan (H.O., Y.M.)

1,4,5,6-Tetrahydro-2-methyl-4-pyrimidinecarboxylic acid (ectoine) functions as a compatible osmolyte in the moderate halophile Halomonas elongata OUT30018. Ectoine is biosynthesized by three successive enzyme reactions from aspartic beta -semialdehyde. The genes encoding the enzymes involved in the biosynthesis, ectA, ectB, and ectC, encoding L-2,4-diaminobutyric acid acetyltransferase, L-2,4-diaminobutyric acid transaminase, and L-ectoine synthase, respectively, have been previously cloned. To investigate the function of ectoine as a compatible solute in plant cells, the three genes were individually placed under the control of the cauliflower mosaic virus 35S promoter and introduced together into cultured tobacco (Nicotiana tabacum L.) cv Bright Yellow 2 (BY2) cells. The transgenic BY2 cells accumulated a small quantity of ectoine (14-79 nmol g-1 fresh weight) and showed increased tolerance to hyperosmotic shock (900 mOsm). Furthermore, the transgenic BY2 cells exhibited a normal growth pattern even under hyperosmotic conditions (up to 530 mOsm), in which the growth of the untransformed BY2 (wild type) cells was obviously delayed. These results suggest that genetically engineered synthesis of ectoine results in the increased hyperosmotic tolerance of cultured tobacco BY2 cells despite the low level of accumulation of the solute.

1 This work was supported by the "Research for the Future" Program of the Japan Society for the Promotion of Science (JSPS-RFTF96R16001) and by a JSPS Research Fellowship for Young Scientists to H.N.

* Corresponding author; e-mail kazz{at}bs.aist-nara.ac.jp; fax 81-743-72-5469.

© 2000 American Society of Plant Physiologists


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