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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
 -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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