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JXB Advance Access published online on May 31, 2005
Journal of Experimental Botany, doi:10.1093/jxb/eri195
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© The Author [2005]. Published by Oxford University Press [on behalf of the Society for Experimental Biology]. All rights reserved. For Permissions, please e-mail: journals.permissions@oupjournals.org
Received March 25, 2005
Accepted April 8, 2005

RESEARCH PAPER

Effects of free proline accumulation in petunias under drought stress

Mika Yamada 1, Hiromasa Morishita 2, Kaoru Urano 3, Noriko Shiozaki 1, Kazuko Yamaguchi-Shinozaki 4, Kazuo Shinozaki 3, and Yoshu Yoshiba 1*

1 Central Research Laboratory, c/o Advanced Research Laboratory, Hitachi Ltd., 2520 Akanuma, Hatoyama-cho, Hiki-gun, Saitama 350-0395, Japan
2 Research Resources Center, Brain Science Institute, RIKEN, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan
3 Laboratory of Plant Molecular Biology, RIKEN Tsukuba Institute, 3-1-1 Koyadai, Tsukuba, Ibaraki 305-0074, Japan
4 Biological Resources Division, Japan International Research Center for Agricultural Sciences (JIRCAS), Ministry of Agriculture, Forestry and Fisheries, 2-1 Ohwashi, Tsukuba, Ibaraki 305-8686, Japan

* To whom correspondence should be addressed.
Yoshu Yoshiba, E-mail: yoshiba{at}harl.hitachi.co.jp


  Abstract

Petunias (Petunia hybrida cv. ‘Mitchell’) accumulate free proline (Pro) under drought-stress conditions. It is therefore believed that Pro acts as an osmoprotectant in plants subjected to drought conditions. Petunia plants were transformed by {Delta}1-pyrroline-5-carboxylate synthetase genes (AtP5CS from Arabidopsis thaliana L. or OsP5CS from Oryza sativa L.). The transgenic plants accumulated Pro and their drought tolerance was tested. The Pro content amounted to 0.57-1.01% of the total amino acids in the transgenic plants, or 1.5-2.6 times that in wild-type plants grown under normal conditions. The transgenic plant lines tolerated 14 d of drought stress, which confirms that both P5CS transgenes had full functionality. Exogenous L-Pro treatment caused the plants to accumulate Pro; plants treated with 5 mM L-Pro accumulated up to 18 times more free Pro than untreated plants. Exogenous L-Pro restricted the growth of wild-type petunias more than that of Arabidopsis plants. The capacity for free Pro accumulation might depend on the plant species. The growth of petunia plants was influenced not only by the Pro concentration in the plants, but by the ratio of the Pro content to the total amino acids, because the growth of the transgenic petunia plants appeared normal.

Keywords: Drought tolerance; genetic engineering; osmoprotectant; stress response; transgenic petunia.
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