First published online April 2, 2004; 10.1104/pp.103.036970
Plant Physiology 134:1697-1707 (2004)
© 2004 American Society of Plant Biologists
ENVIRONMENTAL STRESS AND ADAPTATION
Tissue-Specific Localization of an Abscisic Acid Biosynthetic Enzyme, AAO3, in Arabidopsis1
Hanae Koiwai,
Kentaro Nakaminami2,
Mitsunori Seo,
Wataru Mitsuhashi,
Tomonobu Toyomasu and
Tomokazu Koshiba*
Department of Biological Sciences, Tokyo Metropolitan University, Hachioji-shi, Tokyo 192–0397, Japan (H.K., T.K); Course of the Science of Bioresources, The United Graduate School of Agricultural Science, Iwate University, Morioka, Iwate 020–8550, Japan (K.N., W.M., T.T.); Plant Science Center, RIKEN (The Institute of Physical and Chemical Research), Yokohama, Kanagawa 230–0045, Japan (M.S.); and Faculty of Agriculture, Yamagata University, Wakaba-cho, Turuoka 997–8555, Japan (W.M., T.T.)
Arabidopsis aldehyde oxidase 3 (AAO3) is an enzyme involved in abscisic acid (ABA) biosynthesis in response to drought stress. Since the enzyme catalyzes the last step of the pathway, ABA production sites may be determined by the presence of AAO3. Here, AAO3 localization was investigated using AAO3 promoter:AAO3-GFP transgenic plants and by an immunohistochemical technique. AAO3-GFP protein exhibited an activity to produce ABA from abscisic aldehyde, and the transgene restored the wilty phenotype of the aao3 mutant. GFP-fluorescence was detected in the root tips, vascular bundles of roots, hypocotyls and inflorescence stems, and along the leaf veins. Intense immunofluorescence signals were localized in phloem companion cells and xylem parenchyma cells. Faint but significant GFP- and immuno-fluorescence signals were observed in the leaf guard cells. In situ hybridization with antisense AAO3 mRNA showed AAO3 mRNA expression in the guard cells of dehydrated leaves. These results indicate that the ABA synthesized in vascular systems is transported to various target tissues and cells, and also that the guard cells themselves are able to synthesize ABA.
1 This work was supported in part by the Ministry of Education, Science, Sports and Culture of Japan (Grant-in-Aid for Scientific Research (B) no. 13490024 to T.K.) and by the Japan Society for the Promotion of Science for Young Scientists (Research Fellowship to H.K.).
2 Present address: National Agricultural Research Center for Hokkaido Region, Hitsujigaoka 1, Toyohira-ku, Sapporo 062–8555, Japan.
Article, publication date, and citation information can be found at www.plantphysiol.org/cgi/doi/10.1104/pp.103.036970.
* Corresponding author; e-mail koshiba-tomokazu{at}c.metro-u.ac.jp; fax 81–426–2565.
Received December 1, 2003;
returned for revision January 29, 2004;
accepted January 30, 2004.
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