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First published online November 18, 2005; 10.1104/pp.105.067884

Plant Physiology 139:1881-1889 (2005)
© 2005 American Society of Plant Biologists

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PLANTS INTERACTING WITH OTHER ORGANISMS

Induction of Thioredoxin Is Required for Nodule Development to Reduce Reactive Oxygen Species Levels in Soybean Roots1

Mi-Young Lee2, Ki-Hye Shin2, Yun-Kyoung Kim, Ji-Yeon Suh, Young-Yun Gu, Mi-Ran Kim, Yoon-Sun Hur, Ora Son, Jin-Sun Kim, Eunsook Song, Myeong-Sok Lee, Kyoung Hee Nam, Keum Hee Hwang, Mi-Kyung Sung, Ho-Jung Kim, Jong-Yoon Chun, Miey Park, Tae-In Ahn, Choo Bong Hong, Suk-Ha Lee, Hong Jae Park, Jong-Sug Park, Desh Pal S. Verma and Choong-Ill Cheon*

Department of Biological Science (M.-Y.L., K.-H.S., Y.-K.K., J.-Y.S., Y.-Y.G., M.-R.K., Y.-S.H., O.S., J.-S.K., E.S., M.-S.L., K.H.N., C.-I.C.) and Department of Food and Nutrition (M.-K.S.), Sookmyung Women's University, Seoul 140–742, Korea; Bio-Food and Drug Research Center, Konkuk University, Choongjoo 380–701, Korea (K.H.H.); Seegene, Seoul 138–050, Korea (H.-J.K., J.-Y.C.); School of Biological Sciences (M.P., T.-I.A., C.B.H.) and College of Agriculture and Life Sciences (S.-H.L.), Seoul National University, Seoul 151–742, Korea; National Institute of Agricultural Biotechnology, Rural Development Administration, Suwon 441–707, Korea (H.J.P., J.-S.P.); and Ohio State Biotechnology Center, Ohio State University, Columbus, Ohio 43210 (D.P.S.V.)

Nodules are formed on legume roots as a result of signaling between symbiotic partners and in response to the activities of numerous genes. We cloned fragments of differentially expressed genes in spot-inoculated soybean (Glycine max) roots. Many of the induced clones were similar to known genes related to oxidative stress, such as thioredoxin and {beta}-carotene hydroxylase. The deduced amino acid sequences of full-length soybean cDNAs for thioredoxin and {beta}-carotene hydroxylase were similar to those in other species. In situ RNA hybridization revealed that the thioredoxin gene is expressed on the pericycle of 2-d-old nodules and in the infected cells of mature nodules, suggesting that thioredoxin is involved in nodule development. The thioredoxin promoter was found to contain a sequence resembling an antioxidant responsive element. When a thioredoxin mutant of yeast was transformed with the soybean thioredoxin gene it became hydrogen peroxide tolerant. These observations prompted us to measure reactive oxygen species levels. These were decreased by 3- to 5-fold in 7-d-old and 27-d-old nodules, coincident with increases in the expression of thioredoxin and {beta}-carotene hydroxylase genes. Hydrogen peroxide-producing regions identified with cerium chloride were found in uninoculated roots and 2-d-old nodules, but not in 7-d-old and 27-d-old nodules. RNA interference-mediated repression of the thioredoxin gene severely impaired nodule development. These data indicate that antioxidants such as thioredoxin are essential to lower reactive oxygen species levels during nodule development.


1 This work was supported by the Technology Development Program of the Ministry of Agriculture and Forestry (grant no. 305005–4); the BioGreen 21 Program, Rural Development Administration (grant no. 20050501034831); and the Crop Functional Genomics Center of the 21st Century Frontier Research Program funded by the Ministry of Science and Technology of the Republic of Korea (grant no. CG1542).

2 These authors contributed equally to the paper.

The author responsible for distribution of materials integral to the findings presented in this article in accordance with the policy described in the Instructions for Authors (www.plantphysiol.org) is: Choong-Ill Cheon (ccheon{at}sookmyung.ac.kr).

Article, publication date, and citation information can be found at www.plantphysiol.org/cgi/doi/10.1104/pp.105.067884.

* Corresponding author; ccheon{at}sookmyung.ac.kr; fax 82–2–2077–7322.

Received June 30, 2005; returned for revision September 15, 2005; accepted September 19, 2005.




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