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First published online August 28, 2003; 10.1104/pp.103.024810 Plant Physiology 133:560-570 (2003) © 2003 American Society of Plant Biologists Molecular Cloning and Functional Analysis of a Novel Type of Bowman-Birk Inhibitor Gene Family in Rice1Peking-Yale Joint Center for Plant Molecular Genetics and Agro-Biotechnology, National Laboratory of Protein Engineering and Plant Genetic Engineering, College of Life Sciences, Peking University, Beijing 100871, China (L.-J.Q., J.C., M.L., N.P., H.O., D.L., J.W., G.Z., X.Z., X.C., H.G., Z.C.); China Agriculture University, Beijing 100094, China (Z.C.); Institute of Crop Breeding and Cultivation, Chinese Academy of Agricultural Sciences, Beijing 100094, China (Z.L.); and Institute of Agro-Biotechnology, Yunnan Provincial Academy of Agricultural Sciences, Kunming 650223, China (C.L.)
Bowman-Birk inhibitor (BBI) genes encode serine protease inhibitors that have repetitive cysteine-rich domains with reactive sites for the trypsin or chymotrypsin family. We have identified seven BBI genes from japonica rice (Oryza sativa subsp. japonica var Teqing). All of the genes identified were found in a single cluster on the southern end of the long arm of rice chromosome 1. Four of the seven BBI genes have two repetitive cysteine-rich domains, whereas one has a truncated domain with only one reactive site. We have also identified three novel BBI genes, each of which possesses three repetitive domains instead of two. In situ hybridization analyses indicated that the accumulation of rice BBI transcripts was differentially regulated in germinating embryos and also in the leaves, roots, and flower organs at later developmental stages. Different members of the rice BBI gene family displayed different expression patterns during rice seed germination, and wounding induced the expression of rice BBI transcripts. The three-domain BBIs had higher expression levels than the two-domain BBIs. It was also found that the mRNA of rice BBI genes was present in abundant amounts in scutellar epithelium and aleurone layer cells. RBBI3-1, one of the three-domain RBBI, exhibited in vitro trypsin-inhibiting activity but no chymotrypsin-inhibiting activity. Overexpression of RBBI2-3 in transgenic rice plants resulted in resistance to the fungal pathogen Pyricularia oryzae, indicating that proteinase inhibitors confer resistance against the fungal pathogen in vivo and that they might play a role in the defense system of the rice plant.
Article, publication date, and citation information can be found at www.plantphysiol.org/cgi/doi/10.1104/pp.103.024810.
1 This work was supported by the National High-Tech Program of China (project no. GN 863-01-101-02-02 and 2001AA222261 to L.-J.Q.), by the National Natural Science Foundation (project no. GN 39830020 to H.G.), by the State Key Basic Research and Development Plan (project no. GN G19990
2 These authors contributed equally to the paper.
3 Present address: Department of Plant Sciences, University of Oxford, Oxford OX1 3RB, UK
* Corresponding author; e-mail zlchen{at}pku.edu.cn; fax 86-10-6275-1841.
Received April 5, 2003;
returned for revision April 24, 2003;
accepted June 4, 2003.
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