Abstract
Gibberellins (GAs) are a class of phytohormones that regulate many aspects of plant growth and development processes including stem elongation, flowering, and seed germination. A novel GA-enhanced gene, designated as OsGAE1, was identified using microarray analysis of GA-regulated genes. OsGAE1 expressed in a dose- and time course-dependent manner with minimum expression at 1 µM GA3 and maximum expression at 50 µM GA3 starting from 30 min and peaked at 24 h after GA3 treatment. OsGAE1 expression was up-regulated by GA3 at transcript level while no significant effect was observed for other hormones. OsGAE1 was expressed in Escherichia coli with N-terminal His6 tag and the recombinant protein migrated at 38 kDa, slightly larger than the predicted 29 kDa, during SDS-PAGE. Anti-OsGAE1 antibodies immunoreacted with a protein of 40 kDa in rice leaf sheath. OsGAE1 expressed mainly in growing leaf sheath and callus compared to leaf and root. In situ hybridization and OsGAE1 promoter analysis revealed that OsGAE1 expressed in shoot apex meristem and young primary leaves. Northern blot, Western blot, and GUS activities revealed that OsGAE1 is up-regulated by GA3. Transgenic rice expressing OsGAE1 in antisense orientation exhibited severely affected vegetative and reproductive growth. The transgenic plants were 55–70% short compared to control. These results suggest that OsGAE1 is differentially expressed in rice␣leaf sheath in relation to GA3 and it encodes a functional protein which is involved in GA-regulated growth and development of rice.
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Abe M, Takahashi T, Komeda Y (1999) Cloning and characterization of an L1 layer-specific gene in Arabidopsis thaliana. Plant Cell Physiol 40:571–580
Abe M, Takahashi T, Komeda Y (2001) Identification of a cis-regulatory element for L1 layer-specific gene expression, which is targeted by an L1-specific homeodomain protein. Plant J 26:487–494
Aharoni A, Vorst O (2002) DNA microarrays for functional plant genomics. Plant Mol Biol 48:99–118
Ashikari M, Wu J, Yano M, Sasaki T, Yoshimura A (1999) Rice gibberellin-insensitive dwarf mutant gene Dwarf 1 encodes the alpha-subunit of GTP-binding protein. Proc Natl Acad Sci USA 96:10284–10289
Bailey GS (1985) In: Walker JM (ed) Methods in molecular biology. Humana Press pp 295–300
Ben-Nissan G, Lee JY, Borohov A, Weiss D (2004) GIP, a Petunia hybrida GA-induced cysteine-rich protein: a possible role in shoot elongation and transition to flowering. Plant J 37:229–238
Cassab GI, Varner JE (1998) Cell wall proteins. Annu Rev Plant Physiol Plant Mol Biol 39:321–353
Chomoczynski P, Sacchi N (1987) Single-step method of RNA isolation by acid guanidium thiocyanate–phenol–chloroform extraction. Anal Biochem 162:156–159
Cosgrove DJ (1999) Enzymes and other agents that enhance cell wall extensibility. Annu Rev Plant Physiol Plant Mol Biol 50:391–417
Cosgrove DJ (2001) Wall structure and wall loosening. A look backwards and forwards. Plant Physiol 125:131–134
Dong C, Thomas S, Becker D, Lorz H, Whitford R, Sutton T, Able JA, Langridge P (2005) WM5: isolation and characterization of a gene expressed during early meiosis and shoot meristem development in wheat. Funct Plant Biol 32:249–258
Hedden P, Proebsting WM (1999) Genetic analysis of gibberellin biosynthesis. Plant Physiol 119:365–370
Higo K, Ugawa Y, Iwamoto M, Korenaga T (1999) Plant cis-acting regulatory DNA elements (PLACE) database. Nucl Acids Res 27:297–300
Hoad GV (1995) Transport of hormones in the phloem of higher plants. Plant Growth Reg 16:173–182
Hood EE, Helmer GL, Fraley RT, Chilton MD (1986) The hypervirulence of Agrobacterium tumefaciens A281 is encoded in a region of pTiBo542 outside of T-DNA. J Bacteriol 168:1291–1301
Hooley R (1994) Gibberellins: perception, transduction and responses. Plant Mol Biol 26:1529–1555
Itoh H, Ueguchi-Tanaka M, Sato Y, Ashikari M, Matsuoka M (2002) The gibberellin signaling pathway is regulated by the appearance and disappearance of SLENDER RICE1 in nuclei. Plant Cell 14:57–70
Itoh H, Tatsumi T, Sakamoto T, Otomo K, Toyomasu T, Kitano H, Ashikari M, Ichihara S, Matsuoka M (2004) A rice semi-dwarf gene, Tan-Ginbozu (D35), encodes the gibberellin biosynthesis enzyme, ent-kaurene oxidase. Plant Mol Biol 54:533–547
Iwasaki Y, Kato T, Kaidoh T, Ishikawa A, Asahi T (1997) Characterization of the putative alpha subunit of a heterotrimeric G Protein in rice. Plant Mol Biol 34:563–572
Jan A, Yang G, Nakamura H, Ichikawa H, Kitano H, Matsuoka M, Matsumoto H, Komatsu S (2004) Characterization of a xyloglucan endotransglucosylase gene that is up-regulated by gibberellin in rice. Plant Physiol 136:3640–3681
Jefferson RA (1987) Assaying chimeric genes in plants: the GUS gene fusion system. Plant Mol Biol Rep 5:387–405
Jones RL, Phillips IDJ (1966) Organs of gibberellin synthesis in light-grown sunflower plants. Plant Physiol 41:1381–1386
Kaneko M, Itoh H, Inukai Y, Sakamoto T, Ueguchi-Tanaka M, Ashikari M, Matsuoka M (2003) Where do gibberellin biosynthesis and gibberellin signaling occur in rice plants? Plant J 35:104–115
Katsumi M, Foard DE, Phinney BO (1983) Evidence for the translocation of gibberellin A3 and gibberellin-like substances in grafts between normal, dwarf1 and dwarf5 seedlings of Zea mays L. Plant Cell Physiol 24:379–388
van der Knaap E, Kim JH, Kende H (2000) A novel gibberellin-induced gene from rice and its potential regulatory role in stem growth. Plant Physiol 122:695–704
Kobayashi M, Yamaguchi I, Murofushi N, Ota Y, Takahashi N (1988) Fluctuation and localization of endogenous gibberellins in rice. Agric Biol Chem 52:1189–1194
Komatsu S, Hirano H (1992) 80 kDa mouse sperm protein as a substrate of protein kinase C. Chem Pharm Bull 40:2780–2782
Komatsu S, Hirano H (1993) Protein kinase activity and protein phosphorylation in rice (Oryza sativa L.) leaf. Plant Sci 94:127–137
Kouchi H, Hata S (1993) Isolation and characterization of novel nodulin cDNAs representing genes expressed at early stages of soybean nodule development. Mol Gen Genet 238:106–119
Lang A (1970) Gibberellins: structure and metabolism. Annu Rev Plant Physiol 21:537–570
Liang CP, Lee YC, Liu YC (1992) Deletion studies to reveal the basis for size discrepancy in proliferating cell nuclear antigen. Electrophoresis 13:346–353
Liszkay A, Kenk B, Schopfer P (2003) Evidence for the involvement of cell wall peroxidase in the generation of hydroxyl radicals mediating extension growth. Planta 217:658–667
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497
Nakagawa M, Shimamoto K, Kyozuka J (2002) Overexpression of RCN1 and RCN2, rice TERMINAL FLOWER 1/CENTRORADIALIS homologs, confers delay of phase transition and altered panicle morphology in rice. Plant J 29:743–750
Ogawa M, Hanada A, Yamauchi Y, Kuwahara A, Kamiya Y, Yamaguchi S (2003) Gibberellin biosynthesis and response during Arabidopsis seed germination. Plant Cell 15:1591–1604
Peng J, Carol P, Richards DE, King KE, Cowling RJ, Murphy GP, Harberd NP (1997) The Arabidopsis GAI gene defines a signaling pathway that negatively regulates gibberellin responses. Genes Dev. 11:3194–3205
Phillips A (1998) Gibberellins in Arabidopsis. Plant Physiol Biochem 36:115–124
Potts WC, Reid JB, Murfet IC (1982) Internode length in Pisum. I. The effect of the Le/le gene difference on endogenous gibberellin-like substances. Physiol Plant 55:323–328
Proebsting WM, Hedden P, Lewis MJ, Croker SJ, Proebsting LN (1992) Gibberellin concentration and transport in genetic lines of pea: effect of grafting. Plant Physiol 100:1354–1360
Shen SH, Sharma A, Komatsu S (2003) Characterization of proteins responsive to gibberellin in the leaf-sheath of rice (O. sativa L.) seedling using proteome analysis. Biol Pharm Bull 26:129–136
Shibaoka H (1994) Plant hormone induced changes in the orientation of cortical microtubules: alterations in the cross linking between microtubules and the plasma membrane. Annu Rev Plant Physiol 45:527–544
Silverstone AL, Ciampaglio CN, Sun T-P (1998) The Arabidopsis RGA gene encodes a transcriptional regulator repressing the gibberellin signal transduction pathway. Plant Cell 10:155–169
Skriver K, Olsen FL, Rogers JC, Mundy J (1991) Cis-acting DNA elements responsive to gibberellin and its antagonist abscisic acid. Proc Natl Acad Sci USA 88:7266–7270
Stader JA, Silhavy TJ (1990) Engineering Escherichia coli to secrete heterologous gene products. Methods Enzymol 185:166–187
Thelen JJ, Muszynski MG, Miernyk JA, Randall DD (1998) Molecular analysis of two pyruvate dehydrogenase kinases from maize. J Biol Chem 27:26618–26623
Toki S (1997) Rapid and efficient Agrobacterium-mediated transformation in rice. Plant Mol Biol Rep 15:16–21
Tusnady GE, Simon I (2001) The HMMTOP transmembrane topology prediction server. Bioinformatics 17:849–850
Ueguchi-Tanaka M, Ashikari M, Nakajima M, Itoh H, Kato E, Kobayashi M, Chow T, Hsing YC, Kitano H, Yamaguchi I, Matsuoka M (2005) GIBBERELLIN-INSENSITIVE DWARF 1 (GID1) encodes a soluble receptor for gibberellin. Nature 437:693–698
Uozu S, Tanaka-Ueguchi M, Kitano H, Hattori K, Matsuoka M (2000) Characterization of XET-related genes of rice. Plant Physiol 122:853–859
Yamauchi Y, Ogawa M, Kuwahara A, Hanada A, Kamiya Y, Yamaguchi S (2004) Activation of gibberellin biosynthesis and response pathways by low temperature during imbibition of Arabidopsis thaliana seeds. Plant Cell 16:367–378
Yang GX, Jan A, Shen SH, Yazaki J, Ishikawa M, Shimatani Z, Kishimoto N, Kikuchi S, Matsumoto H, Komatsu S (2004) Microarray analysis of brassinosteroids-and gibberellin regulated gene expression in rice seedlings. Mol Genet Genomics 271:468–478
Yazaki J, Kishimoto N, Nakamura K, Fujii F, Shimbo K, Otsuka Y, Wu J, Yamamoto K, Sakata K, Sasaki T, Kikuchi S (2000) Embarking on rice functional genomics via cDNA microarray: use of 3′ UTR probes for specific gene expression analysis. DNA Res 7:367–370
Acknowledgments
The authors are grateful to Dr T. Asano for his valuable discussion. We are also thankful to Dr M. Nakamura and Dr H. Ichikawa for their valuable help. Special thanks are extended to Mr H. Takasaki for his help in MS/MS analysis. We are grateful Dr E.E. Hood for providing Agrobacterium strain EHA105. Special thanks are due to Dr Y. Ohashi for letting us use her microscopic facilities. The full-length cDNA was provided by NIAS DNA Bank. This work was supported by the Program of BRAIN and Rice Genome Project from the Ministry of Agriculture, Forestry and Fisheries, Japan.
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Jan, A., Kitano, H., Matsumoto, H. et al. The rice OsGAE1 is a novel gibberellin-regulated gene and involved in rice growth. Plant Mol Biol 62, 439–452 (2006). https://doi.org/10.1007/s11103-006-9030-1
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DOI: https://doi.org/10.1007/s11103-006-9030-1