GAMYB controls different sets of genes and is differentially regulated by microRNA in aleurone cells and anthers.

GAMYB is a component of gibberellin (GA) signaling in cereal aleurone cells, and has an important role in flower development. However, it is unclear how GAMYB function is regulated. We examined the involvement of a microRNA, miR159, in the regulation of GAMYB expression in cereal aleurone cells and flower development. In aleurone cells, no miR159 expression was observed with or without GA treatment, suggesting that miR159 is not involved in the regulation of GAMYB and GAMYB-like genes in this tissue. miR159 was expressed in tissues other than aleurone, and miR159 over-expressors showed similar but more severe phenotypes than the gamyb mutant. GAMYB and GAMYB-like genes are co-expressed with miR159 in anthers, and the mRNA levels for GAMYB and GAMYB-like genes are negatively correlated with miR159 levels during anther development. Thus, OsGAMYB and OsGAMYB-like genes are regulated by miR159 in flowers. A microarray analysis revealed that OsGAMYB and its upstream regulator SLR1 are involved in the regulation of almost all GA-mediated gene expression in rice aleurone cells. Moreover, different sets of genes are regulated by GAMYB in aleurone cells and anthers. GAMYB binds directly to promoter regions of its target genes in anthers as well as aleurone cells. Based on these observations, we suggest that the regulation of GAMYB expression and GAMYB function are different in aleurone cells and flowers in rice.

[1]  Danny J. Llewellyn,et al.  Gibberellin Signaling in Barley Aleurone Cells. Control of SLN1 and GAMYB Expression , 2002, Plant Physiology.

[2]  F. Gubler,et al.  The Role of GAMYB Transcription Factors in GA-Regulated Gene Expression , 2003, Journal of Plant Growth Regulation.

[3]  Javier F. Palatnik,et al.  Specific effects of microRNAs on the plant transcriptome. , 2005, Developmental cell.

[4]  S. Iwahori,et al.  The Conserved KNOX Domain Mediates Specificity of Tobacco KNOTTED1-Type Homeodomain Proteins , 1999, Plant Cell.

[5]  T. Komari,et al.  Efficient transformation of rice (Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA. , 1994, The Plant journal : for cell and molecular biology.

[6]  M. Rothenberg,et al.  Characterization of the rice (Oryza sativa) actin gene family , 1990, Plant Molecular Biology.

[7]  D. Weigel,et al.  GAMYB-like genes, flowering, and gibberellin signaling in Arabidopsis. , 2001, Plant physiology.

[8]  H. Kitano,et al.  Rice plant development: from zygote to spikelet. , 2005, Plant & cell physiology.

[9]  D. Bartel,et al.  Computational identification of plant microRNAs and their targets, including a stress-induced miRNA. , 2004, Molecular cell.

[10]  Masatomo Kobayashi,et al.  Fluctuation and Localization of Endogenous Gibberellins in Rice , 1988 .

[11]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[12]  Huanming Yang,et al.  A Draft Sequence of the Rice Genome (Oryza sativa L. ssp. indica) , 2002, Science.

[13]  M. Matsuoka,et al.  Loss-of-function mutations of the rice GAMYB gene impair alpha-amylase expression in aleurone and flower development. , 2004, The Plant cell.

[14]  D. Choi,et al.  Expansins: ever-expanding numbers and functions. , 2001, Current opinion in plant biology.

[15]  R. Stracke,et al.  The R2R3-MYB gene family in Arabidopsis thaliana. , 2001, Current opinion in plant biology.

[16]  T. Gojobori,et al.  The genome sequence and structure of rice chromosome 1 , 2002, Nature.

[17]  M. Chilton,et al.  The hypervirulence of Agrobacterium tumefaciens A281 is encoded in a region of pTiBo542 outside of T-DNA , 1986, Journal of bacteriology.

[18]  Patrick Achard,et al.  Modulation of floral development by a gibberellin-regulated microRNA , 2004, Development.

[19]  Patrick S Schnable,et al.  Laser-capture microdissection, a tool for the global analysis of gene expression in specific plant cell types: identification of genes expressed differentially in epidermal cells or vascular tissues of maize. , 2003, The Plant cell.

[20]  S. Arai,et al.  Molecular cloning and gibberellin-induced expression of multiple cysteine proteinases of rice seeds (oryzains). , 1991, The Journal of biological chemistry.

[21]  M. Matsuoka,et al.  The Gibberellin Signaling Pathway Is Regulated by the Appearance and Disappearance of SLENDER RICE1 in Nuclei Article, publication date, and citation information can be found at www.plantcell.org/cgi/doi/10.1105/tpc.010319. , 2002, The Plant Cell Online.

[22]  F. Gubler,et al.  The Arabidopsis GAMYB-Like Genes, MYB33 and MYB65, Are MicroRNA-Regulated Genes That Redundantly Facilitate Anther Development , 2005, The Plant Cell Online.

[23]  D. Yamauchi,et al.  Molecular Dissection of the Gibberellin/Abscisic Acid Signaling Pathways by Transiently Expressed RNA Interference in Barley Aleurone Cells Article, publication date, and citation information can be found at www.plantcell.org/cgi/doi/10.1105/tpc.003376. , 2002, The Plant Cell Online.

[24]  Masatomo Kobayashi,et al.  Loss-of-function of a Rice Gibberellin Biosynthetic Gene, GA20 oxidase (GA20ox-2), Led to the Rice ‘Green Revolution’ , 2002 .

[25]  D. Twell,et al.  Novel anther-specific myb genes from tobacco as putative regulators of phenylalanine ammonia-lyase expression. , 2001, Plant physiology.

[26]  K. Sutoh,et al.  Two cis-acting elements necessary and sufficient for gibberellin-upregulated proteinase expression in rice seeds. , 2003, The Plant journal : for cell and molecular biology.

[27]  K. Washio Functional Dissections between GAMYB and Dof Transcription Factors Suggest a Role for Protein-Protein Associations in the Gibberellin-Mediated Expression of the RAmy1A Gene in the Rice Aleurone , 2003, Plant Physiology.

[28]  Masatomo Kobayashi,et al.  GIBBERELLIN INSENSITIVE DWARF1 encodes a soluble receptor for gibberellin , 2005, Nature.

[29]  Yuji Kamiya,et al.  Genome-wide profiling of stored mRNA in Arabidopsis thaliana seed germination: epigenetic and genetic regulation of transcription in seed. , 2005, The Plant journal : for cell and molecular biology.

[30]  J. Casaretto,et al.  Crosstalk Between Gibberellin and Abscisic Acid in Cereal Aleurone , 2003, Journal of Plant Growth Regulation.

[31]  Yujun Zhang,et al.  Sequence and analysis of rice chromosome 4 , 2002, Nature.

[32]  R. Kalla,et al.  A role for HvGAMYB in anther development. , 2003, The Plant journal : for cell and molecular biology.

[33]  F. Gubler,et al.  Gibberellin-responsive elements in the promoter of a barley high-pI alpha-amylase gene. , 1992, The Plant cell.

[34]  M. Rothenberg,et al.  Structural characterization of a rice actin gene , 1990, Plant Molecular Biology.

[35]  Ayuko Kuwahara,et al.  Gibberellin Biosynthesis and Response during Arabidopsis Seed Germination Online version contains Web-only data. Article, publication date, and citation information can be found at www.plantcell.org/cgi/doi/10.1105/tpc.011650. , 2003, The Plant Cell Online.

[36]  M. Matsuoka,et al.  Where do gibberellin biosynthesis and gibberellin signaling occur in rice plants? , 2003, The Plant journal : for cell and molecular biology.

[37]  W. Cleveland Robust Locally Weighted Regression and Smoothing Scatterplots , 1979 .

[38]  J. Kawai,et al.  Collection, Mapping, and Annotation of Over 28,000 cDNA Clones from japonica Rice , 2003, Science.

[39]  Javier F. Palatnik,et al.  Control of leaf morphogenesis by microRNAs , 2003, Nature.

[40]  Takuji Sasaki,et al.  The map-based sequence of the rice genome , 2005, Nature.

[41]  M. Matsuoka,et al.  slender Rice, a Constitutive Gibberellin Response Mutant, Is Caused by a Null Mutation of the SLR1 Gene, an Ortholog of the Height-Regulating Gene GAI/RGA/RHT/D8 , 2001, Plant Cell.

[42]  F. Gubler,et al.  Target genes and regulatory domains of the GAMYB transcriptional activator in cereal aleurone. , 1999, The Plant journal : for cell and molecular biology.

[43]  Ho,et al.  Hormonal regulation of a cysteine proteinase gene, EPB-1, in barley aleurone layers: cis- and trans-acting elements involved in the co-ordinated gene expression regulated by gibberellins and abscisic acid. , 1999, The Plant journal : for cell and molecular biology.

[44]  R. Kalla,et al.  Gibberellin-regulated expression of a myb gene in barley aleurone cells: evidence for Myb transactivation of a high-pI alpha-amylase gene promoter. , 1995, The Plant cell.