IMB1, a bromodomain protein induced during seed imbibition, regulates ABA- and phyA-mediated responses of germination in Arabidopsis.

We report the characterization of a plant gene encoding a member of the BET subgroup of bromodomain proteins, a novel class of putative transcription factors. Imbibition-inducible 1 (IMB1) appears to be a nuclear protein as suggested by subcellular localization in onion epidermal cells using an IMB1-yellow fluorescent protein (YFP) fusion protein. In Arabidopsis thaliana, IMB1 is expressed at very low levels in dry seeds, but is markedly induced during seed imbibition. In addition, IMB1 transcript levels are down regulated during germination. Seeds of a loss-of-function mutant allele, imb1, show impaired cotyledon greening during germination in abscisic acid (ABA) and express higher levels of ABI5 protein than the wild type. Moreover, imb1 seeds are deficient in the phytochrome A (phyA)-mediated very-low-fluence response of germination. Microarray analysis revealed that genes included in different functional categories, such as cell-wall metabolism or plastid function, are repressed in imbibed imb1 seeds. Mutant imb1 plants appear normal, indicating that IMB1 is involved in regulating a specific developmental stage. Taken together, these results show that IMB1 plays a role in the promotion of seed germination by both negatively and positively regulating the ABA and phyA transduction pathways, respectively. In imbibed seeds, IMB1 modulates the transcription of a battery of genes, providing clues on its mode of action.

[1]  J. Botto,et al.  Modes of action of phytochromes , 1998 .

[2]  Jinrong Peng,et al.  Gibberellin regulates Arabidopsis seed germination via RGL2, a GAI/RGA-like gene whose expression is up-regulated following imbibition. , 2002, Genes & development.

[3]  Jonathan D. G. Jones,et al.  Patterns of gene action in plant development revealed by enhancer trap and gene trap transposable elements. , 1995, Genes & development.

[4]  N. Chua,et al.  A GFP-mouse talin fusion protein labels plant actin filaments in vivo and visualizes the actin cytoskeleton in growing pollen tubes. , 1998, The Plant journal : for cell and molecular biology.

[5]  J. Casal,et al.  Phytochromes and seed germination , 1998, Seed Science Research.

[6]  R. Sharrock,et al.  Patterns of Expression and Normalized Levels of the Five Arabidopsis Phytochromes1 , 2002, Plant Physiology.

[7]  J. Casal Review article. Modes of action of phytochromes , 1998 .

[8]  I B Dawid,et al.  The Drosophila fsh locus, a maternal effect homeotic gene, encodes apparent membrane proteins. , 1989, Developmental biology.

[9]  J Chory,et al.  From seed germination to flowering, light controls plant development via the pigment phytochrome. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[10]  J. Botto,et al.  Phytochrome A Mediates the Promotion of Seed Germination by Very Low Fluences of Light and Canopy Shade Light in Arabidopsis , 1996, Plant physiology.

[11]  K. Bradford,et al.  A gibberellin-regulated xyloglucan endotransglycosylase gene is expressed in the endosperm cap during tomato seed germination. , 2002, Journal of experimental botany.

[12]  L. Hennig,et al.  Phytochrome E controls light-induced germination of Arabidopsis. , 2002, Plant physiology.

[13]  C. Allis,et al.  Translating the Histone Code , 2001, Science.

[14]  T. Shinomura,et al.  Action spectra for phytochrome A- and B-specific photoinduction of seed germination in Arabidopsis thaliana. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[15]  Fred Winston,et al.  The bromodomain: a chromatin-targeting module? , 1999, Nature Structural Biology.

[16]  L. Lopez-Molina,et al.  A postgermination developmental arrest checkpoint is mediated by abscisic acid and requires the ABI5 transcription factor in Arabidopsis , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[17]  B. Séraphin,et al.  The yeast BDF1 gene encodes a transcription factor involved in the expression of a broad class of genes including snRNAs. , 1994, Nucleic acids research.

[18]  Nam-Hai Chua,et al.  ABI5 acts downstream of ABI3 to execute an ABA-dependent growth arrest during germination. , 2002, The Plant journal : for cell and molecular biology.

[19]  R. Tsien,et al.  Fluorescent indicators for Ca2+based on green fluorescent proteins and calmodulin , 1997, Nature.

[20]  R. Reiter,et al.  Genetic linkage of the Arabidopsis genome: methods for mapping with recombinant inbreds and random amplified polymorphic DNAs (RAPDs). , 1992 .

[21]  Daniel J. Cosgrove,et al.  Loosening of plant cell walls by expansins , 2000, Nature.

[22]  V. Sundaresan,et al.  Analysis of Flanking Sequences from Dissociation Insertion Lines: A Database for Reverse Genetics in Arabidopsis , 1999, Plant Cell.

[23]  S. Mayfield,et al.  Interorganellar crosstalk: new perspectives on signaling from the chloroplast to the nucleus , 2001, Genome Biology.

[24]  S. Buratowski,et al.  Bromodomain factor 1 corresponds to a missing piece of yeast TFIID. , 2000, Genes & development.

[25]  D. Faller,et al.  RING3 kinase transactivates promoters of cell cycle regulatory genes through E2F. , 2000, Cell growth & differentiation : the molecular biology journal of the American Association for Cancer Research.

[26]  Dawid Ib,et al.  The maternal-effect gene fsh is essential for the specification of the central region of the Drosophila embryo. , 1990 .

[27]  J. Lippincott-Schwartz,et al.  A Bromodomain Protein, MCAP, Associates with Mitotic Chromosomes and Affects G2-to-M Transition , 2000, Molecular and Cellular Biology.

[28]  D. Faller,et al.  You bet-cha: a novel family of transcriptional regulators. , 2001, Frontiers in bioscience : a journal and virtual library.

[29]  M. Delseny,et al.  Isolation of total RNA from Arabidopsis thaliana seeds. , 1999, Analytical biochemistry.

[30]  F. Chen,et al.  Expression of an expansin is associated with endosperm weakening during tomato seed germination. , 2000, Plant physiology.

[31]  T. Hall,et al.  Plant chromatin: development and gene control. , 2002, BioEssays : news and reviews in molecular, cellular and developmental biology.

[32]  S. Clough,et al.  Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. , 1998, The Plant journal : for cell and molecular biology.