GLK transcription factors regulate chloroplast development in a cell-autonomous manner.

In a number of land plants, Golden2-like (GLK) genes encode a pair of partially redundant nuclear transcription factors that are required for the expression of nuclear photosynthetic genes and for chloroplast development. As chloroplast biogenesis depends on close co-operation between the nuclear and plastid compartments, GLK gene function must be dependent on tight intracellular control. However, the extent to which GLK-mediated chloroplast development depends on intercellular communication is not known. Here we used sector analysis to show that GLK proteins operate cell-autonomously in leaf mesophyll cells. To establish whether GLK proteins are able to influence adjacent cell layers, we used tissue-specific promoters to restrict GLK gene expression to the epidermis and to the phloem. GLK genes driven by the Arabidopsis epidermal FIDDLEHEAD (FDH) and MERISTEM LAYER1 (AtML1) promoters failed to rescue the pale-green Atglk1 Atglk2 mutant phenotype, confirming the suggestion that GLK proteins can only influence chloroplast development intracellularly. An exception to this rule was seen in lines in which GLK genes were expressed in the phloem. However, we believe that the partial complementation of the mutant phenotype that was observed resulted from phloem unloading, as opposed to inherent properties of GLK proteins. We conclude that GLK proteins act in a cell-autonomous manner to coordinate and maintain the photosynthetic apparatus within individual cells. Significantly, this suggests that GLK proteins provide a means to fine-tune photosynthesis according to the differential requirements of cells within the leaf.

[1]  A. Gleave A versatile binary vector system with a T-DNA organisational structure conducive to efficient integration of cloned DNA into the plant genome , 1992, Plant Molecular Biology.

[2]  P. Huijser,et al.  Functional conservation and maintenance of expression pattern of FIDDLEHEAD-like genes in Arabidopsis and Antirrhinum , 2004, Plant Molecular Biology.

[3]  J. Langdale,et al.  The Maize Golden2 Gene Defines a Novel Class of Transcriptional Regulators in Plants , 2001, Plant Cell.

[4]  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.

[5]  John R. Evans,et al.  Profiles of light absorption and chlorophyll within spinach leaves from chlorophyll fluorescence , 2002 .

[6]  David Jackson,et al.  Intercellular trafficking of a KNOTTED1 green fluorescent protein fusion in the leaf and shoot meristem of Arabidopsis , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[7]  K. Shimamoto,et al.  Light-Regulated and Cell-Specific Expression of Tomato rbcS-gusA and Rice rbcS-gusA Fusion Genes in Transgenic Rice , 1993, Plant physiology.

[8]  R. Porat,et al.  Identification of a meristem L1 layer-specific gene in Arabidopsis that is expressed during embryonic pattern formation and defines a new class of homeobox genes. , 1996, The Plant cell.

[9]  G. Fink,et al.  Regulated degradation of the transcription factor Gcn4. , 1994, The EMBO journal.

[10]  P. Chomczyński,et al.  The single-step method of RNA isolation by acid guanidinium thiocyanate–phenol–chloroform extraction: twenty-something years on , 2006, Nature Protocols.

[11]  A. Bel,et al.  The phloem, a miracle of ingenuity , 2003 .

[12]  G. Jürgens,et al.  Transcriptional regulation of epidermal cell fate in the Arabidopsis embryo , 2007, Development.

[13]  A. Roberts,et al.  Quantification of Plasmodesmatal Endoplasmic Reticulum Coupling between Sieve Elements and Companion Cells Using Fluorescence Redistribution after Photobleaching1[W] , 2006, Plant Physiology.

[14]  C. Kidner,et al.  bundle sheath defective, a mutation that disrupts cellular differentiation in maize leaves , 1994 .

[15]  S. Rogers,et al.  PEST sequences and regulation by proteolysis. , 1996, Trends in biochemical sciences.

[16]  K. Oparka,et al.  Expression of GFP-fusions in Arabidopsis companion cells reveals non-specific protein trafficking into sieve elements and identifies a novel post-phloem domain in roots. , 2004, The Plant journal : for cell and molecular biology.

[17]  E. Truernit,et al.  Cell-to-Cell and Long-Distance Trafficking of the Green Fluorescent Protein in the Phloem and Symplastic Unloading of the Protein into Sink Tissues , 1999, Plant Cell.

[18]  D. Weigel,et al.  The Arabidopsis thaliana MERISTEM LAYER 1 promoter specifies epidermal expression in meristems and young primordia. , 1999, The Plant journal : for cell and molecular biology.

[19]  D. Winge,et al.  Identification of a copper‐induced intramolecular interaction in the transcription factor Mac1 from Saccharomyces cerevisiae , 1998, The EMBO journal.

[20]  B. Scheres,et al.  Mosaic analyses using marked activation and deletion clones dissect Arabidopsis SCARECROW action in asymmetric cell division. , 2004, Genes & development.

[21]  I. Potrykus,et al.  Expression in plants of two bacterial antibiotic resistance genes after protoplast transformation with a new plant expression vector. , 1986, Nucleic acids research.

[22]  S. A. Barnes,et al.  Alteration of the Amount of the Chloroplast Phosphate Translocator in Transgenic Tobacco Affects the Distribution of Assimilate between Starch and Sugar , 1994, Plant physiology.

[23]  J. Langdale,et al.  GOLDEN 2: A Novel Transcriptional Regulator of Cellular Differentiation in the Maize Leaf , 1998, Plant Cell.

[24]  K. Pyke,et al.  A Genetic Analysis of Chloroplast Division and Expansion in Arabidopsis thaliana , 1994, Plant physiology.

[25]  Teva Vernoux,et al.  An Evolutionarily Conserved Mechanism Delimiting SHR Movement Defines a Single Layer of Endodermis in Plants , 2007, Science.

[26]  Shelley Hepworth,et al.  CONSTANS acts in the phloem to regulate a systemic signal that induces photoperiodic flowering of Arabidopsis , 2004, Development.

[27]  J. Langdale,et al.  GLK gene pairs regulate chloroplast development in diverse plant species. , 2002, The Plant journal : for cell and molecular biology.

[28]  P. Lockhart,et al.  Shopping for plastids. , 2007, Trends in plant science.

[29]  D. Weigel,et al.  Cell-cell signaling and movement by the floral transcription factors LEAFY and APETALA1. , 2000, Science.

[30]  U. Grossniklaus,et al.  A Gateway Cloning Vector Set for High-Throughput Functional Analysis of Genes in Planta[w] , 2003, Plant Physiology.

[31]  Jan-Peter Nap,et al.  pBINPLUS: An improved plant transformation vector based on pBIN19 , 1995, Transgenic Research.

[32]  Y. Komeda,et al.  Identification of a cis-regulatory element for L1 layer-specific gene expression, which is targeted by an L1-specific homeodomain protein. , 2001, The Plant journal : for cell and molecular biology.

[33]  G. Edwards,et al.  Proof of C4 photosynthesis without Kranz anatomy in Bienertia cycloptera (Chenopodiaceae). , 2002, The Plant journal : for cell and molecular biology.

[34]  R. J. Porra,et al.  Determination of accurate extinction coefficients and simultaneous equations for assaying chlorophylls a and b extracted with four different solvents: verification of the concentration of chlorophyll standards by atomic absorption spectroscopy , 1989 .

[35]  Sabine Cornelsen,et al.  Evolutionary analysis of Arabidopsis, cyanobacterial, and chloroplast genomes reveals plastid phylogeny and thousands of cyanobacterial genes in the nucleus , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[36]  Paul Dupree,et al.  Expression of photosynthesis gene‐promoter fusions in leaf epidermal cells of transgenic tobacco plants , 1991 .

[37]  P. Huijser,et al.  Characterization of the FIDDLEHEAD Gene of Arabidopsis Reveals a Link between Adhesion Response and Cell Differentiation in the Epidermis , 1999, Plant Cell.

[38]  J. Langdale,et al.  Cellular pattern of photosynthetic gene expression in developing maize leaves. , 1988, Genes & development.

[39]  J. Chory,et al.  Coordination of gene expression between organellar and nuclear genomes , 2008, Nature Reviews Genetics.

[40]  S. Hake,et al.  Shoot meristem size is dependent on inbred background and presence of the maize homeobox gene, knotted1. , 2000, Development.

[41]  P. Quail,et al.  phyA dominates in transduction of red-light signals to rapidly responding genes at the initiation of Arabidopsis seedling de-etiolation. , 2006, The Plant journal : for cell and molecular biology.

[42]  Min Chen,et al.  Chlorophylls, ligands and assembly of light-harvesting complexes in chloroplasts , 2007, Photosynthesis Research.

[43]  Y. Komeda,et al.  Regulation of shoot epidermal cell differentiation by a pair of homeodomain proteins in Arabidopsis , 2003, Development.

[44]  Elizabeth C Moylan,et al.  A Conserved Transcription Factor Mediates Nuclear Control of Organelle Biogenesis in Anciently Diverged Land Plantsw⃞ , 2005, The Plant Cell Online.

[45]  Fabio Fornara,et al.  FT Protein Movement Contributes to Long-Distance Signaling in Floral Induction of Arabidopsis , 2007, Science.

[46]  K. Oparka,et al.  Sieve Elements and Companion Cells—Traffic Control Centers of the Phloem , 1999, Plant Cell.