Dissecting the Role of CHITINASE-LIKE1 in Nitrate-Dependent Changes in Root Architecture1[C][W]

The root phenotype of an Arabidopsis (Arabidopsis thaliana) mutant of CHITINASE-LIKE1 (CTL1), called arm (for anion-related root morphology), was previously shown to be conditional on growth on high nitrate, chloride, or sucrose. Mutants grown under restrictive conditions displayed inhibition of primary root growth, radial swelling, proliferation of lateral roots, and increased root hair density. We found here that the spatial pattern of CTL1 expression was mainly in the root and root tips during seedling development and that the protein localized to the cell wall. Fourier-transform infrared microspectroscopy of mutant root tissues indicated differences in spectra assigned to linkages in cellulose and pectin. Indeed, root cell wall polymer composition analysis revealed that the arm mutant contained less crystalline cellulose and reduced methylesterification of pectins. We also explored the implication of growth regulators on the phenotype of the mutant response to the nitrate supply. Exogenous abscisic acid application inhibited more drastically primary root growth in the arm mutant but failed to repress lateral branching compared with the wild type. Cytokinin levels were higher in the arm root, but there were no changes in mitotic activity, suggesting that cytokinin is not directly involved in the mutant phenotype. Ethylene production was higher in arm but inversely proportional to the nitrate concentration in the medium. Interestingly, eto2 and eto3 ethylene overproduction mutants mimicked some of the conditional root characteristics of the arm mutant on high nitrate. Our data suggest that ethylene may be involved in the arm mutant phenotype, albeit indirectly, rather than functioning as a primary signal.

[1]  Fusuo Zhang,et al.  Inhibition of maize root growth by high nitrate supply is correlated with reduced IAA levels in roots. , 2008, Journal of plant physiology.

[2]  Antony Bacic,et al.  Plant cell wall biosynthesis: genetic, biochemical and functional genomics approaches to the identification of key genes. , 2006, Plant biotechnology journal.

[3]  D. Bush,et al.  Chitinase-Like Protein CTL1 Plays a Role in Altering Root System Architecture in Response to Multiple Environmental Conditions1[C][W][OA] , 2009, Plant Physiology.

[4]  J. Kieber,et al.  Characterization of the response of the Arabidopsis response regulator gene family to cytokinin. , 2000, Plant physiology.

[5]  M. Hauser,et al.  Histochemical analysis of root meristem activity in Arabidopsis thaliana using a cyclin:GUS (β-glucuronidase) marker line , 2000, Plant and Soil.

[6]  G. Engler,et al.  The Arabidopsis 1-Aminocyclopropane-1-Carboxylate Synthase Gene 1 Is Expressed during Early Development. , 1993, The Plant cell.

[7]  P. Doerner,et al.  Technical advance: spatio-temporal analysis of mitotic activity with a labile cyclin-GUS fusion protein. , 1999, The Plant journal : for cell and molecular biology.

[8]  Jan Petrásek,et al.  Cytokinin regulates root meristem activity via modulation of the polar auxin transport , 2009, Proceedings of the National Academy of Sciences.

[9]  T. Kiba,et al.  Hormonal control of nitrogen acquisition: roles of auxin, abscisic acid, and cytokinin. , 2011, Journal of experimental botany.

[10]  I. Cullis,et al.  Comparison of lignin deposition in three ectopic lignification mutants. , 2005, The New phytologist.

[11]  Brian G Forde,et al.  Nitrogen regulation of root branching. , 2006, Annals of botany.

[12]  G. Mouille,et al.  Homogalacturonan methyl-esterification and plant development. , 2009, Molecular plant.

[13]  J. G. Dubrovsky,et al.  Ethylene-auxin interactions regulate lateral root initiation and emergence in Arabidopsis thaliana. , 2008, The Plant journal : for cell and molecular biology.

[14]  Christian Hermans,et al.  How do plants respond to nutrient shortage by biomass allocation? , 2006, Trends in plant science.

[15]  P. Benfey,et al.  Conditional root expansion mutants of Arabidopsis. , 1995, Development.

[16]  J. Kieber,et al.  Biochemical and functional analysis of CTR1, a protein kinase that negatively regulates ethylene signaling in Arabidopsis. , 2003, The Plant journal : for cell and molecular biology.

[17]  D. Gibbs,et al.  Branching out in new directions: the control of root architecture by lateral root formation. , 2008, The New phytologist.

[18]  Zheng-Hua Ye,et al.  Mutation of a Chitinase-Like Gene Causes Ectopic Deposition of Lignin, Aberrant Cell Shapes, and Overproduction of Ethylene Article, publication date, and citation information can be found at www.plantcell.org/cgi/doi/10.1105/tpc.010278. , 2002, The Plant Cell Online.

[19]  M. Pauly,et al.  OLIgo Mass Profiling (OLIMP) of Extracellular Polysaccharides , 2010, Journal of visualized experiments : JoVE.

[20]  M. Tester,et al.  Evidence that L-glutamate can act as an exogenous signal to modulate root growth and branching in Arabidopsis thaliana. , 2006, Plant & cell physiology.

[21]  B. Forde,et al.  Glutamate in plants: metabolism, regulation, and signalling. , 2007, Journal of experimental botany.

[22]  Pei Sun,et al.  Ethylene is involved in nitrate-dependent root growth and branching in Arabidopsis thaliana. , 2009, The New phytologist.

[23]  J. Malamy,et al.  Environmental regulation of lateral root initiation in Arabidopsis. , 2001, Plant physiology.

[24]  Yan Liang,et al.  Response of root branching to abscisic acid is correlated with nodule formation both in legumes and nonlegumes. , 2005, American journal of botany.

[25]  A. Hills,et al.  EZ-Rhizo: integrated software for the fast and accurate measurement of root system architecture. , 2009, The Plant journal : for cell and molecular biology.

[26]  E. Vierling,et al.  Arabidopsis hot Mutants Define Multiple Functions Required for Acclimation to High Temperatures1 , 2003, Plant Physiology.

[27]  E. Vierling,et al.  Mutants of Arabidopsis thaliana defective in the acquisition of tolerance to high temperature stress. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[28]  L. Hogge,et al.  Abscisic acid. ABA immunoassay and gas chromatography/mass spectrometry verification. , 2000, Methods in molecular biology.

[29]  J. Ecker,et al.  Genetic analysis of ethylene signal transduction in Arabidopsis thaliana: five novel mutant loci integrated into a stress response pathway. , 1995, Genetics.

[30]  B. Usadel,et al.  Rapid Structural Phenotyping of Plant Cell Wall Mutants by Enzymatic Oligosaccharide Fingerprinting1 , 2002, Plant Physiology.

[31]  B. Kloareg,et al.  Evolution and diversity of plant cell walls: from algae to flowering plants. , 2011, Annual review of plant biology.

[32]  S. Fry,et al.  Oligosaccharides as Signals and Substrates in the Plant Cell Wall , 1993, Plant physiology.

[33]  T. Baskin,et al.  Two Leucine-Rich Repeat Receptor Kinases Mediate Signaling, Linking Cell Wall Biosynthesis and ACC Synthase in Arabidopsis[W] , 2008, The Plant Cell Online.

[34]  Joseph R. Ecker,et al.  CTR1, a negative regulator of the ethylene response pathway in arabidopsis, encodes a member of the Raf family of protein kinases , 1993, Cell.

[35]  R. Hill,et al.  Turning on gibberellin and abscisic acid signaling. , 2006, Current opinion in plant biology.

[36]  S. Rothstein,et al.  Understanding plant response to nitrogen limitation for the improvement of crop nitrogen use efficiency. , 2011, Journal of experimental botany.

[37]  B. Forde,et al.  Regulation of Arabidopsis root development by nitrate availability. , 2000, Journal of experimental botany.

[38]  L. Dolan,et al.  Structural and genetic analysis of epidermal cell differentiation in Arabidopsis primary roots. , 1997, Development.

[39]  C. Lamb,et al.  Control of root growth and development by cyclin expression , 1996, Nature.

[40]  Staffan Persson,et al.  Identification of genes required for cellulose synthesis by regression analysis of public microarray data sets. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[41]  Sandra Pelletier,et al.  A Receptor-like Kinase Mediates the Response of Arabidopsis Cells to the Inhibition of Cellulose Synthesis , 2007, Current Biology.

[42]  Ping Wu,et al.  AtCYT-INV1, a neutral invertase, is involved in osmotic stress-induced inhibition on lateral root growth in Arabidopsis , 2007, Plant Molecular Biology.

[43]  P. R. van Weeren,et al.  A microtiter plate assay for the determination of uronic acids. , 1998, Analytical biochemistry.

[44]  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.

[45]  P. Albersheim,et al.  3-deoxy-d-manno-2-octulosonic acid (KDO) is a component of rhamnogalacturonan II, a pectic polysaccharide in the primary cell walls of plants , 1985 .

[46]  H. Goodman,et al.  The Arabidopsis Abscisic Acid Response Locus ABI4 Encodes an APETALA2 Domain Protein , 1998, Plant Cell.

[47]  T. Nühse,et al.  Cell Wall Integrity Controls Root Elongation via a General 1-Aminocyclopropane-1-Carboxylic Acid-Dependent, Ethylene-Independent Pathway1[W] , 2011, Plant Physiology.

[48]  N. Eckardt.  Chitin Signaling in Plants: Insights into the Perception of Fungal Pathogens and Rhizobacterial Symbionts , 2008, The Plant Cell Online.

[49]  Markus Langhans,et al.  Role of cytokinin in the regulation of root gravitropism , 2004, Planta.

[50]  J. Kieber,et al.  Localization of the Raf-like Kinase CTR1 to the Endoplasmic Reticulum of Arabidopsis through Participation in Ethylene Receptor Signaling Complexes* , 2003, Journal of Biological Chemistry.

[51]  H. Leyser,et al.  Nitrate and phosphate availability and distribution have different effects on root system architecture of Arabidopsis. , 2002, The Plant journal : for cell and molecular biology.

[52]  D. Straeten,et al.  Regulation of cell length in the Arabidopsis thaliana root by the ethylene precursor 1-aminocyclopropane- 1-carboxylic acid: a matter of apoplastic reactions. , 2005, The New phytologist.

[53]  Herman Höfte,et al.  Classification and identification of Arabidopsis cell wall mutants using Fourier-Transform InfraRed (FT-IR) microspectroscopy. , 2003, The Plant journal : for cell and molecular biology.

[54]  S. Kauppinen,et al.  A xyloglucan-specific endo-beta-1,4-glucanase from Aspergillus aculeatus: expression cloning in yeast, purification and characterization of the recombinant enzyme. , 1999, Glycobiology.

[55]  Ping Wu,et al.  BREVIS RADIX is involved in cytokinin-mediated inhibition of lateral root initiation in Arabidopsis , 2009, Planta.

[56]  T. Lynch,et al.  Direct targets of the transcription factors ABA-Insensitive(ABI)4 and ABI5 reveal synergistic action by ABI4 and several bZIP ABA response factors , 2011, Plant Molecular Biology.

[57]  J. Ecker,et al.  EIN2, a bifunctional transducer of ethylene and stress responses in Arabidopsis. , 1999, Science.

[58]  H. S. Chae,et al.  The eto1, eto2, and eto3 Mutations and Cytokinin Treatment Increase Ethylene Biosynthesis in Arabidopsis by Increasing the Stability of ACS Protein Article, publication date, and citation information can be found at www.plantcell.org/cgi/doi/10.1105/tpc.006882. , 2003, The Plant Cell Online.

[59]  A. Gojon,et al.  Root uptake regulation: a central process for NPS homeostasis in plants. , 2009, Current opinion in plant biology.

[60]  R. Zhong,et al.  Ectopic deposition of lignin in the pith of stems of two Arabidopsis mutants. , 2000, Plant physiology.

[61]  Renze Heidstra,et al.  Cytokinins Determine Arabidopsis Root-Meristem Size by Controlling Cell Differentiation , 2007, Current Biology.

[62]  Sun-Hee Kim,et al.  Arabidopsis hot2 encodes an endochitinase-like protein that is essential for tolerance to heat, salt and drought stresses. , 2007, The Plant journal : for cell and molecular biology.

[63]  S. Persson,et al.  Cellulose synthases and synthesis in Arabidopsis. , 2011, Molecular plant.

[64]  S. Ring,et al.  Determination of aldoses and uronic acid content of vegetable fiber. , 1979, Analytical biochemistry.

[65]  P. Masson,et al.  Loss-of-Function Mutations of ROOT HAIR DEFECTIVE3 Suppress Root Waving, Skewing, and Epidermal Cell File Rotation in Arabidopsis1 , 2005, Plant Physiology.

[66]  M. Ivanchenko,et al.  Ethylene regulates lateral root formation and auxin transport in Arabidopsis thaliana , 2008, The Plant journal : for cell and molecular biology.

[67]  D. Inzé,et al.  An abscisic acid-sensitive checkpoint in lateral root development of Arabidopsis. , 2003, The Plant journal : for cell and molecular biology.

[68]  Staffan Persson,et al.  Phytohormones and the cell wall in Arabidopsis during seedling growth. , 2010, Trends in plant science.

[69]  B. Forde,et al.  An Arabidopsis MADS box gene that controls nutrient-induced changes in root architecture. , 1998, Science.

[70]  C. Foyer,et al.  ABA plays a central role in mediating the regulatory effects of nitrate on root branching in Arabidopsis. , 2002, The Plant journal : for cell and molecular biology.

[71]  J. Chory,et al.  Suppressors of an Arabidopsis thaliana phyB mutation identify genes that control light signaling and hypocotyl elongation. , 1998, Genetics.

[72]  A. Theologis,et al.  Recessive and dominant mutations in the ethylene biosynthetic gene ACS5 of Arabidopsis confer cytokinin insensitivity and ethylene overproduction, respectively. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[73]  R. O’Malley,et al.  Ethylene Stimulates Nutations That Are Dependent on the ETR1 Receptor1[W][OA] , 2006, Plant Physiology.

[74]  J. Malamy,et al.  Osmotic regulation of root system architecture. , 2005, The Plant journal : for cell and molecular biology.

[75]  R. Fall,et al.  A continuous fluorometric assay for pectin methylesterase. , 1996, Analytical biochemistry.

[76]  V. Gewin.  Food: An underground revolution , 2010, Nature.

[77]  T. Lynch,et al.  The Arabidopsis Abscisic Acid Response Gene ABI5 Encodes a Basic Leucine Zipper Transcription Factor , 2000, Plant Cell.

[78]  T. Kuromori,et al.  AtIPT3 is a key determinant of nitrate-dependent cytokinin biosynthesis in Arabidopsis. , 2004, Plant & cell physiology.

[79]  H. V. Van Onckelen,et al.  Arabidopsis mutants with increased organ regeneration in tissue culture are more competent to respond to hormonal signals , 2001, Planta.

[80]  M. Mutwil,et al.  Laying down the bricks: logistic aspects of cell wall biosynthesis. , 2008, Current opinion in plant biology.

[81]  P W Barlow,et al.  Dual pathways for regulation of root branching by nitrate. , 1999, Proceedings of the National Academy of Sciences of the United States of America.