A Link between Ethylene and Auxin Uncovered by the Characterization of Two Root-Specific Ethylene-Insensitive Mutants in Arabidopsis

The plant hormone ethylene participates in the regulation of a variety of developmental processes and serves as a key mediator of plant responses to biotic and abiotic stress factors. The diversity of ethylene functions is achieved, at least in part, by combinatorial interactions with other hormonal signals. Here, we show that ethylene-triggered inhibition of root growth, one of the classical effects of ethylene in Arabidopsis thaliana seedlings, is mediated by the action of the WEAK ETHYLENE INSENSITIVE2/ANTHRANILATE SYNTHASE α1 (WEI2/ASA1) and WEI7/ANTHRANILATE SYNTHASE β1 (ASB1) genes that encode α- and β-subunits of a rate-limiting enzyme of Trp biosynthesis, anthranilate synthase. Upregulation of WEI2/ASA1 and WEI7/ASB1 by ethylene results in the accumulation of auxin in the tip of primary root, whereas loss-of-function mutations in these genes prevent the ethylene-mediated auxin increase. Furthermore, wei2 and wei7 suppress the high-auxin phenotypes of superroot1 (sur1) and sur2, two auxin-overproducing mutants, suggesting that the roles of WEI2 and WEI7 in the regulation of auxin biosynthesis are not restricted to the ethylene response. Together, these findings reveal that ASA1 and ASB1 are key elements in the regulation of auxin production and an unexpected node of interaction between ethylene responses and auxin biosynthesis in Arabidopsis. This study provides a mechanistic explanation for the root-specific ethylene insensitivity of wei2 and wei7, illustrating how interactions between hormones can be used to achieve response specificity.

[1]  Ottoline Leyser,et al.  An Auxin-Dependent Distal Organizer of Pattern and Polarity in the Arabidopsis Root , 1999, Cell.

[2]  P. Masson,et al.  Complex physiological and molecular processes underlying root gravitropism , 2002, Plant Molecular Biology.

[3]  Zhenbiao Yang,et al.  Brassinosteroids Interact with Auxin to Promote Lateral Root Development in Arabidopsis1 , 2004, Plant Physiology.

[4]  D. Inzé,et al.  Superroot, a recessive mutation in Arabidopsis, confers auxin overproduction. , 1995, The Plant cell.

[5]  N. Goto,et al.  Auxin and Ethylene Response Interactions during Arabidopsis Root Hair Development Dissected by Auxin Influx Modulators , 2002, Plant Physiology.

[6]  K. Niyogi Molecular and genetic analysis of anthranilate synthase in Arabidopsis thaliana , 1993 .

[7]  M. Estelle,et al.  The F-box protein TIR1 is an auxin receptor , 2005, Nature.

[8]  J. Alonso,et al.  The Ethylene Signaling Pathway , 2004, Science.

[9]  Michael Sauer,et al.  Efflux-dependent auxin gradients establish the apical–basal axis of Arabidopsis , 2003, Nature.

[10]  R. Last,et al.  Tryptophan biosynthesis and metabolism: biochemical and molecular genetics. , 1995, The Plant cell.

[11]  J. Ecker,et al.  Assignment of 30 microsatellite loci to the linkage map of Arabidopsis. , 1994, Genomics.

[12]  P. Masson,et al.  Mutations in Arabidopsis thaliana genes involved in the tryptophan biosynthesis pathway affect root waving on tilted agar surfaces. , 1998, The Plant journal : for cell and molecular biology.

[13]  D. Inzé,et al.  Auxin Transport Promotes Arabidopsis Lateral Root Initiation , 2001, Plant Cell.

[14]  R. J. Pitts,et al.  Auxin and ethylene promote root hair elongation in Arabidopsis. , 1998, The Plant journal : for cell and molecular biology.

[15]  The Arabidopsis Genome Initiative Analysis of the genome sequence of the flowering plant Arabidopsis thaliana , 2000, Nature.

[16]  P. Larsen,et al.  Enhanced ethylene responsiveness in the Arabidopsis eer1 mutant results from a loss-of-function mutation in the protein phosphatase 2A A regulatory subunit, RCN1. , 2003, The Plant journal : for cell and molecular biology.

[17]  M. Bennett,et al.  Auxin transport: the fountain of life in plants? , 2003, Developmental cell.

[18]  G. Wasteneys,et al.  Ethylene Modulates Root-Wave Responses in Arabidopsis1[w] , 2003, Plant Physiology.

[19]  L. Dolan,et al.  Pointing roots in the right direction: the role of auxin transport in response to gravity. , 1998, Genes & development.

[20]  Anna Stepanova,et al.  Convergence of signaling pathways in the control of differential cell growth in Arabidopsis. , 2004, Developmental cell.

[21]  J. Slovin,et al.  Two genetically discrete pathways convert tryptophan to auxin: more redundancy in auxin biosynthesis. , 2003, Trends in plant science.

[22]  K. Ljung,et al.  The SUR2 gene of Arabidopsis thaliana encodes the cytochrome P450 CYP83B1, a modulator of auxin homeostasis. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[23]  Integrative biology : dissecting cross-talk between plant signalling pathways , 2005 .

[24]  Peter McCourt,et al.  GENETIC ANALYSIS OF HORMONE SIGNALING. , 1999, Annual review of plant physiology and plant molecular biology.

[25]  M. Estelle,et al.  Auxin signaling and regulated protein degradation. , 2004, Trends in plant science.

[26]  B. Bartel AUXIN BIOSYNTHESIS. , 1997, Annual review of plant physiology and plant molecular biology.

[27]  A. Theologis,et al.  Unique and Overlapping Expression Patterns among the Arabidopsis 1-Amino-Cyclopropane-1-Carboxylate Synthase Gene Family Members1[w] , 2004, Plant Physiology.

[28]  E. Liscum,et al.  Genetics of Aux/IAA and ARF action in plant growth and development , 2002, Plant Molecular Biology.

[29]  G. Fink,et al.  Two anthranilate synthase genes in Arabidopsis: defense-related regulation of the tryptophan pathway. , 1992, The Plant cell.

[30]  K. Brown Ethylene and abscission , 1997 .

[31]  G. Hagen,et al.  Aux/IAA proteins repress expression of reporter genes containing natural and highly active synthetic auxin response elements. , 1997, The Plant cell.

[32]  N. Goto,et al.  Auxin is a positive regulator for ethylene-mediated response in the growth of Arabidopsis roots. , 2001, Plant & cell physiology.

[33]  E. Pahlich,et al.  A rapid DNA isolation procedure for small quantities of fresh leaf tissue , 1980 .

[34]  G. Jürgens,et al.  Local, Efflux-Dependent Auxin Gradients as a Common Module for Plant Organ Formation , 2003, Cell.

[35]  N. Graham,et al.  Auxin cross-talk: integration of signalling pathways to control plant development , 2004, Plant Molecular Biology.

[36]  J. Chory,et al.  A role for flavin monooxygenase-like enzymes in auxin biosynthesis. , 2001, Science.

[37]  J. Ecker,et al.  Trp-dependent auxin biosynthesis in Arabidopsis: involvement of cytochrome P450s CYP79B2 and CYP79B3. , 2002, Genes & development.

[38]  M. Schmid,et al.  Genome-Wide Insertional Mutagenesis of Arabidopsis thaliana , 2003, Science.

[39]  A. Bleecker,et al.  A Mutation Altering Auxin Homeostasis and Plant Morphology in Arabidopsis. , 1995, The Plant cell.

[40]  L. Hobbie Auxin: Molecular genetic approaches in Arabidopsis , 1998 .

[41]  P. Masson,et al.  ALTERED RESPONSE TO GRAVITY Is a Peripheral Membrane Protein That Modulates Gravity-Induced Cytoplasmic Alkalinization and Lateral Auxin Transport in Plant Statocytes Article, publication date, and citation information can be found at www.plantcell.org/cgi/doi/10.1105/tpc.015560. , 2003, The Plant Cell Online.

[42]  Ottoline Leyser,et al.  The Arabidopsis F-box protein TIR1 is an auxin receptor , 2005, Nature.

[43]  K. Feldmann,et al.  PCR-based identification of T-DNA insertion mutants. , 1998, Methods in molecular biology.

[44]  P. Naur,et al.  Arabidopsis mutants in the C-S lyase of glucosinolate biosynthesis establish a critical role for indole-3-acetaldoxime in auxin homeostasis. , 2004, The Plant journal : for cell and molecular biology.

[45]  O. Leyser Molecular genetics of auxin signaling. , 2002, Annual review of plant biology.

[46]  J. Ecker,et al.  HOOKLESS1, an Ethylene Response Gene, Is Required for Differential Cell Elongation in the Arabidopsis Hypocotyl , 1996, Cell.

[47]  D. Galbraith,et al.  Systematic reverse genetics of transfer-DNA-tagged lines of Arabidopsis. Isolation of mutations in the cytochrome p450 gene superfamily. , 1998, Plant physiology.

[48]  K. Ljung,et al.  Sites and homeostatic control of auxin biosynthesis in Arabidopsis during vegetative growth. , 2002, The Plant journal : for cell and molecular biology.

[49]  O. Leyser,et al.  Hormonal interactions in the control of Arabidopsis hypocotyl elongation. , 2000, Plant physiology.

[50]  Masashi Yamada,et al.  Sites and Regulation of Auxin Biosynthesis in Arabidopsis Roots , 2005, The Plant Cell Online.

[51]  F. Ausubel,et al.  Five components of the ethylene-response pathway identified in a screen for weak ethylene-insensitive mutants in Arabidopsis , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[52]  G. Fink,et al.  Arabidopsis thaliana auxotrophs reveal a tryptophan-independent biosynthetic pathway for indole-3-acetic acid. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[53]  A. Theologis,et al.  ASC4, a Primary Indoleacetic Acid-responsive Gene Encoding 1-Aminocyclopropane-1-carboxylate Synthase in Arabidopsis thaliana , 1995, The Journal of Biological Chemistry.

[54]  E. Liscum,et al.  The NPH4 Locus Encodes the Auxin Response Factor ARF7, a Conditional Regulator of Differential Growth in Aerial Arabidopsis Tissue , 2000, Plant Cell.

[55]  R. Solano,et al.  ETHYLENE RESPONSE FACTOR1 Integrates Signals from Ethylene and Jasmonate Pathways in Plant Defense Online version contains Web-only data. Article, publication date, and citation information can be found at www.plantcell.org/cgi/doi/10.1105/tpc.007468. , 2003, The Plant Cell Online.

[56]  J. Ecker,et al.  Exploiting the triple response of Arabidopsis to identify ethylene-related mutants. , 1990, The Plant cell.

[57]  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.

[58]  R. Last,et al.  Characterization of tryptophan synthase alpha subunit mutants of Arabidopsis thaliana , 1996, Molecular and General Genetics MGG.

[59]  J. Celenza,et al.  Arabidopsis cytochrome P450s that catalyze the first step of tryptophan-dependent indole-3-acetic acid biosynthesis. , 2000, Proceedings of the National Academy of Sciences of the United States of America.