Regulation of plant development by ethylene Ethylene biosynthesis Expression analysis of ACS Transcriptional regulation of ACS expression Auxin Wounding -IAA +IAA Mutant screens for ethylene pathway genes Wt mutants airC2H4 air C2H4 * Screen for ethylene mutants I. Genes involved in regulation of biosynthesis Roman et al., 1994 * phenotype rescued by inhibitor of ethylene biosynthesis eto mutants – constitutive triple response WT eto2 eto2 eto3 -5 0 5 10 15 20 25 30 35 40 WT eto2-1 ethyleneproductionpl.mg-1 .h-1 - overproduction of ethylene eto2,eto3 dominant mutation results from single amino acid change in the C terminus ACS5,ACS9 eto2 mutation does not affect specific activity of ACS5 eto3 mutation does not affect level of ACS9 mRNA Effect of eto2 mutation on ACS5 protein stability Posttranscritpional regulation of ACS Eto1- recesive mutant with constitutive triple response ETO1 interacts with ACS5 and regulates its activity ETO1 promotes ACS5 degradation through proteasome dependent pathway Model for regulation of ethylene biosynthesis by ETO1 * * Screen for ethylene mutants II. Genes invoved in signalling pathway * * * Roman et al., 1994 * * * * * Roman et al., 1994 Roman et al., 1994 Roman et al., 1994 Roman et al., 1994 ETR1 codes for histidine kinase etr1 ein4 ers2 etr2 Bleecker et al.,1988 Hua et al., 1998 Sakai et al., 1998 His-kinases in Arabidopsis Ethylene signalling – homology to two component system ? Ctr1 – codes for protein kinase of Raf family ETR1 interacts with CTR1 Clark et al., 1998 CTR1 ctr1-1 CTR1 has protein kinase activity CTR1 ctr1-1 -ethylene receptor dominant mutation > ethylene insensitivity -ethylene receptor loss of function mutation > constitutive ethylene response -ctr1 loss of function mutation > constitutive ethylene response EIN2 - contains domain similar to Nramp metal transporters protein. EIN3 codes for transcription factor Chao et al., 1997 GUS::EIN3 GUS::Nla GUS EIN3 protein level is controlled by ethylene EIN3 is rapidly degraded by a proteasome-mediated pathway Guo and Ecker, 2003 EIN3 accumulates in nucleus upon ethylene or MG132 treatment ACC MG132 EIN3 interacts with F-box proteins Guo and Ecker, 2003 Mutation in EBF1 and EBF2 results in hypersensitivity to ethylene and accumulation of EIN3 protein Guo and Ecker, 2003 Overexpression of EBF1 and EBF2 results in ethylene insensitivity and reduced accumulation of EIN3 protein Guo and Ecker, 2003 MODELS Guo and Ecker, 2003 Ethylene signal transduction pathway Ethylene Modulates Stem Cell Division in roots Olga Ortega-Martínez et al., 2007 Ethylene promotes QC cell division. QC cell identity and function are maintained in eto1 mutants Ethylene stimulates formation of additional columella cell layers