Abstract
Ethylene is a gaseous hormone important for adaptation and survival in plants. To further understand the signaling and regulatory network of ethylene, we used a phenotype-based screening strategy to identify chemical compounds interfering with the ethylene response in Arabidopsis thaliana. By screening a collection of 10,000 structurally diverse small molecules, we identified compounds suppressing the constitutive triple response phenotype in the ethylene overproducer mutant eto1-4. The compounds reduced the expression of a reporter gene responsive to ethylene and the otherwise elevated level of ethylene in eto1-4. Structure and function analysis revealed that the compounds contained a quinazolinone backbone. Further studies with genetic mutants and transgenic plants involved in the ethylene pathway showed that the compounds inhibited ethylene biosynthesis at the step of converting S-adenosylmethionine to 1-aminocyclopropane-1-carboxylic acid (ACC) by ACC synthase. Biochemical studies with in vitro activity assay and enzyme kinetics analysis indicated that a representative compound was an uncompetitive inhibitor of ACC synthase. Finally, global gene expression profiling uncovered a significant number of genes that were co-regulated by the compounds and aminoethoxyvinylglycine, a potent inhibitor of ACC synthase. The use of chemical screening is feasible in identifying small molecules modulating the ethylene response in Arabidopsis seedlings. The discovery of such chemical compounds will be useful in ethylene research and can offer potentially useful agrochemicals for quality improvement in post-harvest agriculture.
Highlights
Hook, radial swelling of the hypocotyl, and shortening of the hypocotyl and root [5]
The ethylene signaling pathway has been elucidated by mainly studying genetic mutants in Arabidopsis, additional factors regulating the key components have been revealed by new approaches [18, 19, 23], which suggest the use of a new methodology to study ethylene function
All of the enzymes involved in ethylene biosynthesis, including AdoMet synthetase, ACC synthase, 2 The abbreviations used are: ER, endoplasmic reticulum; ACC, 1-aminocyclopropane-1-carboxylic acid; ACS, ACC synthase; AVG, aminoethoxyvinylglycine; CTR1, constitutive triple response 1; EBS, EIN3-binding sequence; ETO, ethylene overproducer; PLP, pyridoxal 5Ј-phosphate; AdoMet, Sadenosylmethionine; STS, silver thiosulfate; ANOVA, analysis of variance
Summary
Hook, radial swelling of the hypocotyl, and shortening of the hypocotyl and root [5]. Our results demonstrate that the phenotype-based strategy for chemical screening successfully identified small molecules suppressing the ethylene response, most likely by inhibiting ethylene biosynthesis in Arabidopsis.
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