The discovery and development of compounds counteracting ethylene at the receptor level.

The discovery and development of 1-methylcyclopropene (1-MCP) was not done as a single step but rather was found by studying various aspects of ethylene action and the ethylene receptors. It was first noted that 2, 5-norbornadiene seemed to counteract ethylene. Studies showed it was a competitive inhibitor of ethylene responses, and knowledge that ethylene antagonists like ethylene agonists bound to silver in the same order as they were active as inhibitors was obtained. Ring strain appeared to be a primary factor that led to trans-cyclooctene then to diazocyclopentadiene. This same concept allowed for the use of chemical concepts that lead to cyclopropenes. More recent work indicates additional factors can come into play in the development of ethylene antagonists at the receptor level and these are now being utilized to find additional and improved antagonists. 1-MCP is likely to remain a primary means of controlling ethylene responses for the immediate future.

Characterization of an ethylene-induced esterase gene isolated from Citrus sinensis by competitive hybridization.

A simple new method, competitive hybridization, for identification of differentially regulated genes was used to isolate novel genes induced by ethylene in citrus (Citrus sinensis [L.] Osbeck cv. Shamouti) leaves. One of the isolated genes, an ethylene-induced esterase gene (EIE), was further characterized. The deduced protein sequence of this gene shows a similarity to those of several plant alpha/beta hydrolase gene family members, which are known to be involved in secondary metabolism. Northern blot analysis demonstrated that EIE mRNA was induced by ethylene within 4 h and accumulated to a very high level 24 h after the initiation of ethylene treatment. Induction of EIE by ethylene could be counteracted by 1-methylcyclopropene, a potent ethylene perception inhibitor, indicating that the expression of EIE is ethylene-dependent. The bacterially expressed protein of EIE was recognized by antiserum against Pir7b, a naphthol AS esterase induced in rice by the non-host pathogen, Pseudomonas syringae pv. syringae. The EIE protein was identified in ethylene-treated leaves using anti-Pir7b antibodies. An alpha-naphthyl acetate esterase accumulated concomitantly with the increase in EIE protein in ethylene-treated citrus leaves. An enzyme activity assay followed by western analysis confirmed that the esterase was EIE.