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Assay for and enzymatic formation of an ethylene precursor, 1-aminocyclopropane-1-carboxylic acid

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Abstract

A simple and sensitive chemical assay was developed for 1-aminocyclopropane-1-carboxylic acid (ACC), a precursor of ethylene. The assay is based on the liberation of ethylene from ACC at pH 11.5 in the presence of pyridoxal phosphate, MnCl2 and H2O2. This assay was used to detect ACC in extracts of tomato fruits (Lycopersicon esculentum Mill.) and to measure the activity of a soluble enzyme from tomato fruit that converted S-adenosylmethionine (SAM) to ACC. The enzyme had a Km of 13 μM for SAM, and conversion of SAM to ACC was competitively and reversibly inhibited by aminoethoxyvinylglycine (AVG), an analog of rhizobitoxine. The Ki value for AVG was 0.2 μM. The level of the ACC-forming enzyme activity was positively correlated with the content of ACC and the rate of ethylene formation in wild-type tomatoes of different developmental stages. Mature fruits of the rin (non-ripening) mutant of tomato, which only produce low levels of ethylene, contained much lower levels of ACC and of the ACC-forming enzyme activity than wild-type tomato fruits of comparable age.

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Abbreviations

ACC:

1-aminocyclopropane-1-carboxylic acid

AVG:

ammoethoxyvinylglycine, the aminoethoxy analog of rhizobitoxine L-2-amino-4-(2′-aminoethoxy)-trans-3-butenoic acid

SAM:

S-adenosyl-L-methionine

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Author notes

  1. Thomas Boller

    Present address: Botanisches Institut der Universität Basel, Schönbeinstr 6, CH-4056, Basel, Switzerland

Authors and Affiliations

  1. MSU-DOE Plant Research Laboratory, Michigan State University, 48824, East Lansing, MI, USA

    Thomas Boller & Hans Kende

  2. Department of Horticulture, Michigan State University, 48824, East Lansing, MI, USA

    Robert C. Herner

Authors
  1. Thomas Boller
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  2. Robert C. Herner
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  3. Hans Kende
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Michigan Agricultural Experiment Station No. 8876

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Boller, T., Herner, R.C. & Kende, H. Assay for and enzymatic formation of an ethylene precursor, 1-aminocyclopropane-1-carboxylic acid. Planta 145, 293–303 (1979). https://doi.org/10.1007/BF00454455

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  • DOI: https://doi.org/10.1007/BF00454455

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