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Extracts for ACC synthase (ACS) activity assays were prepared according to the method of Boller et al. (1979), and determined by the method of Lizada and Yang (1979) with slight modifications. One g tomato pericarp tissue was homogenized using a cold pestle and mortar in 5mL of extraction buffer which consisting of 4 mM dithiothreitol, 10mM pyridoxal phosphate and 1.0mM EDTA in 100mM pH 8.0 Hepes buffer. After centrifugation of the homogenate at 10,000¡Ág for 20min, the supernatant was used for enzyme assays. All steps in the enzyme extraction were performed at 4◦C. ACC synthase activity was assayed in a 0.8mL reaction mixture containing 0.25mM S-adenosylmethionine, 50mM Hepes buffer and 10mM pyridoxal phosphate with 0.2mL of enzyme extract in test tubes. After incubation of the reaction mixture at 30 ◦C for 1 h, the tubes were sealed with a stopper and rubber septum, 0.1mL HgCl2 (80mM) and 0.1mL 5% NaOH¨CNaClO solution (1:2, v/v) were injected into the tubes, shaken and incubated for 2min. One mL of the gaseous portion was removed and assayed for ethylene as described above. ACS activity was expressed as nmol g−1 FWh−1.
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Extracts for ACC synthase (ACS) activity assays were prepared according to the method of Boller et al. (1979), and determined by the method of Lizada and Yang (1979) with slight modifications. One g tomato pericarp tissue was homogenized using a cold pestle and mortar in 5mL of extraction buffer which consisting of 4 mM dithiothreitol, 10mM pyridoxal phosphate and 1.0mM EDTA in 100mM pH 8.0 Hepes buffer

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After centrifugation of the homogenate at 10,000¡Ág for 20min, the supernatant was used for enzyme assays. All steps in the enzyme extraction were performed at 4◦C. ACC synthase activity was assayed in a 0.8mL reaction mixture containing 0.25mM S-adenosylmethionine, 50mM Hepes buffer and 10mM pyridoxal phosphate with 0.2mL of enzyme extract in test tubes.

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