Inhibitors of ethylene synthesis inhibit auxin-induced stomatal opening in epidermis detached from leaves of Vicia faba L.
Using leaf epidermis from Vicia faba, we tested whether auxin-induced stomatal opening was initiated by auxin-induced ethylene synthesis. Epidermis was dark-incubated in buffered KNO3 containing 0.1 mM alpha-napthalene acetic acid or 1 mM indole-3-acetic acid. Maximum net opening was ca. 4 micron after 6 h. Opening was reversed by 20 microM ABA, 0.1 mM CaCl2. 1-Aminocyclopropane carboxylic acid (ACC) synthase catalyzes synthesis of ACC, the immediate precursor to ethylene. Auxin-induced stomatal opening was fully inhibited by 10 microM 1-aminoethoxyvinylglycine (AVG), an ACC synthase inhibitor. In solutions containing AVG, auxin-induced opening was restored in a concentration-dependent manner by exogenous ACC, but not in control solutions lacking an auxin. ACC-mediated reversal of AVG-inhibition of stomatal opening was inhibited by alpha-aminoisobutyric acid (AIB), an inhibitor of ACC oxidase, the last enzyme in the ethylene biosynthetic pathway, by 10 microM silver thiosulfate (STS), an inhibitor of ethylene action, and by 20 microM ABA, 0.1 mM CaCl2. CoCl2, an inhibitor of ethylene synthesis, also inhibited auxin-induced opening. Both STS and CoCl2 inhibited opening induced by light or by fusicoccin, but neither light- nor fusicoccin-induced opening was inhibited by AVG. These results support the hypothesis that auxin-induced stomatal opening is mediated through auxin-induced ethylene production by guard cells.
Natural Science Division, Pepperdine University, 24255 Pacific Coast Highway, Malibu, CA 90265, USA.
F
F
Merritt
A
A
Kemper
G
G
Tallman
D000594:Amino Acid Oxidoreductases / Q000037:antagonists & inhibitors*; D000598:Amino Acids, Cyclic / Q000378:metabolism; D000621:Aminoisobutyric Acids / Q000494:pharmacology; D003035:Cobalt / Q000494:pharmacology; D005030:Ethylenes / Q000378:metabolism* / Q000494:pharmacology; D005998:Glycine / Q000031:analogs & derivatives / Q000494:pharmacology; D007210:Indoleacetic Acids / Q000378:metabolism* / Q000494:pharmacology; D008190:Lyases / Q000037:antagonists & inhibitors*; D018515:Plant Leaves / Q000187:drug effects / Q000378:metabolism; D019685:Rosales / Q000187:drug effects / Q000378:metabolism*; D013885:Thiosulfates / Q000494:pharmacology
D000598:Amino Acids, Cyclic; D000621:Aminoisobutyric Acids; D005030:Ethylenes; D007210:Indoleacetic Acids; D013885:Thiosulfates; C100049:2-aminoisobutyric acid; C095213:silver thiosulfate; D003035:Cobalt; C023863:1-aminocyclopropane-1-carboxylic acid; C030737:indoleacetic acid; C036216:ethylene; D000594:Amino Acid Oxidoreductases; C059460:1-aminocyclopropane-1-carboxylic acid oxidase; D008190:Lyases; C021773:1-aminocyclopropanecarboxylate synthase; C018021:cobaltous chloride; C088137:aminoethoxyvinylglycine; D005998:Glycine