The concentration of free indoleacetic acid (IAA) is high in cotton (Gossypium hirsutum L.) fruiting forms before anthesis, but is low at and for a few days after anthesis. Amide-linked and ester IAA were measured in fruiting forms at 9, 6, and 3 days before anthesis; at anthesis; and at 2, 4, 7, and 9 days after anthesis to determine if free IAA decreased because it was converted to a conjugated form. Cotton (Gossypium hirsutum L.) fruiting forms rarely abscise during the week before anthesis (12, 22), but are most susceptible to abscission during the week after anthesis (16,22). Abscission rate then decreases with increasing fruit (boll) age until it is essentially zero at 18 d after anthesis (15,16).Because exogenous IAA delays or prevents abscission (2, 7), we measured the concentration of free IAA in fruiting forms from 9 d before until 9 d after anthesis (18). The concentration of free IAA decreased about 80% during the 6 d before anthesis, remained low for the next 4 d, and then increased rapidly between 7 and 9 d after anthesis. Abscission rate increased rapidly after anthesis to a maximum at 5 and 6 d postanthesis. it then decreased as free IAA increased. Because these changes in free IAA were inversely correlated with changes in fruiting-form abscission, we suggested that the high concentration of IAA in large flower buds inhibited abscission before anthesis, whereas the low concentration at and for 4 d after anthesis promoted abscission of young bolls (18).A decrease in the concentration of free IAA could result from decreased synthesis, increased destruction, export, conversion to another form, or a combination of these possible events. We measured the concentrations of ester and amidelinked IAA before, at, and after anthesis to see if the decrease in free IAA was associated with a similar increase in either of the conjugated forms.
MATERIALS AND METHODS
Plant MaterialCotton (Gossypium hirsutum L.), cv Deltapine 61,' was grown in a field at the Western Cotton Research Laboratory in Phoenix as reported earlier (18). Flower buds were harvested 9, 6, and 3 before anthesis; flowers were harvested at anthesis; and bolls were harvested 2, 4, 7, and 9 d after anthesis. All fruiting forms (buds, flowers, and bolls) were harvested the same day to minimize possible differences due to any change in environment. The fruiting forms were rinsed in cold water, quickly frozen at -80°C, and lyophilized. The dry tissue was ground to pass a 40-mesh screen and stored under N2 at -80°C.Analyses of Ester and Amide IAA Samples of 200 or 500 mg each were extracted overnight in 70% acetone that contained antioxidants (19). '4C-IAA was added as an internal standard. The samples plus three 10-mL rinses with 70% acetone were filtered and the volume reduced to about 9 mL by rotary evaporation at 45°C. Ester IAA was hydrolyzed by adding 1 ml of 10 N KOH, mixing, and incubating 1 h at room temperature (3). Amide-linked IAA was hydrolyzed in 7 N NaOH during 3 h at about 100°C (3). All samples were hydrolyzed in a nitrogen a...