ABSCISIC ALCOHOL IS AN INTERMEDIATE IN ABSCISIC-ACID BIOSYNTHESIS IN A SHUNT PATHWAY FROM ABSCISIC ALDEHYDE

It has previously been shown that the abscisic acid (ABA)-deficient flacca and sitiens mutants of tomato are impaired in ABA-aldehyde oxidation and accumulate trans-ABA-alcohol as a result of the biosynthetic block (IB Taylor, RST Linforth, RJ Al-Naieb, WR Bowman, BA Marples [1988] Plant Cell Environ 11:739-745). Here we report that the flacca and sitiens mutants accumulate trans-ABA and trans-ABA glucose ester and that this accumulation is due to trans-ABA biosynthesis. O-18 labeling of water-stressed wild-type and mutant tomato leaves and analysis of [O-18]ABA by tandem mass spectrometry show that the tomato mutants synthesize a significant percentage of their ABA and trans-ABA as [O-18]ABA with two O-18 atoms in the carboxyl group. We further show, by feeding experiments with [H-2(6)]ABA-alcohol and O-18(2), that this doubly-carboxyl-labeled ABA is synthesized from [O-18]ABA-alcohol with incorporation of molecular oxygen. In vivo inhibition of [H-2(6)]ABA-alcohol oxidation by carbon monoxide establishes the involvement of a P-450 monooxygenase. Likewise, carbon monoxide inhibits the synthesis of doubly-carboxyl-labeled ABA in O-18-labeling experiments. This minor shunt pathway from ABA-aldehyde to ABA-alcohol to ABA operates in all plants examined. For the ABA-deficient mutants impaired in ABA-aldehyde oxidation, this shunt pathway is an important source of ABA and is physiologically significant.