Abscisic acid accumulates in detached, wilted leaves of Xanthium strumarium. When these leaves are subsequently rehydrated, phaseic acid, a catabolite of abscisic acid, accumulates. Analysis by ps chromatography-mass spectrometry of phaseic acid isolated from stressed and subsequently rehydrated leaves placed in an atmosphere containing 20% I802 and 80% N2 indicates that one atom of "0 is incorporated in the 6'-hydroxymethyl group of phaseic acid. This suggests that the enzyme that converts abscisic acid to phaseic acid is an oxygenase.Analysis by gas chromatography-mass spectrometry of abscisic acid isolated from stressed leaves kept in an atmosphere containing "02indicates that one atom of "IO is present in the carboxyl group of abscisic acid. Thus, when abscisic acid accumulates in water-stressed leaves, only one of the four oxygens present in the abscisic acid molecule is derived from molecular oxygen. This suggests that either (a) the oxygen present in the 1'-, 4'-, and one of the two oxygens at the 1-position of abscisic acid arise from water, or (b) there exists a stored precursor with oxygen atoms already present in the 1'-and 4'-positions of abscisic acid which is converted to abscisic acid under conditions of water stress.Little is known about the biosynthetic pathway ofABA, except that as a sesquiterpenoid, ABA is ultimately derived from MVA.2 When radioactive MVA was applied to higher plant tissues, the percentage of incorporation into ABA was always very low, and no intermediates have ever been isolated. Some controversy exists as to whether ABA is synthesized from a C-15 precursor, presumably farnesyl pyrophosphate (the direct pathway), or results from the degradation of a C-40 precursor (the indirect pathway), such as the xanthophyll violaxanthin (6). It is known that the stereochemistry of protons in ABA derived from MVA is identical to that found in carotenoids and it should be noted that the terminal ring structure of certain xanthophylls is similar to ABA (6).Oxygen incorporation into carotenes to form xanthophylls is a late step in carotenoid biosynthesis occurring after ring formation. The oxygen atoms in the hydroxyl groups of lutein and the epoxide groups of antheraxanthin and violaxanthin are derived from molecular oxygen (12,13 bons of the ABA molecule should be a late step in the pathway, the oxygens being derived from molecular oxygen.With respect to ABA catabolism, Gillard and Walton (2) have shown with a crude enzyme preparation from Echinocystis lobata that hydroxylation at the 6'-methyl group to give PA via the unstable intermediate 6'-hydroxymethyl-ABA, is inhibited by CO and anaerobic conditions. They concluded that the enzyme involved is very similar to Cyt P-450 monooxygenases found in animals, but it did not meet all the criteria necessary for calling ABA hydroxylating enzyme a Cyt P450 monooxygenase. These criteria are: (a) inhibition of the reaction in the presence of CO, (b) presence in the reduced enzyme preparation ofa CO-binding pigment with a maximum A at 450 nm...