Endogenous Biosynthetic Precursors of (+)-Abscisic Acid. V. Inhibition by Tungstate and its Removal by Cinchonine shows that Xanthoxal is Oxidised by a Molybdo-Aldehyde Oxidase

1997 ◽  
Vol 24 (6) ◽  
pp. 727 ◽  
Author(s):  
H-S. Lee ◽  
B. V. Milborrow

A cell-free preparation from avocado fruit incorporates [14C]mevalonate into ABA. A number of specific inhibitors have been used to probe the system and tungstate ions at 100 µM reduce the 14C in ABA by 80% The inhibitory effect was overcome by the alkaloid cinchonine (2000 µM) which binds tungstate strongly and selectively. More 14C from mevalonate was present in xanthoxal (4600 dpm), less in ABA (340 dpm) when the cell-free system was inhibited by tungstate (100 µM) than in controls (1810 dpm in xanthoxal, 1200 dpm in ABA), which shows that xanthoxal is the substrate for the aldehyde oxidase. Xanthoxic acid, therefore, is the next intermediate and AB-aldehyde is not a normal precursor. The potential for using the tungstate/cinchonine reaction to probe other biosynthetic pathways which require a molybdate ion is discussed.

1998 ◽  
Vol 25 (5) ◽  
pp. 507 ◽  
Author(s):  
B.V. Milborrow ◽  
H.-S. Lee

A cell-free system from avocado fruit which routinely incorporated [14C]mevalonate into ABA (1000 dpm per 5 mL of preparation), and into carotenoids, has now been shown to incorporate [14C]pyruvate even more successfully (1620 dpm). Intact chloroplasts from spinach leaf protoplasts incorporated 2990 dpm of [14C]pyruvate (from 2 x 106 dpm) into ABA compared with 990 dpm from [3-R-5-14C]mevalonate (also from 2 x 106 dpm). The intact chloroplasts also produced [14C]ABA (1575 dpm) when supplied with [14C]isopentenyl diphosphate. This result establishes that the whole pathway of biosynthesis of ABA can occur within chloroplasts. Little [14C]acetate or [14C]alanine was incorporated into ABA by avocado fruit mesocarp. Most of the ABA in leaf tissue now appears to be formed by the triose-pyruvate pathway in chloroplasts and incorporation of [14C]mevalonate occurs after activation in the cytoplasm and importation of a later intermediate into the plastids.


1994 ◽  
Vol 21 (3) ◽  
pp. 327 ◽  
Author(s):  
RD Willows ◽  
AG Netting ◽  
BV Milborrow

RS-[5-18O]mevalonolactone has been synthesised and fed as the free acid via the transpiration stream or through the roots to tomato seedlings, to avocado fruit just prior to the climacteric, to cultures of the hyphomycete, Cercospora rosicola, to excised barley embryos and to an excised barley embryo cell-free system. Small amounts of 18O from [5-18O] mevalonolactone were detected in the abscisic acid from tomato plants and from the barley cell-free system but the mechanism involved is unclear. No 18O was detected in abscisic acid from the other tissues. Mass spectrometry of the pentafluorobenzyl derivative of abscisic acid extracted from tomato plants that had been waterlogged in 2H2O (55 atom %) for 8 or 9 days showed that 40-47% was unlabelled or contained just one 2H atom. The remainder was seen as an envelope of peaks containing from two to 17 2H atoms. When plants waterlogged in 2H2O were subsequently wilted in an atmosphere, 80-90% of the multiply deuteriated abscisic acid was also labelled with 18O while only 43% of the mono/undeuteriated abscisic acid was so labelled. By saponification and re-analysis it was shown that most of the 18O, in the multiply deuteriated category of abscisic acid, was present in the carboxyl group. In the mono/undeuteriated abscisic acid a maximum of 50% was labelled with an 18O atom in the carboxyl group. These experiments led to the conclusion that there were two precursor pools involved in abscisic acid biosynthesis and that neither of these pools consisted of carotenoids.


1974 ◽  
Vol 13 (1) ◽  
pp. 131-136 ◽  
Author(s):  
Barry V. Milborrow

1982 ◽  
Vol 23 (6) ◽  
pp. 803-810
Author(s):  
S Hata ◽  
T Nishino ◽  
N Ariga ◽  
H Katsuki

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