Regulation of alternative oxidase activity during phosphate deficiency in bean roots (Phaseolus vulgaris).
Cyanide-resistant respiration was studied in mitochondria isolated from the roots of bean plants (Phaseolus vulgaris L. cv. Złota Saxa) grown hydroponically up to 16 days on a phosphate-sufficient (+P, control) or phosphate-deficient (-P) medium. Western blotting indicated that the alternative oxidase (AOX) was present only in its reduced (active) form, both in phosphate-sufficient and phosphate-deficient roots, but in the latter, the amount of AOX protein was greater. Addition of pyruvate to the isolation, washing and reaction media made mitochondria from +P roots cyanide-insensitive, similar to mitochondria from -P roots. The doubled activity of NAD-malic enzyme (NAD-ME) in -P compared with +P root mitochondria may suggest increased pyruvate production in -P mitochondria. Lower cytochrome c oxidase (COX) activity and no uncoupler effect on respiration indicated limited cytochrome chain activity in -P mitochondria. In -P mitochondria, the oxygen uptake decreased and the level of Q reduction increased from 60 to 80%. With no pyruvate present (AOX not fully activated), inhibition of the cytochrome pathway resulted in an increased level of the ratio of reduced ubiquinone (Qr) to total ubiquinone (Qt) (Qr/Qt) in +P mitochondria, but did not change Qr/Qt in -P mitochondria. When pyruvate was present, the kinetics for AOX were similar in mitochondria from -P and +P roots. It is suggested that AOX participation in -P respiration may provide an acclimation to phosphate deficiency. Stabilization of the ubiquinone reduction level by AOX might prevent the harmful effect of an increased formation of reactive oxygen species.
Juszczuk
Izabela M.
IM
Institute of Experimental Plant Biology, University of Warsaw, Miecznikowa 1, PL-02-096 Warsaw, Poland Department of Molecular Cell Physiology, Free University, De Boelelaan 1087, NL-1081 HV Amsterdam, The Netherlands.
Wagner
Anneke M.
AM
Rychter
Anna M.
AM