Holden M. J., Crimmins T. J., Chaney R. L.
Department of Biological Sciences, The George Washington University, Washington, DC 20052 (M.J.H., T.J.C.).
Plant Physiol. 1995 Jul;108(3):1093-1098. doi: 10.1104/pp.108.3.1093.
Reduction of Cu2+ by plasma membrane vesicles isolated from tomato (Lycopersicon esculentum Mill.) roots was investigated. Plants were grown in hydroponic culture with complete nutrition for 4 weeks or were deprived of Fe for the last 7 d. Plasma membrane vesicles were prepared by aqueous two-phase partitioning. Reduction of Cu, Fe, and ferricyanide by plasma membrane vesicles was measured. An increase in the activity of all three pyridine-nucleotide-dependent activities was noted in plasma membrane preparations from Fe-deficient, compared to Fe-sufficient, plants. Solubilization and chromatographic separation of two plasma membrane electron transport systems indicated that the Fe-chelate reductase was probably responsible for reduction of Cu. Assays used a variety of Cu chelates, and for each the Cu activity in the assay was determined by the program Geochem PC. The rate of reduction of Cu correlated with the level of Cu activity, and results support the idea that free Cu2+ and not Cu chelates may serve as the true substrate for reduction. Reduction was observed only in assays in which Cu activity was equivalent to Cu-enriched or Cu-toxic soils. These results suggest that reduction of Cu by tomato root may have little or no physiological relevance under conditions experienced by the root in the soil.
研究了从番茄(Lycopersicon esculentum Mill.)根系分离的质膜囊泡对Cu2+的还原作用。将植株在完全营养液的水培条件下培养4周,或在最后7天缺铁培养。通过水相双相分配法制备质膜囊泡。测定了质膜囊泡对Cu、Fe和铁氰化物的还原作用。与铁充足的植株相比,缺铁植株的质膜制剂中所有三种依赖吡啶核苷酸的活性均有所增加。两种质膜电子传递系统的溶解和色谱分离表明,铁螯合物还原酶可能负责Cu的还原。实验使用了多种Cu螯合物,每种实验中Cu的活性由Geochem PC程序测定。Cu的还原速率与Cu活性水平相关,结果支持游离Cu2+而非Cu螯合物可能是真正还原底物的观点。仅在Cu活性相当于富Cu或Cu毒性土壤的实验中观察到还原现象。这些结果表明,在根系在土壤中经历的条件下,番茄根系对Cu的还原作用可能几乎没有生理相关性。