以小麦‘陇春20’为实验材料,用两相法分离根质膜微囊,研究NaCl处理对质膜NADPH氧化酶活性的影响。结果显示:(1)温和胶中酶活性条带的出现依赖于NADPH和Ca^2+,DPI(NADPH氧化酶抑制剂)完全抑制酶活性条带的出现;与0.2%的浓度相比,0.5%和1%的去垢剂Triton X-100或Chapso增溶质膜微囊明显减弱酶活性条带,表明高浓度的去垢剂抑制小麦根质膜NADPH氧化酶活性;(2)与对照相比,NaCl处理明显增强NADPH氧化酶活性温和胶染色出现的酶带;进一步研究发现,未处理质膜微囊超氧阴离子(O2^-·)的产生只有7.55nmol·mg^-1 protein·min^-1而100mmol/L NaCl处理的质膜微囊O2^-·的产生为13.63nmol·mg^-1protein·min^-1。结果表明:质膜蛋白温和胶活性染色出现的酶带可能是小麦根质膜NADPH氧化酶,NaCl处理增强小麦根质膜NADPH氧化酶的活性。
Plasma membrane vesicles in the roots of the wheat variety ‘Longchun 20' was isolated by twophase method and used to study NaC1 effect on plasmalemma NADPH oxidase activity. It was shown that (1) the appearance of the native activity bands depended on NADPH and Ca^2+ and completely inhibited by DPI (an inhibitor of NADPH oxidase) ,and compared with Triton X-100 and or Chapso at 0.2% ,the detergents Triton X-100 and or Chapso remarkably weakened the activity bands at 0.5% and 0.1% ,indicating that the detergents inhibited plasmalemma NADPH oxidase activity in wheat roots ; (2) Compared with the control, NaCl remarkably intensified the enzymatic bands of NADPH oxidase in native gel staining t further study showed that in the untreated plasmalemma vesicles superoxide anion was produced at 7.55 nmol · mg^-1 protein · min^-1 and in the plasmlemma vesicles treated with NaCl superoxide anion was produced at 13.63 nmol · mg^-1 protein · min^-1. These results indicated that the enzymatic bands appeared in the native gel staining were probably plasmalemma NADPH oxidase and NaCl could enhance plasmalemma NADPH oxidase activity in wheat roots.