Kim Y C, Miller C D, Anderson A J
Department of Biology, Utah State University, Logan, Utah 84322-5305, USA.
Appl Environ Microbiol. 2000 Apr;66(4):1460-7. doi: 10.1128/AEM.66.4.1460-1467.2000.
To investigate the role of superoxide dismutases (SOD) in root colonization and oxidative stress, mutants of Pseudomonas putida lacking manganese-superoxide dismutase (MnSOD) (sodA), iron-superoxide dismutase (FeSOD) (sodB), or both were generated. The sodA sodB mutant did not grow on components washed from bean root surfaces or glucose in minimal medium. The sodB and sodA sodB mutants were more sensitive than wild type to oxidative stress generated within the cell by paraquat treatment. In single inoculation of SOD mutants on bean, only the sodA sodB double mutant was impaired in growth on root surfaces. In mixed inoculations with wild type, populations of the sodA mutant were equal to those of the wild type, but levels of the sodB mutant and, to a great extent, the sodA sodB mutant, were reduced. Confocal microscopy of young bean roots inoculated with green fluorescent protein-tagged cells showed that wild type and SOD single mutants colonized well predominantly at the root tip but that the sodA sodB double mutant grew poorly at the tip. Our results indicate that FeSOD in P. putida is more important than MnSOD in aerobic metabolism and oxidative stress. Inhibition of key metabolic enzymes by increased levels of superoxide anion may cause the impaired growth of SOD mutants in vitro and in planta.
为了研究超氧化物歧化酶(SOD)在根部定殖和氧化应激中的作用,构建了恶臭假单胞菌缺乏锰超氧化物歧化酶(MnSOD)(sodA)、铁超氧化物歧化酶(FeSOD)(sodB)或两者皆缺的突变体。sodA sodB突变体在从豆科植物根表面冲洗下来的成分或基本培养基中的葡萄糖上无法生长。sodB和sodA sodB突变体比野生型对百草枯处理在细胞内产生的氧化应激更敏感。在豆科植物上单独接种SOD突变体时,只有sodA sodB双突变体在根表面的生长受到损害。在与野生型混合接种时,sodA突变体的数量与野生型相等,但sodB突变体以及在很大程度上sodA sodB突变体的数量减少。对接种了绿色荧光蛋白标记细胞的幼嫩豆科植物根进行共聚焦显微镜观察表明,野生型和SOD单突变体主要在根尖定殖良好,但sodA sodB双突变体在根尖生长不良。我们的结果表明,恶臭假单胞菌中的FeSOD在有氧代谢和氧化应激中比MnSOD更重要。超氧阴离子水平升高对关键代谢酶的抑制可能导致SOD突变体在体外和植物体内生长受损。