首页|外源超氧化物歧化酶基因MnSOD在玉米中的过量表达及抗逆性的提高

外源超氧化物歧化酶基因MnSOD在玉米中的过量表达及抗逆性的提高

Over-expression of Exotic Superoxide Dismutase Gene MnSOD and Increase in Stress Resistance in Maize

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为研究过量表达的超氧化物歧化酶基因MnSOD对玉米抗逆性的作用,构建了小麦来源MnSOD基因的单子叶植物高效表达载体,用基因枪法转化优良玉米自交系胚性愈伤组织.经潮霉素梯度浓度培养基筛选,从阳性愈伤组织再生获得9个正常结实的植株.其中5株经PCR和Southern印迹检测表现为阳性,表明外源基因已整合到玉米基因组中.提取SOD酶液,非变性聚丙烯酰胺浓度梯度凝胶电泳分离,用H2O2 5 mmol/L抑制FeSOD和Cu/ZnSOD活性,氯化硝基四氮唑蓝染色检测MnSOD酶活性.Southern印迹呈阳性的5个植株,MnSOD酶活性均高于未转基因的对照.甲基紫精氧化损伤处理后,用电解质渗漏率法测定阳性株系的叶片渗透液的电导率.结果表明,转基因株系的抗氧化损伤能力显著高于对照.
To evaluate the effect of over-expressed Mnsuperoxide dismutase (MnSOD) on stress resistance in maize, an over-expression vector with MnSOD gene of wheat was constructed and used to transform embryonic calli of elite maize inbred line by gene gun. After screening with a concentration gradient of hygromycin, 9 plants were regenerated from the positive calli and reproduced fertile seeds, 5 of which were shown to be positive by PCR and Southern blot analysis. This result indicated that the exotic gene had been integrated in maize genome.SOD enzyme extracts were separated by non-denatured polyacrylamide gel electrophoresis of gradient concentration. After inhibition with 5 mmol/L H2O2 to FeSOD and Cu/ZnSOD, MnSOD enzyme activity was assayed with nitrotetrazolium chloride blue staining. All the 5 plants shown to be positive in Southern blot had higher MnSOD activity than untransformed control. Electric conductivity of leaf exudate was assayed after damage treatment with methyl viologen. The result suggested that the resistance of the transformed plants to oxidative damage was significantly higher than untransformed control.

active oxygensuperoxide dismutasetransgenestress resistancemaize

杜娟、朱祯、李晚忱

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四川农业大学,玉米研究所,四川,雅安625014

四川农业大学,西南作物基因资源与遗传改良教育部重点实验室,四川,雅安625014

中国科学院遗传与发育生物学研究所,北京,100101

活性氧 超氧化物歧化酶 转基因 抗逆性 玉米

国家自然科学基金教育部长江学者和创新团队发展计划

30070476IRT0453

2006

植物生理与分子生物学学报
中国植物生理学会

植物生理与分子生物学学报

CSTPCDCSCD
ISSN:1671-3877
年,卷(期):2006.32(1)
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