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过量生产超氧化物歧化酶和过氧化氢酶增强活性氧清除可延缓木薯贮藏根采后生理劣变。

Enhanced reactive oxygen species scavenging by overproduction of superoxide dismutase and catalase delays postharvest physiological deterioration of cassava storage roots.

机构信息

National Key Laboratory of Plant Molecular Genetics and National Center for Plant Gene Research , Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China.

出版信息

Plant Physiol. 2013 Mar;161(3):1517-28. doi: 10.1104/pp.112.212803. Epub 2013 Jan 23.

Abstract

Postharvest physiological deterioration (PPD) of cassava (Manihot esculenta) storage roots is the result of a rapid oxidative burst, which leads to discoloration of the vascular tissues due to the oxidation of phenolic compounds. In this study, coexpression of the reactive oxygen species (ROS)-scavenging enzymes copper/zinc superoxide dismutase (MeCu/ZnSOD) and catalase (MeCAT1) in transgenic cassava was used to explore the intrinsic relationship between ROS scavenging and PPD occurrence. Transgenic cassava plants integrated with the expression cassette p54::MeCu/ZnSOD-35S::MeCAT1 were confirmed by Southern-blot analysis. The expression of MeCu/ZnSOD and MeCAT1 was verified by quantitative reverse transcription-polymerase chain reaction and enzymatic activity analysis both in the leaves and storage roots. Under exposure to the ROS-generating reagent methyl viologen or to hydrogen peroxide (H2O2), the transgenic plants showed higher enzymatic activities of SOD and CAT than the wild-type plants. Levels of malondialdehyde, chlorophyll degradation, lipid peroxidation, and H2O2 accumulation were dramatically reduced in the transgenic lines compared with the wild type. After harvest, the storage roots of transgenic cassava lines show a delay in their PPD response of at least 10 d, accompanied by less mitochondrial oxidation and H2O2 accumulation, compared with those of the wild type. We hypothesize that this is due to the combined ectopic expression of Cu/ZnSOD and CAT leading to an improved synergistic ROS-scavenging capacity of the roots. Our study not only sheds light on the mechanism of the PPD process but also develops an effective approach for delaying the occurrence of PPD in cassava.

摘要

木薯(Manihot esculenta)贮藏根的产后生理劣变(PPD)是由于活性氧(ROS)爆发迅速导致的,这导致由于酚类化合物的氧化而使血管组织变色。在这项研究中,通过共表达ROS 清除酶铜/锌超氧化物歧化酶(MeCu/ZnSOD)和过氧化氢酶(MeCAT1)来探索ROS 清除与 PPD 发生之间的内在关系。通过 Southern-blot 分析证实了整合有表达盒 p54::MeCu/ZnSOD-35S::MeCAT1的转基因木薯植物。通过定量逆转录聚合酶链反应和酶活性分析,在叶片和贮藏根中均验证了 MeCu/ZnSOD 和 MeCAT1 的表达。在暴露于ROS 生成试剂甲基紫精或过氧化氢(H2O2)时,与野生型植物相比,转基因植物的 SOD 和 CAT 酶活性更高。与野生型相比,转基因系的丙二醛、叶绿素降解、脂质过氧化和 H2O2 积累水平显着降低。收获后,与野生型相比,转基因木薯系的贮藏根 PPD 反应至少延迟了 10 天,并且线粒体氧化和 H2O2 积累较少。我们假设这是由于 Cu/ZnSOD 和 CAT 的异位表达导致根的协同 ROS 清除能力得到改善。我们的研究不仅阐明了 PPD 过程的机制,而且还开发了一种有效延缓木薯 PPD 发生的方法。

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