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一种假定的细胞外 Cu/Zn 超氧化物歧化酶的克隆及其在入侵性和本地粉虱中超氧化物歧化酶功能差异的研究。

Cloning of a putative extracellular Cu/Zn superoxide dismutase and functional differences of superoxide dismutases in invasive and indigenous whiteflies.

出版信息

Insect Sci. 2015 Feb;22(1):52-64. doi: 10.1111/1744-7917.12100.

DOI:10.1111/1744-7917.12100
PMID:25759872
Abstract

Superoxide dismutases (SODs) are a group of important antioxidant defense enzymes. In this study, a putative extracellular Cu/Zn superoxide dismutase (ecCuZnSOD) complementary DNA was cloned and characterized from the whitefly, Bemisia tabaci. Quantitative polymerase chain reaction analysis showed that the expression level of BtecCuZnSOD was more than 10-fold higher in the invasive Middle East Asia Minor 1 (MEAM1) than in the native Asia II 3 species of the B. tabaci species complex. After exposure to low temperature (4 °C), the expression of Bt-ecCuZnSOD gene was significantly up-regulated in MEAM1 but not in Asia II 3. Furthermore, the expression level of B. tabaci intracellular CuZnSOD (Bt-icCuZnSOD), Bt-ecCuZnSOD and mitochondrial MnSOD (Bt-mMnSOD) was compared after transferring MEAM1 and Asia II 3 whiteflies from favorable (cotton) to unfavorable host plants (tobacco). On cotton, both CuZnSOD genes were expressed at a higher level in MEAM1 compared with Asia II 3. Interestingly, after transferring onto tobacco, the expression of Bt-ecCuZnSOD was significantly induced in Asia II 3 but not in MEAM1. On the other hand, while Bt-mMnSOD was expressed equally in both species on cotton, Bt-mMnSOD messenger RNA was up-regulated in MEAM1 on tobacco. Consistently, enzymatic activity assays of CuZnSOD and MnSOD demonstrated that CuZnSOD might play an important protective role against oxidative stress in Asia II 3, whereas MnSOD activation was critical for MEAM1 whiteflies during host adaptation. Taken together, our results suggest that the successful invasion of MEAM1 is correlated with its constitutive high activity of CuZnSOD and inducible expression of MnSOD under stress conditions.

摘要

超氧化物歧化酶(SODs)是一组重要的抗氧化防御酶。本研究从粉虱中克隆并鉴定了一种假定的细胞外 Cu/Zn 超氧化物歧化酶(ecCuZnSOD)。定量聚合酶链反应分析显示,入侵的中东小亚 1 型(MEAM1)比其所在的 B. tabaci 种复合体的本地亚洲 2 型 3 种的 BtecCuZnSOD 表达水平高出 10 倍以上。暴露于低温(4°C)后,MEAM1 中 Bt-ecCuZnSOD 基因的表达显著上调,但亚洲 2 型 3 种中没有。此外,比较了 MEAM1 和亚洲 2 型 3 粉虱从有利(棉花)转移到不利宿主植物(烟草)后 B. tabaci 细胞内 CuZnSOD(Bt-icCuZnSOD)、Bt-ecCuZnSOD 和线粒体 MnSOD(Bt-mMnSOD)的表达水平。在棉花上,与亚洲 2 型 3 相比,MEAM1 中两种 CuZnSOD 基因的表达水平均较高。有趣的是,转移到烟草上后,亚洲 2 型 3 中 Bt-ecCuZnSOD 的表达显著诱导,但 MEAM1 中没有。另一方面,虽然 Bt-mMnSOD 在棉花上两种物种中的表达水平相等,但在烟草上 MEAM1 中的 Bt-mMnSOD 信使 RNA 上调。酶活性测定表明,CuZnSOD 可能在亚洲 2 型 3 中对氧化应激起到重要的保护作用,而 MnSOD 的激活对于 MEAM1 粉虱在适应宿主时至关重要。总之,我们的结果表明,MEAM1 的成功入侵与其在应激条件下 CuZnSOD 的固有高活性和 MnSOD 的诱导表达有关。

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