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NaCl和NaHCO₃胁迫下叶绿体OsCu/Zn-SOD过表达水稻的耐受性分析

Tolerance analysis of chloroplast OsCu/Zn-SOD overexpressing rice under NaCl and NaHCO3 stress.

作者信息

Guan Qingjie, Liao Xu, He Mingliang, Li Xiufeng, Wang Zhenyu, Ma Haiyan, Yu Song, Liu Shenkui

机构信息

Key Laboratory of Saline-alkali Vegetation Ecology Restoration (SAVER), Ministry of Education, Alkali Soil Natural Environmental Science Center (ASNESC), Northeast Forestry University, Harbin, China.

Lab of Soybean Molecular Biology and Molecular Breeding, Northeast Institute of Geography and Agroecology, Chinese Academy of Sciences, Nangang District, Harbin City, Heilongjiang, China.

出版信息

PLoS One. 2017 Oct 11;12(10):e0186052. doi: 10.1371/journal.pone.0186052. eCollection 2017.

Abstract

The 636-bp-long cDNA sequence of OsCu/Zn-SOD (AK059841) was cloned from Oryza sativa var. Longjing11 via reverse transcription polymerase chain reaction (RT-PCR). The encoded protein comprised of 211 amino acids is highly homologous to Cu/Zn-SOD proteins from tuscacera rice and millet. Quantitative RT-PCR revealed that in rice, the level of OsCu/Zn-SOD gene expression was lowest in roots and was highest in petals and during the S5 leaf stage. Moreover, the expression level of OsCu/Zn-SOD gene expression decreased during the L5 leaf stage to maturity. The level of OsCu/Zn-SOD gene expression, however, was increased under saline-sodic stress and NaHCO3 stress. Germination tests under 125, 150, and 175 mM NaCl revealed that OsCu/Zn-SOD-overexpressing lines performed better than the non-transgenic (NT) Longjing11 lines in terms of germination rate and height. Subjecting seedlings to NaHCO3 and water stress revealed that OsCu/Zn-SOD-overexpressing lines performed better than NT in terms of SOD activity, fresh weight, root length, and height. Under simulated NaHCO3 stress, OsCu/Zn-SOD-overexpressing lines performed better than NT in terms of survival rate (25.19% > 6.67%) and yield traits (average grain weight 20.6 > 18.15 g). This study showed that OsCu/Zn-SOD gene overexpression increases the detoxification capacity of reactive oxygen species in O. sativa and reduces salt-induced oxidative damage. We also revealed the regulatory mechanism of OsCu/Zn-SOD enzyme in saline-sodic stress resistance in O. sativa. Moreover, we provided an experimental foundation for studying the mechanism of OsCu/Zn-SOD enzymes in the chloroplast.

摘要

通过逆转录聚合酶链反应(RT-PCR)从水稻品种龙井11中克隆了长度为636 bp的水稻铜/锌超氧化物歧化酶(OsCu/Zn-SOD,AK059841)的cDNA序列。所编码的由211个氨基酸组成的蛋白质与来自粳稻和谷子的铜/锌超氧化物歧化酶蛋白高度同源。定量RT-PCR显示,在水稻中,OsCu/Zn-SOD基因的表达水平在根中最低,在花瓣中最高,且在S5叶期也最高。此外,OsCu/Zn-SOD基因的表达水平在L5叶期至成熟期下降。然而,在盐碱胁迫和NaHCO₃胁迫下,OsCu/Zn-SOD基因的表达水平升高。在125、150和175 mM NaCl条件下的发芽试验表明,过表达OsCu/Zn-SOD的株系在发芽率和株高方面比非转基因(NT)龙井11株系表现更好。对幼苗施加NaHCO₃和水分胁迫表明,过表达OsCu/Zn-SOD的株系在超氧化物歧化酶活性、鲜重、根长和株高方面比NT表现更好。在模拟NaHCO₃胁迫下,过表达OsCu/Zn-SOD的株系在存活率(25.19% > 6.67%)和产量性状(平均粒重20.6 > 18.15 g)方面比NT表现更好。本研究表明,OsCu/Zn-SOD基因的过表达提高了水稻中活性氧的解毒能力,减少了盐诱导的氧化损伤。我们还揭示了OsCu/Zn-SOD酶在水稻抗盐碱胁迫中的调控机制。此外,我们为研究叶绿体中OsCu/Zn-SOD酶的机制提供了实验基础。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ea5/5636109/321bae74d6af/pone.0186052.g001.jpg

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