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从黄连中过表达一个新型 E3 泛素连接酶基因可增强转基因烟草的耐旱性。

Overexpression of a novel E3 ubiquitin ligase gene from Coptis chinensis Franch enhances drought tolerance in transgenic tobacco.

机构信息

College of Medical Technology, State Key Laboratory of Characteristic Chinese Medicine Resources in Southwest China, Chengdu University of Traditional Chinese Medicine, Chengdu, 611137, China.

出版信息

Z Naturforsch C J Biosci. 2020 Nov 26;75(11-12):417-424. doi: 10.1515/znc-2019-0211.

Abstract

Drought stress has a significant effect on the growth, physiology and biochemistry of medicinal plants. SDIR1 (Salt- and Drought-Induced Ring Finger1), a C3H2C3-type RING-finger E3 ubiquitin ligase gene plays an important role in the stress response of various plants. However, the role of this gene is not clear in Coptis chinensis. In this study, the CcSDIR1 gene was cloned from C. chinensis using RACE and RT-PCR. Sequence analysis revealed that CcSDIR1 had an open reading frame of 840 bp that encodes 279 amino acids with a theoretical molecular weight about 31 kDa and pI value of 5.65 and shared conserved domains with other plants. On comparison with the wild-type plants, overexpression of CcSDIR1 in transgenic tobaccos increased drought tolerance and showed better growth performance. However, lower malondialdehyde contents and high antioxidant enzyme activities were observed in transgenic tobacco plants compared to wild-type plants. In addition, Evans blue staining showed high cell viability of transgenic lines under drought stress. These results suggest that CcSDIR1 regulates various responses to drought stress by increasing antioxidant enzyme activities and reducing oxidative damage. From the study results, the CcSDIR1 gene will be very useful for drought stress research in plants.

摘要

干旱胁迫对药用植物的生长、生理和生物化学有显著影响。SDIR1(盐和干旱诱导的环指 1)是一种 C3H2C3 型 RING-finger E3 泛素连接酶基因,在各种植物的应激反应中起着重要作用。然而,该基因在黄连中的作用尚不清楚。本研究采用 RACE 和 RT-PCR 从黄连中克隆出 CcSDIR1 基因。序列分析表明,CcSDIR1 有一个 840bp 的开放阅读框,编码 279 个氨基酸,理论分子量约为 31kDa,pI 值为 5.65,与其他植物具有保守结构域。与野生型植物相比,过表达 CcSDIR1 的转基因烟草提高了耐旱性,并表现出更好的生长性能。然而,与野生型植物相比,转基因烟草植物的丙二醛含量较低,抗氧化酶活性较高。此外,Evans 蓝染色显示,在干旱胁迫下,转基因系具有较高的细胞活力。这些结果表明,CcSDIR1 通过增加抗氧化酶活性和减少氧化损伤来调节对干旱胁迫的各种反应。从研究结果来看,CcSDIR1 基因将非常有助于植物的干旱胁迫研究。

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