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《柏木 CfICE1 基因的克隆与抗寒性分析》

Cloning and cold-resistance analyses of CfICE1 gene in Cryptomeria fortunei.

机构信息

Key Laboratory of Forest Genetics & Biotechnology of Ministry of Education, Co-Innovation Center for Sustainable Forestry in Southern China, College of Forestry, Nanjing Forestry University, Nanjing, 210037, China.

出版信息

Plant Physiol Biochem. 2021 May;162:456-467. doi: 10.1016/j.plaphy.2021.03.020. Epub 2021 Mar 13.

Abstract

Cryptomeria fortunei is a conifer species that can attain a height of ~70 m and is cultivated for its timber as well as its ornamental value. It is a subtropical plant that prefers a warm and humid environment. Therefore, low temperature (LT) affects its growth, development, productivity and ecological distribution. Inducer of C-repeat binding factor (CBF) expression 1 (ICE1) plays an important role in the response to cold/freezing stress in plants through the CBF regulation pathway. To date, there is no research on homologue of ICE1 in C. fortunei. In this study, we first isolated the CfICE1 transcript from C. fortunei. The CfICE1 coding sequence was 1728 nucleotides encoding a 575-aa protein and contained a serine-rich motif, a basic helix-loop-helix-Zipper (bHLH-Zip), an ACT domain and a nuclear localization signal (NLS), which were conserved in ICE1 homologous genes. Phylogenetic analysis showed that CfICE1 and all dicots ICE1 proteins were clustered together. CfICE1 had transcriptional activity in yeast cells, was predominantly located in the nucleus and highly expressed in tender needles and roots. 35S::CfICE1 transgenic Arabidopsis thaliana could increase antioxidant enzyme activities and photosynthesis and reduce the malondialdehyde content compared to the wild-type to better cope with LT. Under LT, CfICE1 expression was higher; the C. fortunei clone with stronger cold resistance (CR) significantly upregulated the expression of CfICE1 compared to the weaker clone. In conclusion, these results suggest that CfICE1 plays an active role in CR, which provides a theoretical basis for breeding for CR in C. fortunei.

摘要

柳杉是一种针叶树种,可长到约 70 米高,因其木材和观赏价值而被栽培。它是一种亚热带植物,喜欢温暖潮湿的环境。因此,低温(LT)会影响其生长、发育、生产力和生态分布。诱导 C-repeat binding factor(CBF)表达 1(ICE1)通过 CBF 调节途径在植物对冷/冻胁迫的反应中起着重要作用。迄今为止,在柳杉中还没有关于 ICE1 同源物的研究。在这项研究中,我们首先从柳杉中分离出 CfICE1 转录本。CfICE1 编码序列为 1728 个核苷酸,编码 575 个氨基酸的蛋白质,包含一个富含丝氨酸的基序、一个碱性螺旋-环-螺旋-Zipper(bHLH-Zip)、一个 ACT 结构域和一个核定位信号(NLS),这些结构在 ICE1 同源基因中保守。系统发育分析表明,CfICE1 和所有双子叶植物 ICE1 蛋白聚集在一起。CfICE1 在酵母细胞中具有转录活性,主要定位于细胞核内,在嫩针和根中高表达。与野生型相比,35S::CfICE1 转基因拟南芥可以增加抗氧化酶活性和光合作用,降低丙二醛含量,从而更好地应对 LT。在 LT 下,CfICE1 的表达更高;与较弱的克隆相比,具有较强抗寒性(CR)的柳杉克隆显著上调 CfICE1 的表达。综上所述,这些结果表明 CfICE1 在 CR 中发挥积极作用,为柳杉的 CR 育种提供了理论依据。

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