拟南芥 MhZEP 基因对盐碱胁迫的分子克隆和功能鉴定

IF 3.4 3区 生物学 Q1 PLANT SCIENCES Physiology and Molecular Biology of Plants Pub Date : 2024-09-05 DOI:10.1007/s12298-024-01495-w
Yongjuan Dong, Lei Du, Zhongxing Zhang, Jiao Cheng, Yanlong Gao, Xiaoya Wang, Yuxia Wu, Yanxiu Wang
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引用次数: 0

摘要

土壤盐碱化是限制植物生长和发育的主要环境因素之一。玉米黄质环氧化物酶(ZEP)在 ABA 生物合成和黄绿素循环中发挥作用,在植物应对各种环境胁迫中起着重要作用。通过实时定量 PCR(qRT-PCR)研究发现,MhZEP 对盐碱胁迫有响应,在盐碱胁迫 48 h 时表达量最高,是 0 h 时的 14.53 倍。随后,在拟南芥中进一步研究了 MhZEP(ID:103403091)的功能特性。MhZEP 包含一个完整的开放阅读框,长度为 1998 bp,编码 665 个氨基酸,等电点为 7.18。系统发生树分析表明,MhZEP 与 Glycine max 的同源性最高,亲缘关系最密切。与野生型相比,转基因植株在盐碱胁迫下生长较好,MhZEP-OE品系的叶绿素含量、类胡萝卜素含量、酶活性(POD、SOD、CAT和APX)和K+含量均高于野生型,而叶绿素沉降和Na+含量低于野生型,表明其具有较强的抗逆性。通过qRT-PCR检测了MhZEP-OE中盐碱胁迫相关基因的表达水平,发现MhZEP通过调控类胡萝卜素合成基因(MhPSY、MhZDS、MhLYCB和MhVDE)和ABA生物合成基因(MhNCED5、MhABI1和MhCYP707A2)的表达,提高了大叶黄杨对盐碱胁迫的耐受性。通过调节 Na+转运体基因(MhCHX15 和 MhSOS1)和 K+转运体基因(MhHKT1;1、MhNHX1 和 MhSKOR1)的表达,增加细胞质中的钾钠比,以维持离子平衡。此外,H+-ATPase 基因(MhAHA2 和 MhAHA8)的表达也有所增加,从而减轻了盐碱胁迫造成的氧化损伤。总之,MhZEP在植物抵抗盐碱胁迫的过程中发挥了重要作用,这为进一步研究其在苹果中的功能奠定了基础。
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Molecular cloning and functional characterization in response to saline-alkali stress of the MhZEP gene in Arabidopsis thaliana

Soil salinization is one of the major environmental factors that restrict plant growth and development. Zeaxanthin epoxidase (ZEP) functions in ABA biosynthesis and the xanthophyll cycle and has a vital role in plant responses to various environmental stresses. It was found by quantitative real-time PCR (qRT-PCR) that MhZEP responded to saline-alkali stress and showed the highest expression at 48 h of saline-alkali stress, which was 14.53-fold of 0 h. The MhZEP gene was cloned from the apple rootstock begonia (Malus halliana Koehne) and its protein physicochemical properties were analyzed. Subsequently, the functional characterization of MhZEP (ID: 103403091) was further investigated in Arabidopsis thaliana. The MhZEP contained a complete open reading frame with a length of 1998 bp, and encoded 665 amino acids with an isoelectric point of 7.18. Phylogenetic tree analysis showed that MhZEP was the most homologous and closely related to Glycine max. Compared with wild-type, transgenic plants grew better under saline-alkali stress and the MhZEP-OE line showed higher chlorophyll content, carotenoid content, enzyme activities (POD, SOD, CAT and APX) and K+ content, whereas they had lower chlorosis and Na+ content than the wild type (WT), which indicated that they had strong resistance to stress. The expression levels of saline-alkali stress-related genes in A. thaliana MhZEP-OE were examined by qRT-PCR, and it was found that the MhZEP improved the tolerance of A. thaliana to saline-alkali stress tolerance by regulating the expression of carotenoid synthesis genes (MhPSY, MhZDS, MhLYCB and MhVDE) and ABA biosynthesis genes (MhNCED5, MhABI1 and MhCYP707A2). And the potassium-sodium ratio in the cytoplasm was increased to maintain ionic homeostasis by modulating the expression of Na+ transporter genes (MhCHX15 and MhSOS1) and K+ transporter genes (MhHKT1;1, MhNHX1 and MhSKOR1). Moreover, the expression of H+-ATPase genes (MhAHA2 and MhAHA8) was increased to reduce the oxidative damage caused by saline-alkali stress. In summary, MhZEP acted as an essential role in plant resistance to saline-alkali stress, which lays the foundation for further studies on its function in apple.

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来源期刊
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期刊介绍: Founded in 1995, Physiology and Molecular Biology of Plants (PMBP) is a peer reviewed monthly journal co-published by Springer Nature. It contains research and review articles, short communications, commentaries, book reviews etc., in all areas of functional plant biology including, but not limited to plant physiology, biochemistry, molecular genetics, molecular pathology, biophysics, cell and molecular biology, genetics, genomics and bioinformatics. Its integrated and interdisciplinary approach reflects the global growth trajectories in functional plant biology, attracting authors/editors/reviewers from over 98 countries.
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