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| Kiwifruit (Actinidia chinensis ‘Hongyang’) cytosolic ascorbate peroxidases (AcAPX1 and AcAPX2) enhance salinity tolerance in Arabidopsis thaliana |
| GUO Xiu-hong1*, HE Yan2*, ZHANG Yu3, WANG Yi2, HUANG Sheng-xiong3, LIU Yong-sheng1, 2, 3, LI Wei2 |
1 Key Laboratory for Bio-resource and Eco-environment of Ministry of Education, College of Life Sciences/State Key Laboratory of Hydraulics and Mountain River Engineering, Sichuan University, Chengdu 610064, P.R.China
2 School of Horticulture, Anhui Agricultural University, Hefei 230036, P.R.China
3 School of Food and Biological Engineering, Hefei University of Technology, Hefei 230009, P.R.China
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摘要
高等植物中，抗坏血酸过氧化物酶（APX）在清除活性氧的过程中发挥重要作用。中华猕猴桃因富含维生素C而成为具有重要经济和营养价值的园艺作物，其APX的相关研究及报道甚少。本研究分离鉴定出中华猕猴桃‘红阳’的两个细胞质APX基因（AcAPX1和AcAPX2）。两个基因的时空表达模式研究发现，两者分别在叶和根中表达量相对较高。氯化钠处理猕猴桃的根可以提高二者的转录水平。利用GFP融合蛋白的亚细胞定位分析显示两个蛋白均定位于细胞质中。两个基因的his标签重组蛋白成功得以原核表达，并测定出酶活。最后，两个基因在拟南芥中过表达可在盐胁迫处理下提高维生素C和谷胱甘肽的含量。我们的研究揭示了中华猕猴桃细胞质APX可保护猕猴桃免受环境不良刺激。
Abstract Ascorbate peroxidase (APX) plays a key role in scavenging reactive oxygen species (ROS) in higher plants. However, there is very little information available on the APXs in kiwifruit (Actinidia), which is an economically and nutritionally important horticultural crop with exceptionally high ascorbic acid (AsA) accumulation. This study aims to identify and characterize two cytosolic APX genes (AcAPX1 and AcAPX2) derived from A. chinensis ‘Hongyang’. The constitutive expression pattern was determined for both AcAPX1 and AcAPX2, and showed relatively higher expression abundances of AcAPX1 in leaf and AcAPX2 in root. Transcript levels of AcAPX1 and AcAPX2 were increased in kiwifruit roots treated with NaCl. Subcellular localization assays using GFP-fusion proteins in Arabidopsis protoplasts showed that both AcAPX1 and AcAPX2 are targeted to the cytosol. Recombinant AcAPX1 or AcAPX2 proteins were successfully expressed in the prokaryotic expression system and their individual ascorbate peroxidase activities were determined. Finally, constitutive over-expression of AcAPX1 or AcAPX2 could dramatically increase total AsA, glutathione level and salinity tolerance under NaCl stress in Arabidopsis thaliana. Our findings revealed that cytosolic AcAPX1/2 may play an important protective role in the responses to unfavorable environmental stimuli in kiwifruit.
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Received: 04 October 2020
Accepted: 18 February 2021
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Fund: This research was funded by the National Natural Science Foundation of China (31972474), the Natural Science Research Program of Universities of Anhui Province, China (K1832004), the Leading Talent Group Funding of Anhui Province, China (WRMR-2020-75), the Natural Science Foundation of Anhui Province, China (19232002) and the Anhui Agriculture University Shennong Scholar Project, China (RC321901).
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| About author: GUO Xiu-hong, E-mail: guoxiuhong@126.com; HE Yan, E-mail: 1304609580@qq.com; Correspondence LI Wei, E-mail: childelee@vip.126.com; LIU Yong-sheng, E-mail: liuyongsheng1122@ahau.edu.cn
* These authors contributed equally to this study. |
Cite this article:
GUO Xiu-hong, HE Yan, ZHANG Yu, WANG Yi, HUANG Sheng-xiong, LIU Yong-sheng, LI Wei.
2022.
Kiwifruit (Actinidia chinensis ‘Hongyang’) cytosolic ascorbate peroxidases (AcAPX1 and AcAPX2) enhance salinity tolerance in Arabidopsis thaliana. Journal of Integrative Agriculture, 21(4): 1058-1070.
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