Overexpression of the spinach S -nitrosoglutathione reductase ( SoGSNOR ) in tobacco resulted in enhanced nitrate stress
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ORIGINAL ARTICLE
Overexpression of the spinach S‑nitrosoglutathione reductase (SoGSNOR) in tobacco resulted in enhanced nitrate stress tolerance Manqi Wang1 · Yanyan Dong1 · Jinping Yan1 · Qinqin Han1 · Kunzhi Li1 · Huini Xu1 Received: 1 June 2020 / Accepted: 4 August 2020 © Springer Nature B.V. 2020
Abstract S-nitrosylation, the addition of nitric oxide (NO) moiety to a reactive cysteine thiol, to form an S-nitrosothiol (SNO), is emerging as a prototypic redox-based post-translational modification. S-nitrosoglutathione reductase (GSNOR) is thought to be the major regulator of total cellular SNO levels in plants. However, its role in excess nitrate stress has not been investigated in spinach. In this study, a spinach GSNOR gene (GenBank Accession No. KR381778) was amplified and designated as SoGSNOR. The transcript and protein level of SoGSNOR were reduced after excess nitrate treatment for 24 h. Addition of NO donor to the nitrate solution decreased the SoGSNOR expression, while supplementation inhibitor of nitrate reductase and nitric oxide synthase increased its expression. Overexpression of SoGSNOR in tobacco increased the germination rate of transgenic seeds, compared to the wild type (WT) under nitrate stress. Higher photosynthetic rate, transpiration rate, stomatal conductance, water use efficiency and expression level of some stress-related genes were detected in the transgenic seedlings than the WT under nitrate stress. The transgenic tobacco seedlings have lower malondialdehyde content, reactive oxygen species (ROS) fluorescence, and higher activities and transcript level of superoxide dismutase, catalase, peroxidase under nitrate stress. SoGSNOR transgenic tobacco plants have lower NR activity and protein level, higher GSNOR and non-symbiotic class 1 hemoglobin (nsHb) protein level than the WT plants, leading to lower NO accumulation and SNOs contents under nitrate stress. These results suggested that overexpression of SoGSNOR increased nitrate stress tolerance of tobacco by regulating ROS and RNS metabolism. Key message A spinach S-nitrosoglutathione reductase (SoGSNOR) gene was isolated from spinach root. The expression level and activities of SoGSNOR were reduced by excess nitrate treatment. Overexpression of SoGSNOR in tobacco increased nitrate stress tolerance by regulating ROS and RNS metabolism. Keywords Nitrate stress · S-nitrosoglutathione reductase · Nitric oxide · Spinach
Introduction
Communicated by Degao Liu. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s11240-020-01906-2) contains supplementary material, which is available to authorized users. * Huini Xu [email protected] 1
Faculty of Life Science and Technology, Kunming University of Science and Technology, Jingming South Street, Kunming 650224, Yunnan, People’s Republic of China
Nitric oxide (NO) is an important free radical that is considered to be a general plant signal. NO regulates both normal developmental processes and biotic and abiotic stress responses (Jasid et al. 200
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