Molecular mechanisms of potato (Solanum tuberosum L.) transcription factor StbZIP1 in regulating saline-alkaline stress response through enhanced antioxidant capacity

IF 5.2 2区 农林科学 Q1 AGRICULTURE, MULTIDISCIPLINARY
Shujuan Jiao, Xiongliang Hu, Yong Wang, Ruyan Zhang, Xingxing Wang, Yuan Lu, Weina Zhang, Yuhui Liu, Shuhao Qin, Yichen Kang
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Abstract

Background

Saline-alkali stress severely impacts global crop productivity, while basic leucine zipper (bZIP) transcription factors (TFs) are known regulators of abiotic stress responses, the specific mechanisms of StbZIP1 in potato saline-alkaline tolerance remains unclear.

Methods

We cloned StbZIP1 from tetraploid potato ‘Favorita’, analyzed its sequence characteristics, and generated overexpression lines. StbZIP1-overexpressing (OE) and wild-type (WT) plants were subjected to saline-alkaline stress (NaCl: NaHCO3 = 1:1) to assess physiological and molecular responses.

Results

StbZIP1 encodes a 16.61 kDa protein with conserved bZIP domains. Secondary structure prediction revealed that the protein comprises 55.48% α-helix and 44.52% random coil, consistent with the structural characteristics of typical bZIP family features. Physicochemical characterization revealed StbZIP1 was a highly hydrophilic protein (GRAVY index: −0.882) with no transmembrane region, and it harbors 27 predicted phosphorylation sites. Subcellular localization analysis using GFP-tagged StbZIP1 via confocal microscopy confirmed its exclusive nuclear localization, classifying it as a nuclear-targeted transcription factor. Under saline-alkaline stress, WT plants displayed severe wilting and complete desiccation of lower leaves, whereas StbZIP1-OE plants exhibited delayed wilting, no death and retained greener apical leaves. Quantitative analysis revealed that StbZIP1-OE plants showed a 33%–50% increase in chlorophyll content compared to WT (p < 0.01). Notably, StbZIP1-OE plants exhibited a more pronounced increase in Pro content (28% ~ 46%) higher than WT, while their MDA content was significantly reduced compared to WT. Furthermore, the activities of antioxidant enzymes (SOD, POD, and APX) were markedly elevated in StbZIP1-OE plants, showing increases of 81% ~ 100%, 81% ~ 104%, and 20% ~ 43%, respectively, relative to WT. Analysis of stress-related gene expression showed that after 12 d of saline-alkaline stress, the OE plants exhibited significantly increased expression of all six genes (StNCED, StRD29B, StABI5, StP5CS, StSOD, and StCAT) compared with WT (p < 0.05).

Conclusions

StbZIP1 positively regulates saline-alkaline tolerance by enhancing antioxidant capacity, providing a reference for the further cultivation of new stress-resistant potatoes.

Graphical Abstract

马铃薯转录因子StbZIP1通过增强抗氧化能力调控盐碱胁迫反应的分子机制
盐碱胁迫严重影响全球作物产量,虽然碱性亮氨酸拉链(bZIP)转录因子(TFs)是已知的非生物胁迫响应的调节因子,但StbZIP1在马铃薯盐碱耐受性中的具体机制尚不清楚。方法从马铃薯“Favorita”四倍体中克隆StbZIP1,分析其序列特征,建立过表达系。以stbzip1过表达(OE)和野生型(WT)植物为研究对象,研究了盐碱胁迫(NaCl: NaHCO3 = 1:1)下的生理和分子反应。结果stbzip1编码一个具有保守bZIP结构域的16.61 kDa蛋白。二级结构预测表明,该蛋白由55.48%的α-螺旋和44.52%的随机螺旋组成,符合典型bZIP家族的结构特征。理化性质分析表明,StbZIP1为高度亲水性蛋白(卤汁指数:−0.882),无跨膜区,具有27个预测磷酸化位点。通过共聚焦显微镜对gfp标记的StbZIP1进行亚细胞定位分析,证实了其核特异性定位,将其归类为核靶向转录因子。在盐碱胁迫下,WT植株表现为严重的萎蔫,下部叶片完全干燥,而StbZIP1-OE植株表现为延迟萎蔫,无死亡,顶端叶片较绿。定量分析显示,StbZIP1-OE植株叶绿素含量较WT增加33% ~ 50% (p < 0.01)。值得注意的是,StbZIP1-OE植株Pro含量较WT显著升高(28% ~ 46%),而MDA含量较WT显著降低,抗氧化酶(SOD、POD和APX)活性显著升高,分别较WT升高81% ~ 100%、81% ~ 104%和20% ~ 43%。胁迫相关基因表达分析表明,在盐碱胁迫12 d后,与WT相比,OE植株的StNCED、StRD29B、StABI5、StP5CS、StSOD和StCAT 6个基因的表达均显著增加(p < 0.05)。结论stbzip1基因通过增强抗氧化能力正向调节马铃薯的耐盐碱性,为进一步培育新型抗逆性马铃薯提供参考。图形抽象
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来源期刊
Chemical and Biological Technologies in Agriculture
Chemical and Biological Technologies in Agriculture Biochemistry, Genetics and Molecular Biology-Biotechnology
CiteScore
6.80
自引率
3.00%
发文量
83
审稿时长
15 weeks
期刊介绍: Chemical and Biological Technologies in Agriculture is an international, interdisciplinary, peer-reviewed forum for the advancement and application to all fields of agriculture of modern chemical, biochemical and molecular technologies. The scope of this journal includes chemical and biochemical processes aimed to increase sustainable agricultural and food production, the evaluation of quality and origin of raw primary products and their transformation into foods and chemicals, as well as environmental monitoring and remediation. Of special interest are the effects of chemical and biochemical technologies, also at the nano and supramolecular scale, on the relationships between soil, plants, microorganisms and their environment, with the help of modern bioinformatics. Another special focus is the use of modern bioorganic and biological chemistry to develop new technologies for plant nutrition and bio-stimulation, advancement of biorefineries from biomasses, safe and traceable food products, carbon storage in soil and plants and restoration of contaminated soils to agriculture. This journal presents the first opportunity to bring together researchers from a wide number of disciplines within the agricultural chemical and biological sciences, from both industry and academia. The principle aim of Chemical and Biological Technologies in Agriculture is to allow the exchange of the most advanced chemical and biochemical knowledge to develop technologies which address one of the most pressing challenges of our times - sustaining a growing world population. Chemical and Biological Technologies in Agriculture publishes original research articles, short letters and invited reviews. Articles from scientists in industry, academia as well as private research institutes, non-governmental and environmental organizations are encouraged.
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