马铃薯StDREB4基因响应盐和干旱胁迫的功能研究

doi:10.13560/j.cnki.biotech.bull.1985.2025-1062

摘要/Abstract

摘要：

目的研究马铃薯CBF/DREB1家族成员与其耐盐性和耐旱性之间的关系，为培育优异抗逆性能的马铃薯新种质提供参考。方法利用RT-qPCR技术检测马铃薯CBF/DREB1家族成员在盐和干旱胁迫下的表达模式，筛选关键基因，通过构建其过表达和敲低表达的马铃薯转基因株系，检测该基因在盐和干旱胁迫应答中的功能，同时通过亚细胞定位检测和互作蛋白筛选，分析该基因的作用机制。结果基于RT-qPCR检测结果，选择StDREB4基因为研究对象进行功能分析。生长表型和理化指标检测结果显示，过表达株系具有更明显表型和指标变化。盐胁迫下，过表达StDREB4会降低植株根长、鲜重、根系活力、光合能力、抗氧化能力和渗透调节物质含量，以及提高MDA含量，从而降低马铃薯耐盐性。然而，干旱胁迫下，过表达StDREB4的植株相关指标优于野生型株系，能够增强马铃薯的耐旱性。此外，StDREB4定位于细胞核，且具有转录激活活性，利用酵母双杂筛选到3个重要的互作蛋白，包括EBF蛋白，DSK2A蛋白和PSMB3蛋白。结论过表达StDREB4可能通过泛素降解途径参与乙烯信号传导过程，调节多种理化指标参数，从而降低马铃薯耐盐性，增强耐旱性。

关键词: 马铃薯, 盐胁迫, 干旱胁迫, 理化参数, 酵母双杂交

Abstract:

Objective Studying the relationship between the CBF/DREB1 family members and the salt tolerance and drought tolerance may provide scientific references for breeding new potato germplasms with excellent stress resistance. Method The expression patterns of potato CBF/DREB1 family members under salt and drought stresses were detected by RT-qPCR technology, and the key gene was screened. Potato transgenic lines with overexpression and knockdown expression were constructed to detect the function of the key gene in response to salt and drought stresses. Meanwhile, the action mechanism of the key gene was analyzed by detecting subcellular localization and screening the interaction proteins. Result Based on the RT-qPCR detection results, the StDREB4 gene was selected as the research object for functional analysis. The results of growth phenotype and physicochemical index detection showed that the overexpressing lines had more obvious phenotypic and index changes. Under salt stress, the overexpression of StDREB4 reduced the root length, fresh weight, root activity, photosynthetic capacity, antioxidant capacity and osmotic regulatory substance content of plants, as well as increased MDA content, thereby reducing the tolerance of potatoes to salt. However, under drought stress, the relevant indicators of plants overexpressing StDREB4 were superior to those of wild-type lines, which enhanced the tolerance of potatoes to drought. In addition, StDREB4 was located in the nucleus and had transcriptional activation activity. Three important interacting proteins, including EBF protein, DSK2A protein and PSMB3 protein, were screened out using yeast two hybrid technology. Conclusion The overexpression of StDREB4 may be involved in the ethylene signaling process through the ubiquitin degradation pathway, regulating multiple physicochemical index parameters, thereby reducing the tolerance of potatoes to salt and enhancing its tolerance to drought.

Key words: Solanum tuberosum, salt stress, drought stress, physical and chemical parameters, yeast two hybrid

李万, 武亚倩, 吴芳, 赵永平. 马铃薯StDREB4基因响应盐和干旱胁迫的功能研究[J]. 生物技术通报, 2026, 42(6): 53-63.

LI Wan, WU Ya-qian, WU Fang, ZHAO Yong-ping. The Functional Study of StDREB4 Gene in Response to Salt and Drought Stress in Potato[J]. Biotechnology Bulletin, 2026, 42(6): 53-63.

图/表 9

图1 盐和干旱胁迫下StDREBs的表达模式A：盐胁迫。B：干旱胁迫。内参基因为Ef1α基因。定量数据由3个生物学重复和3个技术重复组成。“*”表示在P0.05水平上差异显著

Fig. 1 Expression patterns analysis of StDREBs under salt and drought stressesA: Salt stress. B: Drought stress. The internal reference gene is the Ef1α gene. The quantitative data consist of three biological replicates and three technical replicates. “*” indicates significant differences at the P0.05 level

图2 StDREB4对马铃薯在盐和干旱胁迫下生长的影响A：生长表型。B：株高增长、根长和鲜重积累。“*”表示与Desiree相比，转基因株系的参数值具有显著差异（P0.05），下同

Fig. 2 Effects of StDREB4 on the growth of potatoes under salt and drought stressesA: Growth phenotype. B: Plant height growth, root length and fresh weight accumulation. “*” indicates that the parameter values of transgenic lines are significantly different from those of Desiree (P0.05), the same below

图3 StDREB4对马铃薯在盐和干旱胁迫下光合特性指标的影响

Fig. 3 Effects of StDREB4 on the photosynthetic characteristic indicators of potatoes under salt and drought stresses

图4 StDREB4对马铃薯在盐和干旱胁迫下根系活力和MDA含量的影响

Fig. 4 Effects of StDREB4 on the root vitality and MDA content of potatoes under salt and drought stresses

图5 StDREB4对马铃薯在盐和干旱胁迫下抗氧化能力的影响

Fig. 5 Effects of StDREB4 on the antioxidant capacity of potatoes under salt and drought stresses

图6 StDREB4对马铃薯在盐和干旱胁迫下渗透调节物质含量的影响

Fig. 6 Effects of StDREB4 on the osmotic regulating substances contents of potatoes under salt and drought stresses

图7 StDREB4对马铃薯在盐和干旱胁迫下叶绿体色素含量的影响图中数据为原数据以10为底的对数

Fig. 7 Effects of StDREB4 on the chloroplast pigment contents of potatoes under salt and drought stressesThe data in the figure was taken as the logarithm of the original data with base 10

图8 StDREB4的亚细胞定位

Fig. 8 Subcellular localization of StDREB4

图9 StDREB4的自激活检测和互作蛋白筛选A：StDREB4转录激活活性分析。B：显色反应。黑色箭头表示代表性显色菌斑。C：PCR检测。M：DNA标记，从上至下依次为5 000、3 000、2 000、1 000、750、500、250和100 bp。D：StDREB4与互作蛋白的“点对点”验证。1-4表示不同浓度，依次为稀释1、10、100、1 000倍

Fig. 9 Self-activation detection and interacting proteins screening of StDREB4A: Analysis of transcriptional activation activity of StDREB4. B: Color reaction. The black arrows indicate representative chromogenic colony. C: PCR testing. M: DNA marker, from top to bottom, they are 5 000, 3 000, 2 000, 1 000, 750, 500, 250 and 100 bp, respectively. D: “Point-to-point” verification of StDREB4 and interacting proteins. 1 to 4 refer to different concentrations, which were diluted 1, 10, 100, and 1 000 times in sequence

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