钙调素结合蛋白StIQD20介导马铃薯耐盐性的功能研究

doi:10.13560/j.cnki.biotech.bull.1985.2025-1346

摘要/Abstract

摘要：

目的揭示马铃薯StIQD20基因在盐胁迫应答中的功能，明确其增强马铃薯耐盐性的内在机制，为马铃薯耐盐分子育种提供新的基因资源和遗传材料。方法克隆StIQD20基因，构建过表达载体，转化二倍体野生型马铃薯‘C65’获得阳性植株，对比分析不同盐胁迫下过表达植株与野生型植株的表型性状、生理生化指标，进行Ca²⁺表型恢复实验与盐胁迫相关基因表达量测定。结果获得5棵过表达植株，RT-qPCR检测表明过表达植株StIQD20基因表达量为野生型植株的1.5-2.9倍；盐胁迫下过表达植株的株高、鲜重及叶片数均高于野生型，在盐胁迫条件下，施加100 μmol/L CaCl₂较50 μmol/L能有效地缓解幼苗受害程度；不同盐胁迫浓度处理下，过表达植株的抗氧化酶活性与脯氨酸含量显著高于野生型，丙二醛含量显著低于野生型；过表达植株中盐胁迫响应基因SnRK2s、ABF4、ABI3、MYB2和DREB2A的表达水平均高于野生型，ABI5基因的表达量相对较低。结论 StIQD20通过调控Ca²⁺信号及抗氧化防御系统，促进渗透调节物质积累并调控相关基因表达，从而显著增强马铃薯的耐盐性。

关键词: 马铃薯, StIQD20, 愈伤组织转化体系, 盐胁迫, Ca²⁺

Abstract:

Objective This study aimed to elucidate the function of the potato StIQD20 gene in response to salt stress, clarify its intrinsic mechanism for enhancing potato salt tolerance, and provide new gene resources and genetic materials for molecular breeding of salt-tolerant potatoes. Method The StIQD20 gene was cloned, and an overexpression vector was constructed. The vector was transformed into the diploid wild-type potato ‘C65’ to obtain positive plants. Phenotypic traits and physiological and biochemical indicators of the overexpression plants and wild-type plants under different salt stress conditions were compared and analyzed. A Ca²⁺ phenotypic recovery experiment and expression level measurement of salt stress-related genes were conducted. Result Five overexpression plants were obtained. RT-qPCR analysis showed that the expression level of the StIQD20 gene in overexpression plants was 1.5-2.9 times higher than that in wild-type plants. Under salt stress, plant height, fresh weight, and number of blades of overexpression plants were higher than those of wild-type plants. The application of 100 μmol/L CaCl₂ was more effective than 50 μmol/L in alleviating seedling damage under salt stress conditions. Under different salt stress concentrations, the antioxidant enzyme activities and proline content in overexpression plants were significantly higher than those in wild-type plants, while malondialdehyde (MDA) content was significantly lower. The expression levels of salt stress-responsive genes SnRK2s, ABF4, ABI3, MYB2, and DREB2A in overexpression plants were higher than those in wild-type plants, while the expression level of the ABI5 gene was relatively lower. Conclusion StIQD20 significantly enhances salt tolerance in potato by modulating Ca²⁺ signaling and the antioxidant defense system, promoting the accumulation of osmoregulatory substances, and modulating the expression of related genes.

Key words: potato, StIQD20, callus transformation system, salt stress, Ca²⁺

冯瑞云, 李京昊, 王紫腾, 宋宇鹏, 侯雅静, 段永红, 宋倩娜, 梅超. 钙调素结合蛋白StIQD20介导马铃薯耐盐性的功能研究[J]. 生物技术通报, 2026, 42(10): 207-215.

FENG Rui-yun, LI Jing-hao, WANG Zi-teng, SONG Yu-peng, HOU Ya-jing, DUAN Yong-hong, SONG Qian-na, MEI Chao. Functional Study of the Calmodulin-binding Protein StIQD20 in Mediating Salt Tolerance in Potato[J]. Biotechnology Bulletin, 2026, 42(10): 207-215.

图/表 7

图1 马铃薯StIQD20过表达载体构建与基因表达分析A：1代表含目的基因StIQD20的pMDC85质粒；2代表Asc I-Pac I酶切质粒；M代表10 000 bp DNA marker。B：M代表2 000 bp DNA marker；N代表阴性对照；P代表阳性对照；1-6代表阳性菌斑。C：M代表2 000 bp DNA marker；CW代表野生型；P代表重组载体质粒；8、9、13、14、18代表过表达阳性植株。D：pMDC85-StIQD20重组载体图。E：C65代表二倍体野生型植株；OEs代表过表达阳性植株；显著性差异通过Duncan新复极差法确定；不同字母表示差异显著（P<0.05）；误差线代表平均值±SD，n=3，下同

Fig. 1 Construction of the overexpression vector of StIQD20 in potato and analysis of gene expressionA: 1 represents the pMDC85 plasmid containing the target gene StIQD20; 2 represents the plasmid digested by Asc I-Pac I; M represents the 10 000 bp DNA marker. B: M represents the 2 000 bp DNA marker; N represents the negative control; P represents the positive control; 1-6 represent the positive colonies. C: M represents the 2 000 bp DNA marker; CW represents the wild type; P represents the recombinant vector plasmid; 8, 9, 13, 14, 18 represent the overexpressing positive plants. D: The recombinant vector map of pMDC85-StIQD20. E: C65 represent a diploid wild-type plant. OEs represent the overexpressing positive plants; significant differences among groups were identified using Duncan’s new multiple range test; values followed by different lowercase letters are significantly different (P<0.05); error bars in the figures denote average value ± the standard deviation derived from three independent biological replicates, n=3, the same below

图2 马铃薯StIQD20基因过表达阳性植株遗传转化过程

Fig. 2 The genetic transformation process of positive plants overexpressing StIQD20 gene in potato

图3 盐胁迫处理30 d后野生型植株与过表达阳性植株表型

Fig. 3 Phenotypes between wild-type plants and overexpressing positive plants after 30 days of salt stress treatment

图4 盐胁迫下野生型植株与过表达阳性植株生长指标

Fig. 4 Growth indicators of wild-type plants and overexpressing positive plants under salt stress

图5 不同CaCl2浓度对盐胁迫下马铃薯幼苗生长发育的影响

Fig. 5 Effects of different CaCl2 concentrations on the growth and development of potato seedlings under salt stress

图6 不同氯化钠浓度下植株中抗氧化指标的变化

Fig. 6 Changes in antioxidant indicators in plants under different NaCl concentrations

图7 过表达StIQD20阳性植株中盐胁迫相关基因的表达量分析

Fig. 7 Analysis of the expression levels of salt stress-related genes in StIQD20-overexpressing positive plants

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