褪黑素增强红小豆芽期耐盐碱能力的机制解析

doi:10.7668/hbnxb.20196393

摘要/Abstract

摘要：

土壤中过量的盐碱会抑制红小豆植株的正常生长发育,造成产量和品质的下降。芽期是植物生命周期中最脆弱的阶段之一,提高芽期的耐盐碱性对于确保整个生长周期内作物的健康生长尤为重要。探究褪黑素对盐碱胁迫下芽期红小豆的缓解机制,选用红小豆品种龙小豆5号作为试验材料。试验设置了以蒸馏水为对照(CK),以及3个处理组,分别为混合盐碱溶液SA和褪黑素溶液MT以及MT+SA(在SA基础上添加浓度为100 μmol/L的褪黑素溶液)。与SA相比,MT+SA处理能够显著促进红小豆芽期的生长。红小豆的抗氧化酶活性、可溶性蛋白以及可溶性糖含量也显著增加。红小豆中ROS含量、MDA含量、H₂O₂含量显著降低。在转录组学分析方面,SA与对照(CK)对比、MT+SA与SA对比,分别产生了9 111,4 662个差异表达基因(DEGs)。CK vs SA处理组中有4 487个上调基因和4 624个下调基因;MT+SA与SA处理组中有1 800个上调基因和2 862个下调基因。对差异基因进行GO富集分析发现,主要富集在代谢过程、生物调节、转录调节活性、转运活性、催化活性等方面;通过KEGG富集分析确定的主要通路为map00940、map00941和map00943。对这3个通路中识别出的差异基因进行qRT-PCR验证,结果显示,qRT-PCR与RNA-seq的表达趋势一致,从而证实了RNA-seq数据的可靠性。综上所述,基于表型、生理、转录组数据以及qRT-PCR数据的研究结果,发现褪黑素对芽期红小豆在抵御盐碱胁迫时有显著作用。

关键词: 芽期红小豆, 盐碱胁迫, 外源褪黑素, 表型指标, 生理指标, 转录组数据

Abstract:

Excessive salt-alkali in the soil will inhibit the growth of adzuki bean plants,resulting in a decrease in yield and quality.The bud stage is one of the most vulnerable stages in the plant life cycle,and improving salt and alkali tolerance during the bud stage is particularly important to ensure the healthy growth of crops throughout the entire growth cycle.This study focuses on exploring the alleviation mechanism of melatonin on sprouting red beans under salt alkali stress,using the red bean variety Longxiaodou 5 as the experimental material.The experiment was conducted with distilled water as the control(CK)and three treatment groups,namely SA(mixed saline alkali solution),MT(melatonin solution),and MT+SA(melatonin solution with a concentration of 100 μmol/L added to SA).Compared with SA,MT+SA treatment can significantly promote the growth of red bean sprouts.The antioxidant enzyme activity,soluble protein,and soluble sugar content of red beans were significantly enhanced.The content of ROS,MDA,and H₂O₂ of red beans were significantly reduced.In terms of transcriptome analysis,a comparison between SA and CK,and a comparison between MT+SA and SA,resulted in 9 111 and 4 662 differentially expressed genes(DEGs),respectively.There were 4 624 upregulated genes and 4 487 downregulated genes in the SA vs CK treatment group;there were 1 800 upregulated genes and 2 862 downregulated genes in the MT+SA and SA treatment groups.GO enrichment analysis of differentially expressed genes revealed that they were mainly enriched in metabolic processes,biological regulation,transcriptional regulation activity,transport activity,catalytic activity,and other aspects;the main pathways identified through KEGG enrichment analysis were map00940,map00941,and map00943.The differential genes identified in these three pathways were validated by qRT-PCR,and the results showed that the expression trend of qRT-PCR was consistent with that of RNA-seq,thus confirming the reliability of RNA-seq data.In summary,based on phenotype,physiology,transcriptome data,and qRT-PCR data,this study found that melatonin has a significant effect on the resistance of sprouting red beans to salt alkali stress.

Key words: Adzuki bean at bud stage, Salt and alkali stress, Exogenous melatonin, Phenotypicindex, Physiological index, Transcriptome data

中图分类号:

徐博, 段柯韩, 陈博源, 余德臻, 张琦, 杜吉到. 褪黑素增强红小豆芽期耐盐碱能力的机制解析[J]. 华北农学报, 2026, 41(4): 132-139. doi: 10.7668/hbnxb.20196393.

XU Bo, DUAN Kehan, CHEN Boyuan, YU Dezhen, ZHANG Qi, DU Jidao. Mechanistic Analysis of Melatonin-Enhanced Salt-Alkali Tolerance in Adzuki Bean Seedlings[J]. Acta Agriculturae Boreali-Sinica, 2026, 41(4): 132-139. doi: 10.7668/hbnxb.20196393.

http://www.hbnxb.net/CN/Y2026/V41/I4/132

图/表 10

图1 外源褪黑素对盐碱胁迫下芽期红小豆生长指标的影响图中不同字母代表差异显著(P<0.05)。图2—5,9同。

Fig.1 The effect of exogenous melatonin on growth indicators of sprouting adzuki bean under saline alkali stress Different letters in the figure represent significant differences(P<0.05).The same as Fig.2—5,9.

图2 外源褪黑素对盐碱胁迫下芽期红小豆抗氧化酶活性影响

Fig.2 Effect of exogenous melatonin on antioxidant enzyme activity of adzuki bean during germination under salt alkali stress

图3 外源褪黑素对盐碱胁迫下芽期红小豆ROS含量影响

Fig.3 The effect of exogenous melatonin on ROS content in sprouting adzuki beans under salt alkali stress

图4 外源褪黑素对盐碱胁迫下芽期红小豆渗透调节物质含量的影响

Fig.4 The effect of exogenous melatonin on the content of osmoregulatory substances in sprouting adzuki beans under salt alkali stress

图5 外源褪黑素对盐碱胁迫下芽期红小豆细胞膜的影响

Fig.5 The effect of exogenous melatonin on the cell membrane of adzuki bean during germination under salt alkali stress

图6 芽期红小豆表达差异分析

Fig.6 Statistical analysis of differential expression in adzuki bean during the germination stage

图7 SA vs CK和MT+SA vs SA差异表达基因GO注释分类 1.结构分子活性;2.蛋白质具有分子传导活性;3.分子功能调节活性;4.ATP依赖性活性;5.转录调节活性;6.转运活性;7.催化活性;8.结合活性;9.蛋白质复合物;10.细胞解剖实体;11.生殖过程;12.内稳定过程;13.多细胞生物过程;14.生物种间相互作用的生物过程;15.发育过程;16.定位;17.刺激反应;18.生物调节;19.代谢过程;20.细胞进程。

Fig.7 Annotation classification of SA vs CK and MT+SA vs SA differentially expressed genes 1.Structural molecule activity;2.Molecular transducer activity;3.Molecular function regulator activity;4.ATP-dependent activity;5.Transcription regulator activity;6.Transporter activity;7.Catalytic activity;8.Binding;9.Protein-containing complex;10.Cellular anatomical entity;11.Multicellular organismal process;12.Reproductive process;13.Homeostatic process;14.Biological process involved in interspecies interaction between organisms;15.Developmental process;16.Localization;17.Response to stimulus;18.Biological regulation;19.Metabolic process;20.Cellular process.

图8 SA vs CK和MT+SA vs SA差异基因KEGG分析 1.异黄酮生物合成;2.苯丙素生物合成;3.马达蛋白;4.类黄酮生物合成;5.MAPK信号通路-植物;6.植物-病原体相互作用;7.α-亚麻酸代谢;8.半乳糖代谢;9.色氨酸代谢;10.吲哚生物碱生物合成;11.泛醌和其他萜类醌生物合成;12.黄酮和黄酮醇生物合成;13.氰基氨基酸代谢;14.玉米素生物合成;15.单萜生物合成;16.丁酸代谢产物;17.萜类骨生物合成;18.ABC运输;19.缬氨酸、亮氨酸和异亮氨酸降解;20.亚油酸。

Fig.8 Circular plot of KEGG analysis of differentially expressed genes in SA vs CK and MT+SA vs SA 1.Isoflavonoid biosynthesis;2.Phenylpropanoid biosynthesis;3.Motor proteins;4.Flavonoid biosynthesis;5.MAPK signaling pathway-plant;6.Plant-pathogen interaction;7.Alpha-linolenic acid metabolism;8.Galactose metabolism;9.Tryptophan metabolism;10.Indole alkaloid biosynthesis;11.Ubiquinone and other terpenoid-quinone biosynthesis;12.Flavone and flavonol biosynthesis;13.Cyanoamino acid metabolism;14.Zeatin biosynthesis;15.Monoterpenoid biosynthesis;16.Butanoate metabolism;17.Terpenoid backbone biosynthesis;18.ABC transporters;19.Valine,leucine and isoleucine degradation;20.Linoleic acid metabolism.

图9 红小豆基因在盐碱胁迫以及褪黑素处理下的表达模式分析

Fig.9 Analysis of expression patterns of adzuki bean genes under salt alkali stress and melatonin treatment

图10 褪黑素分子与POD、4CL、CAD蛋白在第一位点进行对接

Fig.10 Docking of melatonin molecules with POD,4CL,and CAD proteins at the first site

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