盐胁迫诱导木豆幼苗形态、生理及黄酮代谢的多层次响应特征

doi:10.7525/j.issn.1673-5102.2026.04.013

摘要/Abstract

摘要：

为系统阐明不同浓度盐胁迫条件下木豆（Cajanus cajan）幼苗的生长响应特征及其抗氧化酶活性与黄酮类代谢物积累的动态变化规律，采用生长一致的木豆幼苗作为材料，设置0、50、100、150、200 mmol⋅L^-1 NaCl处理，在处理后1、3、5、7 d进行表型观察及根系扫描分析，测定根长、表面积、投影面积、体积及平均直径等根系形态指标，同时测定超氧化物歧化酶（SOD）和过氧化物酶（POD）活性，并在150 mmol⋅L^-1 NaCl处理条件下选取不同时间点样品，利用超高效液相色谱-串联质谱（UPLC-MS/MS）技术对黄酮类代谢物进行定性与定量分析，并结合相关基因表达变化进行分析。结果表明：盐胁迫对木豆幼苗生长具有显著的浓度依赖性抑制作用。随着盐浓度升高及处理时间延长，幼苗表现为主根伸长受阻、侧根发育受限及地上部生长减缓，且在处理3 d后抑制效应明显增强。根系形态指标整体随时间呈增长趋势，但在100 mmol⋅L^-1及更高盐浓度处理下增长幅度明显降低，50 mmol⋅L^-1处理在部分观测时期对根系生长表现出一定促进作用；根系平均直径变化幅度较小，但在中高盐浓度条件下略有增加。SOD活性在中等盐浓度下显著升高并趋于稳定，POD活性随盐浓度增加及处理时间延长持续上升。在150 mmol⋅L^-1 NaCl处理条件下，根系黄酮类代谢物呈现阶段性积累特征，木豆芪酸质量浓度整体呈上升趋势，异牡荆素质量浓度波动变化明显，染料木苷质量浓度呈诱导积累趋势，染料木素质量浓度在处理后期显著增加。基因表达分析结果显示，CcPAL2、Cc4CL4及CcCHS3表达量降低，CcC4H1表达显著上调，CcCHI2与CcIFS1表达量升高。研究表明，盐胁迫条件下，木豆幼苗生长受到抑制并表现出明显的时间与浓度效应，抗氧化酶活性及黄酮类代谢物含量随胁迫过程呈动态变化，黄酮生物合成通路关键基因表达存在差异性调控。

关键词: 木豆, 盐胁迫, 根系形态构型, 黄酮类次生代谢产物

Abstract:

To systematically elucidate the growth response characteristics of pigeon pea（Cajanus cajan） seedlings under different concentrations of salt stress and the dynamic changes in antioxidant enzyme activities and flavonoid metabolite accumulation， uniformly grown pigeon pea seedlings were used as plant materials. The seedlings were treated with 0， 50， 100， 150， and 200 mmol⋅L^-1 NaCl solution. Phenotypic observation and root scanning analysis were conducted at 1， 3， 5， and 7 days after treatment. Root morphological parameters， including root length， surface area， projected area， volume， and average diameter， were measured. Meanwhile， the activities of superoxide dismutase（SOD） and peroxidase（POD） were determined. In addition， samples collected under 150 mmol⋅L^-1 NaCl treatment at different time points were analyzed using UPLC-MS/MS for qualitative and quantitative profiling of flavonoid metabolites， combined with analysis of related gene expression changes. Results showed that salt stress significantly inhibited the growth of pigeon pea seedlings in a concentration-dependent manner. With increasing salt concentration and prolonged treatment duration， seedlings exhibited inhibited primary root elongation， restricted lateral root development， and reduced shoot growth， with the inhibitory effect becoming more pronounced after 3 days of treatment. Root morphological parameters generally increased over time； however， the magnitude of increase was significantly reduced under 100 mmol⋅L^-1 and higher NaCl treatments， whereas 50 mmol⋅L^-1 NaCl showed a promoting effect on root growth at certain time points. The average root diameter showed relatively small variation under 50 mmol⋅L^-1 NaCl treatment but slight increase under moderate to high salt conditions. SOD activity increased significantly under moderate salt concentration and then stabilized， while POD activity continuously increased with increasing salt concentration and treatment duration. Under 150 mmol⋅L^-1 NaCl treatment， flavonoid metabolites in roots exhibited stage-dependent accumulation patterns. Root stilbene acid mass concentration showed an overall increasing trend， isovitexin mass concentration fluctuated markedly， genistin displayed induced accumulation， and genistein mass concentration significantly increased at later stages of treatment. Gene expression analysis showed that the expression levels of CcPAL2， Cc4CL4， and CcCHS3 were downregulated， while CcC4H1 was significantly upregulated， and CcCHI2 and CcIFS1 were also induced. In conclusion， salt stress inhibited the growth of pigeon pea seedlings and showed clear time- and concentration-dependent effects. Antioxidant enzyme activities and flavonoid metabolite levels dynamically changed during the stress process， and the key genes in the flavonoid biosynthesis pathway exhibited differential regulation under salt stress conditions.

Key words: pigeon pea, salt stress, root system architecture, flavonoid secondary metabolites

中图分类号:

Q945

姚耀源, 杨杰, 张悦新, 付玉杰. 盐胁迫诱导木豆幼苗形态、生理及黄酮代谢的多层次响应特征[J]. 植物研究, 2026, 46(4): 728-739.

Yaoyuan YAO, Jie YANG, Yuexin ZHANG, Yujie FU. Morpho-Physiological and Metabolic Responses and Expression Profiling of the CcTCP Gene Family in Pigeon Pea under Salt Stress[J]. Bulletin of Botanical Research, 2026, 46(4): 728-739.

图/表 7

表1

图1

图2

图3

图4

图5

图6

参考文献 29

[1]	Hassani A， Azapagic A， Shokri N.Global predictions of primary soil salinization under changing climate in the 21st century［J］.Nature Communications，2021，12：6663.
[2]	Shokri N， Hassani A， Sahimi M.Multi-scale soil salinization dynamics from global to pore scale：a review［J］.Reviews of Geophysics，2024，62（4）：e2023RG000804.
[3]	Zhou H P， Shi H F， Yang Y Q，et al.Insights into plant salt stress signaling and tolerance［J］.Journal of Genetics and Genomics，2024，51（1）：16-34.
[4]	Singh D.Juggling with reactive oxygen species and antioxidant defense system：a coping mechanism under salt stress［J］.Plant Stress，2022，5：100093.
[5]	Wang H Q， Sun J， Ren H，et al.Inhibiting reactive oxygen species production mitigates endoplasmic reticulum damage in florets of developing maize ears under heat stress［J］.The Plant Journal，2025，122（5）：e70243.
[6]	Lu X， Chen G P， Ma L，et al.Integrated transcriptome and metabolome analysis reveals antioxidant machinery in grapevine exposed to salt and alkali stress［J］.Physiologia Plantarum，2023，175（3）：e13950.
[7]	Zhang X Y， He P Y， Guo R Y，et al.Effects of salt stress on root morphology，carbon and nitrogen metabolism，and yield of Tartary buckwheat［J］.Scientific Reports，2023，13：12483.
[8]	Zhang L， Zhang Z J， Fang S Z，et al.Integrative analysis of metabolome and transcriptome reveals molecular regulatory mechanism of flavonoid biosynthesis in Cyclocarya paliurus under salt stress［J］.Industrial Crops and Products，2021，170：113823.
[9]	Wang C L， Wei X T， Wang Y M，et al.Metabolomics and transcriptomic analysis revealed the response mechanism of maize to saline-alkali stress［J］.Plant Biotechnology Journal，2025，23（12）：5397-5416.
[10]	Su L T， Lv A M， Wen W W，et al.MsMYB206-MsMYB450-MsHY5 complex regulates alfalfa tolerance to salt stress via regulating flavonoid biosynthesis during the day and night cycles［J］.The Plant Journal，2025，121（2）：e17216.
[11]	Zheng X Y， Zhang X J， Zeng F K.The regulatory network composed of phytohormones，transcription factors and non-coding RNAs is involved in the flavonoids biosynthesis of fruits［J］.Horticultural Plant Journal，2026，12（3）：497-508.
[12]	Liu C， Tai Y L， Luo J J，et al.Integrated multi-omics analysis provides insights into genome evolution and phosphorus deficiency adaptation in pigeonpea （Cajanus cajan）［J］.Horticulture Research，2022，9：uhac107.
[13]	Awana M， Jain N， Samota M K，et al.Protein and gene integration analysis through proteome and transcriptome brings new insight into salt stress tolerance in pigeonpea （Cajanus cajan L.）［J］.International Journal of Biological Macromolecules，2020，164：3589-3602.
[14]	Du T T， Fan Y X， Cao H Y，et al.Transcriptome analysis revealed key genes involved in flavonoid metabolism in response to jasmonic acid in pigeon pea （Cajanus cajan （L.） Millsp.）［J］.Plant Physiology and Biochemistry，2021，168：410-422.
[15]	Yang W L， Li N， Fan Y X，et al.Transcriptome analysis reveals abscisic acid enhancing drought resistance by regulating genes related to flavonoid metabolism in pigeon pea［J］.Environmental and Experimental Botany，2021，191：104627.
[16]	Meng D， Dong B Y， Niu L L，et al.The pigeon pea CcCIPK14-CcCBL1 pair positively modulates drought tolerance by enhancing flavonoid biosynthesis［J］.The Plant Journal，2021，106（5）：1278-1297.
[17]	Giannopolitis C N， Ries S K.Superoxide dismutases：Ⅰ.occurrence in higher plants［J］.Plant Physiology，1977，59（2）：309-314.
[18]	Liu C X， Gu C Y， Huang W，et al.Targeted UPLC-MS/MS high-throughput metabolomics approach to assess the purine and pyrimidine metabolism［J］.Journal of Chromatography B，2019，1113：98-106.
[19]	Livak K J， Schmittgen T D.Analysis of relative gene expression data using real-time quantitative PCR and the 2^-ΔΔCT method［J］.Methods，2001，25（4）：402-408.
[20]	Khan F， Siddiqi T O，Mahmooduzzafar，et al.Morphological changes and antioxidant defence systems in soybean genotypes as affected by salt stress［J］.Journal of Plant Interactions，2009，4（4）：295-306.
[21]	Lan J Q， Qin G J.The regulation of CIN-like TCP transcription factors［J］.International Journal of Molecular Sciences，2020，21（12）：4498.
[22]	Gill S S， Tuteja N.Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants［J］.Plant Physiology and Biochemistry，2010，48（12）：909-930.
[23]	Passardi F， Penel C， Dunand C.Performing the paradoxical：how plant peroxidases modify the cell wall［J］.Trends in Plant Science，2004，9（11）：534-540.
[24]	Mittova V， Tal M， Volokita M，et al.Salt stress induces up-regulation of an efficient chloroplast antioxidant system in the salt-tolerant wild tomato species Lycopersicon pennellii but not in the cultivated species［J］.Physiologia Plantarum，2002，115（3）：393-400.
[25]	Gharsallah C， Fakhfakh H， Grubb D，et al.Effect of salt stress on ion concentration，proline content，antioxidant enzyme activities and gene expression in tomato cultivars［J］.AoB Plants，2016，8：plw055.
[26]	Demiral T， Türkan İ.Comparative lipid peroxidation，antioxidant defense systems and proline content in roots of two rice cultivars differing in salt tolerance［J］.Environmental and Experimental Botany，2005，53（3）：247-257.
[27]	Mittler R， Zandalinas S I， Fichman Y，et al.Reactive oxygen species signalling in plant stress responses［J］.Nature Reviews Molecular Cell Biology，2022，23（10）：663-679.
[28]	Miransari M， Smith D L.Alleviating salt stress on soybean （Glycine max （L.） Merr.）-Bradyrhizobium japonicum symbiosis，using signal molecule genistein［J］.European Journal of Soil Biology，2009，45（2）：146-152.
[29]	Wang Y B， Liu W， Li W，et al.Integrative analysis of metabolome and transcriptome reveals regulatory mechanisms of flavonoid biosynthesis in soybean under salt stress［J］.Frontiers in Plant Science，2024，15：1415867.

基因名称 Gene name	上游引物（5′→3′） Forward primer（5′→3′）	下游引物（5′→3′） Reverse primer（5′→3′）
qCcActin	AAGCTCAGTCCAAGAGAGGT	ACATGGCGGGGGTATTGAAG
qCcPAL2	TGAACAACGGGACCGACAGT	AAAACCTGATGAGTTCCTTCTGC
qCc4CL4	GCTGACGCACAAGAGTCTGATG	CAGCAACGCCCTAATCTCAA
qCcC4H1	CCTATGGTCCCGTTTTCCTACT	TGAGTGGAGGACATGTGTGGC
qCcCHS3	ACGCCAGGCAGGACATAGTA	ACTTTGGTTGACCCCATTCC
qCcCHI2	CCTCCGCCAAGACCTATTTC	CGAGTGATTGGATGGCTTGA
qCcIFS1	AGCCCTTCGCCACCTACCTAA	GGGAATAGTGGAGAAGGGGTTT