NaCl胁迫下鹰嘴紫云英根系基因差异表达及相关通路分析

doi:10.11686/cyxb2024185

摘要/Abstract

摘要：

土壤盐渍化严重影响植物的生长和产量，限制农业生态经济的发展。鹰嘴紫云英作为优良生态草种、高效绿肥作物和优质蛋白饲草，探究其对NaCl胁迫适应性的分子机制，对提高鹰嘴紫云英的耐盐性，促进其在盐渍土中的栽培具有重要意义。本研究对NaCl胁迫下不同时间点（0、12、48和72 h）的鹰嘴紫云英根系进行转录组学分析，以探究鹰嘴紫云英对NaCl胁迫的响应机制。结果表明，与0 h相比，NaCl胁迫处理12、48和72 h后鹰嘴紫云英根系中分别有46051、45653和42869个差异表达基因，包括上调基因8027、10053和11042个，下调基因38024、35600和31827个。GO富集和KEGG富集显示，鹰嘴紫云英根系主要通过氧化还原酶活性、类黄酮生物合成和苯丙素生物合成等通路来响应NaCl胁迫。通过趋势分析，发现苯丙素生物合成和类黄酮生物合成通路的差异基因在胁迫处理48 h上调表达或恢复到初始量。C2H2、C3H、NAC、MYB、WRKY和bZIP等转录因子在胁迫处理不同时间点下均有表达，可能与鹰嘴紫云英的耐盐性密切相关。本研究结果为进一步探究鹰嘴紫云英耐盐机制提供了基础数据，也为后续提高鹰嘴紫云英的耐盐性提供了理论支撑。

关键词: 鹰嘴紫云英, 根系, 转录组, NaCl胁迫, 差异表达基因, 代谢通路

Abstract:

Soil salinization seriously affects the growth and yield of plants， and limits the development of agricultural and ecological economies. Astragalus cicer is a versatile ecological grass species， an efficient green manure crop， and a superior protein forage. Exploring the molecular mechanism of its adaptability to NaCl stress is of great significance for enhancing its salt tolerance and advancing its cultivation in saline soil. To investigate its responses to NaCl stress， transcriptomic analyses were performed on the roots of A. cicer at different time points （0， 12， 48， and 72 hours） during an NaCl treatment. The results showed that， compared with 0 hour， after 12， 48， and 72 hours of NaCl stress， there were， respectively， 46051， 45653， and 42869 differentially expressed genes （DEGs） in the roots of A. cicer， including 8027， 10053， and 11042 up-regulated DEGs and 38024， 35600， and 31827 down-regulated DEGs， respectively， at the three time points. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses showed that the pathways responding to NaCl stress in the roots of A. cicer were oxidoreductase activity， flavonoid biosynthesis， and phenylpropanoid biosynthesis. A trend analysis showed that the DEGs in phenylpropanoid biosynthesis and flavonoid biosynthesis pathway were still up-regulated or had returned to their baseline levels at 48 hours of NaCl treatment. Genes encoding C2H2， C3H， NAC， MYB， WRKY， and bZIP transcription factors were expressed at different time points during the NaCl treatment， suggesting that these transcription factors are closely related to salt tolerance in A. cicer. The results of this study provide basic data for further exploration of the mechanism of salt tolerance of A. cicer， and also provide theoretical support for breeding to improve salt tolerance.

Key words: Astragalus cicer, roots, transcriptome, NaCl stress, differentially expressed genes, metabolic pathway

马婷, 陈奋奇, 王勇, 哈雪, 李亚君, 马晖玲. NaCl胁迫下鹰嘴紫云英根系基因差异表达及相关通路分析[J]. 草业学报, 2025, 34(4): 104-123.

Ting MA, Fen-qi CHEN, Yong WANG, Xue HA, Ya-jun LI, Hui-ling MA. Differentially expressed genes and related pathways in root systems of Astragalus cicer under NaCl stress[J]. Acta Prataculturae Sinica, 2025, 34(4): 104-123.

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名称Name	描述Description	上游引物Forward primer	下游引物Reverse primer
CHI	查尔酮异构酶Chalcone isomerase	TGCACCACCGAGGAAATAGG	GGGTATGGCAGCAACAATGG
CHR1	查尔酮还原酶1 Chalcone reductase 1	AGCTCTTGGGAACAAACGTG	AGCTAGCAGGGGACCAAATG
F3'H	类黄酮3'-羟化酶Flavonoid 3'-hydroxylase	CGTCCAGCACCAAAGGGTAT	GCCATAGCTCGTGACCCAAA
CHS	查尔酮合成酶Chalcone synthase	CCAGGTGGTCCTGCAATTCT	GAGGACACAAGCACTCGACA
CHS1A-like	查尔酮合成酶1A-like Chalcone synthase 1A-like	TGTCACACATGCGTCTGAACT	CCTGCCACTGTCTTGGCAAT
CYP450	细胞色素P450 Cytochrome P450	ACGCATTTTGCGCCAATGAT	TGGCCACCCAAGTTCTACAC
C4H	反肉桂酸4单加氧酶Trans-cinnamate 4-monooxygenase	TGCGCACACACACAAAAGAG	CTCCACATGAGGGACCAACC
PYL	脱落酸感受器Abscisic acid receptor	TGATGATGAACGCCACGTCA	TCCATGCGGAACATCAACCA
bZIP	bZIP转录因子bZIP transcription factor	TATGGTCCTAGTTTGGCGGC	ACAACACCAGCTCTCACCAA
Actin	肌动蛋白Actin	TGTTCCCTGGCATTGCTGAT	CTTTCTCTCGGGTGGTGCAA

名称Name	描述Description	上游引物Forward primer	下游引物Reverse primer
CHI	查尔酮异构酶Chalcone isomerase	TGCACCACCGAGGAAATAGG	GGGTATGGCAGCAACAATGG
CHR1	查尔酮还原酶1 Chalcone reductase 1	AGCTCTTGGGAACAAACGTG	AGCTAGCAGGGGACCAAATG
F3'H	类黄酮3'-羟化酶Flavonoid 3'-hydroxylase	CGTCCAGCACCAAAGGGTAT	GCCATAGCTCGTGACCCAAA
CHS	查尔酮合成酶Chalcone synthase	CCAGGTGGTCCTGCAATTCT	GAGGACACAAGCACTCGACA
CHS1A-like	查尔酮合成酶1A-like Chalcone synthase 1A-like	TGTCACACATGCGTCTGAACT	CCTGCCACTGTCTTGGCAAT
CYP450	细胞色素P450 Cytochrome P450	ACGCATTTTGCGCCAATGAT	TGGCCACCCAAGTTCTACAC
C4H	反肉桂酸4单加氧酶Trans-cinnamate 4-monooxygenase	TGCGCACACACACAAAAGAG	CTCCACATGAGGGACCAACC
PYL	脱落酸感受器Abscisic acid receptor	TGATGATGAACGCCACGTCA	TCCATGCGGAACATCAACCA
bZIP	bZIP转录因子bZIP transcription factor	TATGGTCCTAGTTTGGCGGC	ACAACACCAGCTCTCACCAA
Actin	肌动蛋白Actin	TGTTCCCTGGCATTGCTGAT	CTTTCTCTCGGGTGGTGCAA

样本 Sample	原始读数 Raw reads	原始数据量 Raw bases （Gb）	干净读数 Clean reads	干净数据量 Clean bases （Gb）	Q30 （%）	GC （%）	总对比率 Total mapped （%）
r_0_1	25608576	7.68	24842810	7.45	95.58	42.49	70.68
r_0_2	22654911	6.80	22160067	6.65	95.77	42.37	71.75
r_0_3	23401444	7.02	22736208	6.82	95.45	42.27	70.54
r_12_1	24325553	7.30	23233562	6.97	95.25	42.27	69.73
r_12_2	23471096	7.04	22839301	6.85	95.95	40.19	65.84
r_12_3	21972596	6.59	21124167	6.34	95.28	42.35	70.40
r_48_1	22903659	6.87	21974443	6.59	95.51	41.66	69.90
r_48_2	23716748	7.12	22832712	6.85	95.50	41.74	70.40
r_48_3	23380813	7.01	22673020	6.80	95.88	41.64	70.86
r_72_1	23445027	7.03	22237254	6.67	95.73	41.95	70.89
r_72_2	23476556	7.04	22400966	6.72	95.93	41.81	71.57
r_72_3	27057092	8.12	26181124	7.85	94.97	41.80	67.48

样本 Sample	原始读数 Raw reads	原始数据量 Raw bases （Gb）	干净读数 Clean reads	干净数据量 Clean bases （Gb）	Q30 （%）	GC （%）	总对比率 Total mapped （%）
r_0_1	25608576	7.68	24842810	7.45	95.58	42.49	70.68
r_0_2	22654911	6.80	22160067	6.65	95.77	42.37	71.75
r_0_3	23401444	7.02	22736208	6.82	95.45	42.27	70.54
r_12_1	24325553	7.30	23233562	6.97	95.25	42.27	69.73
r_12_2	23471096	7.04	22839301	6.85	95.95	40.19	65.84
r_12_3	21972596	6.59	21124167	6.34	95.28	42.35	70.40
r_48_1	22903659	6.87	21974443	6.59	95.51	41.66	69.90
r_48_2	23716748	7.12	22832712	6.85	95.50	41.74	70.40
r_48_3	23380813	7.01	22673020	6.80	95.88	41.64	70.86
r_72_1	23445027	7.03	22237254	6.67	95.73	41.95	70.89
r_72_2	23476556	7.04	22400966	6.72	95.93	41.81	71.57
r_72_3	27057092	8.12	26181124	7.85	94.97	41.80	67.48

项目Item	r_12 vs r_0	r_48 vs r_0	r_72 vs r_0	r_48 vs r_12	r_72 vs r_12	r_72 vs r_48
上调 Up-regulated	8027	10053	11042	11821	7633	4653
下调 Down-regulated	38024	35600	31827	5515	3768	4345
总数 Total	46051	45653	42869	17336	11401	8998