Identification of Plasma Membrane H+-ATPase Gene Family in Safflower and Expression Analysis in Response to Low Nitrogen and Low Phosphorus Stress

doi:10.13560/j.cnki.biotech.bull.1985.2025-0057

Abstract

Abstract:

Objective Genome-wide identification and analysis of the plasma membrane H⁺-ATPase (PMA) gene family in safflower (Carthamus tinctorius L.) may provide the basis for investigating this gene family and its function in response to low nitrogen and low phosphorus stress. Method Bioinformatics was used to identify and systematically analyze the members of the CtPMA gene family. Real-time fluorescence quantitative PCR (RT-qPCR) was applied to analyze the tissue-specific expression pattens and the responses to low nitrogen and low phosphorus stress. Result A total of 10 CtPMAs (named CtPMA1-CtPMA10) were identified in safflower, encoding proteins ranging from 785 to 958 aa, with relative molecular weight of 85.23 to 105.52 kD, and isoelectric points of 5.26 to 7.91. All were predicted to be localized to the plasma membrane by the subcellular localization. All members contained conserved domains, including the Cation_ATPase_N domain, the E1_E2 ATPase domain, and the HAD-superfamily hydrolase subfamily ⅢA domain. Cis-acting element analysis revealed a lot of regulatory elements related to growth, development, hormones and stress responses. Gene collinearity indicated that whole-genome duplication and segmental duplication played key roles in the evolution of the CtPMA gene family. RT-qPCR results showed that the CtPMA1 and CtPMA7, which were highly expressed in the roots, were significantly induced by low nitrogen and low phosphorus stress. This finding suggests that CtPMA1 and CtPMA7 may play a crucial role in the process of nitrogen and phosphorus uptake in safflower roots. Conclusion A total of 10 CtPMAs members are identified in the safflower genome.These members demonstrate evolutionary conservation and tissue-specific expression patterns. The highly expressed CtPMA1 and CtPMA7 transcripts in the roots were significantly induced under low nitrogen and phosphorus stress. This suggests that safflower may improve the absorption and utilization efficiency of nitrogen and phosphorus by increasing its CtPMAs transcription level in order to resist nutritional stress.

Key words: Carthamus tinctorius L., plasma membrane H⁺-ATPase gene family, bioinformatics analysis, low nitrogen stress, low phosphorus stress

LA Gui-xiao, ZHAO Yu-long, DAI Dan-dan, YU Yong-liang, GUO Hong-xia, SHI Gui-xia, JIA Hui, YANG Tie-gang. Identification of Plasma Membrane H⁺-ATPase Gene Family in Safflower and Expression Analysis in Response to Low Nitrogen and Low Phosphorus Stress[J]. Biotechnology Bulletin, 2025, 41(8): 220-233.

Figures/Tables 12

References 27

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基因 Gene	正向引物 Forward primer （5'-3'）	反向引物 Reverse primer （5'-3'）	长度Length （bp）
CtPMA1	TTGACCAATCTGCCCTCA	CCTTCTGGAAATGCCCAA	189
CtPMA2	CCACCAGATTGGCAAGACT	GGCATCAGCAGGAACGAT	231
CtPMA3	AGATGCCGCCGTGTTAGT	GTCAAAGCCGACTGGTCAA	117
CtPMA4	AGGGATGGGAGATGGTCTG	GCTTCTTGGTGACTGGTAAGG	169
CtPMA5	TGACGACTGATGATGGTAACC	CAATCTGGAGGCTTTCCC	181
CtPMA6	GCAGCCATTCTCGTTCCA	AGGGTGCTTTGTCACAGGA	144
CtPMA7	GGAATCCTCTCTCGTGGGT	GCAAGCATCTTCTTCGCTC	242
CtPMA8	GGTGCTTCTTCTCATAAACTCG	ACGATGTCTCCTGGAACCA	162
CtPMA9	TGAAGGCATTGATAACCTGC	ACACCAGCGGCAAATACC	246
CtPMA10	GGCGAGTCTATTCCAGTAACC	GATTGGTGTTCTCCACAAGG	148
18S	ACACGGGGAGGTAGTGACAA	CCTCCAATGGATCCTCGTTA	135

基因 Gene	正向引物 Forward primer （5'-3'）	反向引物 Reverse primer （5'-3'）	长度Length （bp）
CtPMA1	TTGACCAATCTGCCCTCA	CCTTCTGGAAATGCCCAA	189
CtPMA2	CCACCAGATTGGCAAGACT	GGCATCAGCAGGAACGAT	231
CtPMA3	AGATGCCGCCGTGTTAGT	GTCAAAGCCGACTGGTCAA	117
CtPMA4	AGGGATGGGAGATGGTCTG	GCTTCTTGGTGACTGGTAAGG	169
CtPMA5	TGACGACTGATGATGGTAACC	CAATCTGGAGGCTTTCCC	181
CtPMA6	GCAGCCATTCTCGTTCCA	AGGGTGCTTTGTCACAGGA	144
CtPMA7	GGAATCCTCTCTCGTGGGT	GCAAGCATCTTCTTCGCTC	242
CtPMA8	GGTGCTTCTTCTCATAAACTCG	ACGATGTCTCCTGGAACCA	162
CtPMA9	TGAAGGCATTGATAACCTGC	ACACCAGCGGCAAATACC	246
CtPMA10	GGCGAGTCTATTCCAGTAACC	GATTGGTGTTCTCCACAAGG	148
18S	ACACGGGGAGGTAGTGACAA	CCTCCAATGGATCCTCGTTA	135

基因名称 Gene name	基因号 Gene ID	蛋白质长度 Protein length （aa）	相对分子质量 Molecular mass （kD）	等电点 Theoretical point	亲水性系数 Hydrophilicity coefficient	不稳定系数 Instability index	亚细胞定位 Subcellular localization
CtPMA1	CtAH01T0012700.1	886	97.78	5.75	0.034	37.94	细胞膜 Plasma membrane
CtPMA2	CtAH03T0170800.1	937	102.76	6.63	0.054	39.45	细胞膜 Plasma membrane
CtPMA3	CtAH05T0036500.1	858	93.95	5.26	0.205	39.82	细胞膜 Plasma membrane
CtPMA4	CtAH05T0146800.1	956	105.47	6.49	0.069	33.32	细胞膜 Plasma membrane
CtPMA5	CtAH05T0252100.1	785	85.23	5.34	0.150	36.06	细胞膜 Plasma membrane
CtPMA6	CtAH06T0267900.1	906	99.49	6.20	0.034	33.52	细胞膜 Plasma membrane
CtPMA7	CtAH09T0032200.1	911	100.36	5.98	0.122	35.71	细胞膜 Plasma membrane
CtPMA8	CtAH09T0161100.1	957	105.49	6.03	0.116	38.33	细胞膜 Plasma membrane
CtPMA9	CtAH11T0113500.1	889	97.10	5.87	0.068	36.90	细胞膜 Plasma membrane
CtPMA10	CtAH12T0151100.1	958	105.52	7.91	0.087	36.00	细胞膜 Plasma membrane

基因名称 Gene name	基因号 Gene ID	蛋白质长度 Protein length （aa）	相对分子质量 Molecular mass （kD）	等电点 Theoretical point	亲水性系数 Hydrophilicity coefficient	不稳定系数 Instability index	亚细胞定位 Subcellular localization
CtPMA1	CtAH01T0012700.1	886	97.78	5.75	0.034	37.94	细胞膜 Plasma membrane
CtPMA2	CtAH03T0170800.1	937	102.76	6.63	0.054	39.45	细胞膜 Plasma membrane
CtPMA3	CtAH05T0036500.1	858	93.95	5.26	0.205	39.82	细胞膜 Plasma membrane
CtPMA4	CtAH05T0146800.1	956	105.47	6.49	0.069	33.32	细胞膜 Plasma membrane
CtPMA5	CtAH05T0252100.1	785	85.23	5.34	0.150	36.06	细胞膜 Plasma membrane
CtPMA6	CtAH06T0267900.1	906	99.49	6.20	0.034	33.52	细胞膜 Plasma membrane
CtPMA7	CtAH09T0032200.1	911	100.36	5.98	0.122	35.71	细胞膜 Plasma membrane
CtPMA8	CtAH09T0161100.1	957	105.49	6.03	0.116	38.33	细胞膜 Plasma membrane
CtPMA9	CtAH11T0113500.1	889	97.10	5.87	0.068	36.90	细胞膜 Plasma membrane
CtPMA10	CtAH12T0151100.1	958	105.52	7.91	0.087	36.00	细胞膜 Plasma membrane