Expression analysis of AP2/ERFs genes in alfalfa regulated by exogenous NO under drought stress

doi:10.11686/cyxb2024277

Abstract

Abstract:

Alfalfa （Medicago sativa） is the most widely grown forage legume crop in the world. APETALA2/ethylene-responsive （AP2/ERF） transcription factors play a key role in plant resistance to abiotic stress. Nitric oxide （NO）， as a signaling molecule in plants， plays an important role in plant drought resistance. In this study， bioinformatics methods were used to identify members of the alfalfa MsAP2/ERF gene family and analyze their response patterns to NO and drought. The MsERF07 gene， which strongly responds to NO regulation， was screened from the MsAP2/ERF gene family for subcellular localization. The results showed that all members of the family contained AP2 domains， and the number of amino acids in their proteins ranged from 176 to 422. Subcellular localization predicted that most proteins were localized in the nucleus. MsERF01 and MsERF11 were closely related and had similar domains. 61.54% of MsAP2/ERF genes contained only exons and also had highly similar conserved motifs. Protein interaction showed that MsERF01 and MsERF11， MsERF05 and MsERF07 were all located at the same node in the protein interaction map. The 13 MsAP2/ERF gene family members were unevenly distributed on 13 chromosomes， and there were 43 cis-regulatory elements related to light response， tissue-specific expression， stress and plant hormones in the promoter sequences of the MsAP2/ERF gene family members. In addition， transcriptome sequencing data analysis of alfalfa showed that the expression of most MsAP2/ERF gene family members increased under the regulation of NO， and further qRT-RCR experimental results showed that exogenous NO promoted the expression of MsAP2/ERF genes under drought stress. The MsERF07 gene was cloned， and subcellular localization results showed that the protein was localized in the nucleus and cell membrane. This study provides a basis for subsequent research on the molecular mechanism of the alfalfa MsERF07 gene action in response to drought stress.

Key words: drought stress, nitric oxide, alfalfa, AP2/ERF

Xiao-Yue WEN, Ying ZHAO, Bao-qiang WANG, Xian WANG, Xiao-lin ZHU, Yi-zhen WANG, Xiao-hong WEI. Expression analysis of AP2/ERFs genes in alfalfa regulated by exogenous NO under drought stress[J]. Acta Prataculturae Sinica, 2025, 34(6): 154-167.

Figures/Tables 10

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登录号 Gene ID	基因名 Gene name	上游引物序列 Forward primer sequence	下游引物序列 Reverse primer sequence
MS.gene057432.t1	MsERF01	AGATTACCAAACAGCCGCCA	TAGCACCACTGCCACGTAAG
MS.gene022984.t1	MsERF02	AACCGAAGAGTGAACAGCCT	TCCTGCAAGGGTTGGTTGAA
MS.gene31024.t1	MsERF03	TCGTAACCCTCCACCAGCTA	CCTTGCAGAAGGAATACCCGA
MS.gene011550.t1	MsERF04	ATGCGTCAATGGGGCAAATG	TTGAGGTAAGCCGAAGAGCC
MS.gene30848.t1	MsERF05	TGCCACTACTCAGGCAAAGG	GGCGGCTATAGTCGTGTCAA
MS.gene072828.t1	MsERF06	CTCTAACAGCCGCCTTGGAA	CAGTAGCCAGCAACACTCCA
MS.gene006341.t1	MsERF07	GCCGGAGAGTGTGTTTGAGA	ACTCCGGCACTGTATCCTCT
MS.gene58366.t1	MsERF08	CCACCGCCGCTTGATTTAAC	ACGTGCGAATGCGTCAAAAA
MS.gene022105.t1	MsERF09	ATGGGCAGCTGAAATACGTGA	TTGTGCCTTTGAATTTGAGTGC
MS.gene030784.t1	MsERF10	TACCGCGGAGTTAGACAACG	ATGGAAGCGCATGTGAGGAA
MS.gene025280.t1	MsERF11	CATCGGATGGGACGACAACA	TCTCCGGCACGTAGAAATCG
MS.gene067817.t1	MsERF12	CAGGAAGATGGCGTTGTTGC	AGCACGGGTCGAAATAGTGT
MS.gene016368.t1	MsERF13	AGGAATCAATGCCGTGACCA	AGGCGGGGTAGTTGTTGTTT

登录号 Gene ID	基因名 Gene name	上游引物序列 Forward primer sequence	下游引物序列 Reverse primer sequence
MS.gene057432.t1	MsERF01	AGATTACCAAACAGCCGCCA	TAGCACCACTGCCACGTAAG
MS.gene022984.t1	MsERF02	AACCGAAGAGTGAACAGCCT	TCCTGCAAGGGTTGGTTGAA
MS.gene31024.t1	MsERF03	TCGTAACCCTCCACCAGCTA	CCTTGCAGAAGGAATACCCGA
MS.gene011550.t1	MsERF04	ATGCGTCAATGGGGCAAATG	TTGAGGTAAGCCGAAGAGCC
MS.gene30848.t1	MsERF05	TGCCACTACTCAGGCAAAGG	GGCGGCTATAGTCGTGTCAA
MS.gene072828.t1	MsERF06	CTCTAACAGCCGCCTTGGAA	CAGTAGCCAGCAACACTCCA
MS.gene006341.t1	MsERF07	GCCGGAGAGTGTGTTTGAGA	ACTCCGGCACTGTATCCTCT
MS.gene58366.t1	MsERF08	CCACCGCCGCTTGATTTAAC	ACGTGCGAATGCGTCAAAAA
MS.gene022105.t1	MsERF09	ATGGGCAGCTGAAATACGTGA	TTGTGCCTTTGAATTTGAGTGC
MS.gene030784.t1	MsERF10	TACCGCGGAGTTAGACAACG	ATGGAAGCGCATGTGAGGAA
MS.gene025280.t1	MsERF11	CATCGGATGGGACGACAACA	TCTCCGGCACGTAGAAATCG
MS.gene067817.t1	MsERF12	CAGGAAGATGGCGTTGTTGC	AGCACGGGTCGAAATAGTGT
MS.gene016368.t1	MsERF13	AGGAATCAATGCCGTGACCA	AGGCGGGGTAGTTGTTGTTT

基因 Gene name	登录号 Gene ID	氨基酸数目 Number of amino acids （aa）	分子量 Molecular weight （Da）	等电点 Isoelectric point	亲水性 GRAVY	亚细胞定位 Subcellular localization
MsERF01	MS.gene057432.t1	217	23830.29	4.58	-0.535	细胞核Nucleus
MsERF02	MS.gene022984.t1	233	26506.06	8.91	-0.758	叶绿体Chloroplast
MsERF03	MS.gene31024.t1	324	35332.93	9.76	-0.572	细胞核Nucleus
MsERF04	MS.gene011550.t1	199	21765.18	5.93	-0.551	细胞核Nucleus
MsERF05	MS.gene30848.t1	243	27717.37	6.55	-0.676	叶绿体Chloroplast
MsERF06	MS.gene072828.t1	231	25918.02	5.71	-0.526	细胞核Nucleus
MsERF07	MS.gene006341.t1	180	20443.92	5.76	-0.687	细胞核Nucleus
MsERF08	MS.gene58366.t1	210	22898.71	9.66	-0.596	叶绿体Chloroplast
MsERF09	MS.gene022105.t1	176	19699.82	7.89	-0.823	细胞核Nucleus
MsERF10	MS.gene030784.t1	288	32496.66	7.01	-0.558	细胞核Nucleus
MsERF11	MS.gene025280.t1	202	22322.82	4.58	-0.446	细胞核Nucleus
MsERF12	MS.gene067817.t1	259	28852.95	5.76	-0.661	细胞核Nucleus
MsERF13	MS.gene016368.t1	422	47454.26	5.54	-0.928	细胞核Nucleus

基因 Gene name	登录号 Gene ID	氨基酸数目 Number of amino acids （aa）	分子量 Molecular weight （Da）	等电点 Isoelectric point	亲水性 GRAVY	亚细胞定位 Subcellular localization
MsERF01	MS.gene057432.t1	217	23830.29	4.58	-0.535	细胞核Nucleus
MsERF02	MS.gene022984.t1	233	26506.06	8.91	-0.758	叶绿体Chloroplast
MsERF03	MS.gene31024.t1	324	35332.93	9.76	-0.572	细胞核Nucleus
MsERF04	MS.gene011550.t1	199	21765.18	5.93	-0.551	细胞核Nucleus
MsERF05	MS.gene30848.t1	243	27717.37	6.55	-0.676	叶绿体Chloroplast
MsERF06	MS.gene072828.t1	231	25918.02	5.71	-0.526	细胞核Nucleus
MsERF07	MS.gene006341.t1	180	20443.92	5.76	-0.687	细胞核Nucleus
MsERF08	MS.gene58366.t1	210	22898.71	9.66	-0.596	叶绿体Chloroplast
MsERF09	MS.gene022105.t1	176	19699.82	7.89	-0.823	细胞核Nucleus
MsERF10	MS.gene030784.t1	288	32496.66	7.01	-0.558	细胞核Nucleus
MsERF11	MS.gene025280.t1	202	22322.82	4.58	-0.446	细胞核Nucleus
MsERF12	MS.gene067817.t1	259	28852.95	5.76	-0.661	细胞核Nucleus
MsERF13	MS.gene016368.t1	422	47454.26	5.54	-0.928	细胞核Nucleus

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