Identification and expression analysis of LACS gene family members in Medicago sativa

doi:10.11686/cyxb2024206

Abstract

Abstract:

Members of the long-chain acyl-coenzyme A （CoA） synthetase （LACS） family， in the acyl-activating enzyme superfamily， play important roles in fatty acid anabolic metabolism. In this study， based on genomic data of Medicago sativa （alfalfa）， LACS gene family members were identified by bioinformatics methods， a phylogenetic tree was constructed， and the physicochemical properties of the putative proteins were determined. The chromosome localization， conserved motifs and gene structure， cis-acting elements， and tissue-specific expression patterns of the LACS genes were analyzed， and a protein-protein interaction （PPI） network was constructed. The transcript profiles of LACS genes under biotic stress were analyzed by qRT-PCR. The results showed that 10 MsLACS family members were present in the alfalfa genome， and were located on five chromosomes. In the phylogenetic tree， the 10 MsLACS were grouped into five branches. Motif3 constituted the conserved AMP-binding domain， and the alfalfa MsLACS genes contained 10-12 motifs. There were differences in gene structure among the 10 MsLACS genes， with the number of exons ranging from 11 to 22， and the number of introns ranging from 0 to 3. MsLACS1-1 and MsLACS3 had no introns. The promoter region of MsLACS contained light response elements， hormone response elements， and abiotic stress response elements. The transcript profiles of MsLACS genes differed among different tissues and showed obvious tissue specificity. Analyses of gene expression by qRT-PCR revealed higher transcript levels of MsLACS genes under drought stress and salt stress than under cold stress. Under cold stress， the transcript levels of MsLACS genes in alfalfa initially increased and then decreased， and were higher in the leaves than in the roots. Under drought and salt stress， MsLACS genes were highly expressed in leaves， with peak transcript levels at 6 h. The PPI network analysis showed that the 10 proteins encoded by MsLACS genes in alfalfa interacted with each other， with 18 lines of interaction among the proteins. These results provide a basis for further research on LACS genes in alfalfa and their applications in breeding for stress resistance.

Key words: Medicago sativa, long-chain acyl-coenzyme A （CoA） synthetase （LACS）, gene family, abiotic stress, expression

Tian-rong LUO, Jian-zhi MA, Ming-yang DU, Jie-cuo DUO, Hui-yan XIONG, Rui-jun DUAN. Identification and expression analysis of LACS gene family members in Medicago sativa[J]. Acta Prataculturae Sinica, 2025, 34(4): 124-136.

Figures/Tables 11

References 27

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基因Gene	上游引物Forward primer （5′-3′）	下游引物Reverse primer （3′-5′）
MsLACS1-1	CATGCTGGGATGGCGTAAAAT	AGTGAAGCACTTTTTGCAACC
MsLACS1-2	TACCATGGGGTCAAAGCTCG	CCAACAGTACCAAGCATGCAC
MsLACS2	TTGGTTTCATACAGCCATGAAAAT	TTCTGTCGGGGACTACCACA
MsLACS3	CGGGGCTGTGGAGTTTGTTA	CATGACCCCCGATCTTCTCA
MsLACS5	GGAACGGGACCTTATCACTCC	GCAGTCAGTAAAAGACAGCTCAT
MsLACS8	ACTGGTGACATTGGGCGATT	CCAGAGACTGACGTGAAGCA
MsLACS9-2	TTCAGGGTTGTTTCAGGCGA	CAGCCGTGTCCAACAGATGA
Msactin	CAAAAGATGGCAGATGCTGAGGAT	CATGACACCAGTATGACGAGGTCG

基因Gene	上游引物Forward primer （5′-3′）	下游引物Reverse primer （3′-5′）
MsLACS1-1	CATGCTGGGATGGCGTAAAAT	AGTGAAGCACTTTTTGCAACC
MsLACS1-2	TACCATGGGGTCAAAGCTCG	CCAACAGTACCAAGCATGCAC
MsLACS2	TTGGTTTCATACAGCCATGAAAAT	TTCTGTCGGGGACTACCACA
MsLACS3	CGGGGCTGTGGAGTTTGTTA	CATGACCCCCGATCTTCTCA
MsLACS5	GGAACGGGACCTTATCACTCC	GCAGTCAGTAAAAGACAGCTCAT
MsLACS8	ACTGGTGACATTGGGCGATT	CCAGAGACTGACGTGAAGCA
MsLACS9-2	TTCAGGGTTGTTTCAGGCGA	CAGCCGTGTCCAACAGATGA
Msactin	CAAAAGATGGCAGATGCTGAGGAT	CATGACACCAGTATGACGAGGTCG

基因 Gene	基因ID号 Gene ID	氨基酸数 Number of amino acids （aa）	分子质量 Molecular mass （kD）	等电点pI	不稳定系数 Instability coefficient	脂溶性系数 Fat solubility coefficient	染色体定位 Chromosomal localization	亚细胞定位 Subcellular localization
MsLACS1-1	MsG0180004774.01.T01	610	68724.49	8.00	27.09	88.48	Chr1	叶绿体Chloroplast
MsLACS1-2	MsG0780041383.01.T01	597	66899.61	5.74	31.70	84.00	Chr7	细胞质Cytoplasm
MsLACS2	MsG0180001276.01.T02	654	73203.98	5.74	40.29	86.90	Chr1	细胞核Cell nucleus
MsLACS3	MsG0480023684.01.T01	584	65477.89	6.66	31.47	87.77	Chr4	细胞质Cytoplasm
MsLACS4	MsG0580024346.01.T01	662	73949.13	6.37	33.60	89.18	Chr5	细胞核Cell nucleus
MsLACS5	MsG0580024344.01.T01	744	83552.61	6.67	34.62	91.67	Chr5	细胞质Cytoplasm
MsLACS6	MsG0180005634.01.T01	692	76661.51	6.35	29.23	88.03	Chr1	细胞质Cytoplasm
MsLACS8	MsG0380016055.01.T02	727	79391.51	7.19	29.26	93.59	Chr3	叶绿体Chloroplast
MsLACS9-1	MsG0380016352.01.T01	860	94959.34	6.30	31.37	98.36	Chr3	叶绿体Chloroplast
MsLACS9-2	MsG0180001085.01.T01	697	76384.23	6.51	30.95	95.24	Chr1	叶绿体Chloroplast

基因 Gene	基因ID号 Gene ID	氨基酸数 Number of amino acids （aa）	分子质量 Molecular mass （kD）	等电点pI	不稳定系数 Instability coefficient	脂溶性系数 Fat solubility coefficient	染色体定位 Chromosomal localization	亚细胞定位 Subcellular localization
MsLACS1-1	MsG0180004774.01.T01	610	68724.49	8.00	27.09	88.48	Chr1	叶绿体Chloroplast
MsLACS1-2	MsG0780041383.01.T01	597	66899.61	5.74	31.70	84.00	Chr7	细胞质Cytoplasm
MsLACS2	MsG0180001276.01.T02	654	73203.98	5.74	40.29	86.90	Chr1	细胞核Cell nucleus
MsLACS3	MsG0480023684.01.T01	584	65477.89	6.66	31.47	87.77	Chr4	细胞质Cytoplasm
MsLACS4	MsG0580024346.01.T01	662	73949.13	6.37	33.60	89.18	Chr5	细胞核Cell nucleus
MsLACS5	MsG0580024344.01.T01	744	83552.61	6.67	34.62	91.67	Chr5	细胞质Cytoplasm
MsLACS6	MsG0180005634.01.T01	692	76661.51	6.35	29.23	88.03	Chr1	细胞质Cytoplasm
MsLACS8	MsG0380016055.01.T02	727	79391.51	7.19	29.26	93.59	Chr3	叶绿体Chloroplast
MsLACS9-1	MsG0380016352.01.T01	860	94959.34	6.30	31.37	98.36	Chr3	叶绿体Chloroplast
MsLACS9-2	MsG0180001085.01.T01	697	76384.23	6.51	30.95	95.24	Chr1	叶绿体Chloroplast

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