Identification and transcript profiling of the CAMTA gene family under abiotic stress in alfalfa

doi:10.11686/cyxb2023240

Abstract

Abstract:

Calmodulin-binding transcription activating factors （CAMTA） are calmodulin-binding proteins that play important roles in hormone signal transduction， developmental regulation， and environmental stress tolerance. In this study， bioinformatic methods were used to identify members of the CAMTA family in alfalfa （Medicago sativa） based on the reference genome of M. sativa “XinjiangDaye”. A phylogenetic analysis was conducted to evaluate relationships among the MsCAMTA genes. The physicochemical properties and conserved domains of the putative proteins were determined. The chromosomal positions of MsCAMTA genes were mapped， and cis-acting elements were identified in their promoter regions. The transcriptional profiles of MsCAMTA genes under various stress conditions were analyzed and validated. Seventeen MsCAMTA genes were identified， and the MsCAMTA family members grouped into three subfamilies in the phylogenetic analysis. Members of the same subfamily had relatively similar gene structures and positions of conserved motifs. The chromosomal localization results showed that members of the MsCAMTA family were unevenly distributed on seven chromosomes. A large number of cis-acting elements that respond to low temperature， salt stress， and plant hormone signals were identified in the promoter regions of these genes. We conducted RT-qPCR analyses to determine the transcript levels of MsCAMTA1， MsCAMTA3， MsCAMTA11， and MsCAMTA12 in alfalfa leaves under salt （300 mmol·L^-1 NaCl）， simulated drought （400 mmol·L^-1 mannitol）， low temperature （10 ℃）， and abscisic acid （100 μmol·L^-1） treatments. All four MsCAMTA candidate genes responded to various stressors to varying degrees. All four were up-regulated under salt stress. The results suggested that MsCAMTA may participate in the responses of alfalfa to various abiotic stresses by integrating multiple stress signals. The results of this study provide a reference for further studies on the functions of MsCAMTA genes in plant response to stress.

Key words: alfalfa, CAMTA gene, bioinformatics analysis, gene expression

Hai-ming KONG, Jia-xing SONG, Jing YANG, Qian LI, Pei-zhi YANG, Yu-man CAO. Identification and transcript profiling of the CAMTA gene family under abiotic stress in alfalfa[J]. Acta Prataculturae Sinica, 2024, 33(5): 143-154.

Figures/Tables 8

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基因Gene	正向引物Forward primer （5′-3′）	反向引物Reverse primer （5′-3′）
MsCAMTA1	GAGGAGGAAGCACGCCAG	GCGGCTGCCATTGCTTTT
MsCAMTA3	GCGGACAGTTGGCTTCCT	GCCCTTTGTGCCCATTGC
MsCAMTA11	TCTTCATTGGGCGGCGTT	AGCTAGGTCGGCTGGTGT
MsCAMTA12	TGGTGCATCTGCTGGAGC	CCAGTCCCTTATGCCCGC
MsActin	GACAATGGAACTGGAATGG	CAATACCGTGCTCAATGG

基因Gene	正向引物Forward primer （5′-3′）	反向引物Reverse primer （5′-3′）
MsCAMTA1	GAGGAGGAAGCACGCCAG	GCGGCTGCCATTGCTTTT
MsCAMTA3	GCGGACAGTTGGCTTCCT	GCCCTTTGTGCCCATTGC
MsCAMTA11	TCTTCATTGGGCGGCGTT	AGCTAGGTCGGCTGGTGT
MsCAMTA12	TGGTGCATCTGCTGGAGC	CCAGTCCCTTATGCCCGC
MsActin	GACAATGGAACTGGAATGG	CAATACCGTGCTCAATGG

基因名称 Gene name	基因号 Gene ID	染色体 Chromosome	不稳定指数 Instability index	疏水性指数 Grand average of hydrophobicity （GRAVY）	脂肪指数 Aliphatic index	蛋白质长度 Protein length （aa）	分子量 Molecular weight （kDa）	等电点 Isoelectric point （pI）	亚细胞定位 Subcellular localization
MsCAMTA1	MS.gene00191.t1	Chr2.1	42.00	-0.418	79.07	915	103.86	6.49	细胞核Nucleus
MsCAMTA2	MS.gene56018.t1	Chr2.3	42.52	-0.418	78.73	914	103.81	6.48	细胞核Nucleus
MsCAMTA3	MS.gene002202.t1	Chr2.3	47.33	-0.522	76.66	913	101.75	5.24	细胞核Nucleus
MsCAMTA4	MS.gene09226.t1	Chr4.1	47.88	-0.596	69.50	960	107.40	5.44	细胞核Nucleus
MsCAMTA5	MS.gene57296.t1	Chr4.1	43.64	-0.443	79.89	1745	196.21	6.12	细胞核Nucleus
MsCAMTA6	MS.gene028443.t1	Chr4.1	45.76	-0.575	70.13	1029	116.11	5.70	细胞核Nucleus
MsCAMTA7	MS.gene90181.t1	Chr4.2	45.67	-0.570	70.51	1029	116.11	5.67	细胞核Nucleus
MsCAMTA8	MS.gene030815.t1	Chr4.2	45.61	-0.619	74.79	1086	123.35	5.77	细胞核Nucleus
MsCAMTA9	MS.gene70367.t1	Chr4.3	45.59	-0.567	70.41	1029	116.08	5.73	细胞核Nucleus
MsCAMTA10	MS.gene023441.t1	Chr4.3	47.56	-0.603	69.40	960	107.42	5.44	细胞核Nucleus
MsCAMTA11	MS.gene061784.t1	Chr4.3	45.61	-0.619	74.79	1086	123.35	5.77	细胞核Nucleus
MsCAMTA12	MS.gene09019.t1	Chr4.4	47.56	-0.603	69.40	960	107.42	5.44	细胞核Nucleus
MsCAMTA13	MS.gene28071.t1	Chr4.4	44.14	-0.420	81.90	1437	161.54	6.30	叶绿体Chloroplast
MsCAMTA14	MS.gene35018.t1	Chr4.4	46.06	-0.572	70.13	1029	116.16	5.70	细胞核Nucleus
MsCAMTA15	MS.gene61520.t1	Chr8.3	39.09	-0.467	80.03	864	98.08	7.14	细胞核Nucleus
MsCAMTA16	MS.gene61521.t1	Chr8.3	40.02	-0.474	80.42	924	105.32	7.37	细胞核Nucleus
MsCAMTA17	MS.gene042294.t1	Chr8.3	39.70	-0.470	80.51	923	105.16	7.15	细胞核Nucleus

基因名称 Gene name	基因号 Gene ID	染色体 Chromosome	不稳定指数 Instability index	疏水性指数 Grand average of hydrophobicity （GRAVY）	脂肪指数 Aliphatic index	蛋白质长度 Protein length （aa）	分子量 Molecular weight （kDa）	等电点 Isoelectric point （pI）	亚细胞定位 Subcellular localization
MsCAMTA1	MS.gene00191.t1	Chr2.1	42.00	-0.418	79.07	915	103.86	6.49	细胞核Nucleus
MsCAMTA2	MS.gene56018.t1	Chr2.3	42.52	-0.418	78.73	914	103.81	6.48	细胞核Nucleus
MsCAMTA3	MS.gene002202.t1	Chr2.3	47.33	-0.522	76.66	913	101.75	5.24	细胞核Nucleus
MsCAMTA4	MS.gene09226.t1	Chr4.1	47.88	-0.596	69.50	960	107.40	5.44	细胞核Nucleus
MsCAMTA5	MS.gene57296.t1	Chr4.1	43.64	-0.443	79.89	1745	196.21	6.12	细胞核Nucleus
MsCAMTA6	MS.gene028443.t1	Chr4.1	45.76	-0.575	70.13	1029	116.11	5.70	细胞核Nucleus
MsCAMTA7	MS.gene90181.t1	Chr4.2	45.67	-0.570	70.51	1029	116.11	5.67	细胞核Nucleus
MsCAMTA8	MS.gene030815.t1	Chr4.2	45.61	-0.619	74.79	1086	123.35	5.77	细胞核Nucleus
MsCAMTA9	MS.gene70367.t1	Chr4.3	45.59	-0.567	70.41	1029	116.08	5.73	细胞核Nucleus
MsCAMTA10	MS.gene023441.t1	Chr4.3	47.56	-0.603	69.40	960	107.42	5.44	细胞核Nucleus
MsCAMTA11	MS.gene061784.t1	Chr4.3	45.61	-0.619	74.79	1086	123.35	5.77	细胞核Nucleus
MsCAMTA12	MS.gene09019.t1	Chr4.4	47.56	-0.603	69.40	960	107.42	5.44	细胞核Nucleus
MsCAMTA13	MS.gene28071.t1	Chr4.4	44.14	-0.420	81.90	1437	161.54	6.30	叶绿体Chloroplast
MsCAMTA14	MS.gene35018.t1	Chr4.4	46.06	-0.572	70.13	1029	116.16	5.70	细胞核Nucleus
MsCAMTA15	MS.gene61520.t1	Chr8.3	39.09	-0.467	80.03	864	98.08	7.14	细胞核Nucleus
MsCAMTA16	MS.gene61521.t1	Chr8.3	40.02	-0.474	80.42	924	105.32	7.37	细胞核Nucleus
MsCAMTA17	MS.gene042294.t1	Chr8.3	39.70	-0.470	80.51	923	105.16	7.15	细胞核Nucleus

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