紫花苜蓿突触结合蛋白家族成员鉴定与非生物胁迫下的表达分析

doi:10.11686/cyxb2025151

摘要/Abstract

摘要：

植物中的突触结合蛋白（SYT）属于钙结合蛋白家族，近年来在植物生物学研究领域备受关注。目前，仅在拟南芥中存在对SYT基因家族成员鉴定的相关报道，而豆科植物紫花苜蓿中该方面的研究仍属空白。本研究以拟南芥SYT基因家族成员为参照，运用生物信息学方法，在紫花苜蓿中成功筛选出21个SYT基因。通过对21个MsSYT、25个MtSYT和 9个AtSYT蛋白进行系统发育分析，明确了它们之间的同源性，MsSYT基因被分成4个亚家族。在对MsSYT基因的分析中发现，其家族成员的理化性质存在相对差异，但是基因结构和蛋白保守基序在各亚家族之间却高度保守。进一步对MsSYT基因的顺式作用元件进行分析，结果显示其中包含光响应元件、胁迫响应元件和激素响应元件，这表明该家族基因在植物调节生长发育和应对非生物胁迫过程中发挥重要作用。同时，染色体定位和共线性分析表明，MsSYT基因在染色体上呈不均匀分布，存在两组非串联重复片段以及两组串联重复片段。实时定量PCR（RT-qPCR）检测结果表明部分MsSYT基因对盐、冷和干旱胁迫有响应，其中MsSYT8d对这3种胁迫均表现出显著响应。本研究首次系统地对紫花苜蓿中的SYT基因进行了鉴定，并分析了其理化性质、进化特征和表达模式，为后续研究MsSYT基因在植物生长发育和非生物胁迫响应过程中的功能特性提供了坚实的理论依据。

关键词: 突触结合蛋白, 紫花苜蓿, 非生物胁迫, 表达分析

Abstract:

Synaptotagmin （SYT） proteins in plants belong to the calcium-binding protein family and have attracted much attention in the field of plant biology research. Members of the SYT family have been identified in Arabidopsis thaliana， but little is known about this gene family in the legume alfalfa （Medicago sativa）. In this study， 21 SYT genes were identified in the alfalfa genome based on searches using A. thalianaSYT gene sequences. The alfalfa SYT genes were then analyzed using bioinformatics methods. Phylogenetic analysis of SYT proteins of M. sativa， M. truncatula， and Arabidopsis （21 MsSYTs， 25 MtSYTs， and nine AtSYTs） clarified their homologous relationships and divided the MsSYT genes into four subfamilies. The putative proteins encoded by MsSYT genes showed some differences in their predicted physicochemical properties， but the gene structure and conserved protein motifs were highly conserved among the subfamilies. Analyses of the promoter sequences of MsSYT genes revealed cis-acting elements including light-responsive elements， stress-responsive elements， and hormone-responsive elements， indicating that MsSYTs plays important roles in plant growth and development and in responses to abiotic stresses. Chromosome localization analyses revealed that MsSYT genes were unevenly distributed among the chromosomes， and collinearity analyses detected two sets of non-tandem repeats and two sets of tandem repeats. Real-time quantitative PCR （RT-qPCR） analyses showed that some MsSYT genes responded to salt stress， cold stress， and drought stress， and MsSYT8d responded to all three stresses. This study is the first description of SYT genes in alfalfa， the physicochemical properties of their putative encoded proteins， and their evolutionary characteristics and transcript profiles. These results provide a solid theoretical basis for further research on the functional characteristics of MsSYTs in plant growth and development and abiotic stress responses.

Key words: synaptotagmin, alfalfa, abiotic stress, expression analysis

张钿, 冷华娟, 崔婧, 何飞, 王雪, 李明娜, 杨青川, 康俊梅. 紫花苜蓿突触结合蛋白家族成员鉴定与非生物胁迫下的表达分析[J]. 草业学报, 2026, 35(4): 158-168.

Tian ZHANG, Hua-juan LENG, Jing CUI, Fei HE, Xue WANG, Ming-na LI, Qing-chuan YANG, Jun-mei KANG. Identification of synaptotagmin gene family members in alfalfa and their transcript profiles under abiotic stresses[J]. Acta Prataculturae Sinica, 2026, 35(4): 158-168.

图/表 10

表1 MsSYT基因家族基本性质分析

Table 1 Analysis of the basic properties of MsSYT gene family

基因登录号 Gene ID	基因名称 Gene name	蛋白长度 Protein length （aa）	分子量 Molecular weight （kDa）	等电点 Isoelectric point （pI）	不稳定系数 Instability index	总平均亲水性 Grand average of hydropathicity
Msa083526.t01	MsSYT1	537	61.44	6.36	29.74	-0.339
Msa000097.t01	MsSYT2	535	60.98	7.61	32.65	-0.250
Msa005119.t01	MsSYT3	544	62.05	8.40	35.48	-0.091
Msa086529.t01	MsSYT4	821	92.00	6.92	40.18	-0.216
Msa183305.t01	MsSYT5	566	63.51	5.96	31.42	-0.095
Msa027845.t01	MsSYT6	1023	114.98	5.77	48.58	-0.267
Msa112806.t01	MsSYT7	764	87.85	6.67	43.22	-0.045
Msa083527.t01	MsSYT8a	945	107.72	6.10	38.93	-0.400
Msa083527.t02	MsSYT8b	645	74.41	5.86	39.96	-0.351
Msa083527.t03	MsSYT8c	717	82.48	6.34	39.31	-0.348
Msa083527.t04	MsSYT8d	645	74.41	5.86	39.96	-0.351
Msa083527.t05	MsSYT8e	717	82.48	6.34	39.31	-0.348
Msa038100.t01	MsSYT9	561	63.29	5.78	34.12	-0.099
Msa157005.t01	MsSYT10	775	89.45	9.20	45.43	-0.261
Msa081502.t01	MsSYT11	770	88.96	9.29	42.32	-0.262
Msa179019.t01	MsSYT12	1005	114.51	9.16	45.19	-0.387
Msa156608.t01	MsSYT13	1323	151.85	8.93	41.08	-0.403
Msa156589.t01	MsSYT14	1013	115.13	9.27	43.45	-0.371
Msa135465.t01	MsSYT15	828	93.71	6.90	45.36	-0.301
Msa056918.t01	MsSYT16a	568	64.18	5.98	33.88	-0.114
Msa056918.t02	MsSYT16b	1045	120.18	8.97	43.06	-0.473

图1 MsSYT基因家族成员的染色体分布

Fig.1 Chromosomal distribution of MsSYT gene family members

图2 紫花苜蓿、拟南芥和蒺藜苜蓿的SYT家族成员系统发育关系

Fig.2 Phylogenetic relationships of SYT gene family in M. sativa， A. thaliana and M. truncatula

图3 MsSYT基因进化关系、保守基序、保守结构域和结构分析UTR： Untranslated region； CDS： Coding sequence.

Fig.3 MsSYT genes phylogenetic relationship， conserved motifs， conserved domains， and gene structural analysis

图4 MsSYT家族成员的顺式作用元件分析

Fig.4 Analysis of cis-acting elements of MsSYT family members

图5 紫花苜蓿MsSYT基因的共线性分析

Fig.5 Collinearity analysis of MsSYT gene in M. sativa

图6 紫花苜蓿与拟南芥、蒺藜苜蓿SYT 基因的共线性分析

Fig.6 Collinearity analysis of SYT genes in M. sativa， A. thaliana and M. truncatula

图7 MsSYT基因在干旱胁迫下的表达模式*： P<0.05； **： P<0.01； ***： P<0.001； ****： P<0.0001；下同The same below.

Fig.7 Expression pattern of MsSYT gene under drought stress

图8 MsSYT基因在盐胁迫下的表达模式

Fig.8 Expression pattern of MsSYT gene under salt stress

图9 MsSYT基因在冷胁迫下的表达模式

Fig.9 Expression pattern of MsSYT gene under cold stress

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