辣椒E2基因家族的鉴定及分析

doi:10.13560/j.cnki.biotech.bull.1985.2024-0044

摘要/Abstract

摘要：

【目的】 泛素结合酶E2（UBC）作为泛素蛋白酶体途径中将泛素转移至靶蛋白的重要中转酶，与植物的生长发育和许多胁迫有关。从辣椒全基因组中鉴定E2基因家族成员，分析相关基因表达模式，为后续研究提供理论基础。【方法】 基于辣椒全基因组信息，利用生物信息学方法对E2基因家族进行鉴定，系统分析该家族成员的基本理化性质、亚细胞定位、染色体定位、基因结构、系统进化关系、顺式作用元件、蛋白质互作网络、非生物胁迫响应与组织器官表达模式。【结果】 辣椒E2家族有48个成员，在11条染色体上呈不均匀地分布，其编码蛋白主要定位于细胞外和细胞核中；通过对辣椒、番茄和拟南芥的比较进化分析将其分为Group I-Group XIII亚族，同一亚族具有相似的蛋白保守基序和基因结构；发现共有26个蛋白之间存在互作；顺式作用元件分析表明，CaUBC基因启动子区含有大量光响应、激素响应元件，上游2 000 bp区域中存在多种与生长发育和抗逆性相关的顺式作用元件；CaUBC参与胁迫响应，在不同非生物胁迫和激素处理下，将其分为3类，其中高温环境下第3类基因为高表达；在组织器官中表现出不同的表达，其中CaUBC38在果实中的表达量变化较为明显，进行RT-qPCR分析发现，50 d相对表达量最高，10 d表达量最低。【结论】 获得48个辣椒E2（CaUBC）基因，揭示了E2家族不同成员在辣椒生长发育和非生物胁迫过程中的表达特征。

关键词: 辣椒, E2基因家族, 泛素蛋白酶体系统, 生物信息学, 表达分析, 非生物胁迫, 组织器官

Abstract:

【Objective】 Ubiquitin-binding enzyme E2（UBC）is an important transit enzyme in the ubiquitin-proteasomal pathway that transfers ubiquitin to target proteins, which is involved in plant growth and development and many stresses. The E2 gene family members were identified from the whole genome of pepper, and the expression patterns of related genes were analyzed, which provided a theoretical basis for subsequent research.【Method】 Based on the genome-wide information of pepper, the E2 family was identified by bioinformatics. The basic physio-chemical properties, subcellular localization, chromosomal localization, gene structure and phylogenetic analysis, cis-acting elements, abiotic stress response, tissue and organ expression patterns, and protein-protein interaction network of the family members were systematically analyzed. 【Result】 There were 48 members in the E2 gene family, which were unevenly distributed on 11 chromosomes, and its encoded proteins were mainly localized in the extracellular and nucleus. After comparative evolutionary analysis of pepper, tomato and Arabidopsis thaliana, they were divided into the Group I-Group XIII subfamily, which had similar conserved protein motifs and gene structures. A total of 26 proteins were found to interact. The cis-element analysis showed that the promoter region of CaUBC gene contained a large number of light-responsive and hormone-responsive elements, and there were a variety of cis-acting elements related to growth, development and stress resistance in the upstream 2 000 bp region. CaUBC was involved in the stress, under different abiotic stresses and hormone treatments, they were divided into three categories, among which the third type of genes were highly expressed in high temperature environment; different expressions showed in tissues and organs, among which the expression of CaUBC38 in fruits changed significantly. RT-qPCR analysis showed that the relative expression was the highest at 50 d and the lowest at 10 d. 【Conclusion】 The 48 members of E2（CaUBC）gene family are systematically identified from the pepper genome. The expression characteristics of CaUBC family members during its growth, development, and abiotic stress responses are revealed.

Key words: Capsicum annuum L., E2 gene family, ubiquitin-proteasome system, bioinformatics, expression analysis, abiotic pressure, tissues and organs

常雪瑞, 王田田, 王静. 辣椒E2基因家族的鉴定及分析[J]. 生物技术通报, 2024, 40(6): 238-250.

CHANG Xue-rui, WANG Tian-tian, WANG Jing. Identification and Analysis of E2 Gene Family in Pepper（Capsicum annuum L.）[J]. Biotechnology Bulletin, 2024, 40(6): 238-250.

图/表 9

表1 E2家族基因表达分析的RT-qPCR所需引物

Table 1 Primers required for RT-qPCR for E2 family gene expression analysis

基因名称 Gene name	上游引物Forward primer（5'-3'）	下游引物Reverse primer（5'-3'）
CaUBC3	AGCGGAGGTAGAGGAAGAGG	GCCACCCTCATAAGGTGTTC
CaUBC8	GCTGGTGGTGTTTTCCAAGT	GCTTCGAGCCATTGATTCAT
CaUBC18	TGCAGGTGGTGTTTTTCAAG	CGAGCCACCGATTCATATTT
CaUBC21	ATAGCCCCTATGCAGGAGGT	CAAGTGGATCGTCTGGGTTT
CaUBC23	CCCAATAAGCCACCAACAGT	ACTGAACATCCGAGCTGCTT
CaUBC38	GCGGAAAGTCCTTATCATGG	GATCAACAGGACCTGCCATT
β-Actin	CCACCTCTTCACTCTCTGCTCT	ACTAGGAAAAACAGCCCTTGGT

图1 辣椒E2基因家族在染色体上的分布

Fig. 1 Distribution of E2 gene family on chromosome in pepper（Capsicum annuum L.）

图2 辣椒（48个）、拟南芥（36个）和番茄（52个）UBC蛋白的系统发育进化树 At：拟南芥；Sl：番茄；Ca：辣椒

Fig. 2 Phylogenetic tree of UBC protein in C. annuum L.（48），A. thaliana L.（36）and S. lycopersicum L.（52） At: Arabidopsis thaliana L.; Sl: Solanum lycopersicum L.; Ca: Capsicum annuum L.

图3 48个CaUBC基因的结构分布模式

Fig. 3 Structure distribution patterns of 48 CaUBC genes

图4 辣椒E2家族顺式作用元件

Fig. 4 Cis-acting elements of the pepper E2 family

图5 E2家族的蛋白质互作网络分析蛋白互作网络中的节点代表由一个蛋白质编码基因座产生的所有蛋白质，不同颜色的节点代表互作关系的不同程度，且随着节点间线条颜色的加深，互作强度也随之增强，较细的线条表明蛋白质间的互作较弱

Fig. 5 Analysis of the protein interaction network of the E2 family The nodes in the protein interaction network indicate all proteins produced by a protein-coding locus, and the nodes of different colors indicate different degrees of interaction, and the intensity of interaction increases with the deepening of the color of the lines between nodes, and the thinner lines indicate that the interaction between proteins is weaker

图6 不同处理下CaUBC基因的表达谱

Fig. 6 Expression profile of CaUBC genes under different treatments

图7 CaUBC基因在不同器官中的表达谱 F1-F9：出现花蕾后；G1-G11：授粉后10、15、20、25、30、35、40、45、50、55、60 d的果实；ST1、ST2：授粉后10 d、15 d的果实；S3-S11：授粉后20、25、30、35、40、45、50、55、60 d的果实；T3-T11：授粉后20、25、30、35、40、45、50、55、60 d的果实；L1-L9：新叶刚现后2、5、10、15、20、25、30、40、50、60 d。下同

Fig. 7 Expression profiles of CaUBC genes in different organs F1-F9: After the appearance of flower buds. G1-G11: Fruits at 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, and 60 d after pollination. ST1 and ST2: Fruits at 10 and 15 d after pollination. S3-S11: Fruits at 20, 25, 30, 35, 40, 45, 50, 55, and 60 d after pollination. T3-T11: Fruits at 20, 25, 30, 35, 40, 45, 50, 55 and 60 d after pollination. L1-L9: 2, 5, 10, 15, 20, 25, 30, 40, 50 and 60 d after the emergence of new leaves. The same below

图8 辣椒E2基因家族8个代表基因的相对表达量 A：CaUBC38在果实发育过程中的相对表达量，CaUBC8在花发育过程中的相对表达量，CaUBC3/18在热胁迫下的相对表达量；B：CaUBC8/21/23/38在热胁迫下的相对表达量

Fig. 8 Relative expressions of eight representative genes of E2 gene family in pepper A: Relative expression of CaUBC38 during fruit development; relative expression of CaUBC8 during flower development; relative expression of CaUBC3/18 under heat stress.B: CaUBC8/21/23/38 under heat stress

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