番茄SlMYB52基因鉴定、亚细胞定位及表达分析

doi:10.3969/j.issn.1004-1524.20240596

浙江农业学报 ›› 2025, Vol. 37 ›› Issue (4): 808-819.DOI: 10.3969/j.issn.1004-1524.20240596

番茄SlMYB52基因鉴定、亚细胞定位及表达分析

狄延翠(), 嵇泽琳, 王媛媛, 娄世浩, 张涛, 国志信, 申顺善, 朴凤植, 杜南山, 董晓星, 董韩^*()

河南农业大学园艺学院,河南郑州 450046

收稿日期:2024-07-04 出版日期:2025-04-25 发布日期:2025-05-09
作者简介:狄延翠( 2000—),女,河南三门峡人,硕士研究生,研究方向为设施蔬菜优质高效生产技术。E-mail:13525866912@163.com
通讯作者: *董韩,E-mail:440069@henau.edu.cn
基金资助:
国家自然科学基金(32202579);河南省科技攻关项目(242102111075);中国博士后科学基金面上项目(2022M711062)

Identification, subcellular localization and expression analysis of tomato SlMYB52 gene

DI Yancui(), JI Zelin, WANG Yuanyuan, LOU Shihao, ZHANG Tao, GUO Zhixin, SHEN Shunshan, PIAO Fengzhi, DU Nanshan, DONG Xiaoxing, DONG Han^*()

College of Horticulture, Henan Agricultural University, Zhengzhou 450046, China

Received:2024-07-04 Online:2025-04-25 Published:2025-05-09

摘要/Abstract

摘要： MYB转录因子在植物逆境应答过程中发挥着重要的调控作用。为了挖掘更多番茄(Solanum lycopersicum L.)MYB转录因子成员信息,本研究从番茄中克隆了SlMYB52基因,采用生物信息学手段对SlMYB52基因进行基因结构、编码蛋白信息、保守结构域、系统进化树及启动子顺式作用元件预测等的分析,利用烟草叶片瞬时转化法分析亚细胞定位,通过qRT-PCR进行组织特异性表达及响应逆境胁迫的分析。结果表明SlMYB52基因全长1 431 bp,编码253个氨基酸,预测相对分子质量为29 035.22,理论等电点9.08,属于不稳定蛋白质、亲水性蛋白质;SlMYB52所编码蛋白质不含跨膜结构,没有潜在的信号肽位点;该蛋白质二级结构以无规卷曲为主,占比59.68%;系统进化树分析显示,SlMYB52与NtMYB4a亲缘关系最近;亚细胞定位结果显示,该SlMYB52蛋白定位在细胞核;对SlMYB52启动子分析,发现其含有大量的逆境响应元件;qRT-PCR结果表明,SlMYB52基因在茎中表达最高,其次是叶,在顶芽中表达量最低;SlMYB52基因的表达受高盐、低温及辣椒疫霉菌侵染的诱导,在干旱胁迫下SlMYB52基因的表达受到明显抑制,说明该基因可能参与逆境胁迫的应答。本研究结果为进一步研究SlMYB52基因的生物学功能及作用机制提供理论依据。

关键词: 番茄, MYB转录因子, 逆境胁迫, 亚细胞定位, 生物信息学分析

Abstract:

MYB transcription factors play an important regulatory role in plant stress response. In order to explore more information about MYB transcription factor members in tomato (Solanum lycopersicum L.), the SlMYB52 gene was cloned from tomato, and its gene structure, coding protein information, conserved domain, phylogenetic tree, and promoter cis-acting element prediction were analyzed using bioinformatics methods, the subcellular localization of SlMYB52 was analyzed using transient transformation in tobacco leaves, and tissue-specific expression and stress response were analyzed using qRT-PCR in this study. The results showed that SlMYB52 gene was 1 431 bp in length, encoding 253 amino acids, with a predicted relative molecular weight of 29 035.22 and a theoretical isoelectric point of 9.08, belonging to unstable proteins and hydrophilic proteins. The protein encoded by SlMYB52 has no transmembrane structure and no potential signal peptide site. The secondary structure of the protein was maily composed of random coli, accounting for 59.68%. Phylogenetic tree analysis showed that the amino acid sequence of SlMYB52 and NtMYB4a was closely related. Subcellular localization results showed that the SlMYB52 protein was located in the nucleus. The analysis of the SlMYB52 promoter revealed that it contained a large number of stress response elements. Moreover, the results of qRT-PCR showed that the expression of SlMYB52 gene was highest in stem, followed by leaves, and lowest in apical bud. Furthermore, the expression of SlMYB52 gene was induced by high salt, low temperature, and Phytophthora capsici infection, and was inhibited by drought stress, indicating that this gene may be involved in stress response. The results of this study provide a theoretical basis for further studying the biological function and mechanism of SlMYB52 gene.

Key words: tomato, MYB transcription factor, stress, subcellular localization, bioinformatics analysis

中图分类号:

S641.2

狄延翠, 嵇泽琳, 王媛媛, 娄世浩, 张涛, 国志信, 申顺善, 朴凤植, 杜南山, 董晓星, 董韩. 番茄SlMYB52基因鉴定、亚细胞定位及表达分析[J]. 浙江农业学报, 2025, 37(4): 808-819.

DI Yancui, JI Zelin, WANG Yuanyuan, LOU Shihao, ZHANG Tao, GUO Zhixin, SHEN Shunshan, PIAO Fengzhi, DU Nanshan, DONG Xiaoxing, DONG Han. Identification, subcellular localization and expression analysis of tomato SlMYB52 gene[J]. Acta Agriculturae Zhejiangensis, 2025, 37(4): 808-819.

图/表 11

参考文献 35

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引物名称 Primer name	引物序列 Primer sequence(5'→3')	注释 Annotation
RT-SlMYB52-F	GCAGAAAGGTACGAGCATGG	qRT-PCR
RT-SlMYB52-R	GTTCTTGAAAGCCCTGCGAA
RT-ACTIN2-F	TGTCCCTATTTACGAGGGTTATGC
RT-ACTIN2-R	CAGTTAAATCACGACCAGCAAGAT
RT-UBI3-F	GCCGACTACAACATCCAGAAGG
RT-UBI3-R	TGCAACACAGCGAGCTTAACC
GFP-SlMYB52-F	ctctcgagctttcgc gagctcATGCCAAGGGTACAACAACAGC	融合绿色荧光蛋白Fused with GFP
GFP-SlMYB52-R	gcccttgctcaccat ggatccGATATTTCCAAGTACATCAATCCAGAA

引物名称 Primer name	引物序列 Primer sequence(5'→3')	注释 Annotation
RT-SlMYB52-F	GCAGAAAGGTACGAGCATGG	qRT-PCR
RT-SlMYB52-R	GTTCTTGAAAGCCCTGCGAA
RT-ACTIN2-F	TGTCCCTATTTACGAGGGTTATGC
RT-ACTIN2-R	CAGTTAAATCACGACCAGCAAGAT
RT-UBI3-F	GCCGACTACAACATCCAGAAGG
RT-UBI3-R	TGCAACACAGCGAGCTTAACC
GFP-SlMYB52-F	ctctcgagctttcgc gagctcATGCCAAGGGTACAACAACAGC	融合绿色荧光蛋白Fused with GFP
GFP-SlMYB52-R	gcccttgctcaccat ggatccGATATTTCCAAGTACATCAATCCAGAA

名称 Name	序列 Sequence	功能 Function	数目 Amount
TGA-element	AACGAC	生长素响应元件Auxin-responsive element	1
TC-rich repeats	ATTCTCTAAC	参与防御和应激反应的顺式作用因子	1
		cis-acting element involved in defense and stress responsiveness
ACE	CTAACGTATT	与光响应性有关的顺式作用元件cis-acting element involved in light responsiveness	1
TCA-element	CCATCTTTTT	参与水杨酸反应的顺式作用元件	1
		cis-acting element involved in salicylic acid responsiveness
ABRE	ACGTG	参与脱落酸反应的顺式元件	2
	AACCCGG	cis-acting element involved in the abscisic acid responsiveness	1
ARE	AAACCA	对厌氧诱导必不可少的顺式调节元件	2
		cis-acting regulatory element essential for the anaerobic induction
AuxRR-core	GGTCCAT	参与生长素反应性的顺式调控元件	1
		cis-acting regulatory element involved in auxin responsiveness
G-box	TACGTG	参与光响应的顺式调节元件	2
		cis-acting regulatory element involved in light responsiveness
TGACG-motif	CGTCA	参与MeJA反应的顺式作用调控元件	3
	TGACG	cis-acting regulatory element involved in the MeJA-responsiveness	3
CAAT-box	CAAAT	启动子和增强子区域中常见的顺式作用元件	14
	CCAAT	Common cis-acting element in promoter and enhancer regions	7
TATA-box	ATATAA	核心启动子元件在转录开始的-30左右	3
	ATATAT	Core promoter element around -30 of transcription start-30左右	3
	ATTATA		6
	TACAAAA		2
	TATA		20
	TATAA		6
	TATAAA		1
	TATAAAT		1
	TATAAATA		1
	TATACA		3
	TATATA		3
	TATATAA		2
	taTATAAAtc		1
	TATATTTATATTT		1
	TATTTAAA		1
GT1-motif	GGTTAA	光响应元件Light responsive element	2
MRE	AACCTAA	MYB结合位点参与光响应性MYB binding site involved in light esponsiveness	2
Box 4	ATTAAT	与光反应有关的保守DNA模块的一部分	5
		Part of a conserved DNA module involved in light responsiveness
TCT-motif	TCTTAC	光响应元件的一部分Part of a light responsive element	1
GATA-motif	GATAGGA	光响应元件的一部分Part of a light responsive element	1
TGA-box	TGACGTAA	生长素反应元件的一部分Part of an auxin-responsive element	1

名称 Name	序列 Sequence	功能 Function	数目 Amount
TGA-element	AACGAC	生长素响应元件Auxin-responsive element	1
TC-rich repeats	ATTCTCTAAC	参与防御和应激反应的顺式作用因子	1
		cis-acting element involved in defense and stress responsiveness
ACE	CTAACGTATT	与光响应性有关的顺式作用元件cis-acting element involved in light responsiveness	1
TCA-element	CCATCTTTTT	参与水杨酸反应的顺式作用元件	1
		cis-acting element involved in salicylic acid responsiveness
ABRE	ACGTG	参与脱落酸反应的顺式元件	2
	AACCCGG	cis-acting element involved in the abscisic acid responsiveness	1
ARE	AAACCA	对厌氧诱导必不可少的顺式调节元件	2
		cis-acting regulatory element essential for the anaerobic induction
AuxRR-core	GGTCCAT	参与生长素反应性的顺式调控元件	1
		cis-acting regulatory element involved in auxin responsiveness
G-box	TACGTG	参与光响应的顺式调节元件	2
		cis-acting regulatory element involved in light responsiveness
TGACG-motif	CGTCA	参与MeJA反应的顺式作用调控元件	3
	TGACG	cis-acting regulatory element involved in the MeJA-responsiveness	3
CAAT-box	CAAAT	启动子和增强子区域中常见的顺式作用元件	14
	CCAAT	Common cis-acting element in promoter and enhancer regions	7
TATA-box	ATATAA	核心启动子元件在转录开始的-30左右	3
	ATATAT	Core promoter element around -30 of transcription start-30左右	3
	ATTATA		6
	TACAAAA		2
	TATA		20
	TATAA		6
	TATAAA		1
	TATAAAT		1
	TATAAATA		1
	TATACA		3
	TATATA		3
	TATATAA		2
	taTATAAAtc		1
	TATATTTATATTT		1
	TATTTAAA		1
GT1-motif	GGTTAA	光响应元件Light responsive element	2
MRE	AACCTAA	MYB结合位点参与光响应性MYB binding site involved in light esponsiveness	2
Box 4	ATTAAT	与光反应有关的保守DNA模块的一部分	5
		Part of a conserved DNA module involved in light responsiveness
TCT-motif	TCTTAC	光响应元件的一部分Part of a light responsive element	1
GATA-motif	GATAGGA	光响应元件的一部分Part of a light responsive element	1
TGA-box	TGACGTAA	生长素反应元件的一部分Part of an auxin-responsive element	1

番茄SlMYB52基因鉴定、亚细胞定位及表达分析

Identification, subcellular localization and expression analysis of tomato SlMYB52 gene

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