神农香菊CiMYB4基因分析及其抗旱功能鉴定

doi:10.11686/cyxb2025045

摘要/Abstract

摘要：

干旱是影响菊花地理分布和生长发育的一个重要环境因子，MYB作为植物重要的转录因子家族广泛参与非生物胁迫的调控过程。本研究通过对课题组前期从神农香菊中克隆获得的CiMYB4基因以及启动子序列进行生物信息学分析，并测定干旱胁迫处理后过表达CiMYB4烟草和野菊株系的生理指标，初步解析CiMYB4在干旱胁迫中的功能。研究结果表明：CiMYB4开放阅读框全长846 bp，编码281个氨基酸，属于R2R3-MYB亚族，定位在细胞核中。CiMYB4启动子序列中存在7个与干旱胁迫相关的顺式作用元件，其中4个为脱落酸（ABA）响应元件，3个为STRE干旱胁迫响应元件。在干旱处理7和14 d时，过表达CiMYB4烟草和野菊株系的丙二醛（MDA）含量均显著低于野生型（P<0.05），而脯氨酸（Pro）含量以及超氧化物歧化酶（SOD）、过氧化氢酶（CAT）活性均显著高于野生型（P<0.05）。综合上述结果可知，CiMYB4在干旱胁迫中具有正向调控功能，是培育抗旱性强的菊花新品种潜在的基因资源。

关键词: 神农香菊, CiMYB4, 启动子, 抗旱

Abstract:

Drought is an important environmental factor affecting the geographical distribution and growth of chrysanthemums （Chrysanthemum×morifolium）. The MYB transcription factors， as a significant family of plant transcription factors， are widely involved in the regulation of responses to abiotic stresses. In this research， the CiMYB4 gene and its promoter sequence were cloned from Chrysanthemum indicum var. aromaticum and bioinformatics analyses were conducted. Additionally， the physiological indices of transgenic tobacco （Nicotiana tabacum） and C. indicum lines overexpressing CiMYB4 were measured during a drought stress treatment to provide preliminary insights into its function in the drought response. The results showed that the length of the CiMYB4 open reading frame was 846 bp， encoding a protein consisting of 281 amino acids. The protein was identified as a member of the R2R3-MYB subfamily and was predicted to localize to the nucleus. The promoter region of CiMYB4 contained seven cis-elements associated with drought stress， including four abscisic acid response elements and three STRE drought response elements. At 7 and 14 days of a drought treatment， the malondialdehyde content was significantly lower in CiMYB4-overexpressingtobacco and CiMYB4-overexpressingchrysanthemum plants than in their respective wild-type controls （P<0.05）， whereas the proline content and the activities of superoxide dismutase and catalase were significantly higher than those of the wild type controls （P<0.05）. Together， these results demonstrate that CiMYB4 plays a positive regulatory role in drought stress responses and it is a potential genetic resource for developing new chrysanthemum cultivars with enhanced drought tolerance.

Key words: Chrysanthemum indicum var. aromaticum, CiMYB4, promoter, drought resistance

陈斌, 刘晏廷, 陈胜艳, 薛晴, 李梦雨, 王霁佳, 孙颖, 何淼. 神农香菊CiMYB4基因分析及其抗旱功能鉴定[J]. 草业学报, 2026, 35(1): 179-191.

Bin CHEN, Yan-ting LIU, Sheng-yan CHEN, Qing XUE, Meng-yu LI, Ji-jia WANG, Ying SUN, Miao HE. Bioinformatics analysis of CiMYB4 in Chrysanthemum indicum var. aromaticum and functional characterization of its role in drought resistance[J]. Acta Prataculturae Sinica, 2026, 35(1): 179-191.

图/表 11

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引物名称Primer name	引物序列Primer sequence （5′-3′）
CiMYB4-2001-F （P-F）	GATTACGCCAAGCTTTTGTGGAATCAAACATGCAAAAC
CiMYB4-1764 （P1-F）	GATTACGCCAAGCTTCTATAAAATATGCATAACGAAAAAT
CiMYB4-1299 （P2-F）	GATTACGCCAAGCTTAGAAATTATTATGTAAAACCTAG
CiMYB4-954 （P3-F）	GATTACGCCAAGCTTTTCATTTCCTCTTTATTGTCATTTC
CiMYB4-417 （P4-F）	GATTACGCCAAGCTTTTCATAAGCAAAATAGATTCCCT
CiMYB4-2001-R （P-R）	ACCACCCGGGGATCCCATATTTAGACTTTCTTCTTTG

引物名称Primer name	引物序列Primer sequence （5′-3′）
CiMYB4-2001-F （P-F）	GATTACGCCAAGCTTTTGTGGAATCAAACATGCAAAAC
CiMYB4-1764 （P1-F）	GATTACGCCAAGCTTCTATAAAATATGCATAACGAAAAAT
CiMYB4-1299 （P2-F）	GATTACGCCAAGCTTAGAAATTATTATGTAAAACCTAG
CiMYB4-954 （P3-F）	GATTACGCCAAGCTTTTCATTTCCTCTTTATTGTCATTTC
CiMYB4-417 （P4-F）	GATTACGCCAAGCTTTTCATAAGCAAAATAGATTCCCT
CiMYB4-2001-R （P-R）	ACCACCCGGGGATCCCATATTTAGACTTTCTTCTTTG

类别 Category	名称 Name	核心序列 Core sequence	功能 Function	数量 Amount
结构元件Structure elements	TATA-box	TATA	核心启动子元件Core promoter element	50
结构元件Structure elements	CAAT-box	CAA（A）T	启动子和增强子区域调控元件Regulatory element in promoter and enhancer regions	35
激素响应元件Phytohormone responsive elements	ABRE	ACGTG	脱落酸响应元件Abscisic acid responsive element	3
	AAGAA-motif	GTAAAGAAA	脱落酸响应元件Abscisic acid responsive element	1
	TGA-element	AACGAC	生长素响应元件Auxin acid responsive element	1
胁迫响应元件Abiotic stress responsive elements	LTR	AAAGCC	低温响应元件Low temperature responsive element	1
	STRE	AGGGG	干旱胁迫响应元件Drought stress responsive element	3
	TC-rich repeats	ATTTTCTT	防卫和胁迫响应元件Defense and stress responsive element	2
光响应元件Light responsive elements	G-box	（T）CACGTG（T）	光响应元件Light responsive element	2
	GATA-motif	AAGGATAAGG	光响应元件Light responsive element	1
	GT1-motif	GGTTAA	光响应元件Light responsive element	3
	I-box	CCTTATCCT	光响应元件Light responsive element	1
	Box 4	ATTAAT	光响应元件Light responsive element	2
	chs-CMA1a	TTACTTAA	光响应元件Light responsive element	1
	MRE	AACCTAA	光响应元件Light responsive element	1
生长发育调控元件Development-related elements	CAT-box	GCCACT	分生组织表达元件Meristem expression element	1
功能未知元件Function unknown elements	MYB CORE	TAACCA、CAACAG	MYB结合位点MYB binding site	7
	MYC CORE	CATTTG、CAATTG	MYC结合位点MYC binding site	3
	A-box	CCGTCC	未知功能元件Function unknown element	2
	AT~TATA-box	TATATA	未知功能元件Function unknown element	1

类别 Category	名称 Name	核心序列 Core sequence	功能 Function	数量 Amount
结构元件Structure elements	TATA-box	TATA	核心启动子元件Core promoter element	50
结构元件Structure elements	CAAT-box	CAA（A）T	启动子和增强子区域调控元件Regulatory element in promoter and enhancer regions	35
激素响应元件Phytohormone responsive elements	ABRE	ACGTG	脱落酸响应元件Abscisic acid responsive element	3
	AAGAA-motif	GTAAAGAAA	脱落酸响应元件Abscisic acid responsive element	1
	TGA-element	AACGAC	生长素响应元件Auxin acid responsive element	1
胁迫响应元件Abiotic stress responsive elements	LTR	AAAGCC	低温响应元件Low temperature responsive element	1
	STRE	AGGGG	干旱胁迫响应元件Drought stress responsive element	3
	TC-rich repeats	ATTTTCTT	防卫和胁迫响应元件Defense and stress responsive element	2
光响应元件Light responsive elements	G-box	（T）CACGTG（T）	光响应元件Light responsive element	2
	GATA-motif	AAGGATAAGG	光响应元件Light responsive element	1
	GT1-motif	GGTTAA	光响应元件Light responsive element	3
	I-box	CCTTATCCT	光响应元件Light responsive element	1
	Box 4	ATTAAT	光响应元件Light responsive element	2
	chs-CMA1a	TTACTTAA	光响应元件Light responsive element	1
	MRE	AACCTAA	光响应元件Light responsive element	1
生长发育调控元件Development-related elements	CAT-box	GCCACT	分生组织表达元件Meristem expression element	1
功能未知元件Function unknown elements	MYB CORE	TAACCA、CAACAG	MYB结合位点MYB binding site	7
	MYC CORE	CATTTG、CAATTG	MYC结合位点MYC binding site	3
	A-box	CCGTCC	未知功能元件Function unknown element	2
	AT~TATA-box	TATATA	未知功能元件Function unknown element	1