玉米HSFs转录因子家族在干旱胁迫下的表达特性及功能

doi:10.13560/j.cnki.biotech.bull.1985.2025-0659

摘要/Abstract

摘要：

目的鉴定分析干旱胁迫下玉米HSFs家族成员的结构和功能，为解析玉米耐旱性分子机制奠定基础。方法从玉米基因组数据库（MaizeGDB）中下载蛋白序列，基于Pfam数据库（PF00447）的HSF结构域，通过HMMER 3.0和BLASTP比对筛选玉米HSFs家族成员，利用ExPASy、MEGA X、GSDS、MEME、MCScanX、PlantCARE和TBtools等生物信息学工具分析预测蛋白理化性质、染色体定位、基因结构、共线性、保守基序和顺势元件等信息。基于转录组数据和实时荧光定量PCR分析，了解HSFs家族成员基因在干旱胁迫下的表达模式，挖掘响应干旱胁迫的HSFs基因。结果玉米中共鉴定到28个HSFs基因，编码的氨基酸数量为257‒528 aa，等电点为4.70‒9.53；不均匀地分布在10条染色体上，并且在第1染色体分布最密集；启动子区富含ABRE、MYB等逆境响应元件；共线性分析显示，玉米与水稻之间具有较强的保守性。RNA-seq和荧光定量PCR显示，有4个基因存在显著差异，同时，过表达ZmHSF16可显著提高拟南芥的抗旱性（存活率提高38%‒43%），并通过降低细胞膜损伤（相对电导率减少30%）维持细胞稳态。结论揭示了ZmHSFs家族在干旱胁迫下的表达特性，并验证了ZmHSF16的抗旱功能。

关键词: 玉米, 热激转录因子, 干旱胁迫, 表达模式, 生物信息学

Abstract:

Objective To identify and analyze the structure and function of HSFs family members in maize (Zea mays L.)under drought stress, laying a foundation for understanding the molecular mechanism of maize drought tolerance. Method Protein sequences were downloaded from the Maize Genome Database (MaizeGDB). Based on the HSF domain of the Pfam database (PF00447), the maize HSFs family members were screened through HMMER 3.0 and BLASTP alignment. Bioinformatics tools such as ExPASy, MEGA X, GSDS, MEME, MCScanX, PlantCARE, and TBtools were used to analyze and predict information such as the physicochemical properties of proteins, chromosome localization, gene structure, collinearity, conserved mosequences, and homeotropic elements. Based on transcriptome data and real-time fluorescence quantitative PCR analysis, the expression patterns of HSFs family member genes under drought stress were revealed and explore HSFs genes that respond to drought stress. Result A total of 28 HSFs genes were identified in maize, with the number of amino acids encoded ranging from 257 to 528 aa and isoelectric points ranging from 4.70 to 9.53. It was unevenly distributed on 10 chromosomes, with the densest distribution on chromosome 1. The promoter region was rich in adverse response elements such as ABRE and MYB. Collinearity analysis showed that there was a strong conservativeness between maize and rice (Oryza sativa). RNA-seq and fluorescence quantitative PCR showed that there were significant differences in four genes. Meanwhile, the overexpression of ZmHSF16 significantly enhanced the drought resistance of Arabidopsis thaliana (survival rate increased by 38%‒43%), and maintained cell homeostasis by reducing cell membrane damage (relative conductivity decreased by 30%). Conclusion The expression characteristics of the ZmHSFs family under drought stress are revealed, and the drought resistance function of ZmHSF16 is verified.

Key words: maize, heat shock transcription factor, drought stress, expression pattern, bioinformatics

张冬岭, 张寅生, 王建军, 叶飞宇, 卢子涵, 马晨晨, 柳华峰, 胡德升, 邓亚洲, 曹丽茹. 玉米HSFs转录因子家族在干旱胁迫下的表达特性及功能[J]. 生物技术通报, 2026, 42(2): 178-187.

ZHANG Dong-ling, ZHANG Yin-sheng, WANG Jian-jun, YE Fei-yu, LU Zi-han, MA Chen-chen, LIU Hua-feng, HU De-sheng, DENG Ya-zhou, CAO Li-ru. Expression Characteristics and Functions of HSFs Transcription Factors Family in Maize under Drought Stress[J]. Biotechnology Bulletin, 2026, 42(2): 178-187.

图/表 11

表1 定量引物序列

Table 1 Quantitative primer sequences

引物名称 Primer name	正向引物 Forward primer （5′‒3′）	反向引物 Reverse primer （5′‒3′）
HSF16	ATGCAAGAGCTTGTCAGGCT	CAACGAAGCAGCAGCACTTT
HSF7	GAGCTTGGGAAGGGAAGCAT	CCTCTCCACCACCAGTCAAC
HSF25	TGAACGAAGGGATGAAGGGC	ACTACACGTCTGCACTCTGC
HSF26	CACCCACCATGGAGAAGGAC	TGGTCTGGATCCCTCCTCAG

表2 ZmHSFs蛋白理化性质

Table 2 Physicochemical properties of ZmHSFs protein

基因IDGene ID	基因Gene	蛋白长度 Protein length （aa）	等电点pI	分子量 Molecular weight （Da）	不稳定指数 Instability index	疏水指数 GRAVY	带负电残基数 Asp+Glu	带正电残基数 Arg+Lys
Zm00001eb353710	ZmHSF1	449	5.15	49 978	58.13	-0.586	62	42
Zm00001eb293250	ZmHSF2	469	5.41	51 624	60.51	-0.533	62	48
Zm00001eb017880	ZmHSF3	331	9.11	37 173	61.83	-0.783	40	45
Zm00001eb100770	ZmHSF4	298	9.13	32 270	55.75	-0.556	36	41
Zm00001eb056980	ZmHSF5	371	5.12	41 282	48.30	-0.604	59	47
Zm00001eb160080	ZmHSF6	398	5.84	43 463	42.50	-0.461	52	47
Zm00001eb047760	ZmHSF7	357	4.99	40 502	59.15	-0.722	61	46
Zm00001eb328900	ZmHSF8	323	6.57	35 561	59.80	-0.623	36	31
Zm00001eb399800	ZmHSF91	407	4.97	45 318	53.67	-0.590	55	36
Zm00001eb242480	ZmHSF10	508	4.96	56 062	55.94	-0.536	77	49
Zm00001eb358700	ZmHSF11	348	8.09	37 410	53.05	-0.302	35	37
Zm00001eb100410	ZmHSF12	394	7.81	41 743	66.21	-0.372	36	37
Zm00001eb159390	ZmHSF13	331	5.94	35 884	60.93	-0.345	40	35
Zm00001eb198620	ZmHSF14	383	6.23	41 003	58.83	-0.515	47	43
Zm00001eb239380	ZmHSF15	257	5.85	27 837	46.60	-0.402	33	30
Zm00001eb064860	ZmHSF16	500	5.01	54 420	59.29	-0.507	59	44
Zm00001eb384760	ZmHSF17	267	6.99	28 329	42.41	-0.228	30	30
Zm00001eb371000	ZmHSF18	433	5.25	48 648	51.50	-0.842	69	51
Zm00001eb009170	ZmHSF19	417	5.09	46 816	55.59	-0.693	57	39
Zm00001eb301280	ZmHSF20	375	4.7	42 044	53.81	-0.716	63	37
Zm00001eb060670	ZmHSF21	359	5.57	40 587	53.80	-0.835	53	41
Zm00001eb314890	ZmHSF22	298	9.53	32 258	53.74	-0.567	35	43
Zm00001eb037600	ZmHSF23	379	6.27	40 646	57.77	-0.434	45	42
Zm00001eb319350	ZmHSF24	394	5	41 468	54.72	-0.433	50	35
Zm00001eb004670	ZmHSF25	384	5.3	43 268	71.81	-0.759	62	51
Zm00001eb241300	ZmHSF26	528	5.57	58 139	51.23	-0.714	74	56
Zm00001eb428800	ZmHSF27	318	5.7	33 948	45.96	-0.707	50	45
Zm00001eb210530	ZmHSF28	497	5.06	54 105	63.64	-0.484	58	44
平均值 Average		384.75	6.08	42 206.51	55.59	‒‒	‒‒	‒‒

图1 玉米、拟南芥、水稻HSFs系统进化树分析

Fig. 1 Phylogenetic tree analysis of HSFs in maize (Z. mays), A.thaliana and rice (O. sativa)

图2 玉米HSFs家族基因结构

Fig. 2 Gene structure of maize HSFs family

图3 玉米HSFs家族基因motif

Fig. 3 Motif of HSFs family gene in Zea mays L.

图4 玉米HSFs家族基因染色体位置

Fig. 4 Chromosome locations of HSFs family genes in maize

图5 玉米、水稻和拟南芥HSFs基因家族种间共线性分析

Fig. 5 Interspecific collinearity analysis of HSFs gene families in maize, rice and Arabidopsis

图6 玉米HSFs基因家族顺式作用元件分析

Fig. 6 Analysis of cis-acting elements of maize HSFs gene family

图7 ZmHSFs基因家族在不同组织的表达水平1：第7‒8节间；2：授粉后18 d的胚；3：授粉后38 d的胚；4：授粉后16 d的胚乳；5：授粉后27 d的胚乳；6：根；7：叶；8：花粉；9：花丝；10：雌穗；11：叶耳原基2‒4 mm；12：叶耳原基6‒8 mm；13：授粉后27 d的种皮/糊粉层

Fig. 7 Expressions of ZmHSFs gene family in different tissues1: Internode 7‒8. 2: Embryo 18 DAP. 3: Embryo 38 DAP. 4: Endosperm l6 DAP. 5: Endosperm 27 DAP. 6: Root. 7: Leaf. 8: Pollen. 9: Silk. 10: Female spike. l1: Ear primordium 2‒4 mm. 12: Ear primordium 6‒8 mm. 13: Pericarp/Aleurone 27 DAP

图8 ZmHSFs基因家族在干旱胁迫下的表达水平A：ZmHSFs基因家族在干旱胁迫下RNA-seq数据分析；B：相关基因的表达量检测。**：P<0.01。下同

Fig. 8 Expressions of ZmHSFs gene family under drought stressA: Analysis of RNA-seq data of ZmHSFs gene family under drought stress. B: Detection of gene expressions. **: P<0.01. The same below

图9 ZmHSF16增强拟南芥的抗旱性A：ZmHSF16在WT和6个独立的ZmHSF16过表达系中的表达水平；B：正常和干旱胁迫条件下WT和过表达ZmHSF16株系的表型分析；C、D：干旱胁迫处理5 d后WT和过表达ZmHSF16株系存活率和电导率

Fig. 9 ZmHSF16 enhances drought resistance in ArabidopsisA: The expressions of ZmHSF16 in WT and six independent ZmHSF16 overexpression lines. B: Phenotypic analysis of WT, ZmHSF16 overexpressing materials under normal conditions and drought stress conditions. C, D: Survival rate and conductivity of WT, ZmHSF16-overexpressing materials after 5 d of drought stress treatment

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