Expression Characteristics and Functions of HSFs Transcription Factors Family in Maize under Drought Stress

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

Abstract

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

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.

Figures/Tables 11

References 29

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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

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

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	‒‒	‒‒	‒‒

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	‒‒	‒‒	‒‒