Expression Pattern Analysis of PdbHMGs Genes in Populus davidiana×P. bolleana under Abiotic Stress

doi:10.7525/j.issn.1673-5102.2023.06.015

Abstract

Abstract:

In order to investigate the function of HMG（High mobility group proteins） genes under abiotic stresses in Populus davidiana×P. bolleana， the HMG genes were cloned and the stress tolerant function of HMGs was identified， and the theoretical foundation for using the gene in tree breeding through genetic engineering was laid. Seven homologous genes of ArabidopsisHMGs（named PdbHMG1-7） were found by homologous sequence alignment， and sequence analysis of PdbHMG1-7 was performed using bioinformatics tools respectively， the expression patterns of PdbHMG1-7 gene in leaves and roots of Populus davidiana×P.bolleana under different stresses were investigated by qRT-PCR.Results The ORF length of PdbHMG1-7 were 456-2 004 bp respectively， the amino acid residues encoded by PdbHMG1-7 genes were 151-667 aa， and the molecular weight of the encoded protein were 16.72-75.54 kDa， and the theoretical isoelectric point were 5.35-9.36， and the seven PdbHMG were hydrophilic proteins. The results of multiple sequence alignment and phylogenetic tree analysis showed that all 7 PdbHMG genes contained HMG-box conserved domains and belonged to HMGB subfamily. The analysis results of cis-acting elements contained in the promoter showed that the promoter sequence of PdbHMG genes contained a variety of elements in response to hormones and stress. The expression pattern analysis showed that the expression levels of seven PdbHMGs in roots and leaves under NaCl， PEG₆₀₀₀， ABA and CdCl₂ stress treatment changed in at least one stress treatment time point respectively. In leaves， PdbHMG2 were induced under the stress of NaCl， PEG₆₀₀₀， ABA and CdCl₂， and almost all of them were significantly up-regulated respectively， but PdbHMG7 was down-regulated. Under NaCl and CdCl₂ stress， PdbHMG1， PdbHMG3 and PdbHMG6 were almost significantly up-regulated during the whole stress process respectively. The expressions of PdbHMG3 in leaves were also significantly up-regulated at most time points under NaCl and CdCl₂ stress. In roots， the expression of PdbHMG3 and PdbHMG5 was significantly up-regulated in the whole process of NaCl treatment. However， PdbHMG6 was not significantly expressed during NaCl stress except for 9-12 h， the expression of PdbHMG6 was significantly down-regulated at other stress time points. During ABA treatment， PdbHMG6 was down-regulated in roots. Conclusion The results showed that seven PdbHMG genes could respond to at least one stress treatment in one organ， but the types and mechanisms of involved stress response might be different. This study provided a reference for further studying the function of PdbHMGs family genes in Populus davidiana×P. bolleana.

Key words: HMG, Populus davidiana×P.bolleana, abiotic stress, expression analysis

CLC Number:

Q943.2

Chunyao WANG, Xiaojin LEI, Zhongyuan LIU. Expression Pattern Analysis of PdbHMGs Genes in Populus davidiana×P. bolleana under Abiotic Stress[J]. Bulletin of Botanical Research, 2023, 43(6): 932-942.

Figures/Tables 13

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基因 Genes	引物 Primers	序列（5'→3'） Sequences（5'→3'）
PdbHMG1	PdbHMG1-F	GCCTCCTAGTGCTTTCTTT
PdbHMG1	PdbHMG1-R	CATTATCGTCATCGTCACCAT
PdbHMG2	PdbHMG2-F	CAGGAAAGGCAGCTAAGGAT
PdbHMG2	PdbHMG2-R	CTCATATTCAACCTTCCTCT
PdbHMG3	PdbHMG3-F	AGTGTGCCTGCCAACAAT
PdbHMG3	PdbHMG3-R	ATAGCTCACCAATCATCCT
PdbHMG4	PdbHMG4-F	AAGGAAGTGATTACAGCCT
PdbHMG4	PdbHMG4-R	AATGGCACCTTCTTCTAGGGTTGC
PdbHMG5	PdbHMG5-F	ACAACTAACAGGGGTGGT
PdbHMG5	PdbHMG5-R	AGACGGATCAGGACAATCCCT
PdbHMG6	PdbHMG6-F	AGGCAGATGTTGGTGCTGGAGGT
PdbHMG6	PdbHMG6-R	ACAACAAAGGTGTGTGGCT
PdbHMG7	PdbHMG7-F	AGCTGTAGCCTTGATGCTAAGAT
PdbHMG7	PdbHMG7-R	AGAGTCCGGGTTCGCTTCCT
Pdbactin	Pdbactin-F	GCTGAGAGATTCCGTTGCCCTG
Pdbactin	Pdbactin-R	GGCGGTGATCTCCTTGCTCATT

基因名称 Gene name	基因登录号 Gene ID	编码序列 Coding sequence/bp	内含子 Intron	外显子 Exon	氨基酸数 Number of amino acids/aa	相对分子量 Molecular Weight/kDa	理论等电点 Isoelectric point	定位预测 Location prediction
PdbHMG1	ON357372	531	6	7	176	19.90	5.35	细胞核 Nuclear
PdbHMG2	ON357373	456	6	7	151	16.72	6.06	细胞核 Nuclear
PdbHMG3	ON357374	1 404	5	6	467	51.89	8.16	细胞核 Nuclear
PdbHMG4	ON357375	954	5	6	317	35.86	9.36	细胞核 Nuclear
PdbHMG5	ON357376	990	5	6	329	37.71	9.28	细胞核 Nuclear
PdbHMG6	ON357377	2 004	18	19	667	75.54	8.26	细胞质 Cytoplasmic
PdbHMG7	ON357378	588	7	8	196	21.95	6.26	细胞核 Nuclear

基因名称 Gene name	植物激素响应元件 Phytohormone response element							非生物胁迫响应元件 Abiotic stress response element				共计 Amount
基因名称 Gene name	ABRE	TGA-element	TCA-element	CGTCA-motif	TGACG-motif	P-box	ERE	STRE	MBS	ARE	WUN-motif	共计 Amount
共计Amount	14	3	4	9	10	6	10	10	3	15	3	87
PdbHMG1	2	1	1	0	1	0	0	1	0	1	0	7
PdbHMG2	7	1	0	4	4	1	2	1	0	2	1	23
PdbHMG3	2	0	0	0	0	2	2	4	2	2	0	14
PdbHMG4	1	1	2	2	2	1	1	1	0	2	0	13
PdbHMG5	0	0	1	1	1	0	1	0	0	2	1	7
PdbHMG6	1	0	0	0	0	1	0	1	1	1	0	5
PdbHMG7	1	0	0	2	2	1	4	2	0	5	1	18

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