Function Analysis of the Transcription Factor PsnbZIP1 of Populus simonii×P. nigra in Response to Salt Stress

doi:10.7525/j.issn.1673-5102.2023.02.014

Abstract

Abstract:

To reveal the function of transcription factor PsnbZIP1 from Populus simonii × P. nigra under abiotic stresses， P.simonii × P. nigra was used as material， and the PsnbZIP1 transcription factor gene was cloned with an ORF length of 432 bp， and the molecular mechanism of PsnbZIP1 under salt stress was analyzed. The expression pattern of PsnbZIP1 in P. simonii × P. nigra was analyzed under 150 mmol·L^-1 NaCl by q-PCR， it was found that the expression of the gene increased rapidly. The PsnbZIP1 was predicted to be a hydrophilic unstable protein without trans-membrane structure but with signal peptide by bioinformatics analysis. The sub-cellular localization of the gene was observed by Agrobacterium mediated transient expression in tobacco. The results showed that the gene was a nuclear localization protein. It was confirmed by Y2HGold yeast competent cells that the gene had no transcriptional activation activity in yeast. The analysis of the promoter sequence of the PsnbZIP1 gene showed that the promoter contained auxin response elements， ABA response elements， light response elements and seed-specific cis-acting regulatory elements respectively， which might play a role in plant growth and development and response to stress.The promoter also included the MYB binding site and MYBHv1 binding site involved in drought induction， indicating that the gene was likely to interact with some of MYB involved in drought induction.

Key words: Populus simonii × P.nigra, bZIPtranscription factor, salt stress, bioinformatics analysis

CLC Number:

S722.3+6

Shixian LIAO, Yuting WANG, Liben DONG, Yongmei GU, Fenglin JIA, Tingbo JIANG, Boru ZHOU. Function Analysis of the Transcription Factor PsnbZIP1 of Populus simonii×P. nigra in Response to Salt Stress[J]. Bulletin of Botanical Research, 2023, 43(2): 288-299.

Figures/Tables 12

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引物 Primer	引物序列（5′→3′） Sequence（5′→3′）
PsnbZIP1-F	CTCACACAACAACATTTACG
PsnbZIP1-R	ACAGCATAACCAGAGAAATC
qRT-PsnbZIP1-F	TGAAAAGTTGAAGTTGGCAG
qRT-PsnbZIP1-R	AACTTTGAACATCCCAGAAG
Actin-F	ACCCTCCAATCCAGACACTG
Actin-R	TTGCTGACCGTATGAGCAAG
pBI121-PsnbZIP1-GFP-F	GTCGACATGCCACCATCCTTTGCAAA
pBI121-PsnbZIP1-GFP-R	ACTAGTAACTTTGAACATCCCAGAAG
pBi121-GFP-F1	CCATCGTTGAAGATGCCTCTGC
pBi121-GFP-R1	CAACGCTGATCAATTCCACAG
PsnbZIP1-BD-F	CATATGATGCCACCATCCTTTGCAAA
PsnbZIP1-BD-R	GAATTCCTAAACTTTGAACATCCCAG
BD-F	GTAATACGACTCACTATAGGGCGA
BD-R	TTTCGTTTTAAAACCTAAGAGTC
Pro-PsnbZIP1-F	GGTGCCCAGTAATTACCC
Pro-PsnbZIP1-R	GCAAAGGATGGTGGCATTTT
PsnbZIP1-BD-N-F	GAATTCATGCCACCATCCTTTGCAAAG
PsnbZIP1-BD-N-R	GGATCCAGCAAAATGCCTTTGCCAAAG
PsnbZIP1-BD-C-F	GAATTCCTCGAATCAGACAACGAAGTC
PsnbZIP1-BD-C-R	GGATCCCTAAACTTTGAACATCCCAGA

胁迫时间 /h	相对表达量 Relative expression levels
胁迫时间 /h	根 Root	茎 Stem	叶 Leaf
0	1.000a	1.000a	1.000a
3	（18.835±0.450）b	（16.992±0.325）b	（10.931±0.326）b
6	（38.487±0.394）b	（22.681±0.397）b	（24.564±0.382）b
12	（89.583±0.421）b	（79.792±0.382）b	（43.637±0.153）b
24	（75.107±0.387）b	（217.259±0.298）b	（52.146±0.159）b
48	（126.280±0.207）b	（156.977±0.280）b	（47.963±0.326）b

指标 Indicator	位点 Site	得分 Score
原始剪切位点的分值（Max. C） Score for original splice site （Max. C）	20	0.323
综合剪切位点的分值（Max. Y） Scores for integrated splicing site （Max. Y）	20	0.481
信号肽的分值（Max. S） Signal Peptide Score （Max. S）	13	0.804
信号肽分值的平均值（Mean. S） Mean value of signal peptide score （Mean. S）	1-19	0.724
Mean. S和Max. Y的加权平均值（D） Weighted mean of Mean. S and Max. Y（D）	1-19	0.612

作用元件 Acting element	序列 Sequence	功能 Function
G-box	TACGTG/CACGTG	参与光反应性的顺式作用调节元件 cis-acting regulatory element involved in light responsiveness
ABRE	CGTACGTGCA/CACGTG	参与脱落酸响应性的顺式作用元件 cis-acting element involved in the abscisic acid responsiveness
TGA-element	AACGAC	生长素应答元件 Auxin-responsive element
MRE	AACCTAA	涉及光响应的 MYB 结合位点 MYB binding site involved in light responsiveness
TATA-box	TATA	转录开始-30左右的核心启动子元件 Core promoter element around-30 of transcription start
CCAAT-box	CAACGG	MYBHv1 结合位点 MYBHv1 binding site MYBHv1
MBS	CAACTG	参与干旱诱导的 MYB 结合位点 MYB binding site involved in drought-inducibility
4cl-CMA1b	ATTCCGATAAACT	光响应元件 Light responsive element
Box4	ATTAAT	参与光响应性的保守 DNA 模块的一部分 Part of a conserved DNA module involved in light responsiveness
ATCT-motif	AATCTAATCC	参与光响应性的保守 DNA 模块的一部分 Part of a conserved DNA module involved in light responsiveness
RY-element	CATGCATG	参与种子特异性调控的顺式作用调控元件 cis-acting regulatory element involved in seed-specific regulation
IBox	AAGATAAGGCT	光响应元件的一部分 Part of a light responsive element
ARE	AAACCA	厌氧诱导所必需的顺式作用调节元件 cis-acting regulatory element essential for the anaerobic induction
CAAT-box	CCAAT	启动子和增强子区域的共同顺式作用元件 Common cis-acting element in promoter and enhancer regions
AE-box	AGAAACAA	光响应模块的一部分 Part of a module for light response

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