Response Analysis of Transcription Factor PsnbHLH162 Gene in Populus simonii × P. nigra under Salt Stress and Low Temperature Stress

doi:10.7525/j.issn.1673-5102.2023.02.015

Abstract

Abstract:

To reveal the role of transcription factor PsnbHLH162 of Populus simonii × P. nigra under abiotic stresses， and to explore the signal transduction process of genes in plants， the signal transduction process of this gene in plants is explored to provide the theoretical basis for the acquisition of excellent stress-resistant tree species in the future. In this experiment， PsnbHLH162 gene was cloned and obtained from P. simonii × P. nigra as material， and the target gene and promoter were analyzed by bioinformatics； and the function of genes in response to abiotic stresses was analyzed by fluorescence quantitative PCR. The results showed that the cDNA of PsnbHLH162 gene was 537 bp， and the N-terminal of the gene contained a highly conserved HLH domain， and the encoded protein might be a stable hydrophilic protein without trans-membrane region， localized in the nucleus and had no transcriptional activation activity. The promoter region contained a variety of ABA-responsive， auxin-responsive， light-responsive and circadian elements， confirming that this gene participated in abiotic stress response. The results of fluorescence quantitative PCR showed that under salt stress， the gene expression in root tissue was the highest compared with stem and leaf tissue； under low temperature stress， gene expression in stem tissue was the highest compared with leaf and root tissue. It was found that the expression of PsnbHLH162 could be induced by salt and low temperature in plants.

Key words: Populus simonii × P. nigra, PsnbHLH 162, bioinformatics, low temperature stress, salt stress

CLC Number:

S722.3+6

Senyao LIU, Fenglin JIA, Qing GUO, Gaofeng FAN, Boru ZHOU, Tingbo JIANG. Response Analysis of Transcription Factor PsnbHLH162 Gene in Populus simonii × P. nigra under Salt Stress and Low Temperature Stress[J]. Bulletin of Botanical Research, 2023, 43(2): 300-310.

Figures/Tables 11

Table 1

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Table2

Analysis of the cis-acting elements on the promoter of PsnbHLH162

作用元件 Acting element	序列（5′→3′） Sequence（5′→3′）	功能 Function
G-box	TACGTG/CACGTG	光响应性中涉及的顺式作用调节元件 cis-acting regulatory element involved in light responsiveness
ABRE	CGTACGTGCA/CACGTG	顺式作用元素参与脱落酸反应性 cis-acting element involved in the abscisic acid responsiveness
TGA-element	AACGAC	生长素响应相关 Auxin-responsive element
MRE	AACCTAA	参与光响应MYB结合位点 MYB binding site involved in light responsiveness
TATA-box	TATA	转录开始约-30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	光响应相关 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	种子特异性调控中涉及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	顺式作用调节元件对厌氧诱导至关重要 cis-acting regulatory element essential for the anaerobic induction
CAAT-box	CCAAT	促进子和增强子区域中的常见顺式作用元素 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
A-box	CCGTCC	顺式作用调节元件 cis-acting regulatory element
AuxRR-core	GGTCCAT	参与生长素反应的顺式作用调节元件 cis-acting regulatory element involved in auxin responsiveness
Box II	TGGTAATAA	光响应元件的一部分 Part of a light responsive element
CGTCA-motif	CGTCA	MeJA响应性中涉及的顺式作用调节元件 cis-acting regulatory element involved in the MeJA-responsiveness
GATA-motif	GATAGGA	光响应元件的一部分 Part of a light responsive element
O2-site	GATGATGTGG	参与玉米蛋白代谢调节的顺式作用调节元件 cis-acting regulatory element involved in zein metabolism regulation
TCA-element	CCATCTTTTT	顺式作用元素参与水杨酸反应性 cis-acting element involved in salicylic acid responsiveness
TCCC-motif	TCTCCCT	光响应元件的一部分 Part of a light responsive element
TCT-motif	TCTTAC	光响应元件的一部分 Part of a light responsive element
TGACG-motif	TGACG	MeJA响应性中涉及的顺式作用调节元件 cis-acting regulatory element involved in the MeJA-responsiveness
circadian	CAAAGATATC	参与昼夜节律控制的顺式作用调节元件 cis-acting regulatory element involved in circadian control

Table2

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引物名称 Primer name	引物序列（5′→3′） Primer sequence（5′→3′）	引物大小 Primer size /bp
PsnbHLH162 F1	ATGGAGAAGAGCAACAGTGAGA	22
PsnbHLH162 R1	TTAGGAAATCAGTTCTTGCAATC	23
PsnbHLH162 F2	TCTAGAATGGAGAAGAGCAACAGTGAG	27
PsnbHLH162 R2	GTCGACCGGAAATCAGTTCTTGCAATC	27
pBI121 F	GTAAGGGATGACGCACAATC	20
pBI121 R	GTCTTGTAGTTGCCGTCGTC	20
PsnbHLH 162 BDF	CCTGCATATGGCATGGCGGAGGAGTTTC	28
PsnbHLH 162 BDR	GGCCTCCATGGCCCTGAAAGTACCACC	27
BD F	GTAATACGACTCACTATAGGGCGA	24
BD R	TTTCGTTTTAAAACCTAAGAGTC	23
PsnbHLH 162 F3	GGCTTGAGATTGCCGGTGATTG	22
PsnbHLH 162 R3	GCACTCTTGAAGTCTCCAC	19
ACT F	ACCCTCCAATCCAGACACTG	18
ACT R	TTGCTGACCGTATGAGCAAG	18
QDbHLH162 F	GAGCAAGCCACATAAAAGGATAAC	24
QDbHLH162 R	CACTGTTGCTCTTCTCCATCTTG	23

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