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植物研究 ›› 2023, Vol. 43 ›› Issue (2): 300-310.doi: 10.7525/j.issn.1673-5102.2023.02.015

• 分子生物学 • 上一篇    下一篇

小黑杨转录因子PsnbHLH162基因在盐和低温胁迫下应答分析

刘森尧, 贾丰璘, 国庆, 樊高锋, 周博如, 姜廷波()   

  1. 林木遗传育种国家重点实验室,东北林业大学,哈尔滨 150040
  • 收稿日期:2022-04-19 出版日期:2023-03-20 发布日期:2023-03-07
  • 通讯作者: 姜廷波 E-mail:tbjiang@yahoo.com
  • 作者简介:刘森尧(1997—),男,硕士研究生,主要从事林木遗传育种方面的研究。
  • 基金资助:
    黑龙江省应用技术研究与开发计划(GA20B401)

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

Senyao LIU, Fenglin JIA, Qing GUO, Gaofeng FAN, Boru ZHOU, Tingbo JIANG()   

  1. State Key Laboratory of Tree Genetics and Breeding,Northeast Forestry University,Harbin 150040
  • Received:2022-04-19 Online:2023-03-20 Published:2023-03-07
  • Contact: Tingbo JIANG E-mail:tbjiang@yahoo.com
  • About author:LIU Senyao(1997—),male,graduate student,specialized in research on forest genetics and breeding.
  • Supported by:
    Application Technology Research and Development Program of Heilongjiang Province(GA20B401)

摘要:

为揭示小黑杨(Populus simonii × P.nigra)在面对非生物胁迫时,转录因子PsnbHLH162在植物体内发挥的作用,同时探究该基因在植物体内的信号转导过程,进而为未来获取优良的抗逆树种提供理论基础。以小黑杨为原材料,克隆获取PsnbHLH162基因,对目的基因和启动子进行生物信息学分析;之后用150 mmol·L-1 NaCl、 4 ℃低温分别对野生型小黑杨进行胁迫处理,利用荧光定量PCR,分析基因响应非生物胁迫的功能。结果显示:PsnbHLH162 cDNA基因片段长537 bp,基因N端内含1个高度保守的HLH结构域。该基因表达蛋白是不含跨膜区域的稳定的亲水性蛋白,其定位在细胞核内且没有转录激活活性。启动子区域内含多种ABA应答、生长素应答、光应答和circadian元件,证实此基因参加非生物胁迫应答。荧光定量PCR结果表明在盐胁迫下,与茎、叶组织相比,基因在根组织的表达量最高;在低温胁迫下,与叶、根组织相比,基因在茎组织的表达量最高。发现野生植株内,PsnbHLH162能被盐、低温诱导表达。

关键词: 小黑杨, PsnbHLH162, 生物信息学, 低温胁迫, 盐胁迫

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

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