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‘84K’杨组氨酸激酶基因PaHK3a的表达及功能分析

鲁俊倩 武舒 钟姗辰 张伟溪 苏晓华 张冰玉

鲁俊倩, 武舒, 钟姗辰, 张伟溪, 苏晓华, 张冰玉. ‘84K’杨组氨酸激酶基因PaHK3a的表达及功能分析[J]. 北京林业大学学报, 2021, 43(2): 46-53. doi: 10.12171/j.1000-1522.20200070
引用本文: 鲁俊倩, 武舒, 钟姗辰, 张伟溪, 苏晓华, 张冰玉. ‘84K’杨组氨酸激酶基因PaHK3a的表达及功能分析[J]. 北京林业大学学报, 2021, 43(2): 46-53. doi: 10.12171/j.1000-1522.20200070
Lu Junqian, Wu Shu, Zhong Shanchen, Zhang Weixi, Su Xiaohua, Zhang Bingyu. Expression and function analysis of histidine kinase gene PaHK3a of poplar ‘84K’[J]. Journal of Beijing Forestry University, 2021, 43(2): 46-53. doi: 10.12171/j.1000-1522.20200070
Citation: Lu Junqian, Wu Shu, Zhong Shanchen, Zhang Weixi, Su Xiaohua, Zhang Bingyu. Expression and function analysis of histidine kinase gene PaHK3a of poplar ‘84K’[J]. Journal of Beijing Forestry University, 2021, 43(2): 46-53. doi: 10.12171/j.1000-1522.20200070

‘84K’杨组氨酸激酶基因PaHK3a的表达及功能分析

doi: 10.12171/j.1000-1522.20200070
基金项目: 国家自然基金项目(31770710),转基因生物新品种培育重大课题(2018ZX08020002)
详细信息
    作者简介:

    鲁俊倩。主要研究方向:林木遗传育种。Email:lujq115@163.com 地址:100091北京市海淀区青龙桥街道中国林业科学研究院国家重点实验室

    责任作者:

    张冰玉,博士,研究员。主要研究方向:林木遗传育种。Email:byzhang@caf.ac.cn 地址:同上

  • 中图分类号: S792.11

Expression and function analysis of histidine kinase gene PaHK3a of poplar ‘84K’

  • 摘要:   目的  干旱、高盐等逆境胁迫严重影响了植物的生长发育。研究表明,组氨酸激酶在植物逆境响应中起重要作用。本研究对银腺杨‘84K’组氨酸激酶基因PaHK3a在不同组织的表达模式分析,检测了其对生长素、细胞分裂素等植物激素处理下及人工干旱、盐碱等非生物胁迫下的表达,结合干旱、盐碱条件下丙二醛(MDA)及保护酶活性等生化指标,对该基因的功能进行了初步鉴定,为杨树抗逆分子育种研究奠定基础。  方法  以‘84K’杨无菌苗为材料,通过实时定量PCR(qRT-PCR)技术分析PaHK3a基因在不同组织的表达模式。对‘84K’杨无菌苗进行浓度为10 mmol/L植物激素处理(ABA、6-BA、IBA、GA3及水杨酸(SA))及非生物胁迫处理(42 ℃高温、0 ℃低温、200 mmol/L NaCl和5% PEG6000),采用qRT-PCR技术分析PaHK3a基因对不同植物激素及非生物胁迫的表达响应;进一步对温室‘84K’杨进行自然干旱处理(6、8、10 d)、200 mmol/L NaCl(2、4、6 d)处理,测定不同胁迫时间点叶片PaHK3a基因的表达,以及超氧化物歧化酶(SOD)、过氧化物酶(POD)活性及MDA含量,并分析PaHK3a基因表达与生理指标的相关性,初步鉴定杨树PaHK3a基因的功能。  结果  qRT-PCR结果显示,PaHK3a基因在叶片中表达量最高,根部中等,茎段最低。与正常条件下相比,在高温、低温、NaCl及PEG模拟干旱处理时,PaHK3a基因表达量与对照相比明显增高,分别为对照的2.63、1.49、1.54、1.58倍。用IBA诱导处理时,基因表达量与对照相比差异不大,而在6-BA、ABA、GA3及SA处理时,基因表达量与对照相比均呈现显著下调。在温室干旱、盐碱胁迫处理过程中,PaHK3a基因表达均显著高于对照,呈现先上升后下降的表达模式,MDA含量也呈现类似的趋势,而SOD活性则随处理时间的延长而持续升高,POD活性在干旱胁迫时先上升后下降,而高盐胁迫时呈上升趋势。生理指标与PaHK3a基因表达量相关系分析发现,在干旱和盐胁迫下,PaHK3a基因表达量与叶片MDA含量、SOD活性和POD活性均呈正相关。  结论  PaHK3a基因在‘84K’杨根茎叶中均有表达,且叶中表达量最高;PaHK3a基因表达受细胞分裂素6-BA、GA3及ABA及SA等植物激素的负调控,同时,受温度胁迫、盐胁迫、水分胁迫等非生物胁迫正调控;温室人工干旱盐碱胁迫过程中,PaHK3a基因表达量升高,且与叶片MDA含量、SOD活性、POD活性均具有正相关性。研究结果初步显示,杨树PaHK3a基因参与杨树植物激素激素信号响应,并在抗逆境胁迫过程中发挥重要调控作用。

     

  • 图  1  PaHK3a基因在‘84K’杨根、茎和叶中的表达情况

    *表示差异显著,P < 0.05;**表示差异极显著,P < 0.01。下同。* means significant difference, P < 0.05; ** means very significant difference, P < 0.01. The same below.

    Figure  1.  Expression of PaHK3a in roots, stems and leaves of ‘84K’ poplar

    图  2  非生物胁迫处理下‘84K’杨叶片PaHK3a基因的表达差异

    CK. 对照;HT. 42 ℃高温处理;LT. 0 ℃低温处理。下同。CK, control; HT, 42 ℃ high temperature treatment; LT, 0 ℃ low temperature treatment. The same below.

    Figure  2.  Expression differences of PaHK3a in ‘84K’leaves under abiotic stresses

    图  3  不同植物激素处理下‘84K’杨叶片PaHK3b基因的表达差异

    Figure  3.  Expression differences of PaHK3a in ‘84K’ leaves under several plant hormone treatments

    图  4  ‘84K’杨温室干旱(A)及盐胁迫(B)下叶片PaHK3a基因表达量

    Figure  4.  Expression of PaHK3a in leaves of ‘84K’ under drought (A) and salt (B) stresses in greenhouse

    图  5  ‘84K’杨温室干旱(A)及盐(B)胁迫下叶片MDA的含量

    Figure  5.  MDA contents in leaves of ‘84K’ under drought (A) and salt (B) stresses in greenhouse

    图  6  ‘84K’杨温室干旱(A)及盐胁迫(B)下叶片SOD活性

    Figure  6.  SOD activity in leaves of ‘84K’ under drought (A) and salt (B) stresses in greenhouse

    图  7  ‘84K’杨温室干旱(A)及盐胁迫(B)下叶片POD活性

    Figure  7.  POD activity in leaves of ‘84K’ under drought (A) and salt (B) stresses in greenhouse

    表  1  qRT-PCR引物序列

    Table  1.   Primer sequences of qRT-PCR

    引物名称
    Primer name
    引物序列(5′→3′)
    Primer sequence (5′→3′)
    产物长度
    Product length/bp
    PaHK3a-q-F CTCAGTTTCTTGCTACAGTTTCCC 243
    PaHK3a-q-R ACATCATCCATTATTGCCCTCA
    Actin-F AAACTGTAATGGTCCTCCCTCCG 193
    Actin-R GCATCATCACAATCACTCTCCGA
    下载: 导出CSV

    表  2  干旱、盐胁迫下PaHK3a基因表达与生理指标相关性分析

    Table  2.   Correlation analysis between expression of PaHK3a and physiological indexes under drought and salt stresses

    生理指标
    Physiological index
    干旱胁迫
    Drought stress
    盐胁迫
    Salt stress
    MDA0.9010.994**
    SOD0.7910.734
    POD0.954*0.690
    下载: 导出CSV
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出版历程
  • 收稿日期:  2020-03-16
  • 修回日期:  2020-06-09
  • 网络出版日期:  2020-12-26
  • 刊出日期:  2021-02-24

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