Cloning and Expression Analysis of BRASSINAZOLE RESISTANTANT1 (BZR1) Gene from Fraxinus mandshurica

doi:10.7525/j.issn.1673-5102.2021.05.013

Abstract

Abstract:

To explore the expression characteristics of BZR1 gene under abiotic stress and hormone induction of brassinosteroid（BR） signaling on the growth and development of Fraxinus mandshurica and response to environmental stress，the target gene was cloned from F. mandshurica by PCR and its molecular structures were analyzed by bioinformatics. The expression patterns of FmBZR1 under low temperature， salt stress and ABA， IAA and GA₃ hormones were investigated by real-time fluorescence quantitative PCR. Bioinformatics analysis showed that the FmBZR1 gene was 984bp， encoding 327 amino acids. FmBZR1 was a hydrophilic protein， and had high homology with BZR1 protein of Olea europaea var. sylvestris. The results of abiotic stress and hormone induction showed that the expression of FmBZR1 gene was significantly different from that of control group. The highest expression level of FmBZR1 gene was at 6 h after low temperature treatment and 24 h after salt treatment， which were 1.98 times and 10.13 times higher than that of the control group. After stress with ABA， IAA and GA₃ for 3 h， the expression of FmBZR1 gene was the lowest， which was 0.52 times， 0.41 times and 0.50 times of the control group； after treatment GA₃ for 24 h， the expression level of FmBZR1 gene was the highest， which was 6.23 times of the control group. FmBZR1 gene played a key role in the development and stress process of F. mandshurica.

Key words: Fraxinus mandshurica, FmBZR1, gene cloning, stress treatment, expression analysis

CLC Number:

S792.41

Xin-Tong JI, Lei YU, Ya-Guang ZHAN. Cloning and Expression Analysis of BRASSINAZOLE RESISTANTANT1 (BZR1) Gene from Fraxinus mandshurica[J]. Bulletin of Botanical Research, 2021, 41(5): 744-752.

Figures/Tables 11

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References 27

1	Zhu J Y，Sae-Seaw J，Wang Z Y. Brassinosteroid signalling［J］.Development，2013，140（8）：1615-1620.
2	刘亚洁，安黎哲.低温胁迫下油菜素内酯对高山离子芥悬浮细胞膜系统的影响［J］.四川大学学报：自然科学版，2020，57（4）：797-803.
	Liu Y J，An L Z.Effect of brassinosteroids on the membrane system of Chorisporabungeana suspension-cultured cells under chilling stress［J］.Journal of Sichuan University：Natural Science Edition，2020，57（4）：797-803.
3	He J X，Gendron J M，Sun Y，et al.BZR1 is a transcriptional repressor with dual roles in brassinosteroid homeostasis and growth responses［J］.Science，2005，307（5715）：1634-1638.
4	Kim T W，Guan S H，Sun Y，et al.Brassinosteroid signal transduction from cell-surface receptor kinases to nuclear transcription factors［J］.Nature Cell Biology，2009，11（10）：1254-1260.
5	Tang W Q，Yuan M，Wang R J，et al.PP2A activates brassinosteroid-responsive gene expression and plant growth by dephosphorylating BZR1［J］.Nature Cell Biology，2011，13（2）：124-131.
6	Yu X F，Li L，Zola J，et al.A brassinosteroid transcriptional network revealed by genome-wide identification of BESI target genes in Arabidopsis thaliana［J］.The Plant Journal，2011，65（4）：634-646.
7	Li J M.Regulation of the nuclear activities of brassinosteroid signaling［J］.Current Opinion in Plant Biology，2010，13（5）：540-547.
8	Yang X R，Bai Y，Shang J X，et al.The antagonistic regulation of abscisic acid-inhibited root growth by brassinosteroids is partially mediated via direct suppression of ABSCISIC ACID INSENSITIVE 5 expression by BRASSINAZOLE RESISTANT 1［J］.Plant，Cell & Environment，2016，39（9）：1994-2003.
9	Tang Y Y，Liu H H，Guo S Y，et al.OsmiR396d affects gibberellin and brassinosteroid signaling to regulate plant architecture in rice［J］.Plant Physiology，2018，176（1）：946-959.
10	Dong H X，Liu J，He G H，et al.Photoexcited phytochrome B interacts with brassinazole resistant 1 to repress brassinosteroid signaling in Arabidopsis［J］.Journal of Integrative Plant Biology，2020，62（5）：652-667.
11	Guo Y F，Shan W，Liang S M，et al.MaBZR1/2 act as transcriptional repressors of ethylene biosynthetic genes in banana fruit［J］.Physiologia Plantarum，2019，165（3）：555-568.
12	Yin Y L，Qin K Z，Song X W，et al.BZR1 transcription factor regulates heat stress tolerance through FERONIA receptor-like kinase-mediated reactive oxygen species signaling in tomato［J］.Plant & Cell Physiology，2018，59（11）：2239-2254.
13	Saha G，Park J I，Jung H J，et al.Molecular characterization of BZR transcription factor family and abiotic stress induced expression profiling in Brassica rapa［J］.Plant Physiology and Biochemistry，2015，92：92-104.
14	Lv J X，Li Y B，Liu Z Y，et al.Response of BpBZR genes to abiotic stress and hormone treatment in Betula platyphylla［J］.Plant Physiology and Biochemistry，2020，151：157-165.
15	Srivastava M，Srivastava A K，Orosa-Puente B，et al.SUMO conjugation to BZR1 enables brassinosteroid signaling to integrate environmental cues to shape plant growth［J］.Current Biology，2020，30（8）：1410-1423.e3.
16	林士杰，张忠辉，谢朋，等.中国水曲柳基因资源的保护与利用［J］.中国农学通报，2009，25（24）：158-162.
	Lin S J，Zhang Z H，Xie P，et al.Conservation and application of the genetic resource of Fraxinus mandshurica in China［J］.Chinese Agricultural Science Bulletin，2009，25（24）：158-162.
17	尹立辉，孙亚峰.水曲柳研究进展和展望［J］.长春大学学报，2006，16（3）：72-75.
	Yin L H，Sun Y F.Advances and prospectives of Fraxinus mandshurica Rupr.［J］.Journal of Changchun University，2006，16（3）：72-75.
18	Liang N S，Yu L，Liu C H，et al.Molecular cloning and expression under abiotic stresses and hormones of the ethylene response factor Ⅶ gene FmRAP2.12 from Fraxinus mandshurica［J］.Journal of Forestry Research，2019，30（4）：1289-1300.
19	刘悦，于磊，赵兴堂，等.水曲柳FmNAC072基因克隆及表达模式分析［J］.西部林业科学，2019，48（6）：161-168.
	Liu Y，Yu L，Zhao X T，et al.Cloning and expression analysis of FmNAC072 gene from Fraxinus mandshurica［J］.Journal of West China Forestry Science，2019，48（6）：161-168.
20	任小龙，詹亚光，梁雪，等.水曲柳花发育过程中AG、SOC1基因表达的qRT-PCR分析［J］.植物研究，2015，35（4）：612-617.
	Ren X L，Zhan Y G，Liang X，et al.qRT-PCR Analysis of gene expression of AG and SOC1 during flower development of Fraxinus mandshurica Rupr.［J］.Bulletin of Botanical Research，2015，35（4）：612-617.
21	董军生，张军，周素锐，等.电阻抗法测定杨树无性系抗寒性［J］.西北林学院学报，2014，29（3）：105-108.
	Dong J S，Zhang J，Zhou S R，et al.Measurement of cold resistance of poplar clones by electrical impedance spectroscopy［J］.Journal of Northwest Forestry University，2014，29（3）：105-108.
22	Sun Y，Fan X Y，Cao D M，et al.Integration of brassinosteroid signal transduction with the transcription network for plant growth regulation in Arabidopsis［J］.Developmental Cell，2010，19（5）：765-777.
23	Li H，Ye K Y，Shi Y T，et al.BZR1 positively regulates freezing tolerance via CBF-dependent and CBF-independent pathways in Arabidopsis［J］.Molecular Plant，2017，10（4）：545-559.
24	栗露露，殷文超，牛梅，等.油菜素甾醇调控水稻盐胁迫应答的作用研究［J］.植物学报，2019，54（2）：185-193.
	Li L L，Yin W C，Niu M，et al.Functional analysis of brassinosteroids in salt stress responses in rice［J］.Chinese Bulletin of Botany，2019，54（2）：185-193.
25	Yu H Q，Feng W Q，Sun F A，et al.Cloning and characterization of BES1/BZR1 transcription factor genes in maize［J］.Plant Growth Regulation，2018，86（2）：235-249.
26	Bai M Y，Shang J X，Oh E，et al.Brassinosteroid，gibberellin and phytochrome impinge on a common transcription module in Arabidopsis［J］.Nature Cell Biology，2012，14（8）：810-817.
27	Wang X X，Chen X D，Wang Q J，et al.MdBZR1 and MdBZR1-2like transcription factors improves salt tolerance by regulating gibberellin biosynthesis in apple［J］.Frontiers in Plant Science，2019，10：1473.

引物名称 Primer name	引物序列（5′-3′） Primer sequence	用途 Usages
FmBZR1-F	5′-ATGTGGGAAGCTGGAGGATCA-3′	PCR扩增引物 Primers for PCR amplification primer
FmBZR1-R	5′-TTACATCCGGGTGTTTCCACTT-3′	PCR扩增引物 Primers for PCR amplification primer
qFmBZR1-F	5′-ATGCAGGGTTAAGGGCACAAGG-3′	荧光定量PCR引物 Primers for qRT-PCR
qFmBZR1-R	5′-ACCATCAGGCTCCACGATCCAA-3′	荧光定量PCR引物 Primers for qRT-PCR
Tu-F	5′-AGGACGCTGCCAACAACTTT-3′	荧光定量PCR内参基因引物 Primers for qRT-PCR internal reference gene
Tu-R	5′-TTGAGGGGAAGGGTAAATAGTG-3′	荧光定量PCR内参基因引物 Primers for qRT-PCR internal reference gene

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