The identification and functional analysis of BZR1 genes in yellowhorn

doi:10.12302/j.issn.1000-2006.202403036

JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2025, Vol. 49 ›› Issue (2): 12-22.doi: 10.12302/j.issn.1000-2006.202403036

Special Issue: 推进乡村全面振兴视域下的多功能油用树种文冠果研究

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The identification and functional analysis of BZR1 genes in yellowhorn

XU Huihui(), BAN Zhuo, WANG Chenxue, BI Quanxin, LIU Xiaojuan^*(), WANG Libing

State Key Laboratory of Tree Genetics and Breeding, Research Institute of Forestry, Chinese Academy of Forestry, Key Laboratory of Tree Breeding and Cultivation of National Forestry and Grassland Administration, State Key Laboratory of Tree Genetics and Breeding, Beijing 100091, China

Received:2024-03-27 Accepted:2024-06-25 Online:2025-03-30 Published:2025-03-28
Contact: LIU Xiaojuan E-mail:xuhuihui0206@163.com;liuxiaojuan@caf.ac.cn

Abstract

Abstract:

【Objective】This study aimed to systematically characterize the BZR1 transcription factor family in yellowhorn (Xanthoceras sorbifolium) and elucidate its functional roles in abiotic stress response, thereby providing insights for breeding stress-resistant cultivars.【Method】Genome-wide identification of XsBZR1 genes was performed using bioinformatics tools. Subcellular localization of XsBZR1 proteins was validated via 35S::XsBZR1-eYFP fusion constructs. Expression profiles under abiotic stresses (low temperature, salt, drought) and abscisic acid (ABA) treatment were analyzed by qRT-PCR. Functional validation was conducted by overexpressing XsBZR1- 9 in Arabidopsis thaliana and assessing salt tolerance phenotypes.【Result】Nine XsBZR1 genes (XsBZR1-1 to XsBZR1-9) were identified and distributed unevenly across six chromosomes. Phylogenetic and synteny analyses revealed close evolutionary relationships with dicotyledonous BZR1 orthologs. Promoter regions harbored abundant cis-regulatory elements associated with light responsiveness (48.9%), hormone signaling (35.0%) and stress adaptation (24.6%). The nuclear localization of all XsBZR1 proteins was experimentally confirmed. Differential expression patterns were observed under stress conditions: low temperature (4 ℃) rapidly induced eight XsBZR1 genes (1.3- to 6.0-fold upregulation at 3 h; P<0.05), except XsBZR1-7. Salt stress (150 mmol/L NaCl) suppressed XsBZR1-3/4/5/7 but strongly upregulated XsBZR1-1 (26.1-fold) and XsBZR1-9 (19.6-fold) at 9 h (P<0.001). Drought stress (mass fraction 25% PEG6000) elevated XsBZR1- 3/7/8/9 expression (>1.9-fold at 9 h), while others remained stable. ABA treatment (100 μmol/L) universally induced XsBZR1 genes, with XsBZR1-8 showing a 35.4-fold increase (P<0.001).Transgenic Arabidopsis overexpressing XsBZR1-9 exhibited enhanced salt tolerance, with taproot lengths twice that of the wild-type under 100 mmol/L NaCl (P<0.01).【Conclusion】The XsBZR1 gene family plays pivotal roles in yellowhorn’s abiotic stress response, with XsBZR1-9 demonstrating significant potential for improving salt tolerance. These findings advance the molecular understanding of stress adaptation mechanisms in woody plants and provide targets for precision breeding.

Key words: yellowhorn(Xanthoceras sorbifolium), BZR1 transcription factor, abiotic stress tolerance, gene overexpression, salt stress response, XsBZR1- 9

CLC Number:

S722

XU Huihui, BAN Zhuo, WANG Chenxue, BI Quanxin, LIU Xiaojuan, WANG Libing. The identification and functional analysis of BZR1 genes in yellowhorn[J]. JOURNAL OF NANJING FORESTRY UNIVERSITY, 2025, 49(2): 12-22.

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基因名称 gene name	正向引物序列(5'—) forward primer sequence	反向引物序列(3'—) reverse primer sequence	用途 application
XsBZR1- 1	CTCGGTGGCAATGAAGTT	GCAGAGAAGTTGGTTGTTG	qRT-PCR引物
XsBZR1- 2	AGAAGAGAAGGAGAGGACAA	ACGATGAAGTTACCATAGCA
XsBZR1- 3	ATGCGATGAGTCTGATACAT	TCCATTCCTTCCATTCCTAC
XsBZR1- 4	TGTGGAGCGAATGGATATAG	AACTGGAGCACTGATGGA
XsBZR1- 5	AGAAGACGGCACCACTTA	AGGACTCGGATTGTAAGATG
XsBZR1- 6	CTGGTGGTGGAGGAGATT	CCGCCGTATAAGTAGAGTG
XsBZR1- 7	GAGGCTGGTTGGATTGTT	CGCACTGATGTTCGTAGA
XsBZR1- 8	AATGTGGTGGATGAGAAGAA	CTTGAAGCCTGGCGAATA
XsBZR1- 9	CCTGTAGAGCGAATGGATAT	AACTGGAGCACTGATGGA
XsActin	AGAGATTCCGTTGCCCAGAA	CCACCACTGAGCACAATGTT
AtActin	TTACCCGATGGGCAAGTC	GCTCATACGGTCAGCGATAC
XsBZR1- 1	ATGATTACAATCAGCAACAT	AGCCAGCAGATCGCCCACTA	XsBZR1 基因克隆引物
XsBZR1- 2	ATGGCAACAGATATGCAGAA	CACCTGGAGATCAAGAACTG
XsBZR1- 3	ATGACGTCTGATGGGGCGAC	ACCCTGAGTCTTCCCAGTTC
XsBZR1- 4	ATGACGTCAGGATCGAGACT	GCCAGAAAGCCGCTGCCTAC
XsBZR1- 5	ATGACGTCGGGTACGAGAAT	TCTGGTTTTAGAGTTTCCCA
XsBZR1- 6	ATGTTTCCAATCAGAAAATT	TATTGTACGGCGTGGAGGAG
XsBZR1- 7	ATGACAGCGGGAGGATCAGG	TCCGCGCGTCTTGGTACTAC
XsBZR1- 8	ATGGGGAAAGAGAATGTGGT	GTCTTCATCTCCTGATTGAT
XsBZR1- 9	ATGACGTCAGGATCGAGGTT	CCTGGTCCTTGAGCTCCCAA

基因名称 gene name	基因ID gene ID	蛋白序列长度/aa length	分子质量 /ku molecular weight	等电点 pI	总平均疏水指数 GRAVY	保守结构域 domain	亚细胞号 subcellular localization	染色体号 chromosome No.
XsBZR1-1	Xsorbifolium001562.1	653	73.73	5.62	-0.403	BES1_N Glyco_hydro_14	细胞核	1
XsBZR1-2	Xsorbifolium002147.2	702	78.46	5.40	-0.364	BES1_N Glyco_hydro_14	细胞核	1
XsBZR1-3	Xsorbifolium010436.1	316	34.45	8.39	-0.601	BES1_N	细胞核	6
XsBZR1-4	Xsorbifolium011852.2	341	36.84	8.59	-0.579	BES1_N	细胞核	7
XsBZR1-5	Xsorbifolium012965.1	327	34.96	8.60	-0.627	BES1_N	细胞核	7
XsBZR1- 6	Xsorbifolium013243.1	226	23.89	10.21	-0.365	BES1_N	细胞核	7
XsBZR1- 7	Xsorbifolium015469.1	328	35.46	8.58	-0.547	BES1_N	细胞核	9
XsBZR1- 8	Xsorbifolium019507.1	136	15.89	9.59	-1.113	BES1_N	细胞核	12
XsBZR1- 9	Xsorbifolium024342.1	325	35.05	8.94	-0.619	BES1_N	细胞核	15

The identification and functional analysis of BZR1 genes in yellowhorn

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