文冠果BZR1基因家族鉴定及功能分析

doi:10.12302/j.issn.1000-2006.202403036

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南京林业大学学报（自然科学版） ›› 2025, Vol. 49 ›› Issue (2): 12-22.doi: 10.12302/j.issn.1000-2006.202403036

所属专题：推进乡村全面振兴视域下的多功能油用树种文冠果研究

• 专题报道：推进乡村全面振兴视域下的多功能油用树种文冠果研究（执行主编尹佟明李维林） • 上一篇下一篇

文冠果BZR1基因家族鉴定及功能分析

许慧慧(), 班卓, 王晨雪, 毕泉鑫, 刘肖娟^*(), 王利兵

中国林业科学研究院林业研究所,国家林业和草原局林木培育重点实验室, 林木遗传育种全国重点实验室,北京 100091

收稿日期:2024-03-27 接受日期:2024-06-25 出版日期:2025-03-30 发布日期:2025-03-28
通讯作者: *刘肖娟(liuxiaojuan@caf.ac.cn),助理研究员。
作者简介:
许慧慧(xuhuihui0206@163.com)。
基金资助:
国家自然科学基金项目(32271838)

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

摘要/Abstract

摘要：

【目的】探究文冠果(Xanthoceras sorbifolium)Brassinazole Resistance 1(BZR1)基因家族成员的特征及其在非生物胁迫响应中的作用,为文冠果XsBZR1基因功能的研究和抗逆新品种的选育提供理论参考。【方法】利用生物信息学方法鉴定并系统分析文冠果XsBZR1基因家族;构建35S::XsBZR1-eYFP融合蛋白,对XsBZR1进行亚细胞定位分析;通过实时荧光定量PCR技术分析XsBZR1基因在非生物胁迫下的表达模式;构建XsBZR1- 9基因的过表达载体并转化到拟南芥(Arabidopsis thaliana)中,观察转基因株系与野生型植株在盐胁迫处理下的生长情况。【结果】①在文冠果基因组中共鉴定出9个BZR1基因,分别命名为XsBZR1-1—XsBZR1-9,这些基因不均匀地分布在6条染色体上。②系统进化和共线性分析表明,XsBZR1蛋白与双子叶植物的BZR 1转录因子亲缘关系更为密切。③XsBZR1基因的启动子区域具有大量的光响应、激素响应元件以及胁迫应答响应元件。④亚细胞定位结果和预测结果一致,9个XsBZR1均定位于细胞核。⑤qRT-PCR分析表明,9个XsBZR1基因在不同的非生物胁迫下表现出不同的表达模式。除XsBZR1-7外,其余XsBZR1基因在低温胁迫下3 h时迅速上调表达;盐胁迫下XsBZR1的表达量呈现较大的差异性,XsBZR1-3/4/5/7的表达受到盐胁迫的显著抑制,而XsBZR1-1和XsBZR1-9在盐胁迫处理9 h时表达量提高到约20倍;XsBZR1-3/7/8/9在干旱处理9 h时表达量提高到2倍以上,而其余XsBZR1在干旱处理下的表达水平变化不大;9个XsBZR1均受到ABA的诱导表达,其中XsBZR1-8在ABA处理9 h时表达量提高到35倍。⑥在拟南芥中过表达XsBZR1-9发现,转基因植株在盐胁迫处理后的主根长度显著高于野生型植株。【结论】XsBZR1家族成员参与了文冠果非生物胁迫响应,过表达XsBZR1-9显著提高植物对盐胁迫的耐受性,这为进一步分析XsBZR1基因在文冠果抗逆机制中的作用奠定了基础。

关键词: 文冠果, BZR1转录因子, 非生物胁迫, 基因过表达, 盐胁迫响应, XsBZR1-9

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

中图分类号:

S722

许慧慧,班卓,王晨雪,等. 文冠果BZR1基因家族鉴定及功能分析[J]. 南京林业大学学报（自然科学版）, 2025, 49(2): 12-22.

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 (Natural Science Edition), 2025, 49(2): 12-22.DOI: 10.12302/j.issn.1000-2006.202403036.

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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

文冠果BZR1基因家族鉴定及功能分析

The identification and functional analysis of BZR1 genes in yellowhorn

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