利用原生质体瞬时系统与RNA-seq筛选玉米BES1/BZR1-1下游靶基因及其初步验证靶基因及其初步验证
Identification and Preliminary Validation of Downstream Target Genes of Maize BES1/BZR1-1 via Protoplast Transient Expression System Combined with RNA-seq
宁瑞瑶 1李素贞 2陈茹梅 2郭晋杰1
作者信息
- 1. 河北农业大学农学院
- 2. 中国农业科学院生物技术研究所
- 折叠
摘要
油菜素内酯（BR）信号通路关键转录因子BES1/BZR1家族在植物生长发育中发挥重要调控作用。为筛选玉米BES1/BZR1-1（Zm00001d046305）转录因子的下游靶基因，本研究以玉米自交系B73为材料，分离其原生质体并开展基于原生质体瞬时表达系统的RNA测序（PER-seq）。通过对差异表达基因（DEGs）的分析，共鉴定到1146个显著DEGs，其中上调568个，下调578个。GO功能富集分析表明，这些基因主要参与催化活性、代谢过程及激素响应等生物学过程。KEGG通路分析显示，DEGs显著富集于代谢途径、植物激素信号转导以及苯并噁嗪类生物合成等通路。为进一步验证，本研究基于PER-seq数据，筛选出log2FC > 3且包含E-box（CANNTG）；BRRE（CGTGT/CG）；茉莉酸响应元件（CGTCA/TGACG）及脱落酸响应元件ABRE五种顺式作用元件的5个DEGs作为候选靶点，P450、AP2、BETA、ERF055和bZIP均与抗逆性相关。本研究为深入揭示ZmBES1/BZR1-1在玉米中的生物学功能及分子调控机制提供了理论依据。
Abstract
The BES1/BZR1 family, key transcription factors in the brassinosteroid （BR） signaling pathway, plays an important regulatory role in plant growth and development.To screen downstream target genes of the maize BES1/BZR1-1 （Zm00001d046305） transcription factor, this study used maize inbred line B73 as experimental material, isolated its protoplasts, and performed RNA sequencing based on a protoplast transient expression system （PER-seq）. Through the analysisIdentification and Preliminary Validation of Downstream Target Genes of Maize BES1/BZR1-1 via Protoplast Transient Expression System Combined with RNA-seq of differentially expressed genes, a total of 1,146 significantly differentially expressed genes were identified, including 568 up-regulated genes and 578 down-regulated genes. Gene Ontology （GO） functional enrichment analysis indicated that these genes were mainly involved in biological processes such as catalytic activity, metabolic processes, and hormone response. Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway analysis showed that the differentially expressed genes were significantly enriched in pathways including metabolic pathways, plant hormone signal transduction, and benzoxazinoid biosynthesis. For further verification, based on PER-seq data, five differentially expressed genes with top-ranked log2 fold change （log2FC > 3） and containing five key cis-acting elements （E-box（CANNTG）; BRRE（CGTGT/CG）; Jasmonic Acid Responsive Element（CGTCA/TGACG）and Abscisic Acid Responsive Element （ABRE）） simultaneously were selected as candidate target genes.P450, AP2, BETA, ERF055, and bZIP are related to stress tolerance. This study provides a theoretical basis for further revealing the biological function and molecular regulatory mechanism of BZR/BES1-1 in maize.关键词
玉米/油菜素内酯/BES1/BZR1转录因子/原生质体瞬时表达/RNA测序（RNA-seq）/下游靶基因Key words
Maize/Brassinosteroid/BES1/BZR1 transcription factors/Protoplast transient expression/RNA sequencing (RNA-seq)/Downstream target genes引用本文复制引用
宁瑞瑶,李素贞,陈茹梅,郭晋杰.利用原生质体瞬时系统与RNA-seq筛选玉米BES1/BZR1-1下游靶基因及其初步验证靶基因及其初步验证[EB/OL].(2026-05-11)[2026-05-12].http://www.paper.edu.cn/releasepaper/content/202605-31.学科分类
农作物/生物科学研究方法、生物科学研究技术
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