Transcription factors play critical roles in the regulation of gene expression during maize kernel development.The maize endosperm,a large storage organ,accounting for nearly 90%of the dry weight of mature kernels,ser...Transcription factors play critical roles in the regulation of gene expression during maize kernel development.The maize endosperm,a large storage organ,accounting for nearly 90%of the dry weight of mature kernels,serves as the primary site for starch storage.In this study,we identify an endosperm-specific EREB gene,ZmEREB167,which encodes a nucleus-localized EREB protein.Knockout of ZmEREB167 significantly increases kernel size and weight,as well as starch and protein content,compared with the wild type.In situ hybridization experiments show that ZmEREB167 is highly expressed in the BETL as well as PED regions of maize kernels.Dual-luciferase assays show that ZmEREB167 exhibits transcriptionally repressor activity in maize protoplasts.Transcriptome analysis reveals that a large number of genes are up-regulated in the Zmereb167-C1 mutant compared with the wild type,including key genetic factors such as ZmMRP-1 and ZmMN1,as well as multiple transporters involved in maize endosperm development.Integration of RNA-seq and ChIP-seq results identify 68 target genes modulated by ZmEREB167.We find that ZmEREB167 directly targets OPAQUE2,ZmNRT1.1,ZmIAA12,ZmIAA19,and ZmbZIP20,repressing their expressions.Our study demonstrates that ZmEREB167 functions as a negative regulator in maize endosperm development and affects starch accumulation and kernel size.展开更多
近年来,ERF(Ethylene-responsive element binding factor)家族转录因子已成为植物抗逆、抗病的分子机制和作物分子育种研究的热点。本研究以电子克隆、同源扩增和RACE(Rapid-amplification of cDNA ends)技术相结合的方法从海岛棉中分...近年来,ERF(Ethylene-responsive element binding factor)家族转录因子已成为植物抗逆、抗病的分子机制和作物分子育种研究的热点。本研究以电子克隆、同源扩增和RACE(Rapid-amplification of cDNA ends)技术相结合的方法从海岛棉中分离出了一条新的ERF族转录因子基因,命名为EREB5。该基因包含一个573 bp长的开放阅读框,预期编码190 aa长的蛋白质分子。序列分析表明该蛋白除含有一个ERF保守域外,还含有一段核定位信号序列、一段富丝氨酸的激活域、多种磷酸化位点和一个土豆抑制子Ⅰ型家族基序等。构建系统发育树分析表明该因子属于ERF亚家族B3亚组。瞬时表达实验证明该因子定位于细胞核内,同时,凝胶阻滞实验结果说明EREB5蛋白和GCC盒具有较强的结合能力。再者,荧光定量PCR结果表明乙烯和黄萎病菌处理可以诱导该基因的表达。这些结果暗示EREB5蛋白很可能在黄萎病抗性机制中扮演者重要角色。展开更多
基金supported by STI 2030-Major Project(2023ZD04069)National Key Research and Development Program of China(2023YFD1202900)+3 种基金The National Science Fund for Distinguished Young Scholars(32425041)The“Breakthrough”Science and Technology Project of Tongliao(TL2024TW001)Science and Technology Demonstration Project of Shandong Province(2024SFGC0402)Pinduoduo-China Agricultural University Research Fund(PC2023A01004).
文摘Transcription factors play critical roles in the regulation of gene expression during maize kernel development.The maize endosperm,a large storage organ,accounting for nearly 90%of the dry weight of mature kernels,serves as the primary site for starch storage.In this study,we identify an endosperm-specific EREB gene,ZmEREB167,which encodes a nucleus-localized EREB protein.Knockout of ZmEREB167 significantly increases kernel size and weight,as well as starch and protein content,compared with the wild type.In situ hybridization experiments show that ZmEREB167 is highly expressed in the BETL as well as PED regions of maize kernels.Dual-luciferase assays show that ZmEREB167 exhibits transcriptionally repressor activity in maize protoplasts.Transcriptome analysis reveals that a large number of genes are up-regulated in the Zmereb167-C1 mutant compared with the wild type,including key genetic factors such as ZmMRP-1 and ZmMN1,as well as multiple transporters involved in maize endosperm development.Integration of RNA-seq and ChIP-seq results identify 68 target genes modulated by ZmEREB167.We find that ZmEREB167 directly targets OPAQUE2,ZmNRT1.1,ZmIAA12,ZmIAA19,and ZmbZIP20,repressing their expressions.Our study demonstrates that ZmEREB167 functions as a negative regulator in maize endosperm development and affects starch accumulation and kernel size.
文摘近年来,ERF(Ethylene-responsive element binding factor)家族转录因子已成为植物抗逆、抗病的分子机制和作物分子育种研究的热点。本研究以电子克隆、同源扩增和RACE(Rapid-amplification of cDNA ends)技术相结合的方法从海岛棉中分离出了一条新的ERF族转录因子基因,命名为EREB5。该基因包含一个573 bp长的开放阅读框,预期编码190 aa长的蛋白质分子。序列分析表明该蛋白除含有一个ERF保守域外,还含有一段核定位信号序列、一段富丝氨酸的激活域、多种磷酸化位点和一个土豆抑制子Ⅰ型家族基序等。构建系统发育树分析表明该因子属于ERF亚家族B3亚组。瞬时表达实验证明该因子定位于细胞核内,同时,凝胶阻滞实验结果说明EREB5蛋白和GCC盒具有较强的结合能力。再者,荧光定量PCR结果表明乙烯和黄萎病菌处理可以诱导该基因的表达。这些结果暗示EREB5蛋白很可能在黄萎病抗性机制中扮演者重要角色。
