ERF转录因子调控杨树叶片形态构建的功能分析  

Functional analysis of ERF transcription factors in regulating leaf morphological construction of poplar

作　　者：王升级 周猎鼎 胡佳 金霞 党慧 赵凯[1] 韩有志[1] WANG Shengji;ZHOU Lieding;HU Jia;JIN Xia;DANG Hui;ZHAO Kai;HAN Youzhi(College of Forestry,Shanxi Agricultural University,Jinzhong,Shanxi 030801,China;Forestry College,Inner Mongolia Agricultural University,Huhhot 010018,China)

机构地区：[1]山西农业大学林学院,山西晋中030801 [2]内蒙古农业大学林学院,呼和浩特010018

出　　处：《植物生理学报》2023年第6期1157-1168,共12页Plant Physiology Journal

基　　金：山西省基础研究计划(自由探索类)项目(20210302123425);山西农业大学“十四五”生物育种工程项目(YZGC140);国家自然科学基金(31800564);山西省博士毕业生、博士后研究人员来晋工作奖励经费科研项目(SXBYKY2022054)。

摘　　要：ERF转录因子在植物生长发育及逆境胁迫应答过程中发挥着重要的调控功能,但关于ERF转录因子调控杨树叶片大小发育的功能及作用机制尚不清楚。本研究通过生物信息学筛选获得杨树ERF_II亚族转录因子15个,根据其氨基酸序列特征进一步分为A和B两组, A组成员第77位为丙氨酸-A,含有Motif 4、Motif 6元件及2个UTR区域;而B组成员第77位为苏氨酸-T或者异亮氨酸-I并含有特异的Motif 5。基因片段重复分析表明杨树ERF_II亚族存在7对基因片段重复;基因共线性分析结果显示8个ERF_II亚族转录因子与11个拟南芥基因存在21对片段重复,说明ERF_II亚族转录因子之间存在基因功能冗余。基因组织特异性表达模式分析表明Potri.002G085600、Potri.005G176000、Potri.006G138700、Potri.018G047300和Potri.006G138800等5个基因在根中表达量最高;Potri.018G038100 (ERF194)在叶中表达量最高;Potri.014-G025200和Potri.002G124000在茎中表达量最低。表达相关性分析结果显示Potri.006G138700、Potri.006G138800、Potri.006G218200和Potri.018G047300等4个基因以Potri.006G138700为中枢存在显著相关性, Potri.002G085600与Potri.005G176000表达存在相关性。基因功能注释分析显示15个ERF_II亚族转录因子基因均在细胞组成中发挥着作用,说明其具有组织特异性。叶片大小及生理指标分析结果显示OX株系叶片明显小于非转基因对照, OX叶片鲜重和干重亦小于WT,但并未达到显著差异,而OX比叶重大于WT,说明OX的叶片厚度的增加弥补了面积减小对其重量的影响;OX株系叶片颜色更深,叶绿素含量明显大于WT;而RNAi与WT差异不明显。说明ERF194的抑制表达对于杨树片叶的发育并无明显影响,其功能可能与ERF_II亚族其他转录因子存在冗余或互补。本研究揭示了ERF_II亚族转录因子成员之间存在基因功能冗余,并且在调控杨树叶片大小生长过程中发挥着重要的作用。尤其是ERF194的过�ERF transcription factors play an important regulatory role in plant growth and development and response to stress,but the function and mechanism of ERF transcription factor in regulating poplar leaf size and development remain unclear.In this study,total 15 transcription factors of subfamily ERF_ll were identified by bioinformatics screening,which were further divided into two groups,A and B,according to their amino acid sequence characteristics.Group A contains alanine-A at position 77 of the amino acid sequence and contains Motif 4,Motif 6 elements and 2 UTR regions,while Group B has threonine-T or isoleucine-l at position 77 and a specific Motif 5.Gene fragment duplication analysis showed that 7 pairs of gene fragment duplication existed in poplar ERF_ll subfamily.The results of collinearity analysis showed that there were 21 pairs of gene repeats between 8 ERF_ll subfamily transcription factors and 11 Arabidopsis genes,indicating that there were gene functional redundancy among the ERF_ll subfamily transcription factors.Functional annotation analysis showed that 15 ERF_ll subfamily transcription factor genes all played a role in cell composition,indicating that they were tissue specific.The expression levels of Potri.002G085600,Potri.005G176000,Potri.006G138700,Potri.018G047300,Potri.006G138800 werethe highest in roots.Potri.018G038100(ERF194)was the most expressed in leaves.Potri.014G025200 and Potri.002G124000 had the lowest expression in stem.The results of expression correlation analysis showed that Potri.006G138700,Potri.006G138800,Potri.006G218200 and Potri.018G047300 were significantly correlated with Potri.006G138700 as the center.Potri.002G085600 was correlated with the expression of Potri.005G176000.The results of leaf size and physiological index analysis showed that 0X was significantly smaller than non-transgenic control.Although OX was also smaller than WT in fresh and dry weight,there was no significant difference.In addition,OX was larger than WT in specific leaf weight,indicating that the in

关 键 词：ERF转录因子 杨树 叶片发育 组织特异性表达 ERF194 

分 类 号：S792.11[农业科学—林木遗传育种]