糜子PmDEP1和PmEP3基因的克隆与表达特征分析  

Cloning and Expression Characteristics Analysis of Millet Genes PmDEP1 and PmEP3

作　　者：许圆梦 毛娇 王梦瑶 王数 任江陵 刘宇涵 刘思辰 乔治军 王瑞云[1] 曹晓宁 XU Yuan-meng;MAO Jiao;WANG Meng-yao;WANG Shu;REN Jiang-ling;LIU Yu-han;LIU Si-chen;QIAO Zhi-jun;WANG Rui-yun;CAO Xiao-ning(College of Agriculture,Shanxi Agricultural University,Taigu 030801;Center for Agricultural Genetic Resources Research,Shanxi Agricul-tural University,Taiyuan 030031;Key Laboratory of Crop Genetic Resources and Germplasm Development in Loess Plateau,Ministry of Agri-culture and Rural Affairs,Taiyuan 030031)

机构地区：[1]山西农业大学农学院,太谷030801 [2]山西农业大学农业基因资源研究中心,太原030031 [3]农业农村部黄土高原作物基因资源与种质创制重点实验室,太原030031

出　　处：《生物技术通报》2025年第2期150-162,共13页Biotechnology Bulletin

基　　金：山西省重点研发项目(2022ZDYF110);中央引导地方科技发展资金(YDZJSX2022A044);谷子高粱产业技术体系谷子糜子生理岗位(CARS-06-14.5-A16);山西省现代农业杂粮产业技术体系(2024CYJSTX03-23)。

摘　　要：【目的】穗型是影响作物产量和机械化收割的重要因素,DEP1(Dense Erect Panicles 1)和EP3(Erect Panicle 3)基因是控制穗型形成的关键基因。旨在探究糜子(Panicum miliaceum L.)PmDEP1和PmEP3基因的结构与表达特征。【方法】以散穗型糜子笤帚糜子(ZM)、密穗型糜子M278和侧穗型糜子M350为材料,克隆获得PmDEP1和PmEP3,并对其开展生物信息学分析。采用RT-qPCR检测不同穗型糜子PmDEP1和PmEP3的表达模式。【结果】序列结果分析显示,PmDEP1 cDNA全长为1044 bp,编码347个氨基酸,蛋白结构域预测为PAT1,二级结构以无规则卷曲为主,三级结构与水稻DEP1蛋白高度相似,预测定位于细胞核,与柳枝稷具有较高的同源性,拥有28个磷酸化位点,不具有跨膜结构和信号肽。PmEP3 cDNA全长1215 bp,编码358个氨基酸,编码的蛋白属于F-box家族,二级结构以无规则卷曲和延伸链为主,三级结构与水稻Os02g0260200蛋白高度相似,预测定位于细胞质,与柳枝稷具有较高的同源性,不含信号肽,拥有32个磷酸化位点,具有少量的跨膜结构。RT-PCR结果显示,PmDEP1和PmEP3在不同时期的不同部位中表达,PmDEP1在拔节期的根中表达量最高,在抽穗期的笤帚糜子和M278中的叶部表达量最高,在M350中的茎部表达量最高。PmEP3在拔节期的叶中表达量最高,在笤帚糜子和M278抽穗期的叶部表达量最高,在M350的穗部表达量最高。【结论】PmDEP1和PmEP3为穗型基因,可能参与调控糜子穗型形成。【Objective】Panicle type is an important factor affecting crop yield and mechanized harvesting.DEP1(Dense Erect Panicles 1)and EP3(Erect Panicles 3)genes are key ones controlling spike type formation.This study aims to explore the structure and expression characteristics of PmDEP1 and PmEP3 ones in proso millet(Panicum miliaceum L.).【Method】PmDEP1 and PmEP3 genes were cloned and subjected to bioinformatics analysis using the scattered ear type millet varieties,broomstick millet(ZM),dense ear type millet M278,and lateral ear type millet M350 as materials.RT-qPCR was used to detect the expression patterns of PmDEP1 and PmEP3 genes in different ear types of proso millet.【Result】Sequence result analysis shows that,the full-length cDNA of PmDEP1 is 1044 bp,encoding 347 amino acids.The protein domain is predicted to be PAT1.The secondary structure is dominated by random coils.The tertiary structure is highly similar with that of rice DEP1 protein.It is predicted to be located in the nucleus and has high homology with switchgrass.It has 28 phosphorylation sites and does not have transmembrane structure and signal peptide.The full-length cDNA of PmEP3 is 1215 bp,encoding 358 amino acids.The encoded protein belongs to the F-box family.The secondary structure is dominated by random coils and extended chains.The tertiary structure is highly similar with that of rice Os02g0260200 protein.It is predicted to be located in the cytoplasm.It has high homology with switchgrass,does not contain signal peptides,has 32 phosphorylation sites,and has a small amount of transmembrane structure.The RT-PCR results shows that PmDEP1 and PmEP3 genes are expressed in different parts of the plant at different stages.PmDEP1 gene is mainly expressed in the roots at the jointing stage,with the highest expression in the leaves of broomstick millet and M278 at the heading stage,and in the stems of M350.The expressions of PmEP3 gene is the highest in the leaves at the jointing stage,in the leaves of broomstick millet and M278 at the headi

关 键 词：糜子 穗型基因 PmDEP1 PmEP3 基因克隆 表达分析 

分 类 号：S516[农业科学—作物学] Q943.2[生物学—植物学]