天女木兰MsGID1同源基因的克隆及表达分析  

Cloning and Expression Analysis of Magnolia sieboldii MsGID1 Homolo-gous Gene

作　　者：李银双 关士鑫 曾莞棋 张晓林[1] 陆秀君[1,2] Li Yinshuang;Guan Shixin;Zeng Wanqi;Zhang Xiaolin;Lu Xiujun(College of Forestry,Shenyang Agricultural University,Shenyang,110866;College of Horticulture,Shenyang Agricultural University,Shenyang,110866)

机构地区：[1]沈阳农业大学林学院,沈阳110866 [2]沈阳农业大学园艺学院,沈阳110866

出　　处：《分子植物育种》2020年第16期5252-5260,共9页Molecular Plant Breeding

基　　金：国家自然科学基金项目(31570621)资助。

摘　　要：为了探究天女木兰种子休眠解除的分子机制,本研究以Illumina高通量测序获得的天女木兰转录组数据为参考,从赤霉素信号途径起始受体基因(GIBBERELLIN INSENSITIVE DWARF1, GID1)入手,克隆获得2个同源基因(MsGID1b和Ms GID1c)。在此基础上,对天女木兰2个GID1同源基因进行生物信息学分析:MsGID1b和MsGID1c全长分别为996 bp和1 032 bp,二者分别编码331和334个氨基酸;所编码的蛋白质相对分子量分别为37.021 kD和38.363 kD;2个GID1同源基因均是不稳的亲水性蛋白且无信号肽;Ms-%GID1b和MsGID1c属于α/β折叠水解酶超家族。通过实时荧光定量PCR探究MsGID1b和Ms GID1c在不同温度对种子吸胀过程表达模式的影响,发现4℃处理基因表达量最高,其中Ms GID1c的相对表达量显著高于MsGID1b,表明其在种子解除休眠中起关键作用。To explore the molecular mechanism of seed dormancy affection of Magnolia sieboldii.In this study,two GID1 homologous genes(MsGID1b and MsGID1c)were cloned based on the transcriptome of Magnolia sieboldii obtained by Illumina high-throughput sequencing and started from receptor genes(GIBBERELLIN IN-%SENSITIVE DWARF1,%GID1)of gibberellin signaling pathway.On this basis,bioinformatics analysis of Mag-%nolia sieboldii GID1 homologous gene revealed that MsGID1b and MsGID1c have full length of 996 bp and 1032 bp,encoding 331 and 334 amino acids with 37.021 kD and 38.363 kD molecular weight respectively.MsGID1b and MsGID1c were a unstable hydrophilic protein without signal peptide and belong to theα/β-hy-drolase fold superfamily.Through real-time quantitative PCR,the influence of MsGID1b and MsGID1c at dif-ferent temperatures on the expression pattern of seed imbibition were explored,and it was found that the gene expression MsGID1b and MsGID1c was highest at 4℃,among which the relative expression level of MsGID1c was significantly higher than MsGID1b.It suggested that MsGID1c plays a major regulatory role in seed dor-mancy-breaking process.

关 键 词：天女木兰(Magnolia sieboldii) GID1同源基因 基因克隆 表达分析 

分 类 号：Q943.2[生物学—植物学]