天女木兰MsAHG1和MsAHG3基因的克隆及表达分析  

Cloning and Expression Analysis of MsAHG1 and MsAHG3 Gene in Magnolia sieboldii

作　　者：孙宏涛 王萌[1] 张若晰 沈光贤 张晓林[1,2] Sun Hongtao;Wang Meng;Zhang Ruoxi;Shen Guangxian;Zhang Xiaolin(College of Forestry,ShenyangAgricultural University,Shenyang,110161;Key Laboratory of Protected Horticulture(Ministry of Education),Shenyang Agricultural University,Shenyang,110161)

机构地区：[1]沈阳农业大学林学院,沈阳110161 [2]沈阳农业大学,设施园艺教育部重点实验室,沈阳110161

出　　处：《分子植物育种》2023年第15期4902-4913,共12页Molecular Plant Breeding

基　　金：国家自然科学基金青年基金项目(No.31600505)资助。

摘　　要：为了探究ABA信号通路中蛋白磷酸酶PP2C调控天女木兰种子休眠解除的分子机制,本研究以天女木兰种子转录组数据为参考,克隆获得了A类PP2C的2个基因(MsAHG1和MsAHG3)。生物信息学分析结果表明,MsAHG1、MsAHG3基因全长分别为1269、1209 bp,编码422、402个氨基酸,均属于PP2C家族;所编码的蛋白质相对分子量分别为44.88543、43.35603 kD,均属于不稳定的亲水蛋白;所编码氨基酸的同源性与系统进化分析显示,天女木兰的AHG1、AHG3蛋白分别与沉水樟的AHG1、AHG3蛋白亲缘关系较近。通过实时荧光定量PCR探究MsAHG1、MsAHG3基因在天女木兰不同组织、种子休眠解除过程及吸胀过程的相对表达量,发现MsAHG1和MsAHG3基因在干种子中表达量最高,且MsAHG3基因表达量极显著高于MsAHG1基因;MsAHG3基因在种子休眠解除和吸胀过程中的表达量均高于MsAHG1基因,表明MsAHG3基因在天女木兰种子休眠解除中发挥关键作用。In order to explore the molecular mechanism of dormant release in Magnolia sieboldii seed,two PP2C genes(MsAHG1 and MsAHG3)were cloned from PP2C protein phosphatase in ABA signaling pathway based on transcriptome data of Magnolia sieboldii seed.The results of bioinformatics analysis showed that the full length of MsAHG1 and MsAHG3 genes were 1269 and 1209 bp,encoding 422 and 402 amino acids,and they belonged to the PP2C family.The relative molecular weights of the encoded proteins were 44.88543 and 43.35603 kD,respectively,which were unstable hydrophilic proteins.The homology and phylogenetic analysis of the encoded amino acids showed that AHG1 and AHG3 proteins of Magnolia sieboldii were closely related to those of Cinnamomum micranthum,respectively.The relative expression levels of MsAHG1 and MsAHG3 genes in different tissues,seed dormancy release process and imbiption process were investigated by real-time quantitative PCR.It was found that the expression levels of MsAHG1 and MsAHG3 genes were highest in dry seeds,and the relative expression limit of MsAHG3 gene was higher than that of MsAHG1 gene.The expression of MsAHG3 gene was higher than that of MsAHG1 gene during seed dormancy release and imbibition,suggesting that MsAHG3 gene played a key role in seed dormancy release.

关 键 词：天女木兰(Magnolia sieboldii) AHG1 AHG3 基因克隆 生物信息学分析 表达分析 

分 类 号：S792.99[农业科学—林木遗传育种]