天女木兰PIN基因家族的鉴定及在种子萌发过程中的表达  

Identification of PIN Genes Family and Their Expression During Seed Germination in Magnolia sieboldii K.Koch

作　　者：梅梅 王斯彤 刘怡菲[1] 虞家昕 张芷瑞 陆秀君[4] MEI Mei;WANG Sitong;LIU Yifei;YU Jiaxin;ZHANG Zhirui;LU Xiujun(Liaoning Academy of Forestry Science,Shenyang 110032,China;Liaoning Bingla Mountain Forest Ecosystem Positioning Observation Station,Xifeng Li-aoning 112400,China;Liaoning Forestry Development Service Center,Shenyang 110122,China;College of Forestry,Shenyang Agricultural University,Shenyang 110161,China)

机构地区：[1]辽宁省林业科学研究院,沈阳110032 [2]辽宁冰砬山森林生态系统定位观测研究站,辽宁西丰112400 [3]辽宁省林业发展服务中心,沈阳110031 [4]沈阳农业大学林学院,沈阳110161

出　　处：《沈阳农业大学学报》2025年第1期29-39,共11页Journal of Shenyang Agricultural University

基　　金：辽宁省博士科研启动基金计划项目(2022-BS-051);国家自然科学基金项目(32101508);辽宁省农业科学院院长基金项目(2022BS0704)。

摘　　要：[目的]天女木兰(Magnolia sieboldii K.Koch)是我国重点保护的具有观赏、芳香和药用价值的珍稀濒危野生木本资源植物,有广阔的应用前景。该树种无性繁殖困难,故以种子繁殖为主,但是种子具深休眠特性,种胚发育不完全致使种子萌发和出苗困难,极大限制了开发进程。PIN基因家族编码生长素跨细胞膜外向转运蛋白,与种子发育和萌发有关。[方法]通过分析PIN基因家族结构和功能,探究生长素在种子后熟过程中转运调节机制,为深入研究天女木兰种子发育调控机制提供参考。利用生物信息学技术,基于天女木兰基因组数据,进行全基因组MsPINs基因家族鉴定,研究MsPINs基因家族的分子结构、在种子萌发期的时空表达和基因功能;并通过基因染色体信息获取MsARF5候选靶基因MsPIN1b的启动子序列,利用基因克隆和酵母单杂交等技术,探究MsARF5与MsPIN1b启动子之间的调控关系。[结果]生物信息学和表达分析表明天女木兰基因组中有22个MsPINs基因,分布在13条染色体上;其中9个基因在种子萌发过程中差异表达,MsPIL3、MsPILS2和MsPILS6在休眠解除阶段呈上调表达,MsPIN1a~1c、MsPIN6在种子萌发过程中呈上调表达,MsPILS3在休眠解除过程波动变化;序列分析和酵母单杂交试验表明MsPIN1b的启动子序列有多个MsARF5(生长素响应因子5)识别元件,并能与MsARF5结合。[结论]综上可见,PIN基因家族成员参与天女木兰种子发育和萌发过程。[Objective]Magnolia sieboldii K.Koch is a rare and endangered wild woody plant resource in China that exhibits ornamental,aromatic,and medicinal values,and has broad prospects for development and application.The asexual reproduction of this tree species is challenging,primarily due to the difficulty in sowing propagation.The seeds exhibit deep dormancy characteristics,and incomplete embryo development impedes seed germination and emergence,significantly constraining the development and application process.Auxin efflux carrier proteins,which are linked to seed development and germination,are encoded by the PIN gene family and carry auxin across the cell membrane.[Methods]By analyzing the structure and function of the PIN gene family,this study explores the transport regulation mechanism of auxin during seed after-ripening,providing a reference for in-depth research on the regulatory mechanisms of M.sieboldii K.Koch seed development.Using bioinformatics techniques and based on the genome data of M.sieboldii K.Koch,a genome-wide identification of the MsPINs gene family was conducted,studying the molecular structure,spatiotemporal expression during seed germination,and gene functions of the MsPINs gene.[Results]The results indicate:The M.sieboldii K.Koch genome has 22 MsPINs genes spread across 13 chromosomes,according to bioinformatics and expression research.Nine of these genes exhibit variable expression throughout the germination process.MsPIN1a~1c and MsPIN6 are upregulated during seed germination,MsPILS3 exhibits variable variations during the dormancy release process,and MsPIL3,MsPILS2,and MsPILS6 are upregulated during the dormancy release stage.The promoter sequence of MsPIN1b contains several MsARF5(Auxin Response Factor 5)recognitions,according to sequence analysis and yeast one-hybrid tests.[Conclusion]These findings suggest that M.sieboldii K.Koch seed growth and germination are mediated by members of the PIN gene family.

关 键 词：天女木兰 生长素 MsPIN1b 表达模式 酵母单杂交 

分 类 号：S722[农业科学—林木遗传育种]