水稻OsSPL3启动子克隆及表达分析  

Cloning and expression analysis of OsSPL3 promoter in rice

作　　者：曾慧玲 莫祖意 蒲巧贤 王家澍 范凯 李兆伟 ZENG Huiling;MO Zuyi;PU Qiaoxian;WANG Jiashu;FAN Kai;LI Zhaowei(College of Life Sciences,Fujian Agriculture and Forestry University,Fuzhou 350002,Fujian,China;College of Agriculture,Fujian Agriculture and Forestry University,Fuzhou 350002,Fujian,China)

机构地区：[1]福建农林大学生命科学学院,福建福州350002 [2]福建农林大学农学院,福建福州350002

出　　处：《浙江大学学报（农业与生命科学版）》2023年第3期319-327,共9页Journal of Zhejiang University:Agriculture and Life Sciences

基　　金：福建省自然科学基金面上项目(2021J01098,2022J01142)。

摘　　要：OsSPL转录因子在水稻(Oryza sativa)根系、叶片、花器官、穗等发育与逆境响应过程中起重要作用。本文通过对OsSPL3启动子的分析,探究了OsSPL3转录因子在水稻中的表达模式及其对干旱胁迫的响应方式。利用PLACE和Plant CARE在线软件分析OsSPL3启动子区的顺式作用元件,并构建OsSPL3启动子与β-葡糖醛酸糖苷酶(β-glucuronidase, GUS)基因的重组表达载体,转化‘中花11’(ZH11)水稻愈伤组织,筛选获得阳性转基因植株,且对p OsSPL3-GUS转基因植株的GUS表达活性以及在干旱胁迫与脱落酸(abscisic acid, ABA)处理下的表达方式进行检测。启动子分析结果表明,OsSPL3启动子区除包含必要的转录起始核心元件与光响应元件外,还包括3个MYB参与的干旱诱导元件、3个赤霉素响应元件、2个厌氧诱导必需作用元件、1个低温响应作用元件、1个胚乳表达调控元件、1个玉米醇溶蛋白代谢调控元件和1个分生组织表达相关调控元件。GUS染色结果显示,GUS基因在新生叶片、茎鞘、胚芽鞘等幼嫩组织及根冠、分生区、伸长区等根系旺盛生长部位中表达活性较高。此外,干旱胁迫能明显增强转基因水稻叶片与根系的GUS活性。本研究结果表明,OsSPL3转录因子在水稻种子萌发后的胚芽鞘生长、新叶发生、根系延伸等器官发育与茎鞘伸长等过程中发挥调控作用,同时,OsSPL3转录因子还参与水稻干旱胁迫响应过程。OsSPL transcription factor plays an important role in the development and stress response of rice(Oryza sativa)roots,leaves,floral organs,and ears.In this study,the OsSPL3 promoter was analyzed to explore the expression pattern of OsSPL3 transcription factor in rice and its response to drought stress.The Cis-acting elements in the OsSPL3 promoter region were analyzed by PLACE and Plant CARE online softwares,and the recombinant expression vector of OsSPL3 promoter andβ-glucuronidase(GUS)gene was constructed,which was transformed into ZH11 rice callus,and positive transgenic plants were obtained by screening.The GUS expression activity of pOsSPL3-GUS transgenic plants and the expression patterns under drought stress and abscisic acid(ABA)treatments were detected.The results of promoter analysis showed that in addition to the necessary transcription initiation core elements and light-responsive elements,the OsSPL3 promoter region also included three MYB-involved drought-inducible elements,three gibberellin-responsive elements,two anaerobic induction essential elements,one low temperature response element,one endosperm expression regulatory element,one zein metabolism regulatory element and one meristem expression-related regulatory element.The results of GUS staining showed that the expression activity of GUS gene in young leaves,stem sheaths,coleoptiles and other young tissues was high,as well as in the vigorous growth parts of roots such as root cap,meristem zone,and elongation zone.In addition,the drought stress could significantly enhanced the GUS activity of transgenic rice leaves and roots.It shows that OsSPL3 transcription factor plays a regulatory role in the process of coleoptile growth,new leaf formation,root extension and stem sheath elongation after seed germination,and OsSPL3 transcription factor is also involved in the response process of rice drought stress.

关 键 词：水稻 OsSPL3转录因子 启动子 β-葡糖醛酸糖苷酶 干旱胁迫 

分 类 号：S511.22[农业科学—作物学] Q945.78[生物学—植物学]