拟南芥AtiPGAM2基因参与非生物胁迫的响应  

Response of Arabidopsis AtiPGAM2 Gene to Abiotic Stress

作　　者：李景艳 周家婧 袁媛 苏晓艺 乔文慧 薛岩磊 李国婧[1] 王瑞刚[1] LI Jing-yan;ZHOU Jia-jing;YUAN Yuan;SU Xiao-yi;QIAO Wen-hui;XUE Yan-lei;LI Guo-jing;WANG Rui-gang(Inner Mongolia Key Laboratory of Plants Adversity Adaptation and Genetic Improvement in Cold and Arid Regions,College of Life Sciences,Inner Mongolia Agricultural University,Hohhot 010000;Hohhot No.2 High School,Hohhot 010000;Affiliated Middle School to Inner Mongolia Normal University,Hohhot 010000)

机构地区：[1]内蒙古自治区旱寒区植物逆境适应与遗传修饰改良重点实验室内蒙古农业大学生命科学学院,呼和浩特010000 [2]呼和浩特第二中学,呼和浩特010000 [3]内蒙古师范大学附属中学,呼和浩特010000

出　　处：《生物技术通报》2024年第5期215-224,共10页Biotechnology Bulletin

基　　金：国家自然科学基金项目(32160067);内蒙古自治区高校创新团队发展计划(NMGIRT2222);内蒙古农业大学科研创新团队(BR22-13-05)。

摘　　要：【目的】AtiPGAM2 基因是拟南芥(Arabidopsis thaliana)碱性磷酸酶超家族的成员,参与糖酵解过程。研究AtiPGAM2基因在逆境胁迫下的响应机制,为进一步研究AtiPGAM2的抗逆功能奠定基础。【方法】使用PlantCARE在线软件对AtiPGAM2基因启动子顺式作用元件进行分析。采用实时荧光定量PCR,检测AtiPGAM2在NaCl和ABA胁迫下的响应模式。克隆AtiPGAM2基因转入拟南芥,获得AtiPGAM2基因超表达株系。利用三引物法鉴定Atipgam2突变体。对过表达、突变体和野生型在盐胁迫下的萌发率、绿苗率以及各种非生物胁迫(甘露醇、ABA和MeJA)下的表型,进行统计分析。【结果】其启动子上存在多个光、MeJA、低温、ABA、SA、GAs等非生物胁迫和激素响应元件。荧光定量PCR分析显示,AtiPGAM2在NaCl和ABA胁迫下受到不同程度的诱导。成功获得AtiPGAM2基因过表达和突变体株系。在盐胁迫条件下AtiPGAM2过表达株系萌发率以及绿苗率均高于野生型,突变体表型则相反。甘露醇处理下突变体的萌发率低于野生型。ABA处理48 h内突变体和过表达AtiPGAM2株系的萌发率都低于野生型。MeJA处理下过表达的侧根数量高于野生型,突变体则相反。【结论】AtiPGAM2具有耐盐性,该基因响应甘露醇、ABA 和MeJA处理。【Objective】The AtiPGAM2 gene is a member of the alkaline phosphatase superfamily in Arabidopsis and is involved in the glycolysis process.Studying the response mechanism of AtiPGAM2 gene under stress conditions lays the foundation for further studying the stress resistance function of AtiPGAM2.【Method】This study used PlantCARE online software to analyze the cis-acting elements of the AtiPGAM2 gene promoter.Real time fluorescence quantitative PCR was used to detect the response patterns of AtiPGAM2 under NaCl and ABA stress.The AtiPGAM2 gene was cloned and transferred it into Arabidopsis to obtain an overexpressing strain of the AtiPGAM2 gene.The Atipgam2 mutant was identified using the three primer method.Statistical analysis was conducted on the germination rate,green seedling rate,and phenotypes under various abiotic stresses,including mannitol,ABA,and MeJA,for overexpression,mutants,and wild-type under salt stress.【Result】There are multiple abotic stress and hormone responsive elements on its promoter,such as light,MeJA,low temperature,ABA,SA,GAs,etc.Fluorescence quantitative PCR analysis showed that AtiPGAM2 was induced to varying degrees under NaCl and ABA stress.AtiPGAM2 gene overexpression and mutant strains were obtained successfully.Under salt stress,the germination rate and green seedling rate of AtiPGAM2 overexpressing strains were higher than those of the wild type,while the mutant surface type was the opposite.The germination rate of the mutant under mannitol treatment was lower than that of the wild-type.The germination rates of mutant and overexpressing AtiPGAM2 strains treated with ABA within 48 h were lower than those of the wild-type.Under MeJA treatment,the number of overexpressed lateral roots was higher than that of the wild-type,while the mutant was the opposite.【Conclusion】AtiPGAM2 has the tolerance to salt,and this gene responds to mannitol,ABA,and MeJA treatments.

关 键 词：AtiPGAM2 拟南芥 盐胁迫 非生物胁迫 甘露醇 ABA MEJA 

分 类 号：Q94[生物学—植物学]