FaGST基因过量表达获得拟南芥转基因植株  

Transgenic Arabidopsis thaliana Plants with FaGST Gene Overexpression

作　　者：陈锡 陈莹[1] 刘晓霞 刘重阳 吴溪 姚文琴 王小利 CHEN Xi;CHEN Ying;LIU Xiaoxia;LIU Chongyang;WU Xi;YAO Wenqin;WANG Xiaoli(Guizhou Pratacultural Institute,Guizhou Academy of Agricultural Sciences,Guiyang,Guizhou 550006;College of Life Sciences/Key Laboratory of Plant Resources Conservation and Germplasm Innovation in Mountainous Region,Ministry of Education/The National-local Joint Engineering Research Center for Utilization&Breeding of Plant Resources,Guizhou University,Guiyang,Guizhou 550025,China)

机构地区：[1]贵州省草业研究所,贵州贵阳550006 [2]贵州大学生命科学学院/山地植物资源保护与种质创新教育部重点实验室/喀斯特山区植物资源利用与育种国家地方联合工程研究中心,贵州贵阳550025

出　　处：《贵州农业科学》2022年第7期21-25,共5页Guizhou Agricultural Sciences

基　　金：贵州省科技计划项目“GST基因改良菊苣种质创新及应用研究”[黔科合基础(2019)1302]。

摘　　要：【目的】探明FaGST基因的功能和分子调控机制,为其后期的抗性利用与研究提供科学依据。【方法】利用前期构建的pCAMBIA1300-35S空载体和pCAMBIA1300-35S-FaGST过表达载体,将其转化感受态(DH5α)细胞后导入农杆菌GV3101,利用花序侵染法转化拟南芥,通过潮霉素(Hyp)进行筛选、PCR鉴定,并利用实时荧光定量PCR检测其表达量。【结果】FaGST基因成功导入拟南芥中,获得6株空载体转基因拟南芥植株和8株转FaGST基因拟南芥植株;8株转基因拟南芥植株的FaGST基因表达量为1.31~2.06,较野生型拟南芥植株表达量(1.00)显著或极显著提高。【结论】成功获得拟南芥转基因纯合植株,FaGST基因在拟南芥植株中已过量表达。【Objective】The function and molecular regulation mechanism of FaGST gene is explored to provide the scientific basis for its utilization and research of resistance.【Method】The pCAMBIA1300-35S empty vector and pCAMBIA1300-35S-FaGST overexpression vector are successfully transformed into competence cells(DH5α).The competence cells are transformed into GV3101(Agrobacterium tumefaciens)and the FaGST gene is transformed into Arabidopsis thaliana by the inflorescence infection method.The expression of FaGST gene is detected by hygromycin(Hyp)screening,PCR identification and real-time fluorescence quantitative PCR.【Result】The FaGST gene is transformed into A.thaliana successfully.6 transgenic A.thaliana plants with pCAMBIA1300-35S empty vector and 8 transgenic A.thaliana plants with pCAMBIA1300-35S-FaGST overexpression vector are obtained.The expression of FaGST gene in 8 transgenic A.thaliana plants with pCAMBIA1300-35S-FaGST overexpression vector is 1.31~2.06 and higher than wild A.thaliana plants significantly and very significantly.【Conclusion】The homozygosis transgenic A.thaliana plants with the expression of 1.00 with pCAMBIA1300-35S-FaGST overexpression vector are obtained successfully.The FaGST gene is excessively expressed in the homozygosis transgenic A.thaliana plants with pCAMBIA1300-35S-FaGST overexpression vector.

关 键 词：拟南芥 FaGST 花序侵染 表达量 

分 类 号：S505.53[农业科学—作物学] Q943.2[生物学—植物学]