水稻OsTFL2基因启动子的克隆及功能验证
文章编号：2095-1191（2019）10-2188-08
Cloning and functional identification of rice OsTFL2 gene promoter
CHEN Li1， ZHANG Xiao-dong1， WANG Xin-yi1， WEI Qing-yuan1，2， LI Hu1，
QI Huang-song1， LIU Fang1， GUAN He-xin1， QIU Yong-fu1*
（1College of Agricultural/State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources，
Guangxi University， Nanning 530004， China; 2Cash Crops Research Institute， Guangxi Academy of Agricultural Sciences， Nanning 530007， China）
Abstract：【Objective】The purpose was to clone the promoter sequence of gene OsTFL2 and then to study its structure and function，which would be favor for advanced mechanism analysis of rice flowering and flower development regulated by gene OsTFL2. 【Method】Homologous cloning method was applied to obtained the promoter sequence of OsTFL2 gene， PLACE and PlantCARE analysis were then conducted to analyze its structure and function. Furthermore，it was connected with vector pCAMBIA1301 carrying β-glucuronidase（GUS） gene to construct plant expression vector pCAMBIA1301-Promoter，which was then transferred to the callus tissue of rice variety Nongken 58 through agrobacterium induction. Finally，GUS staining method was conducted to detect the expression characterization and regulation function of the promoter with transgenic plants. 【Result】One 1.8-kb fragment of the upstream promoter sequence of OsTFL2 geneinitiation codon was cloned and it had typical promoter components of eukaryote，such as TATA box and CAAT-box. It also included the special cis-acting regulatory elements Pollen1lelat52（AGAAA），the specific multi-functional transcription factors that related to flowering genetic transcription CACTFTPPCA1（PACT，Y=C/T），CCAAT box1（CCAAT），DOFCORE（AAAG），and GATA box（GATA），the special component of meristem tissue CCGTCC-box，the light responsive elements G-box，Box I，CATT-motif，GATA-motif，and GT1-motif. The result suggested the OsTFL2 may regulate the flower development and flowering through the special elements. Furthermore，a total of 16 positive transgenic rice plants were obtained by PCR detection. The histochemical staining of GUS revealed that the GUS staining could be detected in the out glume，flower， anther，stigma and ovary of rice，but there were no obvious GUS staining in leaf， stem tip and root tip， which indicated the OsTFL2 promoter could be expressed in the out glume， anther and ovary. 【Conclusion】The cloned OsTFL2 promoter has promoting activity and shows tissue specific expression function，it affects expression of OsTFL2 gene to some extents and play a role in regulation of flower development and flowering in rice.