玉米低磷胁迫诱导型强启动子P_(1502)-ZmPHR1的克隆与表达分析  

Cloning and Expression Analysis of Strong Inducible Promoter P_(1502)-ZmPHR1 Responding to Low Phosphorus Stress in Maize

作　　者：杨瑞娟 白建荣[2] 闫蕾[2] 苏亮[2] 王秀红[3] 李锐[2] 张丛卓[2] YANG Rui-Juan;BAI Jian-Rong;YAN Lei;SU Liang;WANG Xiu-Hong;LI Rui;ZHANG Cong-Zhuo(College of Biology Engineering,Shanxi University,Taiyuan 030006,Shanxi,China;Institute of Crop Sciences,Shanxi Academy of Agricultural Sciences,Taiyuan 030031,Shanxi,China;Research Center of Modern Agriculture,Shanxi Academy of Agricultural Sciences,Taiyuan 030031,Shanxi,China)

机构地区：[1]山西大学生物工程学院,山西太原030006 [2]山西省农业科学院作物科学研究所,山西太原030031 [3]山西省农业科学院现代农业研究中心,山西太原030031

出　　处：《作物学报》2018年第7期1000-1009,共10页Acta Agronomica Sinica

基　　金：山西省农业科学院农业科技创新项目(ZDSYS1507)资助~~

摘　　要：玉米ZmPHR1基因是转录因子MYB-CC家族成员,过量表达该基因可促进低磷环境中植物对磷的吸收并改善植物的生长。本研究从磷高效利用型玉米自交系478中获得了ZmPHR1基因的5'上游序列P_(2205)-ZmPHR1,序列长度2205 bp。将P_(2205)-ZmPHR1及它的4种5'端缺失启动子片段与报告基因GUS连接后转化拟南芥。GUS组织化学染色与GUS定量分析结果表明,在0~1502 bp的片段范围内,随着片段长度的增加,它对外界低磷环境的响应逐步增加,在-1502 bp时,响应时间短,表达活性高,持续时间长;超过1502 bp其活性不但没有增高,反而变为下降或无变化。推测0至-972 bp为启动子功能区段,-972 bp至-1502 bp为启动功能增强区段。转P_(1502)-ZmPHR1拟南芥的实时定量PCR结果显示,P_(1502)-ZmPHR1既受外界信号的刺激,也受体内基因产物的影响,有反馈调节的作用,是一个受低磷胁迫诱导的强诱导型启动子,它诱导基因表达的功能受体内外磷浓度的调节。该研究为在低磷环境改良作物提供了理想的启动子和应用依据。ZmPHR1,a member of transcription factor MYB-CC family,enhanced phosphate absorption and improved plant growth when it was overexpressed under low-phosphate conditions.In order to elucidate the transcriptional regulatory mechanism of the promoter of ZmPHR1,its 5′upstream sequence P2205-ZmPHR1 was cloned from P-efficient maize inbred line 478.The full length of P2205-ZmPHR1 and four sequence deletion fragments in different lengths fused with the reporter gene GUS were transformed into Arabidopsis,respectively.The results of histochemical and quantitative fluorometric GUS assay showed that in the interval from 0 to?1502 bp,the response time and activity of sequence deletion fragments were increased with increasing fragment length.However,the activities were decreased or unchanged when the length of fragments were longer than 1502 bp.It was considered that the interval from 0 to?972 bp was function region and that from?972 bp to?1502 bp was function enhancing region.The results of qPCR analysis in transgenic plants at different time points(0,1,2,3,and 4 days)after P-stress treatment showed that the activities of P1502-ZmPHR1 were influenced by both external signals and internal gene products,having feedback regulation.This study indicates that P1502-ZmPHR1 is a strong inducible promoter responding to low phosphorus stress and its functional activity is regulated by both external and internal phosphate concentration.These fundings provide an ideal promoter for crop improvement under low phosphate conditions and a basis for its application.

关 键 词：玉米 低磷胁迫 强诱导型启动子 P1502-ZmPHR1 

分 类 号：Q943.2[生物学—植物学] S513[农业科学—作物学]