CAJ | 학술논문

对前期克隆的水稻GAST基因家族新成员OsGASR4开展研究，利用DNAMAN软件分析水稻OsGASR4进化树；检测GA处理野生型和d-18水稻突变体后OsGASR4表达变化；利用RT-PCR方法分析该基因的时空表达特性；克隆了该基因上游长约2082 bp调控序列，并利用网络工具预测启动子顺式作用元件，构建OsGASR4启动子GUS融合表达载体，通过农杆菌介导的水稻遗传转化。结果表明：OsGASR4蛋白具有典型的GASA保守结构域，启动子里含有多个与激素响应元件、光诱导相关元件以及逆境胁迫响应元件；GA3处理降低了OsGASR4转录水平；RT-PCR检测和GUS染色的结果表明，OsGASR4在水稻茎和幼穗的表达量相对较高。表明OsGASR4可能作为GA信号途径的抑制因子参与水稻茎和穗的早期发育。
대전기극륭적수도GAST기인가족신성원OsGASR4개전연구，이용DNAMAN연건분석수도OsGASR4진화수；검측GA처리야생형화d-18수도돌변체후OsGASR4표체변화；이용RT-PCR방법분석해기인적시공표체특성；극륭료해기인상유장약2082 bp조공서렬，병이용망락공구예측계동자순식작용원건，구건OsGASR4계동자GUS융합표체재체，통과농간균개도적수도유전전화。결과표명：OsGASR4단백구유전형적GASA보수결구역，계동자리함유다개여격소향응원건、광유도상관원건이급역경협박향응원건；GA3처리강저료OsGASR4전록수평；RT-PCR검측화GUS염색적결과표명，OsGASR4재수도경화유수적표체량상대교고。표명OsGASR4가능작위GA신호도경적억제인자삼여수도경화수적조기발육。
OsGASR4, a GAST homologues gene in rice, was investigated in the present study.Phylogenetic analysis of OsGASR4 was performed with DNAMAN and analysis of OsGASR4 expression was detected in the wild-type Nip-ponbare and GA3 synthesis-deficient mutant d-18 treated by GA3 .Expression pattern of OsGASR4 was determined in rice tissues and organs using RT-PCR.The promoter of OsGASR4 was fused with GUS gene to construct expression vector and transformed into rice by Agrobacterium-mediated transformation.The results showed that OsGASR4 is a typical GAST protein with conserved GASA-like domain ( cysteine-rich domain) and the promoter contains multiple components such as light induced, stress response and hormone response elements.OsGASR4 was down-regulated by GA3 .The results of RT-PCR and the histochemical localization of GUS activity showed that a high level of OsGASR4 expression was detected in stem and spike of rice.These results suggest that the transcription level of OsGASR4 is inhibited by GA3 and OsGASR4 might be involved as a negative signaling factor in the development of stems and spikes in rice.