中国农业科学2017，Vol.50Issue(3)：582-590,9.
大豆紫色酸性磷酸酶基因GmPAP4启动子结构与活性分析
Construction and Activity Analysis of the Promoter of Purple Acid Phosphatase Gene GmPAP4 in Soybean
摘要
Abstract
[Objective]GmPAP4 promoter (PAP4-pro) was cloned, and its expression character was analyzed, thus providing a basis for research of regulatory mechanism of GmPAP4.[Method]On the basis of GmPAP4 (GenBank No. HQ162477) cDNA sequence and blasting the soybean reference genome,GmPAP4 promoter sequence was cloned with specific primers. The promoter regulatory elements were predicted through the online database PLACE and PlantCARE.PAP4-pro-GUS was constructed and transformed into Agrobacterium tumefaciens GV3101, which subsequently was transformed into Arabidopsis thaliana with Floral dip method. The T3 transgenic plants were obtained by being screened with kanamycin (kan) and PCR amplification.[Result]GmPAP4 promoter was cloned, and its regulatory elements were analyzed using online databases PLACE and PlantCARE. The results showed that the promoter of GmPAP4 not only contained the core elements, but also contained the following elements: (1) tissue specific regulatory elements including as1 (root specific expression element), Skn-1_motif (endosperm specific expression element); (2): Response elements including TC-rich repeats (stress responsive elements), Box-W3 (fungal response related regulatory element); (3) binding sites including MBS (MYB binding sites of transcription factors),etc. The results of GUS staining showed that GmPAP4 promoter was mainly expressed in roots of transgenic Arabidopsis. The expression and activity of GUS measurement showed that GUS expression and GUS activity in roots of transgenic plants under phytate condition was, respectively, 1.3-fold (P<0.05) and 1.9-fold (P<0.05) than that under normal phosphorus condition.[Conclusion]Soybean GmPAP4 promoter was obtained in this study. According to GUS staining in different tissues and activity analysis under different phosphorus conditions, it was found that the promoter of GmPAP4 was mainly expressed in roots and significantly induced by low phosphorus signal.关键词
大豆/GmPAP4/启动子/组织特异性/植酸磷响应Key words
soybean/GmPAP4/promoter/tissue specificity/response to phytate phosphorus starvation引用本文复制引用
孔佑宾,李喜焕,张彩英..大豆紫色酸性磷酸酶基因GmPAP4启动子结构与活性分析[J].中国农业科学,2017,50(3):582-590,9.基金项目
转基因生物新品种培育科技重大专项(2014ZX0800404B)、河北省自然科学基金(C2014204035) （2014ZX0800404B）
