论文部分内容阅读
利用Affymetrix水稻表达芯片分析了超级稻两优培九母本培矮64S(Oryza sativa L.)在经过低温、干旱、高温等逆境条件胁迫处理后,不同组织器官在不同的生长发育时期全基因组的表达差异。筛选到一个在正常条件和逆境条件下均有较高表达水平的基因Os SG4(Gen Bank登录号:AK068991.1),从水稻日本晴基因组中克隆得到其上游启动子区域Ospz4(1 625 bp),将其与GUS报告基因融合构建植物表达载体p CAMBIA1301P4,并用农杆菌介导法转化台北309获得转基因植株。组织化学染色结果表明,Ospz4启动子可驱动GUS报告基因在转基因水稻植株的叶片,根、茎、颖花、胚乳及胚芽鞘中均有表达。在该启动子的基础上,构建了4个不同长度的5’端缺失片段启动子与GUS报告基因融合的植物表达载体,通过注射烟草(Nicotiana benthamiana)进行瞬时表达分析,结果表明,4个片段均可驱动GUS基因的表达,其中最短片段(492 bp)的表达活性最强。本研究表明Ospz4启动子可用于研究与遗传改良生产,具有重要的理论与实践价值,并有助于该启动子的改造。
Affymetrix rice expression chips were used to analyze the differences of genome-wide expression of different tissues and organs during different growth and development stages after under stress conditions of low temperature, drought and high temperature in Oryza sativa L. super hybrid rice Liangyoupeijiu. Os SG4 (1 625 bp) was cloned from Nipponbare (Nipponbare) genome and cloned into Os SG4 (GenBank accession number: AK068991.1), which has high expression level under both normal and adversity conditions. The recombinant plasmid p CAMBIA1301P4 was constructed by fusion with GUS reporter gene and transformed into Taipei 309 by Agrobacterium tumefaciens method. Histochemical staining showed that the Ospz4 promoter could drive GUS reporter gene expression in the leaves, roots, stems, spikelets, endosperm and coleoptile of transgenic rice plants. Based on this promoter, we constructed four plant expression vectors fused with the GUS reporter gene with 5 ’deletion motifs of different lengths. The transient expression analysis of tobacco with Nicotiana benthamiana showed that four fragments Can drive the expression of GUS gene, of which the shortest fragment (492 bp) expression activity of strongest. This study shows that the Ospz4 promoter can be used for research and genetic improvement of production, has important theoretical and practical value, and contributes to the transformation of the promoter.