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为解析月季失水诱导SNAC类转录因子RhNAC4的转录调控特性,利用染色体步移的方法分离了RhNAC4的启动子,并构建含GUS报告基因的载体转化拟南芥,分析了RhNAC4的启动子表达特性。结果表明:1)在月季中分离得到RhNAC4基因5′端上游1 753bp的启动子序列;2)RhNAC4启动子序列中具有与发育、激素响应、非生物胁迫和生物胁迫等相关的顺式作用元件;3)获得了RhNAC4启动子驱动GUS基因的转基因拟南芥植株,GUS染色表明RhNAC4基因在植株不同的生长发育阶段和花器官中都有表达;4)失水胁迫、赤霉素、1-氨基环丙烷1-羧酸和甘露醇处理能够诱导RhNAC4的启动子活性,而盐和脱落酸处理对启动子活性的影响不显著。总之,月季切花失水胁迫诱导的SNAC类转录因子RhNAC4基因启动子含有逆境相关调控元件,其启动子在植株不同发育阶段和失水、乙烯等条件下具有转录活性。研究结果有助于解析RhNAC4参与月季切花失水胁迫耐性的作用机理。
To analyze the transcriptional regulation of SNAC transcription factor RhNAC4 induced by water loss, RhNAC4 promoter was isolated by chromosome walking and transformed into Arabidopsis thaliana with GUS reporter gene. The expression characteristics of RhNAC4 promoter were analyzed . The results showed as follows: 1) 1 753 bp upstream of 5 ’end of RhNAC4 gene was isolated from rose; 2) cis-acting element related to development, hormone response, abiotic stress and biotic stress were found in RhNAC4 promoter sequence ; 3) Rhizobacteria were obtained from transgenic Arabidopsis plants with GUS gene driven by RhNAC4 promoter. GUS staining showed that RhNAC4 gene was expressed in different stages of plant growth and flower organs; 4) Aminocyclopropane 1-carboxylic acid and mannitol treatment can induce the promoter activity of RhNAC4, while salt and abscisic acid treatment had no significant effect on the promoter activity. In conclusion, the promoter of SNAC transcription factor RhNAC4 gene induced by the loss of cut rose flower possesses stress-related regulatory elements, and its promoter is transcriptionally active at different developmental stages of the plant, such as water loss and ethylene. The results are helpful to analyze the mechanism of RhNAC4 involved in the tolerance of cut rose flower to water stress.