Graduate School of Life and Environmental Sciences, University of Tsukuba, Tennodai 1-1-1, Tsukuba, Ibaraki 305-8572, Japan.
Plant Cell Rep. 2012 Aug;31(8):1415-24. doi: 10.1007/s00299-012-1257-5. Epub 2012 Apr 6.
Fruit-specific promoters have been used as genetic engineering tools for studies on molecular mechanism of fruit development and advance in fruit quality and additional value by increasing functional component. Especially fruit-ripening specific promoters have been well utilized and studied in tomato; however, few studies have reported the development of promoters that act at fruit developing stages such as immature green and mature green periods. In this study, we report novel promoters for gene expression during the green to ripening stages of tomato fruit development. Genes specifically expressed at tomato fruit were selected using microarray data. Subsequent to confirmation of the expression of the selected 12 genes, upstream DNA fragments of the genes LA22CD07, Les.3122.2.A1_a_at and LesAffx.6852.1.S1_at which specifically expressed at fruit were isolated from tomato genomic DNA as promoter regions. Isolated promoter regions were fused with the GUS gene and the resultant constructs were introduced into tomato by agrobacterium-mediated transformation for evaluation of promoter activity in tomato fruit. The two promoters of LA22CD07, and LesAffx.6852.1.S1_at showed strong activity in the fruit, weak activity in the flower and undetectable activity in other tissues. Unlike well-known fruit-ripening specific promoters, such as the E8 promoter, these promoters exhibited strong activity in green fruit in addition to red-ripening fruit, indicating that the promoters are suitable for transgene expression during green to ripening stages of tomato fruit development.
Novel fruit-specific promoters have been identified and are suitable for transgene expression during green to ripening stages of tomato fruit development.
水果特异性启动子已被用作研究水果发育分子机制以及通过增加功能性成分提高水果品质和附加值的基因工程工具。特别是在番茄中,已经很好地利用和研究了成熟特异性启动子;然而,很少有研究报道在果实发育阶段(如未成熟绿色期和成熟绿色期)起作用的启动子的开发。在本研究中,我们报道了番茄果实发育过程中从绿色到成熟阶段基因表达的新启动子。使用微阵列数据选择了在番茄果实中特异性表达的基因。在确认了所选 12 个基因的表达后,从番茄基因组 DNA 中分离出在果实中特异性表达的基因 LA22CD07、Les.3122.2.A1_a_at 和 LesAffx.6852.1.S1_at 的上游 DNA 片段作为启动子区域。分离的启动子区域与 GUS 基因融合,所得构建体通过农杆菌介导的转化导入番茄中,以评估番茄果实中启动子活性。LA22CD07 和 LesAffx.6852.1.S1_at 的两个启动子在果实中表现出强烈的活性,在花中表现出较弱的活性,在其他组织中检测不到活性。与著名的成熟特异性启动子如 E8 启动子不同,这些启动子在绿色果实中除了红色成熟果实外表现出强烈的活性,表明这些启动子适合在番茄果实从绿色到成熟的发育阶段进行转基因表达。
已鉴定出新型果实特异性启动子,适合在番茄果实从绿色到成熟的发育阶段进行转基因表达。