State Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan, Yunnan Agricultural University, Kunming 650201, Yunnan, China.
Post-Doctoral Research Station of Plant Protection as first class discipline, Yunnan Agricultural University, Kunming 650201, Yunnan, China.
Int J Mol Sci. 2020 Mar 11;21(6):1909. doi: 10.3390/ijms21061909.
The regulatory mechanisms of pollen development have potential value for applications in agriculture, such as better understanding plant reproductive regularity. Pollen-specific promoters are of vital importance for the ectopic expression of functional genes associated with pollen development in plants. However, there is a limited number of successful applications using pollen-specific promoters in genetic engineering for crop breeding and hybrid generation. Our previous work led to the identification and isolation of the promoter from rice. In this study, to analyze the effects of different putative regulatory motifs in the promoter, a series of promoter deletions were fused to a reporter gene and then stably introduced into rice and . Histochemical GUS analysis of transgenic plants revealed that p385 (from -385 to -1) specifically mediated maximal GUS expression in pollen tissues. The S region (from -385 to -203) was the key region for controlling the pollen-specific expression of a downstream gene. The E1 (-967 to -606), E2 (-202 to -120), and E3 (-119 to -1) regions enhanced ectopic promoter activity to different degrees. Moreover, the p385 promoter could alter the expression pattern of the 35S promoter and improve its activity when they were fused together. In summary, the p385 promoter, a short and high-activity promoter, can function to drive pollen-specific expression of transgenes in monocotyledon and dicotyledon transformation experiments.
花粉发育的调控机制在农业应用中具有潜在价值,例如更好地了解植物生殖规律。花粉特异性启动子对于植物中与花粉发育相关的功能基因的异位表达至关重要。然而,在利用花粉特异性启动子进行作物遗传改良和杂种生成的基因工程中,成功的应用案例数量有限。我们之前的工作从水稻中鉴定和分离出 启动子。在这项研究中,为了分析 启动子中不同假定调控元件的影响,一系列启动子缺失与报告基因融合,并稳定导入水稻和拟南芥中。对转基因植物的组织化学 GUS 分析显示,p385(从-385 到-1)在花粉组织中特异性介导最大的 GUS 表达。S 区(从-385 到-203)是控制下游基因花粉特异性表达的关键区域。E1(-967 到-606)、E2(-202 到-120)和 E3(-119 到-1)区以不同程度增强了异位启动子的活性。此外,p385 启动子可以改变 35S 启动子的表达模式,并在融合在一起时提高其活性。综上所述,p385 启动子是一个短而活性高的启动子,可以在单子叶和双子叶转化实验中驱动转基因的花粉特异性表达。