Lin Mi, Yan Jingwan, Ali Muhammad Moaaz, Wang Shaojuan, Tian Shengnan, Chen Faxing, Lin Zhimin
Fujian Academy of Agricultural Sciences Biotechnology Institute, Fuzhou 350003, China.
College of Horticulture, Fujian Agriculture and Forestry University, Fuzhou 350002, China.
Biology (Basel). 2022 Jul 22;11(8):1092. doi: 10.3390/biology11081092.
Plant promoters play a vital role in the initiation and regulation of gene transcription. In this study, a rice protein/gene of unknown expression, named , was gained from a rice T-DNA capture line. The semi-quantitative RT-PCR analysis showed that the gene was only expressed in root, glume, and flower, but not in stem, leaf, embryo, and endosperm of japonica rice. The GUS activity analysis of the promoter showed that it was a reverse green tissue expression promoter, except in endosperm. The forward promoter of cannot normally drive the expression of the foreign gene, while the reverse promoter of is a green tissue-specific expression promoter, which can drive the expression of the foreign gene. The region from -2097 to -1543 bp was the key region for controlling the green tissue-specific expression. The regulatory sequences with different lengths from the 2097 bp reverse sequence from the upstream region of the were fused with the reporter gene and stably expressed in rice. Furthermore, transgenic rice plants carrying Cry1Ab encoding endotoxin, regulated by , were resistant to yellow stem borer. The analysis suggested that 10 light responsive elements of tissue-specific expression were found, including ACE, Box4, CAT-box, G-Box, G-box, GATA motif, GC motif, I-box, Sp1, and chs-unit1 M1. In addition, the results of 5' and 3' deletions further speculated that ACE and I-box may be the key elements for determining the green tissue-specific expression of promoter.
植物启动子在基因转录的起始和调控中起着至关重要的作用。在本研究中，从水稻T-DNA捕获系中获得了一个未知表达情况的水稻蛋白/基因，命名为 。半定量RT-PCR分析表明，该基因仅在粳稻的根、颖片和花中表达，而在茎、叶、胚和胚乳中不表达。 启动子的GUS活性分析表明，它是一个除胚乳外的反向绿色组织表达启动子。 的正向启动子不能正常驱动外源 基因的表达，而 的反向启动子是一个绿色组织特异性表达启动子，能够驱动外源 基因的表达。-2097至-1543 bp区域是控制绿色组织特异性表达的关键区域。将来自 上游区域的2097 bp反向序列的不同长度的调控序列与 报告基因融合，并在水稻中稳定表达。此外，由 调控的携带编码 内毒素的Cry1Ab的转基因水稻植株对二化螟具有抗性。分析表明，发现了10个组织特异性表达的光响应元件，包括ACE、Box4、CAT-box、G-Box、G-box、GATA基序、GC基序、I-box、Sp1和chs-unit1 M1。此外，5'和3'缺失的结果进一步推测，ACE和I-box可能是决定 启动子绿色组织特异性表达的关键元件。
