College of Life Science and Engineering, Southwest University of Science and Technology, Mianyang, 621010, China.
Key Laboratory of Application and Safety Control of Genetically Modified Crops, College of Agronomy and Biotechnology, Southwest University, Chongqing, 400715, China.
Sci Rep. 2024 Aug 21;14(1):19412. doi: 10.1038/s41598-024-69955-7.
In the past decade, the exploration of genetic resources in rice has significantly enhanced the efficacy of rice breeding. However, the exploration of genetic resources is hindered by the identification of candidate genes. To expedite the identification of candidate genes, this study examined tapetum programmed cell death-related genes OsiWAK1, OsPDT1, EAT1, TDR, and TIP2 to assess the efficacy of the Dual-Luciferase (Dual-LUC) assay in rapidly determining gene relationships. The study found that, in the Dual-LUC assay, OsiWAK1 and its various recombinant proteins exhibit comparable activation abilities on the EAT1 promoter, potentially indicating a false positive. However, the Dual-LUC assay can reveal that OsiWAK1 impacts both the function of its upstream regulatory factor OsPDT1 and the TDR/TIP2 transcription complex. By rapidly studying the relationship between diverse candidate genes and regulatory genes in a well-known trait via the Dual-LUC assay, this study provides a novel approach to expedite the determination of candidate genes such as genome-wide association study.
在过去的十年中，对水稻遗传资源的探索显著提高了水稻的育种效果。然而，遗传资源的探索受到候选基因鉴定的阻碍。为了加快候选基因的鉴定，本研究检查了与花粉壁程序性细胞死亡相关的基因 OsiWAK1、OsPDT1、EAT1、TDR 和 TIP2，以评估双荧光素酶（Dual-LUC）测定在快速确定基因关系方面的功效。研究发现，在 Dual-LUC 测定中，OsiWAK1 及其各种重组蛋白对 EAT1 启动子表现出可比的激活能力，这可能表明存在假阳性。然而，Dual-LUC 测定可以揭示 OsiWAK1 影响其上游调节因子 OsPDT1 和 TDR/TIP2 转录复合物的功能。通过 Dual-LUC 测定快速研究已知性状中不同候选基因和调节基因之间的关系，本研究为加快全基因组关联研究等候选基因的确定提供了一种新方法。
