Vergish Satyam, Wolf Ryan, Song Wen-Yuan
Department of Plant Pathology, University of Florida, Gainesville, FL, USA.
Santa Fe High School, Alachua, FL, USA.
Bio Protoc. 2024 May 20;14(10):e4987. doi: 10.21769/BioProtoc.4987.
-mediated transient gene expression in Nicotiana benthamiana is widely used to study gene function in plants. One dramatic phenotype that is frequently screened for is cell death. Here, we present a simplified protocol for Agrobacterium-mediated transient gene expression by infiltration. Compared with current methods, the novel protocol can be done without a centrifuge or spectrometer, thereby suitable for K-12 outreach programs as well as rapidly identifying genes that induce cell death. Key features • The protocol simplifies the widely used -mediated transient gene expression assay [1] and can be completed within one week when plants are available. • Rice gene can induce a dramatic and easily identifiable cell death phenotype in • Allows identification of cell death-inducing genes and is suitable for teaching. • Compared to the currently used methods, our protocol omits the use of agroinfiltration buffer, pH meter, temperature-controlled growth chamber, centrifuge, and spectrophotometer. Graphical overview The photo demonstrates the method of agroinfiltration into the abaxial side of leaves using a needleless syringe.
农杆菌介导的本氏烟草瞬时基因表达被广泛用于研究植物基因功能。经常筛选的一种显著表型是细胞死亡。在此，我们展示了一种通过浸润进行农杆菌介导的瞬时基因表达的简化方案。与目前的方法相比，该新方案无需离心机或光谱仪即可完成，因此适用于K - 12推广项目以及快速鉴定诱导细胞死亡的基因。关键特性 • 该方案简化了广泛使用的瞬时基因表达检测方法[1]，当有植物可用时，一周内即可完成。 • 水稻基因可在本氏烟草中诱导出显著且易于识别的细胞死亡表型。 • 可鉴定诱导细胞死亡的基因，适用于教学。 • 与目前使用的方法相比，我们的方案省略了农杆菌浸润缓冲液、pH计、控温生长室、离心机和分光光度计的使用。图形概述 照片展示了使用无针注射器向叶片背面进行农杆菌浸润的方法。
