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Article|01 Nov 2021|OPEN
Expanding the application of a UV-visible reporter for transient gene expression and stable transformation in plants
Guoliang Yuan1,2, Haiwei Lu1, Dan Tang3,4, Md Mahmudul Hassan1,2,5, Yi Li3, Jin-Gui Chen1,2, Gerald A. Tuskan1,2, & Xiaohan Yang1,2,
1Biosciences Division, Oak Ridge National Laboratory, Oak Ridge, TN 37831, USA
2The Center for Bioenergy Innovation, Oak Ridge National Laboratory, Oak Ridge, TN 37831, USA
3Department of Plant Science and Landscape Architecture, University of Connecticut, Storrs, CT 06269, USA
4National Center for Citrus Improvement, College of Horticulture, Hunan Agricultural University, Changsha 410128 Hunan, China
5Department of Genetics and Plant Breeding, Patuakhali Science and Technology University, Dumki, Patuakhali 8602, Bangladesh

Horticulture Research 8,
Article number: 234 (2021)
doi: 10.1038/hortres.2021.234
Views: 16

Received: 20 May 2021
Revised: 25 Jul 2021
Accepted: 01 Aug 2021
Published online: 01 Nov 2021

Abstract

Green fluorescent protein (GFP) has been widely used for monitoring gene expression and protein localization in diverse organisms. However, highly sensitive imaging equipment, like fluorescence microscope, is usually required for the visualization of GFP, limitings its application to fixed locations in samples. A reporter that can be visualized in real-time regardless the shape, size and location of the target samples will increase the flexibility and efficiency of research work. Here, we report the application of a GFP-like protein, called eYGFPuv, in both transient expression and stable transformation, in two herbaceous plant species (Arabidopsis and tobacco) and two woody plant species (poplar and citrus). We observed bright fluorescence under UV light in all of the four plant species without any effects on plant growth or development. eYGFPuv was shown to be effective for imaging transient expression in leaf and root tissues. With a focus on in vitro transformation, we demonstrated that the transgenic events expressing 1x eYGFPuv could be easily identified visually during the callus stage and the shoot stage, enabling early and efficient selection of transformants. Furthermore, whole-plant level visualization of eYGFPuv revealed its ubiquitous stability in transgenic plants. In addition, our transformation experiments showed that eYGFPuv can also be used to select transgenic plants without antibiotics. This work demonstrates the feasibility of utilizing 1x eYGFPuv in studies of gene expression and plant transformation in diverse plants.