Title: Expanding the application of a UV-visible reporter for transient gene expression and stable transformation in plants

Abstract

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 canmore » 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.« less

Authors:

Yuan, Guoliang; Lu, Haiwei ;

• Search DOE PAGES for author "Lu, Haiwei"
• Search DOE PAGES for ORCID "0000-0003-3063-6555"

Tang, Dan; Hassan, Md Mahmudul ;

• Search DOE PAGES for author "Hassan, Md Mahmudul"
• Search DOE PAGES for ORCID "0000-0003-3399-8121"

Li, Yi; Chen, Jin-Gui ;

• Search DOE PAGES for author "Chen, Jin-Gui"
• Search DOE PAGES for ORCID "0000-0002-1752-4201"

Tuskan, Gerald A.; Yang, Xiaohan

• Search DOE PAGES for author "Yang, Xiaohan"
• Search DOE PAGES for ORCID "0000-0001-5207-4210"

Publication Date:

Mon Nov 01 00:00:00 EDT 2021

Research Org.:

Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)

Sponsoring Org.:

USDOE Office of Science (SC), Biological and Environmental Research (BER)

OSTI Identifier:

1828414

Alternate Identifier(s):

OSTI ID: 1831670

Grant/Contract Number:  

AC05-00OR22725

Resource Type:

Published Article

Journal Name:

Horticulture Research

Additional Journal Information:

Journal Name: Horticulture Research Journal Volume: 8 Journal Issue: 1; Journal ID: ISSN 2662-6810

Publisher:

Oxford University Press

Country of Publication:

China

Language:

English

Subject:

59 BASIC BIOLOGICAL SCIENCES