A Generic Split-Gfp-Based Reporter System for Agrobacterium-Mediated Protein Translocation in Plants | [Abstract Book on International Conference on Food and Applied Sciences (ICFAS-23) 3-5 August 23 • 2023]

Author(s):

1. Majid Khan: Molecular and Developmental Genetics, Institute of Biology Leiden, Leiden University, Sylvius Laboratory, Sylviusweg 72, 2333 BE Leiden, Netherlands; Institute of Biotechnology & Genetic Engineering, the University of Agriculture, Peshawar, Khyber Pakhtunkhwa, Pakistan.

2. Ivo Gariboldi: Molecular and Developmental Genetics, Institute of Biology Leiden, Leiden University, Sylvius Laboratory, Sylviusweg 72, 2333 BE Leiden, Netherlands

3. Remko Offringa: Molecular and Developmental Genetics, Institute of Biology Leiden, Leiden University, Sylvius Laboratory, Sylviusweg 72, 2333 BE Leiden, Netherlands

Abstract:

Agrobacterium tumefaciens is well characterized for its ability to transfer DNA to plant and fungal cells, but the fact that it also translocates proteins to its host cells was only revealed more recently. Agrobacterium-mediated protein translocation (AMPT) was first detected by restoration of a resistance marker or GFP reporter following translocation of a Cre recombinase-VirF/VirE2 protein fusion. Later the split-GFP system was used to detect translocation of GFP11-Vir fusions to recipient reporter lines overexpressing GFP1-10. Unfortunately, these translocation reporter systems are not easily applicable to transformation resistant and regeneration recalcitrant plants such as sweet pepper and tulip, for which the generation of reporter lines are difficult. Here, we designed a generic splitGFP-based reporter system for AMPT to be used directly in wild-type plants. In this system, the GFP1-10 part is transiently expressed from a T-DNA that is co-transferred with a fusion protein comprising the GFP11 part and the C-terminal translocation signal of VirF from the same Agrobacterium to any desired wild-type recipient cell. This modified generic protein translocation reporter system was successfully tested in a variety of tissues of different plant species, such as Nicotiana benthamiana, Nicotiana tabacum, Arabidopsis thaliana, Capsicum annum and Tulipa gesneriana. The system reported efficient AMPT to these plant species, and also appeared to be useful for optimization of AMT of tulip, and for the visual selection of transgenic tulip shoots.

Page(s): 133-133

DOI: DOI not available

Published: Journal: Abstract Book on International Conference on Food and Applied Sciences (ICFAS-23) 3-5 August 23, Volume: 0, Issue: 0, Year: 2023

Keywords:

Agrobacterium tumefaciens , Protein translocation , SplitGFP system , Generic

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