Unstable Transgene Expression Affects Long-Term Efficacy of the Arabidopsis Immune Receptor EFR to Confer Quantitative Resistance to Citrus Canker Under Field Conditions

The Arabidopsis thaliana pattern recognition ELONGATION FACTOR-TU RECEPTOR (EFR) recognizes and binds the bacterial pathogen-associated molecular pattern (PAMP) elongation factor thermo unstable (EF-Tu) and its highly conserved derived peptide elf18. Previous work revealed that the transgenic expres...

Full description

Saved in:
Bibliographic Details
Main Authors: Mukesh Jain, Gerald V. Minsavage, Naweena Thapa, Diana M. Horvath, Cyril Zipfel, Gloria A. Moore, Latanya C. Fisher, Vladimir Orbović, Jeffrey B. Jones
Format: Article
Language:English
Published: The American Phytopathological Society 2024-12-01
Series:PhytoFrontiers
Subjects:
Online Access:https://apsjournals.apsnet.org/doi/10.1094/PHYTOFR-06-24-0071-SC
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1850104836715970560
author Mukesh Jain
Gerald V. Minsavage
Naweena Thapa
Diana M. Horvath
Cyril Zipfel
Gloria A. Moore
Latanya C. Fisher
Vladimir Orbović
Jeffrey B. Jones
author_facet Mukesh Jain
Gerald V. Minsavage
Naweena Thapa
Diana M. Horvath
Cyril Zipfel
Gloria A. Moore
Latanya C. Fisher
Vladimir Orbović
Jeffrey B. Jones
author_sort Mukesh Jain
collection DOAJ
description The Arabidopsis thaliana pattern recognition ELONGATION FACTOR-TU RECEPTOR (EFR) recognizes and binds the bacterial pathogen-associated molecular pattern (PAMP) elongation factor thermo unstable (EF-Tu) and its highly conserved derived peptide elf18. Previous work revealed that the transgenic expression of EFR (and chimeric EFR::XA21) in several heterologous plant species, including in members of Solanaceae and in sweet orange, confers broad-spectrum bacterial disease resistance. In this study, stable sweet orange (Citrus × sinensis) and grapefruit (Citrus × paradisi) transformants expressing EFR or EFR::XA21 were generated in an attempt to confer broad spectrum resistance to Xanthomonas citri subsp. citri (Xcc), the causal agent of citrus canker, a serious disease of commercial citriculture. The transgene expression was confirmed via real-time quantitative reverse transcriptase PCR (RT-qPCR). Juvenile transgenic trees were challenged with Xcc (spray and pinprick inoculation) and evaluated for canker susceptibility and disease progression. Three independently transformed transgenic events (out of 19 total) displayed partial resistance to canker when spray-inoculated, and one line showed resistance when wound-inoculated. Surprisingly, when two mature transgenic lines were evaluated under field conditions and exposure to natural infection, both were found to be as susceptible as the wild-type under prevalent pathogen load. RT-qPCR data indicated a gradual decline and significant spatial variability in EFR expression in leaves excised from different branches of the same tree. The data presented here call for a need for evaluating different promoters for stable and long-term EFR expression in woody perennial species. [Figure: see text] Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.
format Article
id doaj-art-b10b47663380427cacad193cf4080c6b
institution DOAJ
issn 2690-5442
language English
publishDate 2024-12-01
publisher The American Phytopathological Society
record_format Article
series PhytoFrontiers
spelling doaj-art-b10b47663380427cacad193cf4080c6b2025-08-20T02:39:15ZengThe American Phytopathological SocietyPhytoFrontiers2690-54422024-12-014449149710.1094/PHYTOFR-06-24-0071-SCUnstable Transgene Expression Affects Long-Term Efficacy of the Arabidopsis Immune Receptor EFR to Confer Quantitative Resistance to Citrus Canker Under Field ConditionsMukesh Jain0Gerald V. Minsavage1Naweena Thapa2Diana M. Horvath3Cyril Zipfel4Gloria A. Moore5Latanya C. Fisher6Vladimir Orbović7Jeffrey B. Jones8Plant Pathology Department, University of Florida, Gainesville, FL 32611, U.S.A.Plant Pathology Department, University of Florida, Gainesville, FL 32611, U.S.A.Plant Pathology Department, University of Florida, Gainesville, FL 32611, U.S.A.2Blades Foundation, Evanston, IL 60201, U.S.A.The Sainsbury Laboratory, Norwich Research Park, University of East Anglia, Norwich, NR4 7UH, U.K.Department of Horticultural Sciences, University of Florida, Gainesville, FL 32611, U.S.A.Department of Horticultural Sciences, University of Florida, Gainesville, FL 32611, U.S.A.Citrus Research and Education Center, University of Florida, Lake Alfred, FL 33850, U.S.A.Plant Pathology Department, University of Florida, Gainesville, FL 32611, U.S.A.The Arabidopsis thaliana pattern recognition ELONGATION FACTOR-TU RECEPTOR (EFR) recognizes and binds the bacterial pathogen-associated molecular pattern (PAMP) elongation factor thermo unstable (EF-Tu) and its highly conserved derived peptide elf18. Previous work revealed that the transgenic expression of EFR (and chimeric EFR::XA21) in several heterologous plant species, including in members of Solanaceae and in sweet orange, confers broad-spectrum bacterial disease resistance. In this study, stable sweet orange (Citrus × sinensis) and grapefruit (Citrus × paradisi) transformants expressing EFR or EFR::XA21 were generated in an attempt to confer broad spectrum resistance to Xanthomonas citri subsp. citri (Xcc), the causal agent of citrus canker, a serious disease of commercial citriculture. The transgene expression was confirmed via real-time quantitative reverse transcriptase PCR (RT-qPCR). Juvenile transgenic trees were challenged with Xcc (spray and pinprick inoculation) and evaluated for canker susceptibility and disease progression. Three independently transformed transgenic events (out of 19 total) displayed partial resistance to canker when spray-inoculated, and one line showed resistance when wound-inoculated. Surprisingly, when two mature transgenic lines were evaluated under field conditions and exposure to natural infection, both were found to be as susceptible as the wild-type under prevalent pathogen load. RT-qPCR data indicated a gradual decline and significant spatial variability in EFR expression in leaves excised from different branches of the same tree. The data presented here call for a need for evaluating different promoters for stable and long-term EFR expression in woody perennial species. [Figure: see text] Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.https://apsjournals.apsnet.org/doi/10.1094/PHYTOFR-06-24-0071-SCcitrus cankerEFREF-TuPAMPPTIXA21
spellingShingle Mukesh Jain
Gerald V. Minsavage
Naweena Thapa
Diana M. Horvath
Cyril Zipfel
Gloria A. Moore
Latanya C. Fisher
Vladimir Orbović
Jeffrey B. Jones
Unstable Transgene Expression Affects Long-Term Efficacy of the Arabidopsis Immune Receptor EFR to Confer Quantitative Resistance to Citrus Canker Under Field Conditions
PhytoFrontiers
citrus canker
EFR
EF-Tu
PAMP
PTI
XA21
title Unstable Transgene Expression Affects Long-Term Efficacy of the Arabidopsis Immune Receptor EFR to Confer Quantitative Resistance to Citrus Canker Under Field Conditions
title_full Unstable Transgene Expression Affects Long-Term Efficacy of the Arabidopsis Immune Receptor EFR to Confer Quantitative Resistance to Citrus Canker Under Field Conditions
title_fullStr Unstable Transgene Expression Affects Long-Term Efficacy of the Arabidopsis Immune Receptor EFR to Confer Quantitative Resistance to Citrus Canker Under Field Conditions
title_full_unstemmed Unstable Transgene Expression Affects Long-Term Efficacy of the Arabidopsis Immune Receptor EFR to Confer Quantitative Resistance to Citrus Canker Under Field Conditions
title_short Unstable Transgene Expression Affects Long-Term Efficacy of the Arabidopsis Immune Receptor EFR to Confer Quantitative Resistance to Citrus Canker Under Field Conditions
title_sort unstable transgene expression affects long term efficacy of the arabidopsis immune receptor efr to confer quantitative resistance to citrus canker under field conditions
topic citrus canker
EFR
EF-Tu
PAMP
PTI
XA21
url https://apsjournals.apsnet.org/doi/10.1094/PHYTOFR-06-24-0071-SC
work_keys_str_mv AT mukeshjain unstabletransgeneexpressionaffectslongtermefficacyofthearabidopsisimmunereceptorefrtoconferquantitativeresistancetocitruscankerunderfieldconditions
AT geraldvminsavage unstabletransgeneexpressionaffectslongtermefficacyofthearabidopsisimmunereceptorefrtoconferquantitativeresistancetocitruscankerunderfieldconditions
AT naweenathapa unstabletransgeneexpressionaffectslongtermefficacyofthearabidopsisimmunereceptorefrtoconferquantitativeresistancetocitruscankerunderfieldconditions
AT dianamhorvath unstabletransgeneexpressionaffectslongtermefficacyofthearabidopsisimmunereceptorefrtoconferquantitativeresistancetocitruscankerunderfieldconditions
AT cyrilzipfel unstabletransgeneexpressionaffectslongtermefficacyofthearabidopsisimmunereceptorefrtoconferquantitativeresistancetocitruscankerunderfieldconditions
AT gloriaamoore unstabletransgeneexpressionaffectslongtermefficacyofthearabidopsisimmunereceptorefrtoconferquantitativeresistancetocitruscankerunderfieldconditions
AT latanyacfisher unstabletransgeneexpressionaffectslongtermefficacyofthearabidopsisimmunereceptorefrtoconferquantitativeresistancetocitruscankerunderfieldconditions
AT vladimirorbovic unstabletransgeneexpressionaffectslongtermefficacyofthearabidopsisimmunereceptorefrtoconferquantitativeresistancetocitruscankerunderfieldconditions
AT jeffreybjones unstabletransgeneexpressionaffectslongtermefficacyofthearabidopsisimmunereceptorefrtoconferquantitativeresistancetocitruscankerunderfieldconditions