Cargando…

Specific Bacterial Pathogen Phytosensing Is Enabled by a Synthetic Promoter-Transcription Factor System in Potato

Phytosensors are genetically engineered plant-based sensors that feature synthetic promoters fused to reporter genes to sense and report the presence of specific biotic and abiotic stressors on plants. However, when induced reporter gene output is below detectable limits, owing to relatively weak pr...

Descripción completa

Detalles Bibliográficos
Autores principales: Persad-Russell, Ramona, Mazarei, Mitra, Schimel, Tayler Marie, Howe, Lana, Schmid, Manuel J., Kakeshpour, Tayebeh, Barnes, Caitlin N., Brabazon, Holly, Seaberry, Erin M., Reuter, D. Nikki, Lenaghan, Scott C., Stewart, C. Neal
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9083229/
https://www.ncbi.nlm.nih.gov/pubmed/35548302
http://dx.doi.org/10.3389/fpls.2022.873480
_version_ 1784703379257163776
author Persad-Russell, Ramona
Mazarei, Mitra
Schimel, Tayler Marie
Howe, Lana
Schmid, Manuel J.
Kakeshpour, Tayebeh
Barnes, Caitlin N.
Brabazon, Holly
Seaberry, Erin M.
Reuter, D. Nikki
Lenaghan, Scott C.
Stewart, C. Neal
author_facet Persad-Russell, Ramona
Mazarei, Mitra
Schimel, Tayler Marie
Howe, Lana
Schmid, Manuel J.
Kakeshpour, Tayebeh
Barnes, Caitlin N.
Brabazon, Holly
Seaberry, Erin M.
Reuter, D. Nikki
Lenaghan, Scott C.
Stewart, C. Neal
author_sort Persad-Russell, Ramona
collection PubMed
description Phytosensors are genetically engineered plant-based sensors that feature synthetic promoters fused to reporter genes to sense and report the presence of specific biotic and abiotic stressors on plants. However, when induced reporter gene output is below detectable limits, owing to relatively weak promoters, the phytosensor may not function as intended. Here, we show modifications to the system to amplify reporter gene signal by using a synthetic transcription factor gene driven by a plant pathogen-inducible synthetic promoter. The output signal was unambiguous green fluorescence when plants were infected by pathogenic bacteria. We produced and characterized a phytosensor with improved sensing to specific bacterial pathogens with targeted detection using spectral wavelengths specific to a fluorescence reporter at 3 m standoff detection. Previous attempts to create phytosensors revealed limitations in using innate plant promoters with low-inducible activity since they are not sufficient to produce a strong detectable fluorescence signal for standoff detection. To address this, we designed a pathogen-specific phytosensor using a synthetic promoter-transcription factor system: the S-Box cis-regulatory element which has low-inducible activity as a synthetic 4xS-Box promoter, and the Q-system transcription factor as an amplifier of reporter gene expression. This promoter-transcription factor system resulted in 6-fold amplification of the fluorescence after infection with a potato pathogen, which was detectable as early as 24 h post-bacterial infection. This novel bacterial pathogen-specific phytosensor potato plant demonstrates that the Q-system may be leveraged as a powerful orthogonal tool to amplify a relatively weak synthetic inducible promoter, enabling standoff detection of a previously undetectable fluorescence signal. Pathogen-specific phytosensors would be an important asset for real-time early detection of plant pathogens prior to the display of disease symptoms on crop plants.
format Online
Article
Text
id pubmed-9083229
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher Frontiers Media S.A.
record_format MEDLINE/PubMed
spelling pubmed-90832292022-05-10 Specific Bacterial Pathogen Phytosensing Is Enabled by a Synthetic Promoter-Transcription Factor System in Potato Persad-Russell, Ramona Mazarei, Mitra Schimel, Tayler Marie Howe, Lana Schmid, Manuel J. Kakeshpour, Tayebeh Barnes, Caitlin N. Brabazon, Holly Seaberry, Erin M. Reuter, D. Nikki Lenaghan, Scott C. Stewart, C. Neal Front Plant Sci Plant Science Phytosensors are genetically engineered plant-based sensors that feature synthetic promoters fused to reporter genes to sense and report the presence of specific biotic and abiotic stressors on plants. However, when induced reporter gene output is below detectable limits, owing to relatively weak promoters, the phytosensor may not function as intended. Here, we show modifications to the system to amplify reporter gene signal by using a synthetic transcription factor gene driven by a plant pathogen-inducible synthetic promoter. The output signal was unambiguous green fluorescence when plants were infected by pathogenic bacteria. We produced and characterized a phytosensor with improved sensing to specific bacterial pathogens with targeted detection using spectral wavelengths specific to a fluorescence reporter at 3 m standoff detection. Previous attempts to create phytosensors revealed limitations in using innate plant promoters with low-inducible activity since they are not sufficient to produce a strong detectable fluorescence signal for standoff detection. To address this, we designed a pathogen-specific phytosensor using a synthetic promoter-transcription factor system: the S-Box cis-regulatory element which has low-inducible activity as a synthetic 4xS-Box promoter, and the Q-system transcription factor as an amplifier of reporter gene expression. This promoter-transcription factor system resulted in 6-fold amplification of the fluorescence after infection with a potato pathogen, which was detectable as early as 24 h post-bacterial infection. This novel bacterial pathogen-specific phytosensor potato plant demonstrates that the Q-system may be leveraged as a powerful orthogonal tool to amplify a relatively weak synthetic inducible promoter, enabling standoff detection of a previously undetectable fluorescence signal. Pathogen-specific phytosensors would be an important asset for real-time early detection of plant pathogens prior to the display of disease symptoms on crop plants. Frontiers Media S.A. 2022-04-25 /pmc/articles/PMC9083229/ /pubmed/35548302 http://dx.doi.org/10.3389/fpls.2022.873480 Text en Copyright © 2022 Persad-Russell, Mazarei, Schimel, Howe, Schmid, Kakeshpour, Barnes, Brabazon, Seaberry, Reuter, Lenaghan and Stewart. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Plant Science
Persad-Russell, Ramona
Mazarei, Mitra
Schimel, Tayler Marie
Howe, Lana
Schmid, Manuel J.
Kakeshpour, Tayebeh
Barnes, Caitlin N.
Brabazon, Holly
Seaberry, Erin M.
Reuter, D. Nikki
Lenaghan, Scott C.
Stewart, C. Neal
Specific Bacterial Pathogen Phytosensing Is Enabled by a Synthetic Promoter-Transcription Factor System in Potato
title Specific Bacterial Pathogen Phytosensing Is Enabled by a Synthetic Promoter-Transcription Factor System in Potato
title_full Specific Bacterial Pathogen Phytosensing Is Enabled by a Synthetic Promoter-Transcription Factor System in Potato
title_fullStr Specific Bacterial Pathogen Phytosensing Is Enabled by a Synthetic Promoter-Transcription Factor System in Potato
title_full_unstemmed Specific Bacterial Pathogen Phytosensing Is Enabled by a Synthetic Promoter-Transcription Factor System in Potato
title_short Specific Bacterial Pathogen Phytosensing Is Enabled by a Synthetic Promoter-Transcription Factor System in Potato
title_sort specific bacterial pathogen phytosensing is enabled by a synthetic promoter-transcription factor system in potato
topic Plant Science
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9083229/
https://www.ncbi.nlm.nih.gov/pubmed/35548302
http://dx.doi.org/10.3389/fpls.2022.873480
work_keys_str_mv AT persadrussellramona specificbacterialpathogenphytosensingisenabledbyasyntheticpromotertranscriptionfactorsysteminpotato
AT mazareimitra specificbacterialpathogenphytosensingisenabledbyasyntheticpromotertranscriptionfactorsysteminpotato
AT schimeltaylermarie specificbacterialpathogenphytosensingisenabledbyasyntheticpromotertranscriptionfactorsysteminpotato
AT howelana specificbacterialpathogenphytosensingisenabledbyasyntheticpromotertranscriptionfactorsysteminpotato
AT schmidmanuelj specificbacterialpathogenphytosensingisenabledbyasyntheticpromotertranscriptionfactorsysteminpotato
AT kakeshpourtayebeh specificbacterialpathogenphytosensingisenabledbyasyntheticpromotertranscriptionfactorsysteminpotato
AT barnescaitlinn specificbacterialpathogenphytosensingisenabledbyasyntheticpromotertranscriptionfactorsysteminpotato
AT brabazonholly specificbacterialpathogenphytosensingisenabledbyasyntheticpromotertranscriptionfactorsysteminpotato
AT seaberryerinm specificbacterialpathogenphytosensingisenabledbyasyntheticpromotertranscriptionfactorsysteminpotato
AT reuterdnikki specificbacterialpathogenphytosensingisenabledbyasyntheticpromotertranscriptionfactorsysteminpotato
AT lenaghanscottc specificbacterialpathogenphytosensingisenabledbyasyntheticpromotertranscriptionfactorsysteminpotato
AT stewartcneal specificbacterialpathogenphytosensingisenabledbyasyntheticpromotertranscriptionfactorsysteminpotato