Cargando…

Rapid in vivo analysis of synthetic promoters for plant pathogen phytosensing

BACKGROUND: We aimed to engineer transgenic plants for the purpose of early detection of plant pathogen infection, which was accomplished by employing synthetic pathogen inducible promoters fused to reporter genes for altered phenotypes in response to the pathogen infection. Toward this end, a numbe...

Descripción completa

Detalles Bibliográficos
Autores principales: Liu, Wusheng, Mazarei, Mitra, Rudis, Mary R, Fethe, Michael H, Stewart, C Neal
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3247077/
https://www.ncbi.nlm.nih.gov/pubmed/22093754
http://dx.doi.org/10.1186/1472-6750-11-108
_version_ 1782220037050859520
author Liu, Wusheng
Mazarei, Mitra
Rudis, Mary R
Fethe, Michael H
Stewart, C Neal
author_facet Liu, Wusheng
Mazarei, Mitra
Rudis, Mary R
Fethe, Michael H
Stewart, C Neal
author_sort Liu, Wusheng
collection PubMed
description BACKGROUND: We aimed to engineer transgenic plants for the purpose of early detection of plant pathogen infection, which was accomplished by employing synthetic pathogen inducible promoters fused to reporter genes for altered phenotypes in response to the pathogen infection. Toward this end, a number of synthetic promoters consisting of inducible regulatory elements fused to a red fluorescent protein (RFP) reporter were constructed for use in phytosensing. RESULTS: For rapid analysis, an Agrobacterium-mediated transient expression assay was evaluated, then utilized to assess the inducibility of each synthetic promoter construct in vivo. Tobacco (Nicotiana tabacum cv. Xanthi) leaves were infiltrated with Agrobacterium harboring the individual synthetic promoter-reporter constructs. The infiltrated tobacco leaves were re-infiltrated with biotic (bacterial pathogens) or abiotic (plant defense signal molecules salicylic acid, ethylene and methyl jasmonate) agents 24 and 48 hours after initial agroinfiltration, followed by RFP measurements at relevant time points after treatment. These analyses indicated that the synthetic promoter constructs were capable of conferring the inducibility of the RFP reporter in response to appropriate phytohormones and bacterial pathogens, accordingly. CONCLUSIONS: These observations demonstrate that the Agrobacterium-mediated transient expression is an efficient method for in vivo assays of promoter constructs in less than one week. Our results provide the opportunity to gain further insights into the versatility of the expression system as a potential tool for high-throughput in planta expression screening prior to generating stably transgenic plants for pathogen phytosensing. This system could also be utilized for temporary phytosensing; e.g., not requiring stably transgenic plants.
format Online
Article
Text
id pubmed-3247077
institution National Center for Biotechnology Information
language English
publishDate 2011
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-32470772011-12-29 Rapid in vivo analysis of synthetic promoters for plant pathogen phytosensing Liu, Wusheng Mazarei, Mitra Rudis, Mary R Fethe, Michael H Stewart, C Neal BMC Biotechnol Research Article BACKGROUND: We aimed to engineer transgenic plants for the purpose of early detection of plant pathogen infection, which was accomplished by employing synthetic pathogen inducible promoters fused to reporter genes for altered phenotypes in response to the pathogen infection. Toward this end, a number of synthetic promoters consisting of inducible regulatory elements fused to a red fluorescent protein (RFP) reporter were constructed for use in phytosensing. RESULTS: For rapid analysis, an Agrobacterium-mediated transient expression assay was evaluated, then utilized to assess the inducibility of each synthetic promoter construct in vivo. Tobacco (Nicotiana tabacum cv. Xanthi) leaves were infiltrated with Agrobacterium harboring the individual synthetic promoter-reporter constructs. The infiltrated tobacco leaves were re-infiltrated with biotic (bacterial pathogens) or abiotic (plant defense signal molecules salicylic acid, ethylene and methyl jasmonate) agents 24 and 48 hours after initial agroinfiltration, followed by RFP measurements at relevant time points after treatment. These analyses indicated that the synthetic promoter constructs were capable of conferring the inducibility of the RFP reporter in response to appropriate phytohormones and bacterial pathogens, accordingly. CONCLUSIONS: These observations demonstrate that the Agrobacterium-mediated transient expression is an efficient method for in vivo assays of promoter constructs in less than one week. Our results provide the opportunity to gain further insights into the versatility of the expression system as a potential tool for high-throughput in planta expression screening prior to generating stably transgenic plants for pathogen phytosensing. This system could also be utilized for temporary phytosensing; e.g., not requiring stably transgenic plants. BioMed Central 2011-11-17 /pmc/articles/PMC3247077/ /pubmed/22093754 http://dx.doi.org/10.1186/1472-6750-11-108 Text en Copyright ©2011 Liu et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Liu, Wusheng
Mazarei, Mitra
Rudis, Mary R
Fethe, Michael H
Stewart, C Neal
Rapid in vivo analysis of synthetic promoters for plant pathogen phytosensing
title Rapid in vivo analysis of synthetic promoters for plant pathogen phytosensing
title_full Rapid in vivo analysis of synthetic promoters for plant pathogen phytosensing
title_fullStr Rapid in vivo analysis of synthetic promoters for plant pathogen phytosensing
title_full_unstemmed Rapid in vivo analysis of synthetic promoters for plant pathogen phytosensing
title_short Rapid in vivo analysis of synthetic promoters for plant pathogen phytosensing
title_sort rapid in vivo analysis of synthetic promoters for plant pathogen phytosensing
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3247077/
https://www.ncbi.nlm.nih.gov/pubmed/22093754
http://dx.doi.org/10.1186/1472-6750-11-108
work_keys_str_mv AT liuwusheng rapidinvivoanalysisofsyntheticpromotersforplantpathogenphytosensing
AT mazareimitra rapidinvivoanalysisofsyntheticpromotersforplantpathogenphytosensing
AT rudismaryr rapidinvivoanalysisofsyntheticpromotersforplantpathogenphytosensing
AT fethemichaelh rapidinvivoanalysisofsyntheticpromotersforplantpathogenphytosensing
AT stewartcneal rapidinvivoanalysisofsyntheticpromotersforplantpathogenphytosensing