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2021
Bezerra, Yane Caroline Dos Anjos; Marques, João Paulo Rodrigues; Stipp, Liliane Cristina Liborio; Attílio, Lísia Borges; Freitas-Astúa, Juliana; Filho, Francisco Assis Alves Mourão
How to drive phloem gene expression? A case study with preferentially expressed citrus gene promoters Journal Article
In: Revista Brasileira de Fruticultura, vol. 43, iss. 4, 2021, ISSN: 0100-2945.
Abstract | Links | BibTeX | Tags: Candidatus Liberibacter spp., disease resistance, genetic transformation, GUS, histolocalization
@article{Bezerra2021,
title = {How to drive phloem gene expression? A case study with preferentially expressed citrus gene promoters},
author = {Yane Caroline Dos Anjos Bezerra and João Paulo Rodrigues Marques and Liliane Cristina Liborio Stipp and Lísia Borges Attílio and Juliana Freitas-Astúa and Francisco Assis Alves Mourão Filho},
url = {http://www.scielo.br/j/rbf/a/tg5tQ8p8CcRK7xKDDcHCTjF/?lang=en},
doi = {10.1590/0100-29452021005},
issn = {0100-2945},
year = {2021},
date = {2021-01-01},
journal = {Revista Brasileira de Fruticultura},
volume = {43},
issue = {4},
publisher = {Sociedade Brasileira de Fruticultura},
abstract = {Abstract New approaches for developing disease-resistant genetically modified organisms have included specific targets for gene expression to enhance the chances for pathogen control. Gene expression driven by phloem-derived Citrus sinensis gene promoters could be evaluated and compared with the expression induced by a strong constitutive promoter in the same tissue, leading to the production of transgenic sweet oranges potentially more resistant to diseases caused by phloem-limited bacteria. ‘Carrizo’ citrange [ (Poncirus trifoliataL.) Raf. x Citrus sinensis (L.) Osbeck] was transformed, via Agrobacterium tumefaciens, with the binary vector pCAMBIA2301 bearing the uidA gene (ß-glucuronidase) driven by the CaMV35S constitutive promoter (CaMV35S::uidA) or by the CsPP2.B1 (CsPP2.B1::uidA) or by the CsVTE2 (CsVTE2::uidA) citrus promoters. In vitro regenerated shoots were grafted onto ‘Rangpur’ lime (C. limonia Osbeck). The genetic transformation was confirmed by Southern blot analyses. uidA gene expression was evaluated by RT-qPCR, and gene histolocalization controlled by these three promoters was accessed by X-GLUC treated stem sections. uidA gene expression exhibited by tissue-specific promoters was overall lower than from the constitutive promoter CaMV35; however, constructs driven by tissue-specific promoters may lead to expression in restricted tissues. CsPP2.B1 and CsVTE2 promoters can be considered adequate for the utilization in gene constructs aiming disease resistance.},
keywords = {Candidatus Liberibacter spp., disease resistance, genetic transformation, GUS, histolocalization},
pubstate = {published},
tppubtype = {article}
}
Abstract New approaches for developing disease-resistant genetically modified organisms have included specific targets for gene expression to enhance the chances for pathogen control. Gene expression driven by phloem-derived Citrus sinensis gene promoters could be evaluated and compared with the expression induced by a strong constitutive promoter in the same tissue, leading to the production of transgenic sweet oranges potentially more resistant to diseases caused by phloem-limited bacteria. ‘Carrizo’ citrange [ (Poncirus trifoliataL.) Raf. x Citrus sinensis (L.) Osbeck] was transformed, via Agrobacterium tumefaciens, with the binary vector pCAMBIA2301 bearing the uidA gene (ß-glucuronidase) driven by the CaMV35S constitutive promoter (CaMV35S::uidA) or by the CsPP2.B1 (CsPP2.B1::uidA) or by the CsVTE2 (CsVTE2::uidA) citrus promoters. In vitro regenerated shoots were grafted onto ‘Rangpur’ lime (C. limonia Osbeck). The genetic transformation was confirmed by Southern blot analyses. uidA gene expression was evaluated by RT-qPCR, and gene histolocalization controlled by these three promoters was accessed by X-GLUC treated stem sections. uidA gene expression exhibited by tissue-specific promoters was overall lower than from the constitutive promoter CaMV35; however, constructs driven by tissue-specific promoters may lead to expression in restricted tissues. CsPP2.B1 and CsVTE2 promoters can be considered adequate for the utilization in gene constructs aiming disease resistance.
2017
Miyata, Luzia Yuriko; Harakava, Ricardo; Attílio, Lísia Borges; Mendes, Beatriz Madalena Januzzi; Lopes, João Roberto Spotti; Coletta-Filho, Helvécio Della; Souza, Alessandra Alves De; Filho, Francisco De Assis Alves Mourão
PHLOEM PROMOTERS IN TRANSGENIC SWEET ORANGE ARE DIFFERENTIALLY TRIGGERED BY Candidatus Liberibacter asiaticus Journal Article
In: Revista Brasileira de Fruticultura, vol. 39, iss. 3, 2017, ISSN: 0100-2945.
Abstract | Links | BibTeX | Tags: Citrus sinensis, Diaphorina citri, genetic transformation, GUS, huanglongbing, quantitative real, time PCR
@article{Miyata2017,
title = {PHLOEM PROMOTERS IN TRANSGENIC SWEET ORANGE ARE DIFFERENTIALLY TRIGGERED BY \textit{Candidatus} Liberibacter asiaticus},
author = {Luzia Yuriko Miyata and Ricardo Harakava and Lísia Borges Attílio and Beatriz Madalena Januzzi Mendes and João Roberto Spotti Lopes and Helvécio Della Coletta-Filho and Alessandra Alves De Souza and Francisco De Assis Alves Mourão Filho},
url = {http://www.scielo.br/j/rbf/a/NFKjGmNsP48jRtP7txqqcFh/?lang=en},
doi = {10.1590/0100-29452017993},
issn = {0100-2945},
year = {2017},
date = {2017-01-01},
journal = {Revista Brasileira de Fruticultura},
volume = {39},
issue = {3},
publisher = {Sociedade Brasileira de Fruticultura},
abstract = {ABSTRACT The use of promoters preferentially expressed in specific plant tissues is a desirable strategy to search for resistance for pathogens that colonize these tissues. The bacterium Candidatus Liberibacter asiaticus (Las), associated with huanglongbing disease (HLB) of citrus, colonizes phloem vessels. Some promoters, besides conferring tissue-specific expression, can also respond to the presence of the pathogen. The objective of the present study was to verify if the presence of Las could modulate the activation of the phloem-specific promoters AtPP2 (Arabidopsis thaliana phloem protein 2), AtSUC2 (A. thaliana sucrose transporter 2) and CsPP2 ( pCitrus phloemrotein 2), known to be expressed in Citrus sinensis phloem. ‘Hamlin’ sweet orange plants (Citrus sinensis L. Osbeck) transformed with the uidA (GUS) reporter gene under the control of AtPP2, AtSUC2 and CsPP2 promoters were infected to evaluate the interdependence between transgene expression and the concentration of Las. Plants were inoculated with Las by Diaphorina citri and eighteen months later, bacterial concentration and uidA expression were determined by qPCR and RT-qPCR, respectively. Reporter gene expression driven by AtSUC2 promoter was strongly and positively correlated with Las concentration. Therefore, this promoter combines desirable features of both tissue-specificity and pathogen-inducibility for the production of transgenic plants tolerant to Las.},
keywords = {Citrus sinensis, Diaphorina citri, genetic transformation, GUS, huanglongbing, quantitative real, time PCR},
pubstate = {published},
tppubtype = {article}
}
ABSTRACT The use of promoters preferentially expressed in specific plant tissues is a desirable strategy to search for resistance for pathogens that colonize these tissues. The bacterium Candidatus Liberibacter asiaticus (Las), associated with huanglongbing disease (HLB) of citrus, colonizes phloem vessels. Some promoters, besides conferring tissue-specific expression, can also respond to the presence of the pathogen. The objective of the present study was to verify if the presence of Las could modulate the activation of the phloem-specific promoters AtPP2 (Arabidopsis thaliana phloem protein 2), AtSUC2 (A. thaliana sucrose transporter 2) and CsPP2 ( pCitrus phloemrotein 2), known to be expressed in Citrus sinensis phloem. ‘Hamlin’ sweet orange plants (Citrus sinensis L. Osbeck) transformed with the uidA (GUS) reporter gene under the control of AtPP2, AtSUC2 and CsPP2 promoters were infected to evaluate the interdependence between transgene expression and the concentration of Las. Plants were inoculated with Las by Diaphorina citri and eighteen months later, bacterial concentration and uidA expression were determined by qPCR and RT-qPCR, respectively. Reporter gene expression driven by AtSUC2 promoter was strongly and positively correlated with Las concentration. Therefore, this promoter combines desirable features of both tissue-specificity and pathogen-inducibility for the production of transgenic plants tolerant to Las.