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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
Em: Revista Brasileira de Fruticultura, vol. 43, iss. 4, 2021, ISSN: 0100-2945.
Resumo | 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.