Differential fine-tuning of gene expression regulation in coffee leaves by CcDREB1D promoter haplotypes under water deficit
Alves, Gabriel Sergio Costa | Torres, Luana Ferreira | Déchamp, Eveline | Breitler, Jean-Christophe | Joët, Thierry | Gatineau, Frédéric | Andrade, Alan Carvalho | Bertrand, Benoît | Marraccini, Pierre | Etienne, Hervé | Embrapa Recursos Genéticos e Biotecnologia = Embrapa Genetic Resources & Biotechnology (CENARGEN) ; Brazilian Agricultural Research Corporation = Empresa Brasileira de Pesquisa Agropecuária (Embrapa) | UMR - Interactions Plantes Microorganismes Environnement (UMR IPME) ; Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Université de Montpellier (UM)-Institut de Recherche pour le Développement (IRD [Occitanie]) | Universidade Federal de Lavras = Federal University of Lavras (UFLA) | Département Systèmes Biologiques (Cirad-BIOS) ; Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad) | Diversité, adaptation, développement des plantes (UMR DIADE) ; Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD [Occitanie]) | Amélioration génétique et adaptation des plantes méditerranéennes et tropicales (UMR AGAP) ; Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Institut National de la Recherche Agronomique (INRA)-Centre international d'études supérieures en sciences agronomiques (Montpellier SupAgro)-Institut national d’études supérieures agronomiques de Montpellier (Montpellier SupAgro) | the Brazilian Consortium on Coffee R&D, Instituto Nacional de Ciência e Tecnologia—Café (Conselho Nacional de Desenvolvimento Cientifico e Tecnologico/Fundação de Amparo à Pesquisa do Estado de Minas Gerais) and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior/Comité Français d’Évaluation de la Coopération Universitaire et Scientifique avec le Brésil (Project 407–2012/University of Lavras and Montpellier SupAgro)
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Показать больше [+] Меньше [-]Английский. Despite the importance of the DREB1D gene (also known as CBF4) in plant responses to water deficit and cold stress, studies analysing its regulation by transgenic approaches are lacking. In the current work, a functional study of three CcDREB1D promoter haplotypes (named HP15, HP16 and HP17) isolated from drought-tolerant and drought-sensitive clones of Coffea canephora was carried out in plants of C. arabica stably transformed by Agrobacterium tumefaciens by analysing their ability to regulate the expression of the uidA reporter gene in response to water deficit mimicked by polyethylene glycol (−2.0 MPa) and low relative humidity treatments. A deletion analysis of their corresponding 5′-upstream regions revealed increased specificity of β-glucuronidase activity in the polyethylene glycol and low relative humidity treatments, with high expression in leaf mesophyll and guard cells in full-length constructs. RT-qPCR assays also revealed that the HP16 haplotype (specific to clone tolerant to water deficit) had stronger and earlier activity compared with the HP15 and HP17 haplotypes. As most of the cis-regulatory elements involved in ABA-dependent and -independent networks, tissue specificity and light regulation are common to these haplotypes, we propose that their organization, as well as the nucleic acid polymorphisms present outside these boxes, may play a role in modulating activities of DREB1D promoters in guard cells.
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