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Projeto de investigação
Grapevine immunity: the innovative role of subtilisin-like proteases
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Deciphering the communication between Vitis vinifera and Plasmopara viticola in plant apoplast : a first picture into host and pathogen proteomes
Publication . Figueiredo, J.; Figueiredo, Andreia; Malhó, Rui
Grapevine (Vitis vinifera L.) is a fruit crop with high economic value due to its final products like wine. However, every year, the vineyards are affected by the attack of different pathogens, leading to a great loss of production. One of the most common grapevine diseases is downy mildew caused by the obligate biotrophic oomycete Plasmopara viticola ((Berk. and Curt.) Berl. & de Toni). Downy mildew is a cryptogamic disease that attacks all the green parts of the plant, leaves, tendrils, branches and bunches, whose development is favoured by mild temperatures and high humidity. In this infection process, the apoplast is the subcellular compartment where there is the greatest contact between the pathogen's infection structures, as well as the molecules secreted by it, and the plant's molecules. A comprehensive knowledge on the grapevine strategies to overcome pathogens, as well as the evolution of pathogen infection mechanisms is paramount for the development of new sustainable disease control strategies and avoid the massive use of pesticides in each grapevine growing season. The apoplast is the first hub of plant-pathogen communication where pathogen effectors are recognized by plant defensive proteins and cell receptors, thus activating signal transduction pathways. As a result of this first contact, the host triggers a defence response that involves the modulation of extra and intracellular proteins, including proteases. On the side of the pathogens, they secrete a myriad of molecules, called effectors, in the plant apoplast that have the function of overcoming the plant's barriers and inducing the disease. Within proteases, subtilases have been the focus of several studies and seems to be involved in plant defence, resistance, and immunity mechanisms. In grapevine, some of them present a constitutive expression, without the plant being under the influence of any stress, quite high in some resistant cultivars, compared to a susceptible cultivar. After inoculation with P. viticola, an expression increase of several subtilases was observed in grapevine leaves. A specific subtilase, VviSBT4.19, showed interesting results that indicate a possible involvement in grapevine resistance to this oomycete. Considering all the above, the aims of this work were: 1) The development of a methodology for extracting the apoplast from grapevine leaves, focusing on two grapevine cultivars with different degrees of tolerance to downy mildew; 2) The study of the apoplast proteome during grapevine-downy mildew interaction, identifying both plant and pathogen proteins; 3) The analysis of VviSBT4.19 gene expression in grapevine leaves infected with other pathogens, namely the fungi Erysiphe necator and Botrytis cinerea, and through the identification of interacting proteins with the VviSBT4.19 during 'Regent'-P. viticola interaction. It is expected that this work will contribute in an active and relevant way to the deep knowledge of the interaction between the grapevine and the downy mildew for the development or improvement of sustainable alternatives for a more environmentally friendly viticulture.
Deep into the apoplast: grapevine and Plasmopara viticola proteomes reveal the secret beneath host and pathogen communication at 6h after contact
Publication . Figueiredo, Joana; Santos, Rita B.; Guerra-Guimarães, Leonor; Leclercq, Céline C.; Renaut, Jenny; Sousa, Lisete; Figueiredo, Andreia
The apoplast is the first hub of plant−pathogen communication where pathogen effectors are recognized by plant defensive proteins and cell receptors, thus activating signal transduction pathways. As a result of this first contact, the host triggers a defense response that involves the modulation of extra- and intracellular proteins. In grapevine−pathogen interactions, little is known about the trafficking between extra- and intracellular spaces. Grapevine is an economically important crop that relies on heavy fungicide use to control several diseases, and a deeper knowledge on the activation of its immune response is crucial to define new control strategies. In this study, we focused on the first 6 h postinoculation with Plasmopara viticola to evaluate grapevine proteome modulation in the apoplast. The in planta P. viticola proteome was also assessed to enable a deeper understanding of plant−pathogen communication. Our results showed that several plant mechanisms are triggered in the tolerant grapevine cultivar Regent after inoculation, such as oomycete recognition, plant cell wall modifications, reactive oxygen species signaling, and secretion of proteins to disrupt oomycete structures. On the other hand, P. viticola proteins related to development and virulence were the most predominant. This pioneer study highlights the early dynamics of cellular communication in grapevine defense that leads to the successful establishment of an incompatible interaction.
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Entidade financiadora
Fundação para a Ciência e a Tecnologia
Programa de financiamento
3599-PPCDT
Número da atribuição
PTDC/BIA-BQM/28539/2017