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      Enhanced Bacterial Wilt Resistance in Potato Through Expression of Arabidopsis EFR and Introgression of Quantitative Resistance from Solanum commersonii

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          Abstract

          Bacterial wilt (BW) caused by Ralstonia solanacearum is responsible for substantial losses in cultivated potato ( Solanum tuberosum) crops worldwide. Resistance genes have been identified in wild species; however, introduction of these through classical breeding has achieved only partial resistance, which has been linked to poor agronomic performance. The Arabidopsis thaliana (At) pattern recognition receptor elongation factor-Tu (EF-Tu) receptor (EFR) recognizes the bacterial pathogen-associated molecular pattern EF-Tu (and its derived peptide elf18) to confer anti-bacterial immunity. Previous work has shown that transfer of AtEFR into tomato confers increased resistance to R. solanacearum. Here, we evaluated whether the transgenic expression of AtEFR would similarly increase BW resistance in a commercial potato line (INIA Iporá), as well as in a breeding potato line (09509.6) in which quantitative resistance has been introgressed from the wild potato relative Solanum commersonii. Resistance to R. solanacearum was evaluated by damaged root inoculation under controlled conditions. Both INIA Iporá and 09509.6 potato lines expressing AtEFR showed greater resistance to R. solanacearum, with no detectable bacteria in tubers evaluated by multiplex-PCR and plate counting. Notably, AtEFR expression and the introgression of quantitative resistance from S. commersonii had a significant additive effect in 09509.6-AtEFR lines. These results show that the combination of heterologous expression of AtEFR with quantitative resistance introgressed from wild relatives is a promising strategy to develop BW resistance in potato.

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          Perception of the bacterial PAMP EF-Tu by the receptor EFR restricts Agrobacterium-mediated transformation.

          Higher eukaryotes sense microbes through the perception of pathogen-associated molecular patterns (PAMPs). Arabidopsis plants detect a variety of PAMPs including conserved domains of bacterial flagellin and of bacterial EF-Tu. Here, we show that flagellin and EF-Tu activate a common set of signaling events and defense responses but without clear synergistic effects. Treatment with either PAMP results in increased binding sites for both PAMPs. We used this finding in a targeted reverse-genetic approach to identify a receptor kinase essential for EF-Tu perception, which we called EFR. Nicotiana benthamiana, a plant unable to perceive EF-Tu, acquires EF-Tu binding sites and responsiveness upon transient expression of EFR. Arabidopsis efr mutants show enhanced susceptibility to the bacterium Agrobacterium tumefaciens, as revealed by a higher efficiency of T-DNA transformation. These results demonstrate that EFR is the EF-Tu receptor and that plant defense responses induced by PAMPs such as EF-Tu reduce transformation by Agrobacterium.
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            Interfamily transfer of a plant pattern-recognition receptor confers broad-spectrum bacterial resistance.

            Plant diseases cause massive losses in agriculture. Increasing the natural defenses of plants may reduce the impact of phytopathogens on agricultural productivity. Pattern-recognition receptors (PRRs) detect microbes by recognizing conserved pathogen-associated molecular patterns (PAMPs). Although the overall importance of PAMP-triggered immunity for plant defense is established, it has not been used to confer disease resistance in crops. We report that activity of a PRR is retained after its transfer between two plant families. Expression of EFR (ref. 4), a PRR from the cruciferous plant Arabidopsis thaliana, confers responsiveness to bacterial elongation factor Tu in the solanaceous plants Nicotiana benthamiana and tomato (Solanum lycopersicum), making them more resistant to a range of phytopathogenic bacteria from different genera. Our results in controlled laboratory conditions suggest that heterologous expression of PAMP recognition systems could be used to engineer broad-spectrum disease resistance to important bacterial pathogens, potentially enabling more durable and sustainable resistance in the field.
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              Coomassie staining as loading control in Western blot analysis.

              In Western blotting, immunodetection of housekeeping proteins is routinely performed to detect differences in electrophoresis loading. The present work describes a much faster and simpler protein staining method, which is compatible with ordinary blocking conditions. In addition, the method can be used after immunodetection with superior linearity compared to ordinary staining methods. After immunoblotting and staining, protein bands can be further identified using peptide mass fingerprinting.
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                Author and article information

                Contributors
                Journal
                Front Plant Sci
                Front Plant Sci
                Front. Plant Sci.
                Frontiers in Plant Science
                Frontiers Media S.A.
                1664-462X
                25 September 2017
                2017
                : 8
                : 1642
                Affiliations
                [1] 1Instituto Nacional de Semillas Canelones, Uruguay
                [2] 2Unidad de Biotecnología, Instituto Nacional de Investigación Agropecuaria Canelones, Uruguay
                [3] 3Departamento de Biociencias, Facultad de Química, Universidad de la República Montevideo, Uruguay
                [4] 4Departamento de Producción Vegetal, Centro Regional Sur, Facultad de Agronomía, Universidad de la República Canelones, Uruguay
                [5] 5The Sainsbury Laboratory, Norwich Research Park Norwich, United Kingdom
                [6] 6Programa de Producción Hortícola, Instituto Nacional de Investigación Agropecuaria Canelones, Uruguay
                Author notes

                Edited by: Carlos Alberto Lopes, Brazilian Agricultural Research Corporation, Brazil

                Reviewed by: Dennis Halterman, Agricultural Research Service (USDA), United States; Arione Da Silva Pereira, Brazilian Agricultural Research Corporation, Brazil

                *Correspondence: Marco Dalla-Rizza, mdallarizza@ 123456inia.org.uy

                This article was submitted to Plant Microbe Interactions, a section of the journal Frontiers in Plant Science

                Article
                10.3389/fpls.2017.01642
                5627020
                29033958
                7b007b40-dc59-444a-9990-26529c1773fb
                Copyright © 2017 Boschi, Schvartzman, Murchio, Ferreira, Siri, Galván, Smoker, Stransfeld, Zipfel, Vilaró and Dalla-Rizza.

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                : 29 May 2017
                : 07 September 2017
                Page count
                Figures: 4, Tables: 2, Equations: 0, References: 60, Pages: 11, Words: 0
                Funding
                Funded by: Agencia Nacional de Investigación e Innovación 10.13039/100008725
                Award ID: POS_NAC_2013_1_11871
                Categories
                Plant Science
                Original Research

                Plant science & Botany
                ralstonia solanacearum,bacterial wilt,solanum tuberosum,solanum commersonii,pattern recognition receptor,efr,quantitative resistance

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