Functional characterization and intracellular localization of a Lotus tenuis NHX antiporter

Lotus tenuis is a perennial forage legume widely scattered in the flooding Pampa region of Argentina. Even when it is considered a salt-tolerant species, the molecular basis underlying this trait still remains unknown. Intracellular compartmentation of Na+ into vacuoles by NHX antiporters is an impo...

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Main Authors: Affinito, María Agostina, Espasandin, Fabiana Daniela, Del Vas, Mariana, Alvarez, Mayra Yanet, Andres, Adriana Noemi, Sansberro, Pedro Alfonso, Diaz Paleo, Antonio Horacio
Format: Artículo
Language:Inglés
Published: Springer 2021
Subjects:
Online Access:http://hdl.handle.net/20.500.12123/10747
https://link.springer.com/article/10.1007%2Fs11240-021-02094-3
https://doi.org/10.1007/s11240-021-02094-3
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author Affinito, María Agostina
Espasandin, Fabiana Daniela
Del Vas, Mariana
Alvarez, Mayra Yanet
Andres, Adriana Noemi
Sansberro, Pedro Alfonso
Diaz Paleo, Antonio Horacio
author_browse Affinito, María Agostina
Alvarez, Mayra Yanet
Andres, Adriana Noemi
Del Vas, Mariana
Diaz Paleo, Antonio Horacio
Espasandin, Fabiana Daniela
Sansberro, Pedro Alfonso
author_facet Affinito, María Agostina
Espasandin, Fabiana Daniela
Del Vas, Mariana
Alvarez, Mayra Yanet
Andres, Adriana Noemi
Sansberro, Pedro Alfonso
Diaz Paleo, Antonio Horacio
author_sort Affinito, María Agostina
collection INTA Digital
description Lotus tenuis is a perennial forage legume widely scattered in the flooding Pampa region of Argentina. Even when it is considered a salt-tolerant species, the molecular basis underlying this trait still remains unknown. Intracellular compartmentation of Na+ into vacuoles by NHX antiporters is an important and well known strategy to reduce ion toxicity during salt stress. In this study, the subcellular localization of Lotus tenuis NHX1 was determined by the transient expression of a LtNHX1-GFP fusion protein in Nicotiana benthamiana agroinfiltrated leaves, confirming that it is a tonoplast-located NHX antiporter. To study LtNHX1 contribution to salinity tolerance, a transgenic Lotus tenuis line was obtained by Agrobacterium-mediated transformation. Overexpression of the transgene was confirmed by PCR and RT-PCR analyses. Compared to wild-type, transgenic plants displayed less sensitivity and grew better under high salt stress, with higher survival rates, shoot length, and dry weight. These results suggest that the activity of the tonoplast-located LtNHX1 antiporter contributes to Lotus tenuis salt tolerance.
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spelling INTA107472021-11-11T12:32:01Z Functional characterization and intracellular localization of a Lotus tenuis NHX antiporter Affinito, María Agostina Espasandin, Fabiana Daniela Del Vas, Mariana Alvarez, Mayra Yanet Andres, Adriana Noemi Sansberro, Pedro Alfonso Diaz Paleo, Antonio Horacio Lotus tenuis Estrés Osmótico Plantas Transgénicas Genética Osmotic Stress Transgenic Plants Genetics Estrés Salino Lotus tenuis is a perennial forage legume widely scattered in the flooding Pampa region of Argentina. Even when it is considered a salt-tolerant species, the molecular basis underlying this trait still remains unknown. Intracellular compartmentation of Na+ into vacuoles by NHX antiporters is an important and well known strategy to reduce ion toxicity during salt stress. In this study, the subcellular localization of Lotus tenuis NHX1 was determined by the transient expression of a LtNHX1-GFP fusion protein in Nicotiana benthamiana agroinfiltrated leaves, confirming that it is a tonoplast-located NHX antiporter. To study LtNHX1 contribution to salinity tolerance, a transgenic Lotus tenuis line was obtained by Agrobacterium-mediated transformation. Overexpression of the transgene was confirmed by PCR and RT-PCR analyses. Compared to wild-type, transgenic plants displayed less sensitivity and grew better under high salt stress, with higher survival rates, shoot length, and dry weight. These results suggest that the activity of the tonoplast-located LtNHX1 antiporter contributes to Lotus tenuis salt tolerance. Instituto de Biotecnología Fil: Affinito, María Agostina. Universidad Nacional del Noroeste de la Provincia de Buenos Aires. Centro de Investigaciones y Transferencia del Noroeste de la Provincia de Buenos Aires; Argentina Fil: Affinito, María Agostina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia del Noroeste de la Provincia de Buenos Aires; Argentina Fil: Affinito, María Agostina. Universidad Nacional del Noroeste de la Provincia de Buenos Aires. Escuela de Ciencias Agrarias, Naturales y Ambientales; Argentina Fil: Espasandin, Fabiana Daniela. Universidad Nacional del Nordeste. Instituto de Botánica del Nordeste; Argentina Fil: Espasandin, Fabiana Daniela. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Botánica del Nordeste; Argentina Fil: Del Vas, Mariana. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; Argentina. Fil: Alvarez, Mayra Yanet. Universidad Nacional del Nordeste. Instituto de Botánica del Nordeste; Argentina Fil: Alvarez, Mayra Yanet. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Botánica del Nordeste; Argentina Fil: Andrés, Adriana Noemi. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Pergamino. Forrajeras; Argentina. Fil: Andrés, Adriana Noemi. Universidad Nacional del Noroeste de la Provincia de Buenos Aires. Escuela de Ciencias Agrarias, Naturales y Ambientales; Argentina Fil: Sansberro, Pedro Alfonso. Universidad Nacional del Nordeste. Instituto de Botánica del Nordeste; Argentina Fil: Sansberro, Pedro Alfonso. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Botánica del Nordeste; Argentina Fil: Diaz Paleo, Antonio Horacio. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Pergamino; Argentina. Fil: Diaz Paleo, Antonio Horacio. Universidad Nacional del Noroeste de la Provincia de Buenos Aires. Escuela de Ciencias Agrarias, Naturales y Ambientales; Argentina 2021-11-11T12:27:31Z 2021-11-11T12:27:31Z 2021-05 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://hdl.handle.net/20.500.12123/10747 https://link.springer.com/article/10.1007%2Fs11240-021-02094-3 0167-6857 1573-5044 https://doi.org/10.1007/s11240-021-02094-3 eng info:eu-repograntAgreement/INTA/PNPA-1126072/AR./Desarrollo de cultivares superiores de especies forrajeras para sistemas ganaderos y agricolo-ganaderos de la Argentina. info:eu-repo/semantics/restrictedAccess application/pdf Springer Plant Cell, Tissue and Organ Culture (PCTOC) 146 : 565–576 (2021)
spellingShingle Lotus tenuis
Estrés Osmótico
Plantas Transgénicas
Genética
Osmotic Stress
Transgenic Plants
Genetics
Estrés Salino
Affinito, María Agostina
Espasandin, Fabiana Daniela
Del Vas, Mariana
Alvarez, Mayra Yanet
Andres, Adriana Noemi
Sansberro, Pedro Alfonso
Diaz Paleo, Antonio Horacio
Functional characterization and intracellular localization of a Lotus tenuis NHX antiporter
title Functional characterization and intracellular localization of a Lotus tenuis NHX antiporter
title_full Functional characterization and intracellular localization of a Lotus tenuis NHX antiporter
title_fullStr Functional characterization and intracellular localization of a Lotus tenuis NHX antiporter
title_full_unstemmed Functional characterization and intracellular localization of a Lotus tenuis NHX antiporter
title_short Functional characterization and intracellular localization of a Lotus tenuis NHX antiporter
title_sort functional characterization and intracellular localization of a lotus tenuis nhx antiporter
topic Lotus tenuis
Estrés Osmótico
Plantas Transgénicas
Genética
Osmotic Stress
Transgenic Plants
Genetics
Estrés Salino
url http://hdl.handle.net/20.500.12123/10747
https://link.springer.com/article/10.1007%2Fs11240-021-02094-3
https://doi.org/10.1007/s11240-021-02094-3
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