Heterologous expression of a chloride transporter gene AoCLCf from Avicennia officinalis enhances salt tolerance of Arabidopsis plants

Plant chloride transporters are pivotal for preserving turgor pressure, pH, and cellular ion balance while adapting to salinity stress. We identified a salt-responsive gene, AoCLCf from Avicennia officinalis, which belongs to the chloride channel (CLC) family, and it shares significant sequence simi...

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Main Authors: Sivamathini Rajappa, Prakash Kumar
Format: Article
Language:English
Published: Elsevier 2025-05-01
Series:Crop Design
Online Access:http://www.sciencedirect.com/science/article/pii/S2772899425000072
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author Sivamathini Rajappa
Prakash Kumar
author_facet Sivamathini Rajappa
Prakash Kumar
author_sort Sivamathini Rajappa
collection DOAJ
description Plant chloride transporters are pivotal for preserving turgor pressure, pH, and cellular ion balance while adapting to salinity stress. We identified a salt-responsive gene, AoCLCf from Avicennia officinalis, which belongs to the chloride channel (CLC) family, and it shares significant sequence similarity with its Arabidopsis counterpart, AtCLCf. Through functional characterization in yeast mutants and Arabidopsis plants, we found that AoCLCf expression was induced primarily in roots under salt stress. Subcellular localization revealed a salt-induced translocation of GFP-AoCLCf from the Golgi apparatus to the plasma membrane. Expression of AoCLCf in the Saccharomyces cerevisiae mutant strain Δgef1 helped to rescue the growth of the mutant at high NaCl concentrations (up to 1.25M). Moreover, constitutive expression of AoCLCf in wild-type Arabidopsis significantly enhanced salt tolerance, as evidenced by increased seed germination rates, and improved seedling growth (greater root and shoot length) under 150 ​mM NaCl treatment. Spectrofluorimetric assays using liposomes embedded with recombinant AoCLCf protein showed that it functions as a chloride channel. These findings underscore the pivotal role of AoCLCf in improving salt stress tolerance through the maintenance of cellular ion homeostasis.
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spelling doaj-art-b470aa09391948caa07f5f1fcd8606bf2025-08-20T03:53:47ZengElsevierCrop Design2772-89942025-05-014210010110.1016/j.cropd.2025.100101Heterologous expression of a chloride transporter gene AoCLCf from Avicennia officinalis enhances salt tolerance of Arabidopsis plantsSivamathini Rajappa0Prakash Kumar1Department of Biological Sciences and Research Centre on Sustainable Urban Farming, National University of Singapore, 14 Science Drive 4, Singapore, 117543Department of Biological Sciences and Research Centre on Sustainable Urban Farming, National University of Singapore, 14 Science Drive 4, Singapore, 117543; NUS Environmental Research Institute, National University of Singapore, #02-01, T-Lab Building, 5A Engineering Drive 1, Singapore, 117411; Corresponding author. Department of Biological Sciences, National University of Singapore, 14 Science Drive 4, Singapore 117543.Plant chloride transporters are pivotal for preserving turgor pressure, pH, and cellular ion balance while adapting to salinity stress. We identified a salt-responsive gene, AoCLCf from Avicennia officinalis, which belongs to the chloride channel (CLC) family, and it shares significant sequence similarity with its Arabidopsis counterpart, AtCLCf. Through functional characterization in yeast mutants and Arabidopsis plants, we found that AoCLCf expression was induced primarily in roots under salt stress. Subcellular localization revealed a salt-induced translocation of GFP-AoCLCf from the Golgi apparatus to the plasma membrane. Expression of AoCLCf in the Saccharomyces cerevisiae mutant strain Δgef1 helped to rescue the growth of the mutant at high NaCl concentrations (up to 1.25M). Moreover, constitutive expression of AoCLCf in wild-type Arabidopsis significantly enhanced salt tolerance, as evidenced by increased seed germination rates, and improved seedling growth (greater root and shoot length) under 150 ​mM NaCl treatment. Spectrofluorimetric assays using liposomes embedded with recombinant AoCLCf protein showed that it functions as a chloride channel. These findings underscore the pivotal role of AoCLCf in improving salt stress tolerance through the maintenance of cellular ion homeostasis.http://www.sciencedirect.com/science/article/pii/S2772899425000072
spellingShingle Sivamathini Rajappa
Prakash Kumar
Heterologous expression of a chloride transporter gene AoCLCf from Avicennia officinalis enhances salt tolerance of Arabidopsis plants
Crop Design
title Heterologous expression of a chloride transporter gene AoCLCf from Avicennia officinalis enhances salt tolerance of Arabidopsis plants
title_full Heterologous expression of a chloride transporter gene AoCLCf from Avicennia officinalis enhances salt tolerance of Arabidopsis plants
title_fullStr Heterologous expression of a chloride transporter gene AoCLCf from Avicennia officinalis enhances salt tolerance of Arabidopsis plants
title_full_unstemmed Heterologous expression of a chloride transporter gene AoCLCf from Avicennia officinalis enhances salt tolerance of Arabidopsis plants
title_short Heterologous expression of a chloride transporter gene AoCLCf from Avicennia officinalis enhances salt tolerance of Arabidopsis plants
title_sort heterologous expression of a chloride transporter gene aoclcf from avicennia officinalis enhances salt tolerance of arabidopsis plants
url http://www.sciencedirect.com/science/article/pii/S2772899425000072
work_keys_str_mv AT sivamathinirajappa heterologousexpressionofachloridetransportergeneaoclcffromavicenniaofficinalisenhancessalttoleranceofarabidopsisplants
AT prakashkumar heterologousexpressionofachloridetransportergeneaoclcffromavicenniaofficinalisenhancessalttoleranceofarabidopsisplants