Evaluating Arabidopsis Primary Root Growth in Response to Osmotic Stress Using an In Vitro Osmotic Gradient Experimental System

The root meristem navigates the highly variable soil environment where water availability limits water absorption, slowing or halting growth. Traditional studies use uniform high osmotic potentials, poorly representing natural conditions where roots gradually encounter increasing osmotic potentials....

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Main Authors: Selene Píriz-Pezzutto, Mauro Martínez-Moré, Maria Sainz, Omar Borsani, Mariana Sotelo-Silveira
Format: Article
Language:English
Published: Bio-protocol LLC 2025-07-01
Series:Bio-Protocol
Online Access:https://bio-protocol.org/en/bpdetail?id=5397&type=0
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author Selene Píriz-Pezzutto
Mauro Martínez-Moré
Maria Sainz
Omar Borsani
Mariana Sotelo-Silveira
author_facet Selene Píriz-Pezzutto
Mauro Martínez-Moré
Maria Sainz
Omar Borsani
Mariana Sotelo-Silveira
author_sort Selene Píriz-Pezzutto
collection DOAJ
description The root meristem navigates the highly variable soil environment where water availability limits water absorption, slowing or halting growth. Traditional studies use uniform high osmotic potentials, poorly representing natural conditions where roots gradually encounter increasing osmotic potentials. Uniform high osmotic potentials reduce root growth by inhibiting cell division and shortening mature cell length. This protocol describes a simple and effective in vitro system using a gradient mixer that generates a vertical gradient in an agar gel based on the principle of communicating vessels, exploiting gravity to generate a continuous mannitol concentration gradient (from 0 to 400 mM mannitol) reaching osmotic potentials of -1,2 MPa. It enables long-term Arabidopsis root growth analysis under progressive water deficit, improving phenotyping and molecular studies in soil-like conditions.
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issn 2331-8325
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publishDate 2025-07-01
publisher Bio-protocol LLC
record_format Article
series Bio-Protocol
spelling doaj-art-2b2ba97cf1904f77a64a702a601ed2d12025-08-20T02:46:01ZengBio-protocol LLCBio-Protocol2331-83252025-07-01151410.21769/BioProtoc.5397Evaluating Arabidopsis Primary Root Growth in Response to Osmotic Stress Using an In Vitro Osmotic Gradient Experimental SystemSelene Píriz-Pezzutto0Mauro Martínez-Moré1Maria Sainz2Omar Borsani3Mariana Sotelo-Silveira4Laboratorio de Bioquímica, Departamento de Biología Vegetal, Facultad de Agronomía, Universidad de la República, Montevideo, UruguayLaboratorio de Bioquímica, Departamento de Biología Vegetal, Facultad de Agronomía, Universidad de la República, Montevideo, UruguayLaboratorio de Bioquímica, Departamento de Biología Vegetal, Facultad de Agronomía, Universidad de la República, Montevideo, UruguayLaboratorio de Bioquímica, Departamento de Biología Vegetal, Facultad de Agronomía, Universidad de la República, Montevideo, UruguayLaboratorio de Bioquímica, Departamento de Biología Vegetal, Facultad de Agronomía, Universidad de la República, Montevideo, UruguayThe root meristem navigates the highly variable soil environment where water availability limits water absorption, slowing or halting growth. Traditional studies use uniform high osmotic potentials, poorly representing natural conditions where roots gradually encounter increasing osmotic potentials. Uniform high osmotic potentials reduce root growth by inhibiting cell division and shortening mature cell length. This protocol describes a simple and effective in vitro system using a gradient mixer that generates a vertical gradient in an agar gel based on the principle of communicating vessels, exploiting gravity to generate a continuous mannitol concentration gradient (from 0 to 400 mM mannitol) reaching osmotic potentials of -1,2 MPa. It enables long-term Arabidopsis root growth analysis under progressive water deficit, improving phenotyping and molecular studies in soil-like conditions.https://bio-protocol.org/en/bpdetail?id=5397&type=0
spellingShingle Selene Píriz-Pezzutto
Mauro Martínez-Moré
Maria Sainz
Omar Borsani
Mariana Sotelo-Silveira
Evaluating Arabidopsis Primary Root Growth in Response to Osmotic Stress Using an In Vitro Osmotic Gradient Experimental System
Bio-Protocol
title Evaluating Arabidopsis Primary Root Growth in Response to Osmotic Stress Using an In Vitro Osmotic Gradient Experimental System
title_full Evaluating Arabidopsis Primary Root Growth in Response to Osmotic Stress Using an In Vitro Osmotic Gradient Experimental System
title_fullStr Evaluating Arabidopsis Primary Root Growth in Response to Osmotic Stress Using an In Vitro Osmotic Gradient Experimental System
title_full_unstemmed Evaluating Arabidopsis Primary Root Growth in Response to Osmotic Stress Using an In Vitro Osmotic Gradient Experimental System
title_short Evaluating Arabidopsis Primary Root Growth in Response to Osmotic Stress Using an In Vitro Osmotic Gradient Experimental System
title_sort evaluating arabidopsis primary root growth in response to osmotic stress using an in vitro osmotic gradient experimental system
url https://bio-protocol.org/en/bpdetail?id=5397&type=0
work_keys_str_mv AT selenepirizpezzutto evaluatingarabidopsisprimaryrootgrowthinresponsetoosmoticstressusinganinvitroosmoticgradientexperimentalsystem
AT mauromartinezmore evaluatingarabidopsisprimaryrootgrowthinresponsetoosmoticstressusinganinvitroosmoticgradientexperimentalsystem
AT mariasainz evaluatingarabidopsisprimaryrootgrowthinresponsetoosmoticstressusinganinvitroosmoticgradientexperimentalsystem
AT omarborsani evaluatingarabidopsisprimaryrootgrowthinresponsetoosmoticstressusinganinvitroosmoticgradientexperimentalsystem
AT marianasotelosilveira evaluatingarabidopsisprimaryrootgrowthinresponsetoosmoticstressusinganinvitroosmoticgradientexperimentalsystem