A Flexible Low Cost Hydroponic System For Assessing Plant Responses To Small Molecules In Sterile Conditions

Carolina C. Monte-Bello, Elias F. Araujo,Marina C. M. Martins,Valeria Mafra, Viviane C. H. Da Silva, Viviane Celente,Camila Caldana

JOVE-JOURNAL OF VISUALIZED EXPERIMENTS(2018)

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摘要
A wide range of studies in plant biology are performed using hydroponic cultures. In this work, an in vitro hydroponic growth system designed for assessing plant responses to chemicals and other substances of interest is presented. This system is highly efficient in obtaining homogeneous and healthy seedlings of the C-3 and C-4 model species Arabidopsis thaliana and Setaria viridis, respectively. The sterile cultivation avoids algae and microorganism contamination, which are known limiting factors for plant normal growth and development in hydroponics. In addition, this system is scalable, enabling the harvest of plant material on a large scale with minor mechanical damage, as well as the harvest of individual parts of a plant if desired. A detailed protocol demonstrating that this system has an easy and low-cost assembly, as it uses pipette racks as the main platform for growing plants, is provided. The feasibility of this system was validated using Arabidopsis seedlings to assess the effect of the drug AZD-8055, a chemical inhibitor of the target of rapamycin (TOR) kinase. TOR inhibition was efficiently detected as early as 30 min after an AZD-8055 treatment in roots and shoots. Furthermore, AZD-8055-treated plants displayed the expected starch-excess phenotype. We proposed this hydroponic system as an ideal method for plant researchers aiming to monitor the action of plant inducers or inhibitors, as well as to assess metabolic fluxes using isotope-labeling compounds which, in general, requires the use of expensive reagents.
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关键词
Environmental Sciences,Issue 138,Hydroponic system,in vitro culture,small molecules,Arabidopsis thaliana,Setaria viridis,pipette tip rack,target of rapamycin inhibitor,AZD-8055
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