Jun 20, 2025
Hydroponic cultivation of Salicornia
- Yveline Pailles1,
- Vanessa Melino2,
- Lina Maria Caceres Leal1,
- Nataliya Kovalchuk1
- 1King Abdullah University of Science and Technology;
- 2University of Newcastle
- Salt Lab KAUST
Protocol Citation: Yveline Pailles, Vanessa Melino, Lina Maria Caceres Leal, Nataliya Kovalchuk 2025. Hydroponic cultivation of Salicornia. protocols.io https://dx.doi.org/10.17504/protocols.io.n2bvjb4rpgk5/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: May 26, 2025
Last Modified: June 20, 2025
Protocol Integer ID: 218915
Keywords: Salicornia, Root, Cultivation, Ebb and flow hydroponics, hydroponic cultivation of salicornia, hydroponic cultivation, salicornia species, salicornia, hydroponic ebb, liquid nutrient fertilizer, harvesting plant tissue, ease of harvesting plant tissue, precise nutrient
Abstract
This method describes how to cultivate Salicornia species in a hydroponic ebb and flow system (flood-drain) using an inert substrate and liquid nutrient fertilizers. This method enables application of very precise nutrient/ionic/osmotic stress and for ease of harvesting plant tissue (shoots and roots) for downstream physiological or molecular assays.
Attachments
Guidelines
Always ensure electrical equipment near hydroponic systems and wet areas is properly grounded, waterproof, and installed with ground-fault circuit interrupters (GFCIs) to prevent electrical shock hazards.
Materials
Seed germination:
- Murashige and Skoog Basal Medium - Sigma-Aldrich
-Sucrose - Sigma-Aldrich
-NaCl
-Preservative for Plant Tissue Culture Media (PPM) - Plant Cell Technology
-Agar - Sigma-Aldrich
-Ethanol Absolute (>99.5%) EMPARTA
-Bleach (household grade)
-TWEEN 20 - TermoFisher
-Gibberellic acid - Sigma-Aldrich
- 1ml pipette tips (sterile)
- 70% (v/v) sodium hypochlorite (bleach)
- 70% (v/v) ethanol
- Tap water
- Strainer
- Petri dishes (100mm x 100mm x 15mm)
- Salicornia species seeds
- Sterile forceps and spatulas
- Parafilm
- Plastic beads (high density polyethylene supplied by companies who produce plastic buckets or crates; ensure that they do not float)
- Squared black pots (3”)
- Cheesecloth
- Commercial Bleach
- Rectangular trays and trays with divisions
- Tray covers
- Pot tags with QR code sticker labels
- Growth chamber
Hydroponics dimensions:
-920mm x 920mm x 970mm (see manufacturing instructions for the frame attached)
Growing phase:
- FloraGrow nutrient stock
- FloraMicro nutrient stock
- Tap water
- EC meter
- pH meter
- 2 L plastic beaker
- 2 L volumetric flask
- Spatulas
- Timers
- Econo-Tray Hydroponic systems: 87 x 87 cm (American Hydroponics)
- Metal support structure for Hydroponic systems: 92 x 92 x 97 cm (custom made)
- Submersible aquarium pumps (ViaAqua Powerhead Pump 360)
- Voltage transformer for aquarium pumps (120V/60Hz)
Salt treatments:
- NaCl
- 2M CaCl2 stock solution
- Tap water
- EC meter
- pH meter
- Long-handled stirring rod
Troubleshooting
Safety warnings
The light conditions should be homogeneous across all hydroponic systems. Consider analyzing the light intensity spatially across the greenhouse to ensure uniformity across tank treatments. This is an example.
Comparable light intensity was achieved in three positions. Tank 4 received the least light and was
therefore not used.
Before start
Set up the growing and light conditions of the room where the experiment will run.
Growth room conditions
Day/night 28/24°C
Using LED supplemental lights
The example below shows a ramp-up scenario.
Light wavelength and distribution settings in the greenhouse.
Seed Germination
2w
Select required number of genotypes.
Work out the number of repetitions (take more seeds than you need) (For example, 6 replicates * 2 treatments * 2 seeds per pot) (note. later thinned to one seedling per pot).
Sterilize the seeds and place them on the germination agar medium following the steps described in protocol 'Salicornia Germination on Agar Plate'.
Citation
LINK
After two weeks the seedlings are ready to move to establishment stage in the hydroponics system. (step 8).
Hydroponic System Set up and Cleaning
1d
Before starting:
Measure ideal water level (with a metered hose or a 20 L container) and put a mark inside the tank to fill to that level. (100 liters final volume).
To prepare the pots:
- Sterilize the pots and plastic beads using 20%(v/v) commercial bleach solution, then rinse thoroughly with clean water.
- Cover the bottom of each pot with a square of cheesecloth to prevent the plastic beads from falling through the drainage holes.
- Fill the pots with plastic beads up to the filling line.
- Place the pots in the trays. Each tray can hold up to 118 small square pots (3”).
Location of the pots in the hydroponics.
Prepare the ebb and flow hydroponic system and ensure the pumps are functioning correctly. Set the timer to run 15 minutes on and 15 minutes off. Run the cycle for one day before moving the seedlings to confirm that everything is working properly.
The nutrient solution reaches the pots with the seedlings during the 15 minute 'on' cycle. When the timer ends, the pump stops and the solution drains.
Hydroponic tank assembled.
The nutrient solution use for the hydroponics is "General Purpose – Mild Vegetative" from Flora Series- General hydroponics.
FloraSeries‱ Feedcharts
Seedling establishment
2w
Prepare the nutrient solution and fill the thank. Add the nutrient solution stock, follow the instructions for General Purpose – Mild Vegetative from Flora Series- General hydroponics, (see step 7).
Once all the pots are positioned on the trays and labeled, take one agar plate with the seedlings and use a spatula to scoop out a portion of agar with one seedling. Bury it in the plastic beads until it is completely covered, with the surface still visible. Place two or three seedlings per pot.
Salicornia spp. seedlings after 10 days growth. Move from agar plates to hydroponics system.
2w
Cover the trays with cling wrap for two days to maintain humidity and prevent contamination. The seedlings reach a length of 1-2 cm, they are strong enough to begin the treatment.
Salt Treatment
6w
Add the nutrient solution stock ('General Purpose – Mild Vegetative' from Flora Series- General hydroponics, see step 7.), plus NaCl stock to reach the desired final concentrations.
It is important to add NaCl incrementally during the initial salinity treatment. For example, cultivating Salicornia spp. with a final target salinity of 200 mM NaCl, apply 75 mM on Day 1 and 125 mM on Day 2.
Note: More salt sensitive plant species may require smaller daily increments of the salt treatment (e.g., increasing in 50 mM NaCl increments per day). Increasing salt incrementally allows for proper osmotic adjustment.
Maintain an activity coefficient of 0.4 by supplementing the NaCl treatment with CaCl2 according to the table below.
| A | B | C | D | E | |
| Final [NaCl] (mM) | Final [CaCl2] (mM) | Ca2+ | Volume of 4M NaCl per litre (ml) | Volume of 2M CaCl2 per litre (ml) | |
| 75 | 1 | 0.4 | 18.75 | 0.5 | |
| 150 | 1.2 | 0.4 | 37.5 | 0.6 | |
| 225 | 1.3 | 0.39 | 56.25 | 0.65 | |
| 300 | 1.4 | 0.4 | 75 | 0.7 | |
| 375 | 1.4 | 0.39 | 93.75 | 0.7 | |
| 450 | 1.5 | 0.41 | 112.5 | 0.75 |
CaCl2 supplement for a NaCl solution, to keep a Ca2+ activity coefficient of 0.4
Monitor the pH and Electrical Conductivity (EC) twice per week until plants are 5 weeks old and then every day thereafter. Optimize the pH between 5.8 and 6.2 and salinity at EC target measured after complete dissolution of NaCl (200 mM).
- Replace the nutrient solution when your pH is out of the range of 5.8-6.2
- Take 50 ml samples of your nutrient solution per week and store samples with date and tank number at 4°C. Samples are collected at critical stages of your experiment for elemental analysis to record nutrient depletion rates.
Change nutrient solution (+NaCl and +CaCl2 to treatment tank) every week or when pH or EC is out of expected range. Mix the solution thoroughly using the long-handled stirring rod.
Assess plant growth responses to salt or collect tissue for further analysis following guidelines from Negrao et al. (2017).
Citation
LINK
Protocol references
S. Negrão, S. M. Schmöckel, M. Tester, Evaluating physiological responses of plants to salinity stress, Annals of Botany, Volume 119, Issue 1, January 2017, Pages 1–11, https://doi.org/10.1093/aob/mcw191