May 25, 2025
Salicornia Germination on Agar Plate
- Lina Maria Caceres Leal1
- 1King Abdullah University of Science and Technology
- Salt Lab KAUST
Protocol Citation: Lina Maria Caceres Leal 2025. Salicornia Germination on Agar Plate. protocols.io https://dx.doi.org/10.17504/protocols.io.kqdg394xpg25/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: September 12, 2022
Last Modified: May 25, 2025
Protocol Integer ID: 69844
Keywords: Salicornia, in vitro, seed germination, halophytes, salicornia germination on agar plate, salicornia germination, seeds of various salicornia species, germination, various salicornia species, seed, collected seed, sterile condition, sterile conditions for different purpose, agar plate, free plant, tissue culture
Abstract
This method describes the steps to germinate seeds of various Salicornia species under sterile conditions for different purposes; germination of wild-collected seeds, tissue culture and where disease/pest-free plants are required for hydronic experiments.
Guidelines
It is advisable to perform all the steps under sterile conditions.
Materials
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
Troubleshooting
Media preparation
4h
Salicornia medium germination.
One liter of medium is enough for approximately 20 Petri dishes (100mm x 100mm x 15mm).
| A | B | C | D | |
| Component | 500 mL | 1000 mL | 2000 mL | |
| Murashige and Skoog Basal Medium (MS with vitamins) M0404-10L (Sigma Aldrich) | 2.2 g | 4.4 g | 8.8 g | |
| Sucrose 3% (w/v) | 15 g | 30 g | 60 g | |
| 100 mM NaCl* | 2.922 g | 5.844 g | 11.688 g | |
| Preservative for Plant Tissue Culture Media (PPM)** | 0.5 mL | 1 mL | 2 mL | |
| Adjust pH 5.8 | ||||
| Agar 0.5% (w/v) | 2.5 g | 5 g | 10 g | |
*NaCl is optional and dependent on the aim of the project.
**PPM is optional; it is used to prevent microbial contamination and recommended for wild-collected seeds.
Store the Petri dishes at room temperature for short-term storage or at 4 °C for long-term storage.
Sterilization of seeds
2w 0d 0h 16m
Place the seeds in a 2 mL Eppendorf tube. Add 1 ml 70% (v/v) Ethanol and mix for 1 minute.
Remove the ethanol by pipetting carefully to avoid discarding the seeds.
1m
Prepare a solution containing 20% (v/v) bleach and 0.2% (v/v) TWEEN 20. Add 1 mL of this solution to the seeds and incubate for 10 minutes, mixing occasionally. After incubation, remove the solution and wash the seeds four times with Milli-Q water. Carefully remove the supernatant by pipetting to avoid discarding the seeds.
15m
Place the sterilized seeds on agar plates. Use forceps to place them one by one, or use a 1000 µL pipette to distribute them onto the agar. Cover the Petri dishes with aluminum foil during the first 24 hours.
If germination is low, add one drop (50 µL) of 500 µM Gibberellic Acid directly onto the seeds and keep them in the dark for 24 hours to enhance germination.
500 µM Gibberellic Acid = 17 mg/100 mL water
Keep the plates in a controlled-environment growth chamber for approximately two weeks. Grow the seeds under a 12-hour light / 12-hour dark photoperiod at 28 °C.
Salicornia bigelovii seeds after two weeks on petri dish.
2w
Transplanting of seedlings
Once the seedlings reach a height of 1 cm on the agar plates, they are ready to be transferred to soil or hydroponics.