Griffitts:Seed scarification and germination
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Alfalfa
Note: Alfalfa seeds don't need to be scarified
- Calculate the number of plants you need
- Weigh out ten times more than the number you need
- Use GT13R Plus
- 1.7 g is approximately 1000 seeds
- Place in a smooth sterile flask
- Add 70% ethanol to substantially cover the seeds
- 70% ethanol is next to the scales
- Place on a shaker at room temperature at 150 rpm for 20 minutes
- Decant the ethanol and wash the seeds once with sterile dH20
- Add Clorox bleach to substantially cover the seeds
- Place on a shaker at room temperature at 150 rpm for 20 minutes
- Decant the bleach and wash the seeds 4 to 5 times with sterile dH20
- Add sterile dH20 to substantially cover the seeds
- Place on a shaker at room temperature at 150 rpm for at least 4 hours (preferably 8)
- It's a good idea to wash the seeds with sterile dH20 once during this time
- Decant the sterile dH20 and add fresh sterile dH20
- Dump into 100×25 mm Petri dishes
- Seeds should be in a monolayer with a small amount of space between them
- Pull off excess water with a pipet
- Tip the plate and give it a few seconds to be sure to get all the excess water
- Wrap plate(s) in Parafilm
- Store upside-down in a drawer for 24 hours
Other Medics
- Place twice as many seeds as you wish to plant into a 50 mL Erlenmeyer flask that has been covered with foil and autoclaved.
- To each of the flasks add 5 ml of H2SO4 (sulfuric acid)
- Place on the shaker for the appropriate time (see table below)
- Remove the sulfuric acid
- Discard it into the sulfuric acid waste container in the fume hood
- Immediately rinse the seeds once with dH20
- Add ~5 mL of bleach
- Place them back onto the shaker for two minutes
- Rinse five times with dH20
- Pour the seeds and some dH20 into small plates
- Parafilm and store in the fridge 24 hours to allow imbibement
- Be sure not to have excessive amounts of water in the plate
- Remove the seeds from the fridge and remove the water
- Parafilm the mini-plates, invert and place at room temperature in the dark for one day
Note: some plants may take a little longer to germinate
species | time |
---|---|
arabica | 15 minutes |
constricta | 10 minutes |
disciformis | 10 minutes |
heyniana | 20 minutes |
italica | 10 minutes |
lesinsii | 10 minutes |
lupulina | 20 minutes |
polymorpha | 20 minutes |
praecox | 10 minutes |
rugosa | 30 minutes |
tenoreana | 10 minutes |
truncatula A7 | 10 minutes |