I have been having contamination issues after seed treatments with GA3 and sowing in moist, unsterilized soil. I was thinking of moving to a sterile environment, but I wasn't able to find much information on the protocols used to plant comparatively large seeds in sterile conditions. Will agar work as a medium, eventually with MS or even just water? Will I need to make a lower density agar and push the larger seeds inside it, or just lay them on the surface?
Dear Alessio,
If you have large seeds, they have enough nutrients to germinate (recalcitrant seed, but not orthodox). They can therefore germinate only in petri dishes on moist filter paper. Prepare the dishes first, then sterilize them with UV and then place the seeds.
Good luck, BS
You can use pasteurized soil and sphagnum moss plus sand and vermiculite. Also, wash the seeds with 3 % hydrogen peroxide
Hi Alessio,
For big-sized seeds, you can use on-sand, and between-sand substrates. Sand most be sieved, washed, autoclaved.
Dear Alessio,
Large seeds do not need a growing medium as Bozena Sera said because they have enough nutrients to germinate. You can use sterile soil, sand or any sterile substrate . If the problem persists, contamination must be in the seed. Disinfect the seed in a 70% alcohol solution for 1 minute followed by 20 to 30 minutes in 20% hypochlorite solution with agitation. If the problem still persists, try to remove the outer seed coat and disinfect again before putting it to germinate. Best luck!
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