Standard Treatment (used for Arabidopsis):
Seeds are surface sterilized with bleach solution (30% commercial bleach + 0.02% Triton X-100) for 15 min, washed 3 times with sterile water, and placed at 4C overnight prior to plating.
Species-Specific Modifications:
- Selaginella – Bulbils collected from leaf tips are surface sterilized using standard treatment solution for 15 min. After plating, bulbils may take 2-3 weeks to germinate and another 1-2 weeks to develop roots. (Note bulbils can be stored at -20C for about 1-2 years.)
- Maize – Seeds can be surface sterilized with half-strength standard treatment solution (15% commercial bleach + 0.01% Triton X-100) for 15 min. MS plates need to be covered in dark for about 2 days to aid germination. After removing the cover, maize seeds require ~2 days to develop roots.
- Rice – No special treatments required. Seeds require ~ 2-3 days to germinate and another 2 days to develop roots.
- Soybean – No special treatments required. Seeds require ~ 1-2 days to germinate and another 1-2 days to develop roots.
- Cucumber – No special treatments required. Seeds require ~ 2 days to germinate and another 2 days to develop roots.
- Tomato – MS plates need to be covered in dark for about 2 days to aid germination. After removing the cover, tomato seeds require ~ 2-3 days to develop roots.
- Medicago – Seeds are treated with sulfuric acid for 8 minutes to aid germination and placed in the dark at room temperature following sterilization.