I have planted some sterilized tomato seeds on MS media, but I have not got any germination. Could you please help me to get the best conditions for that???
The method I used for sterilization was: The seeds were first surface sterilized by soaking in 70% ethanol for 2 minutes and I have washed the seeds in sterile distilled water twice. After that, I added 50% (vlv) bleach with drop of Triton X-100 for 15 minutes, then I have discarded the bleach, Washed the seeds several times in water and I have allowed the seeds to dry in sterile conditions.
After drying the sterilized seeds, I have planted them on MS media and kept the tubes at 15 degree for two days. Then, I transferred them to a controlled room at 26 degree for two weeks. The problem is that the seeds are not able to germinate!! Anyone has any idea what is wrong with my tomato seeds??
Thanks in advance!
Get fresh seed with a labelled date and germination test result. The bleach solution you are employing is very high. Generally 10% commercial bleach .525% sodium hypochlorite is sufficient for 4 minutes. Rinse one minute in sterile distilled water and then drained letting the chlorine vent in a aseptic hood. The incubation temperature would be more optimized at 25 to 28 C. Plant your seeds on plates using MS media and incubate to select only seeds with no contaminants. Store your tomato seeds in refrigerated environment with low controlled humidity. Let know of the success you experience. Best Regards.
Tomato seeds usually germinate within 5 to 10 days when kept in the optimum temperature range of 70 to 80F (21 to 27C). Germination is delayed by lower temperatures and accelerated by higher temperatures. Temperatures below 50F (10C) or above 95F (35C) are detrimental to germination.Jan 1, 2013
Dear Dr. Enas
- put seeds under tap water for 1hour
- rinse seeds in absolute ethanol for 2min
- rinse seeds in 40% Sodium hypochlorite (NaOCl) or 40% bleach for 30 min with shacking
- wash seeds with sterilised water 4 times each time for 3-5 min with shacking
- planted seeds in germination media, incubate 26C / dark for 2-6 days
germination media
4.4 g MS media
15 g sucrose
8g plant agar
- day 7 transfer jars to light
good luck
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