Some species of genus Linum (L. grandiflorum, L. pubescens) have many infections on surface of seeds and in time of germination this seeds this infection gets into seedlings....but when I sterilized this seeds in 70 % ethanol for 10 minutes (according to protocl for sterilize of L. usitatissimum seeds), the seeds was not germinated...
Dear Hanna,
10 minutes in ethanol is too long. First wash the seeds in soap and running water (use a mesh to avoid seed loss), then dry on a paper towel and give maximum of 2 minutes ethanol wash and transfer to sodium hypochlorite (3%) solution with few drops of Tween 20 for about 20 minutes under shaking. Wash 3 times with sterile water and plate under aseptic conditions.
Good luck!
Dear Hanna,
I agree with Ranjith, 10 m seems too long. The method that Dr. Pathirana proposed is good but I always try the simplest way. If it works OK you do not need anymore. You can try washing the seeds in sodium hypochlorite (3 or 4 %) solution for 5 m. and wash 3 times with sterile water. It is simple but it works well in the most cases.
You can wash seeds with 5%sodium hypochlorite for 5 min. Then wash with sterile dist. Water. Or you can use an antifungal solution(0.3%)then wash with water.
It is an antibiotic especially for killing fungi. I got it from a research center. They use it in the sterilization of vegetable and crop seeds before planting.
Hi Hanna
Dr Pathirana' suggestions are OK. Only if fungus contaminations persisted, then you could add an antifungic treatment in the process, with a benomyl solution for instance. A simplest way for seed disinfection is to use a commercial disinfectant as Domestos R as 10 or 20% aqueous dilution gently stirred during 20 to 30 min, then washing with sterile distilled or Millipore R water.
Regards.
Marc Jullien
A beautiful discussion was done over here for seed sterilization. There are 2 main surface sterilizers:-
Sodium hypochlorite: as discussed earlier
Mercuric chloride: It is also a potent surface sterilizer, take seeds in 1% solution of HgCl2 solution for 15-20 seconds. It kills a variety of pathogens remain present on seed surface.
cheers!
Sanjeet
In addition to mentioned methods, gamma irradiation was effective in this concern and in addition improve seeds germination, plant growth and yields. There are many references about sterilization using gamma rays
I follow this:
..."Seeds were surface-sterilized for 20 min in 20 % (v/v) sodium
hypochlorite, rinsed and soaked for 1 h in distilled
water"... The sterilization procedure works fine with my seeds and I have a very good germination index.
Good luck!
Gamma irradiation will sterilise the seeds but will either have mutations in the resulting progeny or the seeds will also die along with the microbes - depending on the dose of irradiation you use. So for aseptic culture of seeds, gamma radiation is not the best way to go.
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