Firstly, check your seed whether they are viable or not. If viable plate the seeds on MSA plate and give them cold treatment at 4 degree for 2-4 day in order to beak dormancy.
Yes, I already tested their germinability . But dot found any seedling.
If the seeds were stored at room temperature the whole time period then the seeds are dead, but if stored at -80C then they should be viable. Also if stored at 4C they are probably dead too. Arabidopsis seeds do not have a long seed life span. Remember even seeds are respiring and only have a limited energy reserve (endosperm).
If i want to treatment seeds by any hormones and apply tissue culture technique, It's possible or not.
Follow Dr Carneiro advice first. You will got a good idea of their viability. If they are dead for the test, you can try the advice of Dr D'Allessandro... and pray.
You can also try nicking (making a small hole in) the seed coat to increase the possibility of gibberelins, sucrose, etc. getting to the embryo.
I wouldn't waste my time in seeds that old if they weren't stored at -80°C. If you have to rescue them anyway, I'd try the treatzolium test as it was suggested above. But I would start thinking in ordering the seeds from NASC / ABRC stock centers or people that used that genotype/s...
It is indeed likely the seeds are dead, but if they are not dead and only severely deteriorated, there is a small chance to rescue the seeds. Of course this is only needed when the seeds are valuable and you cannot obtain the mutant from other sources.
My advice is to incubate your seeds first 24 hours at room temperature in a closed box with 100% RH. The next day imbibe your seeds in water of about 20 -25 °C, definitely not cold water, on filter paper. Check daily for germinated seeds. It may take more time than usual. Do not sterilize the seeds! The background is that during the ageing the membranes will be oxidized. Sterilization will provide further oxidative stress. Oxidized cell membranes are more rigid and much more sensitive to leakage. By incubating first at 100% RH the seed take up some moisture without leakage. Next upon imbibition in water, the water potential difference will be relatively less, so new water will enter more slow, giving less disruption of the weak membranes. Cold water imbibition will make the membranes even more rigid and prone to leakage.
In the past we have done experiments where slow imbibition through PEG priming could improve the germination of aged brassica seeds. However, although Easter is approaching, do not expect to raise dead seeds.
of course it is a long time which definitely will reduce the viability of seed, however, if they had been stored in very suitable storage conditions, and these seeds had been harvested from mother plants which had received very careful and very optimum conditions in terms of nutrition and irrigation, you can have some viable seeds.
I absolutely agree with Steven P.C. Groot and I add a step: sterilize the substrate two times to avoid plagues and fungal problems.
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