Remove the seed with a litle media use a scraper and then put on in the hole little drops of PPM.

Good luck!!

1. Use a long pair of flame-sterilized, and cool-down, forceps (see attached picture, but with gloves on) to remove the not-infected seeds one by one carefully to clean tubes or bottles.

2. If you can, try to remove the rest 6 seeds to 6 different tubes/bottles, just in case one of them are already contaminated (but cannot be seen). You don't want to risk to put them together again.

Take not contaminated seeds one by one carefully in laminar flow cabinet again surface sterlise using increased concentration of surface sterilizing agent or increase time of surface sterilization and transfer to fresh culture tube,

its for sure that if one seed gets contaminated then other seeds also get vulnerable. Take them out, treat with hgcl2 or other agent in desired conc. and reinoculate.

I agree with Luis Carlos Ramos if your seeds had cultured for 10 days.

first of all identfy the cause if it,s fungus contaminated or virus in the ssed or in the tubes then tested the seed   carefully in laminar flow cabinet again surface sterlise using increased concentration of surface sterilizing agent like colerex  use diffent dose with differen  time or increase time .

Actually, you get a lot of answer. So, transfer carefully clean seeds to new bottle. Be care ; do not takke any spores.

But next time, if you have potentail contamination, use small bottles and tput just one seeds per bottle. In the case of contamination you can just discard contaminated bottle. Do not use HgCl2: it is very good strelization agent, but eventually very toxic to plants and can have strong effcet on development.

It is very much easy and require a little skill in isolating the uncontaminated seedlings from the contaminated vessel. Isolate the uncontaminated seeds/ seedlings from the contaminated bottle with sterile instruments and place it on a sterile petridish. Cut and discard the doubtful contaminated portion of the uncontaminated seed/ seedling. subculture in a fresh vessel/media. The operations should be inside the laminar air-flow hood after taking necessary precautions to make the hood sterile.

best wishes.

thank you to all for giving these suggestions. I recovered my seeds.

Hai Anil 

Hope you got good answers and especially from Yuan sir (Thank you yuan sir), but as per my knowledge once you got contamination I don't think so it is fine to use same for further , you please re do the experiments and keep triplicates and before inoculation please follow safety precautions received by experts in our discussion.

Good Luck

thanks anil sir,

sir you are right that if i am using the plants from contaminated sources or from contaminated media then the chances that it will affect my further data for experiments.

So I already started fresh experimen with precautions.

Dear Anil,

You can try once atleast. Anyhow its a chance to save, please take other seeds and sterilized in with 70% ethanol wash and then 0.2% HgCl2 further wash with double distilled autoclaved water subculture these in fresh media. It may help sometimes. In side you can try new fresh experiments in triplicates.

Good luck,

Dr. Aloka

Hai Anil 

Its fine ... All the best for your science. 

Good luck 

Good luck Anil

Removed the non-contaminated seeds and transferred to new tissue culture bottle after sterilized in 0.5 % benlate (fungicide) to 15 minutes.  

decide the major  organism and removal by using as they nature if bacteia or fungi or even the virus, be-sure about the outiclave also and use the uv additional to sterlizer