flasking sterilization qurey

[gidi]

Hi
since my phal pod is getting bigger and seems in good condition i started to look for info about the next steps.
I realy don't understand why should it all be sterilized, what happens in nature that doesn't require sterilization of the seeds, media, and the growing environment, and why can't we apply the same process?

More to that after reading that the flasks should be sterilized either by boiling water, presure cooker or chimical and washing solutions, i fail to see how can you keep it sterilized due to the fact that you need to open the lid to sow the sterilized seed, and you can't prevent contanimated air to flow in and have a new unsterilized atmosphire inside the flask (the flask can be sterilized with his lid closed and achive sterilized atmosphire).

also read that oxygen should be at least 21% such as in the air but higher concentration of oxygen assit to the germination, and lower concentration of oxygen does the opposite

farther more at a certuin level the germination should be moved to a new flask which isn't air tight therefore not sterilized how would i know the moment is right, if i move it to soon from the sterilized environment to the none would it kill my eforts?

[jeffrey6115]

Orchid seed unlike corn or peas have no food source attached to them. In nature orchid seeds need to form a simbiotic relationship with a mycorrhizae bacteria. The bacteria provide the food for the growing protocorm. We need to provide a food source for the seed so we have agar. This is a nutrient gel that orchid seed will germinate and grow on. This agar also grows bacteria and molds very rapidly. This is why everything is done under sterile conditions.
This is also why you have to sterilize seed that has been exposed to the elements. If you use the green pod method the seed is still sealed inside the pod and is there for still sterile. Even though I use a glove box when I sow seed I still lose a small percentage of flasks to mold or bacteria. As for oxygen exchange I do not have any in my mother flasks. There is usually enough oxygen obtained when sowing the seed. I do have breathers in the replate flasks because the rapidly growing plants tend to use more.
Replating is also done in the glove box under sterile conditions. After all equipment has been sprayed to sterilize. (I use a 10% bleach solution) You are still taking sterile plants from one jar and moving them to another. With proper handeling (aseptic technique) there should be no contamination.

[gidi]

but even in green pot method the moment you open the pos the seeds are no longer sterile none the less it's not an action that takes less thana second so mean while you breath, doesn't our breath carry microorganizems?
you wrote "As for oxygen exchange I do not have any in my mother flasks." do you sterilize your flask sealed with rubber cork and when they cool down you inject the seeds with a needle through the rubber?
does the hole the needle created in the rubber seals up by itself or do i need to tape the top surface of the rubber cork?

[jeffrey6115]

I use a glove box for all this. I use a fish tank on it side. I seal the open end with plastic and only have 2 small slits for my gloved hands to go through. All work is done inside. All items to be used are placed inside before sealing off the end.
As for the seed pod. You clean the outside with your bleach solution before opening it. This is also done inside the glovebox. If the inside of the box and all your equipment and even your seed capsule has been properly cleaned it should be realitivly free of bacteria and spores. Since only your gloved hands are in there there is little to no air movement to spread them either.
I completely remove the top of my mother flasks when sowing seed. Some people prefer to inject through the stopper. The stoppers used close very tightly after the needle is removed so there is little chance of contamination.

[gidi]

ahh ok now i got it
thanks for the info