transferring protocorms in liquid medium

[King_of_orchid_growing:)]

I have a flask of seeds that germinated some time during late August to early September.

The protocorms are in liquid media. They're still very, very tiny. You need a jeweler's loupe to see them and still not too much bigger than the tiny seeds.

These guys are growing steadily, but they're slow growing at the moment.

How do you know when to replate?

How would you do such a thing when they're just gonna float around in the media?

[King_of_orchid_growing:)]

I looked through some of my flasks, and I may have another one in the same situation as well.

These are even harder to see because of the coloration of the seeds.

[Des]

Would it not be easier to germinate the seed on the surface of agar jel . That would also allow the seed to absorb O2. I know that some hybridizers vibrate the liquid or have a method of stirring it continuously on a turntable so that the liquid is aerated. Leaving the seed to germinate in fluid might stunt the growth .

[King_of_orchid_growing:)]

I don't have those machines. They're expensive.

The species of Disa I'm flasking, started showing signs of germination when I soaked it in sugar water (a method I'm thinking of employing before I flask the seeds from now on, supposed to wake the fungi up so disinfection is more efficient). So I decided to use a liquid media.

From what I understand, using media at full strength for Disas is not recommended and can be detrimental to their survival. Sources recommend 1/4 strength or 1/8 strength.

Not so sure about the second flask. The second one was an experiment. It's Eulophia welwitschii.

[Pippin]

I like my media on the soft side, but I never tried a liquid medium.

[Ben Belton]

I have separated small protocorms from liquid media and replated them onto regular media using a sterilized coffee filter. The protocorms I was working with were much larger than the seed you are working with it seems, but maybe it is an idea for you.

I put the coffee filter between a couple layers of aluminum foil and autoclaved. In the hood I wiped down the foil with 25% clorox and opened the first layer. Then I wiped down the second layer with Clorox and took the coffee filter out and put it into a funnel which had been set into a beaker (both of which had been wiped down with 25% Clorox) I dumped the media into the filter, scooped the protocorms out, and spread them on the media. As tedious as it sounds (and it was) I didn't have any contamination.

[John D.]

The coffee filter method works, I use it to seperate protocorms from liquid media when treating for chromosome conversion. Doesn't have to be a funnel and beaker but something to keep the filter from flatening out completely when it gets wet makes life easier.

[Royal]

If they're still tiny, you may be able to pipette them. When to replate is a tough question to answer. I usually wait until I see a first leaf forming.

I'm still not sure why it would be necessary to use liquid media (unless you are looking for proliferation). Those that use liquid media normally use a physical support of some sort, like cotton fiber, shredded paper, raft, etc. to suspend the culture toward the surface of the medium. Too much available liquid can cause vitrification and halt growth all together.

[King_of_orchid_growing:)]

Originally Posted by RoyalOrchids

If they're still tiny, you may be able to pipette them. When to replate is a tough question to answer. I usually wait until I see a first leaf forming.

I'm still not sure why it would be necessary to use liquid media (unless you are looking for proliferation). Those that use liquid media normally use a physical support of some sort, like cotton fiber, shredded paper, raft, etc. to suspend the culture toward the surface of the medium. Too much available liquid can cause vitrification and halt growth all together.

I didn't know they used a raft. I've never heard about this method until now.

I just looked at the protocorms, and it looks like growth is starting to pick up a bit. I expect them to really take off during the warmer months.

As to why liquid, I'm just going by what I saw worked almost instantaneously in the early stages of disinfection as well as what was said in a scientific article discussing the germination of summer rainfall Disas. This species is a summer rainfall species.

So far, it seems to work.

I guess I'll just wait until they get a bit bigger. From what I understand, these can take quite a while to get a first leaf.