Media preparation for home orchid flasking. Part 2... Replating Media

[kaiha]

This is a great thread, I am enjoying it. Someday I may try it. I'll just have to get everything together and give it a try.

[korxi]

Hi,

I'm new here, but I've been experimenting a bit with propagation this last year!
In regards to the sterilization problem with the vented flasks I'm using another technique:

1. Drill a hole in the lid of the jar and plug it using silicone. Silicone can handle really high temperatures without melting!
2. Now sterilize like you normally do, leave falsks for a couple of days to make sure they aren't contaminated and repalte (If you want you can leave for a couple of days again, to makes sure there was no contamination while replating)
3. Take a syringe needle, plug the end with some cotton wool, preferably the kind that doesn't absorb water (I'm using regular cotton wool with good results)
Wipe the needle with rubbing alcohol just before punching it through the silicone plug, and there you go - a vented flask .. Please ask if I didn't make any sense

Christian

[John]

Hi Christian.. I'm new here also and just came across your post. Good idea, has this worked well for you? Since I'm new to this, would it be better to insert the needle before you pressure cook it so that everything is sterile? Just thinking if pathagens on the cotton when you insert it they could be drawn into the media. I have used the band aid approach and so far so good (band aid on both sides of the lid) Although I did have four replates develop mold after about three weeks. Don't know if the spores were in the mother flask (shipped in from Malaysia) or if I didn't wait long enough for the bleach to sterialize my glove box or if there was a leak in the jar/lid seal. I went in and retreaved the plants that were not affected, dipped them into Physan solution and put back into new flask. Almost all survived. Just cooked up another two dozen flasks with replate media..expect another shipment from Malaysia in next two weeks. I'm using Hill's Replating Medium from G&S Labs. By the way, where are you located?

John

[korxi]

Hi,

Oh yeah I forgot to mention I soak the cotton wool in rubbing alcohol before i insert it into the syringe needle opening, it is then left for the alcohol to evaporate before wiping and inserting it

I'm situated in Denmark, just started growing orchids a year ago, but started out with propagation right from the beginning.. My first phal. was selfed and I spent the time the pod took to mature to do the research. I chose a completly DIY approach - only using stuff available in the kitchen. '
However I think my medium isn't of the best quality, since I've had very low germination rates on both set of seeds I've sown - or perhaps it's just caused by the fact that it has been complex phal hybrids I've tried to cross and they might not be that fertile?

Christian

[John]

Hey Christian...Great to talk to someone from Denmark! It sounds that you have done a lot of research. As you know there must be hundreds of diff formulas for media. I chose to go with Hill's to start because it was complete and no need to check for Ph levels. I understand that media thats works for Phals may not work so good for Den or Cyms or what ever. The trick will be in finding the right one. I haven't tried to start seed yet but I do know the germination rate of seed varies a lot. If you have the sterilization part down that's a great start. Are you working in a glove box? I think we may have lost Phalguy in this thread but I hope he jumps in with an update on what he's doing.
John

[korxi]

Actually I'm not working in anything but my kitchen and the jars I'm using
Here's how i sow:
1. put the amount of seed you wan't to sow in a 5% sugar sollution for 12 hours - to get any dormant bacteria or fungi to wake up..
2. suck up the seed in a disposable syringe.
3. let the seed settle on the bottom and push out the sugar water (but not the seed)
4. suck up some 3% peroxide and agitate the syringe to get all bubbles forming in there out. wait 30 mins. - the seed are now sterile..
5. put a needle on the syringe and sow by pushing the needle through the silicone plug the lid (mentioned above in regards to replating) and push out the amount of seed you want in each jar/flask. You shouldn't worry about the peroxide since it turns in to water and oxygen, but do try to minimizre the amount in the jars/flasks since I don't think the seeds will germinate that good floating in to much water.

This technique was actually invented by a danish guy named Lars, I found out because I made a "pictorial" on another forum showing the complete process from medium preparation to sowing. I had just found the info all over the internet, but when he saw it he told me that he actually was the originator of that method

At the moment I'm working on a way to use one rather large jar for both germinating the seed and replating, using two different media in the same jar at once - and a way to replate inside the jar without ever opening it
I just came up with the initial idea yesterday so not much work has been put into it yet.. but if anybody is interested I could start a thread or something when i get to work on it?

Christian

[John]

Hi Christian...you are really getting into this. That is an interesting idea, the advantage would be to reduce the chance of contamination. I will be interested to see how access to the plants will be performed and how you keep the media apart. Keep me informed. By the way do you vent the mother flasks or just the replate?

John

[korxi]

Until now (only two tries) I've not replated, I just vented the mother flasks. However I doubled the amount of medium in the flasks and only put in around 30-50 seeds in each. As i wrote above I haven't experienced great germination levels so I'm not worried about crowding, my main concern is that there aren't any complex carbs in my medium and it thereby might not be able to support the seedlings (hopefully the protocorms will develop further..) untill it's time for deflasking. If it doesn't work it's no big deal, these first attempts are just for learning!

Christian

[John]

Hi Christian...Good attitude to take. From the little I have heard, there is no need to vent mother flasks.

John

[charmbutterfly]

I'm looking forward to trying this!!