Green Seed Pod Flasking - Phalaenopsis

[czayta]

This my second attempt at flasking. The first attempt was a dry seed pod that I flasked two weeks ago. There still hasn't been any germination in those flasks yet. Today my Phalaenopsis green seed pod was 111 days old so I decided to cut it up and see what was inside.


I started by sterilizing the outside of the pod with 70% Ethanol for 1 minute.


After that I transferred the pod to a 10% bleach solution with a little bit of dish soap for 20 minutes.


After that I transferred the seed pod into the hood and it dried for 20 minutes. The hood had already been sterilized with 70% ethanol before the pod was put in it on a sterile petri dish.

I cleaned off all of the tools I would be using, and put the flasks into the hood.



I sliced open the Phal seed pod and scraped out the seeds into each flask.


I am unsure of how successful this will be because the seeds are sorta clumpy on the agar. I didn't want to open up the jars again and risk contamination and spread it out.


I am concerned that it might also be too early in the seed pods maturation, even though Phalaenopsis are reocmmended to be green pod harvested between 110-120 days (with 111 days being perferred). When I looked at it under the microscope I didn't see lots of seeds or embryo's like I had hoped. I'm not exactly sure what I am looking for though. (inexperienced eye when it comes to this sorta thing lol)



Closer up 100x


Possible chloraphyl?


The flasks will be stored at room temperature under T5HO lights for 12 hrs a day. I'll let you know how it goes and if anything grows! ^^

Edit: Successful germination in 7 out of the 8 flasks on 10/2/14! 0% contamination thus far.

Edit: 4/12/2015
I have returned from my hiatus with some propagation orchid news.

To recap: I cross pollinated one of my NoID orange Phalaenopsis orchids with pollen from a NoID phalaenopsis purple orchid to make a seed pod. The seed pod developed over a period of 4-6 months and then I will plated the seeds on agar. After 8-10 months (today) in the flask I deflasked the orchids into little pots to grow into mature plants.

I never got a chance to make replate media (been so busy) so these little orchids might have a tough time taking off.

Here is what my flasks look like today, 4/12/15. These flasks will need to be deflasked or replated soon. Lots of healthy roots growing and they look a bit crowded.


A mix of different sizes in this jar.


I will have lots of Phalaenopsis orchids when I deflask all of these lol.



These orchids are still small and would benefit from replate media.


And now we come to the contaminated flasks (2 out of 8). White = mold. This mold randomly sprung up over the last three days. Not sure how it got there, but it happened in two jars at the same time.


This is a second contaminated flask, so these orchids will need to be deflasked before the mold takes over them and suffocates the small plants.


These seedlings still have some moldy agar on their roots.


All the agar has been washed off of the roots.


The biggest one:


The first few Phalaenopais orchids have been deflasked and potted.


Deflasking my Phalaenopsis orchid jars that I propagated last August! Two of the 8 flasks suddenly became contaminated so it was time to move the seedlings out of the flasks.
Little orchids all potted up! I think not many of these will survive since they are so small unfortunately.


Update 05/20/2015: I deflasked the remaining flasks

Protocorm clusters?


Ones too small to be potted


All cleaned up:


Some of the bigger ones:


Plants that I am sending to Cntry