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

[orchidsarefun]

Good Luck !
....also wanted to mention that my propagation guy has his operation in his basement ( cooler ) and does not have a 12hr lights-on situation. Hopefully other people can give input on this too.

[czayta]

Originally Posted by orchidsarefun

Good Luck !
....also wanted to mention that my propagation guy has his operation in his basement ( cooler ) and does not have a 12hr lights-on situation. Hopefully other people can give input on this too.

Oh, interesting. I read that a 12 hr light cycle is good for new flasks and Phal growth. I'll have to do some more research on the appropriate light set up.

[Brooke]

I think you harvested the pod too soon to have viable seeds. We wait until the seed pod starts to yellow on the ends but is not open yet. Almost every Phal we have done has been ready to harvest about six months after pollinating, some take longer.

Our flasks are then put under regular T12 bulbs about 6-8" from the source. I have no experience using the hotter T5, but warmth is needed and recommended to help with germination and growth. We also run the lights 12 on/12 off.

Hopefully you will have a few minute seeds hidden in the cotton. Good luck!

Brooke

[czayta]

Originally Posted by Brooke

I think you harvested the pod too soon to have viable seeds. We wait until the seed pod starts to yellow on the ends but is not open yet. Almost every Phal we have done has been ready to harvest about six months after pollinating, some take longer.

Our flasks are then put under regular T12 bulbs about 6-8" from the source. I have no experience using the hotter T5, but warmth is needed and recommended to help with germination and growth. We also run the lights 12 on/12 off.

Hopefully you will have a few minute seeds hidden in the cotton. Good luck!

Brooke

Oh no! Alright thank you for the advice. I'll keep my fingers crossed that maybe a few seeds were ready.

[orchidsarefun]

Not sure if you have seen this site, it has useful information

Lotte & Thomas Orchids

Keep us updated !

[Ben Belton]

Just some thoughts from someone who has sown many a Phalaenopsis pod.

1. I'd wait until 6 months to harvest if you can. Keep an eye on your pod and when it starts to get lighter green or have yellow tips like Brooke says, pull it then. I've also heard you can tell it is getting close when it starts to get soft. I've had pods turn at 4 months under bright light and have lots of seed and at 7 months and have very little. My pods are rarely consistent.

2. I don't know if it is true but I've been told that if you tease the fluff with the seed out without scraping the pod, you won't transfer virus if the pod parent is virused. I don't scrape the pod. In the end, even if you only have the same amount of seed as a grain of rice, you still have a thousand or more seed which is more than you probably want. So no need to scrape the pod.

3. I sterilize my pods in 20% bleach for 5 minutes. Haven't had any contamination yet. Might save you some time.

4. I agree with Brooke on the amount of light. I actually germinate mine on a shelf near my lights, but not under them. Once they have germinated, I move them to where I have 2 bulb T8 figures. They are placed on the shelf which might be right under or several inches from the source and they do fine.

5. Lastly I've had seed germinate as much as a month after I sowed them so don't give up yet. You still might get something.

In the end, except for giving your pod a bit longer, I think you did everything fine and my ideas are just opinion. I think what you did will work great in the future if you like this process and want to stick with it.

Good Luck!

[czayta]

Originally Posted by orchidsarefun

Not sure if you have seen this site, it has useful information

Lotte & Thomas Orchids

Keep us updated !

This site is great. Thanks! I'll keep you guys updated.

Originally Posted by Ben Belton

Just some thoughts from someone who has sown many a Phalaenopsis pod.

1. I'd wait until 6 months to harvest if you can. Keep an eye on your pod and when it starts to get lighter green or have yellow tips like Brooke says, pull it then. I've also heard you can tell it is getting close when it starts to get soft. I've had pods turn at 4 months under bright light and have lots of seed and at 7 months and have very little. My pods are rarely consistent.

2. I don't know if it is true but I've been told that if you tease the fluff with the seed out without scraping the pod, you won't transfer virus if the pod parent is virused. I don't scrape the pod. In the end, even if you only have the same amount of seed as a grain of rice, you still have a thousand or more seed which is more than you probably want. So no need to scrape the pod.

3. I sterilize my pods in 20% bleach for 5 minutes. Haven't had any contamination yet. Might save you some time.

4. I agree with Brooke on the amount of light. I actually germinate mine on a shelf near my lights, but not under them. Once they have germinated, I move them to where I have 2 bulb T8 figures. They are placed on the shelf which might be right under or several inches from the source and they do fine.

5. Lastly I've had seed germinate as much as a month after I sowed them so don't give up yet. You still might get something.

In the end, except for giving your pod a bit longer, I think you did everything fine and my ideas are just opinion. I think what you did will work great in the future if you like this process and want to stick with it.

Good Luck!

Wow, thank you for such a detailed post. I have 4 more Phal pods growing at the moment, so I will try this advice when those ones are 6 months old or show signs of being ready to be flasked.

When you sterlizie your pod do you scrub it with a tooth brush or anything? The information I was following just said to let it sit in the 10% bleach solution with a stirbar (so it was sorta spinning around in the solution) for 20 minutes. I am going to try the 20% solution next time too.

After it is sterlized, do you let it air dry at all in your glovebox/laminar flow hood?

At this link:
How To Grow Orchids From Seed
The picture on the left looks like all seeds. Could you explain more how you 'tease' out the seeds from the fluff? (Do you have pictures to show what the seeds look like compared to the fluff?)

[alaind52]

Originally Posted by Ben Belton

Just some thoughts from someone who has sown many a Phalaenopsis pod.

1. I'd wait until 6 months to harvest if you can. Keep an eye on your pod and when it starts to get lighter green or have yellow tips like Brooke says, pull it then. I've also heard you can tell it is getting close when it starts to get soft. I've had pods turn at 4 months under bright light and have lots of seed and at 7 months and have very little. My pods are rarely consistent.

2. I don't know if it is true but I've been told that if you tease the fluff with the seed out without scraping the pod, you won't transfer virus if the pod parent is virused. I don't scrape the pod. In the end, even if you only have the same amount of seed as a grain of rice, you still have a thousand or more seed which is more than you probably want. So no need to scrape the pod.

3. I sterilize my pods in 20% bleach for 5 minutes. Haven't had any contamination yet. Might save you some time.

4. I agree with Brooke on the amount of light. I actually germinate mine on a shelf near my lights, but not under them. Once they have germinated, I move them to where I have 2 bulb T8 figures. They are placed on the shelf which might be right under or several inches from the source and they do fine.

5. Lastly I've had seed germinate as much as a month after I sowed them so don't give up yet. You still might get something.

In the end, except for giving your pod a bit longer, I think you did everything fine and my ideas are just opinion. I think what you did will work great in the future if you like this process and want to stick with it.

Good Luck!

Hi to All,
The points above were the same for me. I tried to cross some hybrids of Phalaenopsis. It took 6 months the shortest and 10 months (between Phalaenopsis and Ascocenda) the longest for the pods to start to turn yellow green and to soften a bit. No need to scrap as when cutting the pods, the seeds fell down in the flask. The only issue is that you can sowe too many seeds in a narrow place. It took 3 to 4 weeks to germinate and turn green. No light system, I put the flasks at a window when protocorms are visible. The first replate took place after 2 months. Here is just what I met in my first try.

[Ben Belton]

Originally Posted by czayta

When you sterlizie your pod do you scrub it with a tooth brush or anything?

No, I roll it around in the bleach, but that is all

Originally Posted by czayta

with a stirbar (so it was sorta spinning around in the solution) for 20 minutes.

I just have/had a little homemade lab in my basement. No stir bars.

Originally Posted by czayta

After it is sterlized, do you let it air dry at all in your glovebox/laminar flow hood?

I don't let it dry out. I let it drip enough so that there is no bleach that could potentially drip into my flasks.

Originally Posted by czayta

At this link:
How To Grow Orchids From Seed
The picture on the left looks like all seeds.

Wasn't sure which pic you were referring to. The one pic has the fluff and the seed is in that. The actual seed can be seen on the penny.

Originally Posted by czayta

Could you explain more how you 'tease' out the seeds from the fluff? (Do you have pictures to show what the seeds look like compared to the fluff?)

Sure. See pic attached. The focus isn't so great but you can see that the "fluff" is cottony stuff that grows in the pod. I believe it supports the seeds as they grow. It is easy to hook your spatual into it and pick it out without ever touching the wall of the pod. You can see the seed next to the pin.

With that there, you can easily make many mother flasks and have tons of germinated seed. As an earlier poster implies, with a pod like this, you can tap it over the top of a couple flasks and enough seed will fall out to have more than you will ever want.

When you do tease fluff out like this, I try to spread it out as much as possible, while always limiting how much time I spend with the top open. My flasks usually have a few drop extra of water that will pool in the edge if you tilt the flask to its side. I have a spatula that I have bent the end to a 90 degree angle. I try to touch the flat part to the water and then to the fluff which wets it and helps it adhere to the media. The more you get to stick to the media, the more you will have germinate. Hope this helps. I with your other pods, I think it will be easy to see how the seed and fluff are all together in the pod.