update

Using tissue culture to propagate plants.

Moderator: Ghazanfar Ghori

kerokero
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Real Name: Corey W
Location: Northern VA

Post by kerokero »

Both my aquatic stems are growing well as in the above pics by Mary, and it seems the orchid seed Mary also has pictued has done the same thing... the seeds have a more greenish cast to them now. My Blettia seed has definately germinated as the (thankfully larger) seed has turned into little distinct grows growing upward in many!

Both the AVs have shown plantlet growth, but not the crazy growth on if they had been put "in" the agar, nothing spectacular compared to the normal growth I see when I do this to leaves. I have a feeling when the little suckers get their little roots into the hormone laden substrate that story may change a little. It does seem to show the difference of putting the leaves laid on top (not much influence of substrate - plantlets most often grow from the viens and if these don't have contact with the substrate, the substrate doesn't influence their growth) as putting them partially in (where the veins are in contact with the substrate - which results in a super cloud of plantlets!). With stiff substrate like I had (too much agar in it? or it was just left to set too long?) I'll make more of an effort to slice a spot in it for the leaf and stick it in, instead of me trying to shove it in because I thought it was softer than it was (and damaged the leaf severly in the process).

One thing I am sad about is my jewel orchid has a lot of contamination now. I wasn't sure about growth, but now the growth in the container is definately more white mold than jewel orchid. This was a peice of stem floating in a PPM mixture because of it's likely high contamination. I forgot to swirl it for one day last week, and the next day it had mold - it was that quick and that important! I am either going to have to be more careful about what stock I use to start TCs (bloom and stem starts, cuts of stem that have no active roots or leaf growth on them, etc) rather than the section I started with that had lots of root growth on it that was removed, but left a number of "rough spots" that likely didn't get cleaned well - or I'm going to have to set up one of those swirling tables.

Of course now I'm back to being confused on why we had the aquatic stems on the green (no hormone) substrate, while the AVs and my jewel orchid were on the hormone substrate? At some point it made sense, now it doesn't again. I guess I'm confused over why my jewel orchid stem was on the blue, while the other stems were on the green even tho they grow the same way.

One thing I have noticed with the stems that have nodes... it seems like they have more/faster growth and success when the nodes are in contact with the substrate... which is why Mary's specimen #1 and #2 are doing so much better and producing more explant material faster than #3 which had a much more limited exposure to the substrate. It seems important for maximum benefit of doing TC that you expose your material to the substrate the most efficient way!

Hey look, maybe that can be my first blog post :) I should take pics... see if my camera is up for it.
Best, Corey
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Aaron
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Post by Aaron »

Mary - #1 is Proserpinaca palustris (Mermaid Weed)
seiji64
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Post by seiji64 »

Here are my tissue culture updates.

I didn't push the African Violet leaf edge into the medium and the result appears to be, "No growth for YOU!" *spoken as the Soup Nazi* Although there is an odd opaque gel/liquid on that front corner leaf edge.
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I believe this jar holds a cutting of Ghazanfar's aquatic plant. Again, I didn't push the stem into the medium and maybe should have. It's at least growing, but don't think it is generating plantlets.
Image

This jar holds the Bletilla sp. orchid seeds. After weeks of no apparent growth, I'm finally seeing some! When can I pull these saplings out and plant them?
Image

This is my second culture of Ghazanfar's aquatic plant. I cut the stems too close to the meristems, so I think the bleach did them in from the get go. But the mold is doing quite well.
Image

I also have a jar of Schomburgkia sp. orchid seeds, but those are still showing no signs of life. :(

Doug
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Ben Belton
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Post by Ben Belton »

Hey, your African Violet is still green, so you might just have to be patient. I think it will eventually do something.
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Ghazanfar Ghori
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Post by Ghazanfar Ghori »

For jars that are contaminated - don't open them! Could be harmful to your health.
You should put them in a pressure cooker @ 15 psi for 20 minutes to kill everything in before opening it and cleaning it out.
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Ghazanfar Ghori

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kerokero
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Real Name: Corey W
Location: Northern VA

Post by kerokero »

Good to know Ghazanfar - I didn't think about that.

As for my stuff... jewel orchid is a nice big clump of happy white fungi. One of the AVs looks like Doug's curled little leaf, but has some plantlets showing at the veins... no more than if the plant usually produces likely due to lack of contact with the media. The other AV fragment was the one I highly damaged trying to force it into the media (the idea to cut the media and slide the fragment in didn't occur to me until after I had sealed the jars) and after succumbing to the damage and the fact that I don't think I cut off the bleached ends resulted in it eventually dying. Everything else seems to be chugging along.

Doug - it may take a long time before the Bletilla can be ready (some orchids it can take years). I'd wait until they were filling up a good portion of the jar and need to be broken up, and transfered into larger jars to grow out more. I don't know the ideal size to transplant Bletilla, but you'll want a plant with strong roots so it could take breaking up the plantlets into smaller groups a couple times to get this.

What I've realized with the stem plants with multuple nodes on them is laying on the substrate might be better than sticking one end in - you want to maximize the amount of nodes in contact with the substrate so the most of them grow. Placing the plants correctly on the substrate seems to be important to maximize growth.
Best, Corey
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