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Drosera36

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Hey everyone,

I haven't been on in a while, but I just thought I'd share what I've been working on for a little while.

I started an internship back in December with a professor that is friends with my family, and while I was doing that, I saw that a member here was giving away seeds. We were doing tissue culture in the lab at my internship, so I figured I might as well give it a shot.

It was a success (so far), and here are some pics from today. Just so ya know, they're from a Cymbidium hybrid.

The petri dishes with the seeds & a nutrient solution:
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The little green balls are the protocorms, and the white flecks are seeds that did not germinate.

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Some pics with a low powered microscope:
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There is an ungerminated seed above the two protocorms for comparison.

Here's a pic taken with a camera-compatible microscope (edited with photoshop as the picture was washed out and dull):
d211ea1f.jpg


In case you were wondering why I didn't just plant them in soil, it's because they would not be able to produce any food to grow. When the seeds come out of the pod, they are extremely small and thin with an underdeveloped embryo, as opposed to the fully developed embryos in say beans or acorns. In nature, this embryo would become infected with a symbiotic fungus species that is living near the roots of the parent plant. The fungus would then provide food for the developing embryo as it grows.

As it is difficult to get the specific fungus for the seeds in cultivation, you need to provide the seeds with some sort of nourishment. This is what the nutrient solution is for. This is just a simple mix of sucrose and other nutrients that we made, and that's mixed with agar to make a jelly-like substance.

-Ben
 
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That's very cool. What's unusual about orchid seed isn't the development stage of the embryo, it's the lack of an endosperm, which is the food supply most seeds contain to give an embryo it's start in life.
 
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It doesn't matter whether the botanist knew that bit of orchid seed trivia. What's important is that the botanist taught you top notch tissue culture technique. My plant physiology professor wasn't into it and rushed us through it, so I never grew anything but mold or bacteria or whatever it was.
 

Drosera36

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Yeah that's true. I sorta took it for granted just because I found tissue culture so easy to do, but she seemed surprised that I haven't had any mold on any of my dishes.

-Ben
 

Drosera36

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Well they have a pretty sophisticated set-up there, so it was pretty easy to do stuff.

To sterilize the seeds, we just put them in a little container with bleach for ~5min and shook them. They have a pressure cooker, so that's where the agar solution went in to get sterilized.

After that, we poured the liquid agar solution into dishes in a sterile hood which has a constant airflow to prevent colonization of bacteria & other small creatures.

We didn't have to use tools to get the seeds onto the plates, but if we did, we would use ethanol and a burner to sterilize them, which is kinda cool.

Then, we put this stretchy film stuff around the closed dish to limit airflow (can't remember what it's called, but you can see it in the first 2 pics).

-Ben
 
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Nice! I've got a buttload of all that TC stuff out in my garage I just never got it all going. Dreams are bigger than my space.

In the old days people used to try and sow orchid seed on the parent plants' pot and apparently got some success as the very old orchid books (from before TC) say this is how it's done. That or by using "expired soil from repotted orchids". I wonder how this would work today since parent plants are likely all from TC there would be almost no chance of a plant bringing stray fungal mycellium with it. Perhaps, like the unknown mosses, ferns and mushrooms that occasionally pop up in containers the correct mycellium occasionally just happens upon an orchid in a pot?
 

Drosera36

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the stretchy stuff was parafilm.

Ah yea that's what it's called!

Well I came back today to replate them and separate out some of the bigger guys.

Here's one of the larger ones still in its plate:
DSCN7557.jpg


Here's the biggest one in its own tube; probably its final home before we deflask it:
DSCN7561.jpg


And here's what I did with all the rest of the little ones (there's like 50 of 'em):
DSCN7560.jpg


-Ben
 

Drosera36

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That's impressive. As one who failed miserably the only time I tried anything comparable, I know how easily things can go wrong.

Haha thanks man. So far, nothing's gone wrong with any of the stuff I've done, but like I said before, their setup is pretty high-tech and clean.

-Ben
 

Drosera36

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Hey guys

Well I came to visit the plants again, and they've definitely gotten a lot bigger! Most have roots forming, and the largest one has a really big one. They don't seem to enjoy growing their roots down into the gel, however.

When I went there, I saw that one of my rectangular containers was missing; the professor wasn't there, so I assumed she threw it out, maybe because of mold forming. I'll have to ask her later. I still have way too many seedlings, anyways..

Dave, when should I start deflasking them?

Here's the smaller ones (the box is oriented in the same way as it was in the picture in the above post, so you can compare the growth of the individual protocorms over two months):
DSCN8242.jpg


And here's two of the larger ones:
DSCN8244.jpg


As you can see, they like to grow their roots vertically:
DSCN8249.jpg


Also, not an orchid, but still pretty amazing:
DSCN8245.jpg


It's a Stapelia gigantea, and it smells like rotting fish! haha

-Ben
 

Chomp

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Wow. :0o:
Is the Stapelia gigantea one of those plants that uses its smell to get animals to come to it, and then the animals get its pollen all over themselves and spread it to other plants?
 

Drosera36

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Haha yep, it smells like dead animals so that flys and other insects that enjoy carrion come and pollinate it. Apparently some flies like it so much, that they lay their eggs on it...:p

-Ben
 

Ant

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awesome, its great to see how they go about propagation of orchids by seeds. Cool flower too.
 

Drosera36

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Can anyone tell me what this is??

DSCN8747.jpg


They've been out for some time; however, I had not taken care of them for the several weeks that they were acclimating. I left them for care under the professor, but I wonder if they had been under rather wet conditions.

No fungicides were given to them after removal from the flasks, and I really freakin hope that this isn't related.

Thanks!!
-Ben
 

Drosera36

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Here's some more pictures to help with identification:

DSCN8751.jpg


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There is an unnatural sheen and rippling on the top surface of the leaves, as well as these strange light spots. This condition could only have arisen within the short time of about 4 weeks after they had been taken out of tissue culture.

Could this also be just sunburn? I have no idea as to how much light they were exposed to during those weeks, as they were not under my care.

Thanks so much!
-Ben
 

Drosera36

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Ok well sorry for all these posts, but I've asked a few people and they all seem to think that it's caused by a virus, even the professor. I could try to send in samples to a lab for ID, but really I don't want to go through all that trouble.

There are like 2 small ones that may not be affected, so I'll keep them isolated for a while.

Basically the lab there is right next to this big greenhouse full of neglected orchids, and I found thrips in the baby orchids' soil. So I figure they transmitted some virus to my babies. Well that was a total waste...haha.

Anyways thanks
-Ben
 
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