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By Doomsday
Posts:  621
Joined:  Sun Mar 07, 2010 3:11 pm
#278506
Hey guys,
I'm back from my ~3yr hiatus, and finally got my TC stuff set up again, this August. This time around, I made myself a plastic/pvc glovebox, and have had a few small successes so far. I'm going to share some of my TC culture progress here, so pull up a seat and enjoy the ride. Feel free to comment/ask anything you like, but please keep your arms and legs inside the vehicle at all times, because these plants DO bite.

First up is a dionaea seed callusing up nicely, getting ready to be turned into dozens/hundreds of babies.

Dionaea muscipula "typical" seedling on callusing media:
10/26/16
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10/30/16
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11/2/16
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Next, we have some Drosera nitidula gemmae that have started growing. They haven't been in vitro for very long, though, so I still haven't counted out contamination. There is a slight "wisp" of cloudiness under the surface in one area where the media is a bit messed up from my pipette I believe, but it doesn't appear to be growing like my other bacterial contamination, and isn't touching any of the gemmae.

D. nitidula on plain MS media (hard to get a good pic of these):
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By Greenthumbs Garden
Location: 
Posts:  644
Joined:  Mon Jul 27, 2009 10:15 pm
#278698
Smooter80 wrote:Pretty sweet. This would be a great one for a time lapse. They can grow very fast.
Im with Smooter, a time lapse would be awesome
By Doomsday
Posts:  621
Joined:  Sun Mar 07, 2010 3:11 pm
#278731
Dr GreenThumb wrote:
Smooter80 wrote:Pretty sweet. This would be a great one for a time lapse. They can grow very fast.
Dr GreenThumb wrote: Im with Smooter, a time lapse would be awesome
Hmm, that sounds like it could be pretty awesome. I haven't seen any in-vitro time lapse videos yet.

I just replated the two seeds in that jar, into their own jars, on clear media. Hopefully they remained sterile.
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It's insane how quickly these things grow! I can notice a difference if I look at it in the morning and then again at night. The leaf isn't touching the seed coat anymore, like it was this morning. It's standing up now. I think the plant will probably be completely out of the seed coat within 2-3 more days, tops.
Last edited by Doomsday on Sun Nov 06, 2016 5:04 am, edited 1 time in total.
By Doomsday
Posts:  621
Joined:  Sun Mar 07, 2010 3:11 pm
#278733
Smooter80 wrote:I had one grow a .25"+ radicle the same day it germinated. The cotyledon won't completly come out of the seed. They seem almost useless.
Awesome! It's definitely growing faster than I expected. Can't wait to see some dewy leaves in there! Interesting about the cotyledons, too.
By Doomsday
Posts:  621
Joined:  Sun Mar 07, 2010 3:11 pm
#280221
I just got lights set up, but before that, this guy had a little while to get super etiolated lol... Curious what comes of it.
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Now the setup is much better:
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Got a second droso going. I'm curious to see how it does with the lights, and starting out completely submergd. I think it'll look much more normal than the other:
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D. nitidula:
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By Doomsday
Posts:  621
Joined:  Sun Mar 07, 2010 3:11 pm
#280763
Does anyone have any idea why my dionaea and D. natalensis calli are darkening/browning? Based on the timing, I am thinking it has something to do with the installation of the lights. I put a few callus cultures (the two darkening ones, and one D. regia one that looks fine) in my dark cabinet, and left a few other callus cultures under the lights, to see if there is any changes this week.

Do you think I need to cut off the brown part, to prevent the oxidation/tissue death from spreading, or do you think it will be ok, now that they are in the dark. Also, can they be grown in complete darkness?

Another possible solution to the problem is the addition of antioxidants (citric/ascorbic acid) into the media.

Image
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By xr280xr
Posts:  2807
Joined:  Wed Jun 22, 2011 3:29 pm
#280766
Great shots! That media is much more camera friendly than agar. Interesting to see the VFT callus right out of the seed. Is this your own media recipe?
By Doomsday
Posts:  621
Joined:  Sun Mar 07, 2010 3:11 pm
#280768
xr280xr wrote:Great shots! That media is much more camera friendly than agar. Interesting to see the VFT callus right out of the seed. Is this your own media recipe?
Thanks! I've been using Gellan Gum lately, and I like it so much more than agar. For starters, I can detect contamination very quickly, sometimes within 2-3 days, and it also maintains clearness, even when it is firm enough to knock a vial over without disrupting the gel. I also find that it doesn't get stuck to the glass as easily as the agar, which keeps the sides clearer for pictures.

I'm just using 1/3MS, and testing out different hormones and combinations of them. It is pretty wild to see them germinate and then swell up instead of growing into normal seedlings.

I just want to know why they are turning brown... If it's a light issue, then that's the easiest to solve. If it is a media issue, that's a bit more challenging.

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