My TC adventure

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tokol0sh

 
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My TC adventure

by tokol0sh » Wed Dec 12, 2018 5:08 am

Hi All,
I am looking at restarting my CP collection (between a new dog, chickens, university and a few years of neglect al my CP's died). I have also tried tissue culture at home before (growing potatoes) and that was reasonably successful. I figured I would try putting some CP's in tissue culture and repopulate my garden. I mainly interested in Dionaea, Drosera and Sarracenia varieties.

It has been a while since I last tried TC (~6 years) and am just about to place an order with PhytoTechnology. I am looking for some feedback on my order list (basically, have I overlooked anything). I have also added some growth regulators for experimentation, following this paper: http://sci-hub.tw/10.1023/A:1021203811457

The current order list:


I intend to dilute my MS media down to 1/2 or 1/3, I am a bit unsure on some terminology. Some recipes call for 1/3 MS with full vitamins, others say 1/3 MS with vitamins and others will just say 1/3 MS. If I buy MS with vitamins and dilute it 1/3, do i need to add more vitamins to bring it back to "full" vitamins? If so then would something like this be needed:
(M557) Murashige & Skoog Modified Vitamin Solution (1000x)

I am located in New Zealand, so shipping is pretty expensive. I want to avoid putting in a second order if I can.

Any help appreciated
Byron
Last edited by tokol0sh on Tue Feb 12, 2019 10:26 am, edited 1 time in total.

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Re: Getting Started

by TCGator » Tue Dec 18, 2018 8:04 pm

Thiamine and myo-inositol are the two most common vitamins in TC. I know that thiamine should be available at your local pharmacy/store. What about myo-inositol?

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tokol0sh

 
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Re: Getting Started

by tokol0sh » Fri Dec 21, 2018 11:05 am

Thanks for the reply. I added the vitamins to my order.

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Re: Getting Started

by tokol0sh » Wed Jan 23, 2019 1:06 pm

Just a bit of an update:
All my supplies are in and I have managed to repair an older autoclave from my fathers work (it had a loose connector and a blown fuse).

For my first batch I mixed up 175mL of 1/3 MS with vitamins (I guess the vitamins are at 1/3 now too?). This is enough to do 7 containers with 25mL each. I was going to add PPM but the volume needed was a bit small for me to measure accurately.

I dont have a laminar flow hood so just used an old wooden shipping crate I had around and lined it with plastic and sprayed it down with bleach about (with everything inside) about 30min before starting. Sterilized 14 flytrap seeds individually, I will definitely be using a coffee filter or similar next time...

The seeds were placed in the containers, 2 per container. Some seeds were place on the surface others submerged in the gel. Not sure if that matters?

Now to wait and see if there is any contamination and then if the seeds germinate.

Image


Image


Image

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tokol0sh

 
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Re: Getting Started

by tokol0sh » Mon Feb 11, 2019 8:21 am

About 2.5 weeks in, and no sign of contamination yet. No seeds have germinated yet. Got the seeds about 18" below a "300W" full spectrum LED grow light with a temperature of around 25-30C. Hopefully the seeds do alright...

I also mixed up a new batch of media:
1/3MS with full vitamins (I might have added too much of the MS vitamins solution so it might actually be 1.25
Vitamins)
1mL/L PPM

I made up a sterilization mix with 4% PPM and 1/3MS. This was used to try to sterilize some VFT cuttings, a sarracenia seedling and some drosera seeds. So far this is looking good. The VFT cuttings are still green (they have gone black where they were cut, but I suspect that was because they were cut with scissors and not a sharp scalpel), the sarracenia seedling has grown 2 new pitchers in about 2 weeks and the drosera seeds germinated in about 1 week. So far, I cant see any contamination.

This how that experiment is going:
Image



Here is my grow chamber:
Image

I will keep posting updates as things progress.

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tokol0sh

 
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Re: My TC adventure

by tokol0sh » Sat Feb 23, 2019 4:03 am

Still no sign of germination...
The experimental container containing plant cuttings and drosera seeds had an algal bloom, not sure where it cam from. I added the PPM sterilization mixture to the container and let it soak for a few hours. It seems alright now. The flytrap cuttings look like they might be forming calluses, one of the leaves has a few red bumps forming. The other has a red lump forming between the trap and the leaf.

I was reasonably happy with the PPM sterilization, so I used a 4% mixture to sterilize about 40 more seeds and have placed those into the new media I had prepared. So far so good, no contamination.

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Re: My TC adventure

by Matt » Tue Mar 05, 2019 10:34 pm

Well done! Any updates?
My Growlist
Eat Plants! Leave the meat for our pet plants :)

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Re: My TC adventure

by jasonh » Wed Mar 06, 2019 3:24 am

PPM definitely helps a lot for home TCers! I recently ran out of PPM and solely use NaDCC; needless to say the contamination rate has kicked up a notch. For nepenthes seeds I was relying on 500ppm NaDCC + 4% PPM and not sure if things would work out without it...

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Re: My TC adventure

by tokol0sh » Tue Mar 12, 2019 2:14 pm

As if last week, the VFT seeds still show no sign of germination (1st batch sterilized with bleach, second batch sterilized with PPM). I am away for the next 3 months, so probably wont be able to provide many updates, but I have left the lights on hoping something happens while I am gone. None of the seed cultures show any signs of contamination, which I guess is a good start. The culture vessel with the plant clippings now has what looks like algae (its almost black under my LED grow light and bright green under white light) growing inside. I tried to clean it out with a strong dose of PPM, but it didnt seem to do much. The VFT cuttings looked like they were starting to form calluses but the algae has taken over. I tried to rescue the calluses and the pitcher, but unsure if I successfully cleaned all the algae off. I'll try get some pictures of the current progress soon. When I get back, I will try planting some seeds on sterile paper and in some sterile water and see what happens.

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Re: My TC adventure

by sheppyfire87 » Sun May 19, 2019 12:20 am

Looking forward to an update, everything looks great so far. I cant wait till i can give this a try.
Best of luck!

Sent from my LM-Q710.FG using Tapatalk

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tokol0sh

 
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Re: My TC adventure

by tokol0sh » Thu Jun 20, 2019 4:59 am

I have just got home after being away for 3.5 months.
The VTF seeds have not germinated and in some containers the media has dried out. There is no contamination, so I guess thats a start...

The containers that had plant clippings all have contamination that has taken over, I'm not too surprised by this as I wasn't very careful with sterilization.

Any ideas on how to get the VTF seeds to germinate? I still have a few seeds left to try something else

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Re: My TC adventure

by Copper2 » Thu Jun 20, 2019 5:49 am

Ga3?
Don’t kill the insects. I need them to feed my plants

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tokol0sh

 
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Re: My TC adventure

by tokol0sh » Fri Jun 21, 2019 10:52 pm

Could it be temperature?
The temperature inside my growing enclosure was around 22C when I checked on them when I got home. Ill try heating them up a bit and see if anything happens...

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Re: My TC adventure

by tokol0sh » Fri Aug 30, 2019 5:34 am

After getting my growing enclosure temperature controlled at around 23C I tried with the last of the VFT seeds I have (from the flytrapecare store). Its been about 2 months and nothing has started growing yet.

My outdoor VFT's are just starting to grow again as spring is around the corner, a flower has started to sprout. I will try using the flower to start things off.

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Re: My TC adventure

by bananaman » Fri Aug 30, 2019 11:34 pm

For me, VFT seeds in TC took a fair amount longer to sprout than those germinated normally, using 1/3 MS, 1% sucrose as my media. I don't exactly remember, but it was about a month longer than seeds from the same batch ex vitro. I didn't bother doing anything to boost the temperature -- my house is usually somewhere in the range of 21-24C. If they're from FTS, the seeds shouldn't have that much issue sprouting.

I believe I just did 10% bleach for 10 minutes, followed by 3 rinses of 3 minutes each in sterile water. What was your sterilization protocol?

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