Small tissue culturing success

[dionae]

Thanks NYPN! I hope I can get them deflasked successfully. I'm deflasking some typicals atm. I've lost 4 of 18 so far. I'm going to let the Dentes get a lot larger before deflasking those. Thank you for stopping by!

 

[GregNY]

Freakin awesome job dionae. Very impressive stuff your able to do there. Not knowing much about this in general I still had some concern as to how you would transplant them out of these containers. What method(s) of deflasking do you follow??

 

[dionae]

I've only tried to deflask VFTs once and thats atm. I have them in a tupperware container. For the first week I kept the top on most of the time just opening it a couple times a day to give them fresh air. Then I gradually left the top off longer and longer. Now its a couple-few hours on, couple-few hours off, spraying them with RO water when the top is off every so often.

I'm just a noob so I might not be doing something right. Any suggestions are welcome. Thanks for stopping by GregNY!

 

[Sig]

AMAZING! I was under the impression that TC took a long, long time to get the hang of, with contamination being a huge issue. Whatever you're doing to get success with so many things so fast, keep doing it! It's really cool to see how the different plants grow and develop so quickly.

This pic is AWESOME: herbivorous omnivorous carnivorous capitalistic vft (Which looks like a crocodile, BTW.)

 

[dionae]

Thank you sig. I've had plenty contams...I just dont post pics of them. Thanks for the kind words and thanks for stopping by!

---------- Post added at 06:14 AM ---------- Previous post was at 06:02 AM ----------

Hello everyone! Not much going on with my TCing lately but I have a few things to share pics of. Dentes are getting huge. Dews are ready for a replate. Just flasked up some heliamphora seed.

H. heterodoxa x minor

Dente

Random VFTs including a mutant

D aliciae

D capensis all red

 

[dionae]

New Flasks

Thought i'd share a few pics of some recently flasked and replated stuff.

VFT seedling germinated in vitro. Freshly replated onto new media.

Dente explants separated and put onto new media

Typical VFT explant

S flava rubricorpora x S flava Ornata

N kohola x rokko x

S purpurea

S oreo

 

[Rob]

Wow!!! Very nice cp tc varieties you got going on. What is your sterilization procedure? I would love pics of your tc area

 

[amphirion]

very nice TC work sir....i do hope that you're able to induce a callous with the heliamphora.....that would be amazing. maybe with the nepenthes as well!

 

[RL7836]

Wow!!! Very nice cp tc varieties you got going on. What is your sterilization procedure? I would love pics of your tc area

Great work! I'd be interested in seeing your work area also. What do you do to control airflow & mitigate contamination?

 

[cpsammich]

That's amazing
What happened with the ceph explant?

 

[dionae]

What is your sterilization procedure? I would love pics of your tc area

This time around I just used MS with PPM to sterilize everything. Now, I got a few contams but so far its working out okay. I do not have a LFH and I do all my TCing in my living room.

i do hope that you're able to induce a callous with the heliamphora

The heli seeds got contaminated. I even tried taking them out and resterilizing them without success. Its part of the reason I stopped using bleach and peroxide to sterilize explants.

What do you do to control airflow & mitigate contamination?

Make sure theres no fans on and the air is still as possible, lysol and PPM.

What happened with the ceph explant?

It died. Didnt get any contams just never grew and finally turned black. Waiting on my cephs to start putting out winter leaves then i'll be trying again.

Thanks everyone for stopping by and commenting!

 

[Sig]

Still very impressive! In the living room really counts as "Home tissue culture". Nice work! Have you ever gotten nep seeds to grow out?

 

[dionae]

Thanks Sig! Nope, never had any nep seeds germinate. This is only my second try though. I've mostly just concentrated on VFTs.

 

[dionae]

Small Update

Had a few sarr seeds germinate, along with lots of contamination. Sarracenia seed seem very difficult to sterilize. The flytraps that I replated are really growing fast...they were needing replating bad. Nothing on the nep seeds so far but no contams so my fingers are crossed.

Dente explant

Typical VFT explants, a green and a red

S flava var rubricorpora x S flava var ornata in agar

Same cross in gelzan

 

[fdfederation]

Very nice.

 

[BigBella]

Nice effort all around!

 

[dionae]

Thanks fdf and BB!!!

 

[gill_za]

dionae,

I am reading up on the TC in an effort to learn as much as I can to try it in the near future. I was curious if you could clarify a couple of things a bit:
So you have used Murashige and Skoog media with PPM in it.
What was your sterilization procedure (lysol and PPM)? Could you please describe it a bit?

Thank you very much in advance!

 

[sss]

Wow nice pictures. Do sarr seeds grow faster when grown this way?

 

[dionae]

What was your sterilization procedure (lysol and PPM)?

The lysol is for cleaning surfaces etc. I use 1/3 MS and a PPM solution to sterilize the explants. Last run I used 2ml PPM in 4 oz of 1/3 MS. Not too many contams...actually none on the heli seed and ceph explants which were the last things I plated. Even have a callus forming on a ceph leaf finally. 1ml BAP per liter of media is what I used to initiate the callus.

Do sarr seeds grow faster when grown this way?

Not for me but they divide like crazy.