FlytrapCare Carnivorous Plant Forums

Jens, I know you asked this question a while back on the hometissueculture mail list, and I know that now your flower stalks are forming callus, so I was wondering how long it took for it to happen.

Also, where did the callus start? Did it start on the end of the piece of stalk or somewhere in the middle?

Matt,
Just a question,but what's callus?

Callus is the knobby growth that is exhibited by tissue as it's starting to grow. See this photo:

aka a 'strike'?

Adam wrote:aka a 'strike'?

Yes, similar to a "strike" when you're trying to use flower stalks or leaf pullings to start new plants. "Strikes" are generally just a point of new growth, but in tissue culture, usually a mass of cells is formed and it's just a glob. That's callus. You can tissue culture almost any kind of plant even if they won't vegetatively propagate easily as Dionaea do. Most plants won't "strike" from leaf pullings.

Callus is normally induced through use of hormones called cytokinins. The two most commonly used cytokinins are Kinetin and 6-benzylaminopurine (BAP). I think the photo I posted above from Jens tissue culture was taken after a piece of tissue was transferred from BAP media to hormone free media. After enough callus has formed, you need to transfer the tissue to hormone free media to promote normal growth.

Wow, another transfer, another chance to contaminate Seems like a lot of work!

Hi Matt

it takes about 5 weeks to start callus. I just transfered some stalks to new plain media

Adam wrote:Wow, another transfer, another chance to contaminate Seems like a lot of work!

Adam, you need to have the right equipment, then you dont have to worry about this.
When i started with this i allways make it fast and got many contaminations.
Now i have build a flowhood and put the parts/plants out cut and make other things and put on new medium and dont need to worry about how long time it takes or something....
All who want to make more TC need a flowhood.... It is not so much expensive and you can build it easy at home. It cant go better and easier

Jens

nepaholic wrote:it takes about 5 weeks to start callus. I just transfered some stalks to new plain media

Great, I'll be patient. What temperature do you keep the cultures at? I'd like to see some photos of the flower stalks when you get a chance Jens.

Adam wrote:Wow, another transfer, another chance to contaminate Seems like a lot of work!

I've had extremely good luck working in my 30 gallon aquarium laid on its side. I run a HEPA filter for a day or so in the room I do my work in before I start, then I come in and spray the aquarium down with 91% alcohol. I've yet to get a contamination from air borne stuff. The only contaminations I've gotten have come from tissue that wasn't thoroughly enough sterilized.

Hi Guys

sorry for the late answer. Here some Pictures of callus on a stalk.

Awesome Jens! Thanks!
A couple more questions:
1) How do you cut off your flower stalks? Mine always turn a bit black on the end, but yours don't appear to at all. Also, I know I've asked before, but what was your sterilization technique? Was the one in the photo done with an alcohol dip, followed by 3 to 5 minutes in hydrogen peroxide and then 10 minutes in 10% bleach?
2) How much BAP are you using in the media to induce callus?
3) What temperature do you keep them at?

Matt wrote:Awesome Jens! Thanks!

Thank you Matt

Matt wrote: A couple more questions:
1) How do you cut off your flower stalks? Mine always turn a bit black on the end, but yours don't appear to at all. Also, I know I've asked before, but what was your sterilization technique? Was the one in the photo done with an alcohol dip, followed by 3 to 5 minutes in hydrogen peroxide and then 10 minutes in 10% bleach?

Matt, Yes i allways make a Alcohol dip for Explants about 30-60sec...
5min H2O2 followed by 7-8minutes Bleach and 2times rinse for about 5min each rinse.
After this i cut off the black Ends

Matt wrote:2) How much BAP are you using in the media to induce callus?

I use allways 1ml/l

Matt wrote:3) What temperature do you keep them at?

I have my cultures at about 22-25´C

Regards
Jens

Perfect! Thanks for all of the answers Jens!

Yes, thank you so much Jens for giving the forum advise. Much appreciated!

hb