Nepenthes Tissue Culture

[TranMinh]

Hi amitzauber,
I buy a mix from a garden shop they sold MS mix for growing vegatable. Here is some information for TC
http://www.omnisterra.com/botany/cp/slides/tc/tc.htm
http://en.wikipedia.org/wiki/Murashige_and_Skoog_medium
The T.C can make the plant 10x to 15x grow.
You should find some information in the internet.

[paphioboy]

Tissue culture is not easy to DIY for the average CP grower. To be successful, you must have a completely sterile environment. The reason is that agar is nutrient rich, so even a tiny speck of fungus/bacteria will contaminate the entire container/flask and kill all the seedlings inside. Most commercial labs have at least an autoclave, laminar flow hood with air filter and a shaker. It is much easier to send a piece of tissue to the lab and ask if they will culture it for you for a small fee.

Arvin, the apical meristem does not come from the callus. The apical meristem is taken from the adult plant. When chopped up and under the influence of hormones, it then produces the callus, which grow into many individual plantlets.

[paphioboy]

I was also asking about the practicality or sucess rate with using Apical Meristem from an adult nepenthes cutting, because so far I have read that usually cuttings are already contaminated by a lot of organisms that it is almost impossible to get a sterile sample to be able to do TC.

The apical meristem used should be relatively clean compared to the rest of the plant because it is hidden beneath all the other young leaves. If unsure of contamination, can soak in 5% bleach solution to surface-sterilise it before TC.

[arvin555]

Amitz you should try to order a kit from here:
http://www.kitchenculturekit.com/index.htm

That is where I got my kit, but I haven't had time to try it out yet, and so I am also still researching. Best bet to try first are VFTs and Droseras, Nepenthes apparently is hard to TC from meristem so I am asking help here in the forum about it.

It is a bit complicated as Paphioboy mentioned, but if you are the type that likes a challenge or to experiment that TC is for you. By the way I feel that the generalization that TC can make one seed into a thousand plants is a bit of an exaggeration. Well it is possible, but sure heck will need a long long time to get to a million. However it is true that if you do keep at it, you can grow a lot of clones from one meristem.

Paphio, the current suggestion is 10% bleach, but people still say they still get contamination, oh well, we'll see.

TTFN
Arvin

[paphioboy]

Bleach will only sterilize the surface of the meristem tissue, but if your air is still not clean and contaminated with fungal spores, it is no use..

[Robert]

for a fresh starter understanding TC can be confusing. I found a link from another forum which was very useful.
http://www.world-of-carnivores.com/tcathome.html

[Robert]

Originally Posted by arvin555

the current suggestion is 10% bleach, but people still say they still get contamination, oh well, we'll see.

TTFN
Arvin

10% was the acceptable level. when carrying out TC contamination at every level must be eliminated even after the work surface was sterilised. For example accidental drop of tweezer, forcep or cutter on the work surface they must be dip in ethynol (spirit), then place over the burner thus killing the foreign bodies.

[arvin555]

If I use IPA as a disinfectant I understand it's dangerous to have a burner in the box. Also I understand the PPM really helps a lot in controlling contamination?

Still thinking about where I will be able to do TC and if I will want to make a laminar flow hood or not bother anymore (DIY hood I mean).

I'd like to ask another thing though.. maybe this deserves another thread, but may I ask what is the recommended way of keeping the Tissue cultures? I mean.. a rack, with how much watts florecent lamp? how long photo period? Is it better to cover with glass or plastic to prevent possible contamination? etc etc.

TTFN
Arvin

[arvin555]

http://www.world-of-carnivores.com/tcprep.html