Jump to content

in-vitro culture of Cp's


lesthegringo

Recommended Posts

After my mixed success with germinating Nepenthes seeds in-vitro last year, curiosity has me now wanting to try cloning some plants. I have a number of VFT's and sarracenias, 1 heliamphora nutans and utricularia reniformis to play with to see what I can grow. I have to say that I don't remember seeing sarras, heliamphora or utricularia done this way before.

I will make this clear - I am doing this for fun and not for commercial reasons. If it works, great, if not oh well. I toyed with the idea of more nepenthes but they are so slow growing that I won't get to see them as adult plants as I probably will have to move on in three years. It would be quicker to make new plants from arial rooting the ones I have.

So over the course of the next week I'll prepare some media in some flasks, then hopefully next weekend take some tissue from the adult plants. VFT's should be easy - leaf pullings, but I am not clear whether the trap itself should be removed - anyone who has tried this who has an opinion is invited to comment.

The utricularia will be sections of the roots, and the heliamphora and sarras will be bits of the rhyzome, again unless someone can advise different. It won't be large scale, maybe 20 or 30 small flasks in total.

Should be a laugh, wish me luck!

Cheers

Les

Link to comment
Share on other sites

I would agree with Bux, I only have a little experience with CPs in tissue culture but it is usually easier with seed.

 

I've had some mixed results with Nepenthes seeds too, I think they have usually come into contact with a lot of contaminants before I've received them so have been not that easy to sterilise. They best results I've had are for Darlingtonia seeds from our own plants as you can dip an unopened pod in ethanol and flame it to clean the outside and the seeds inside remain sterile. I have sterilised some of these with bleach as well but usually get no contamination and very high germination rates anyway. I haven't tried this approach with Sarracenia but if you have some seed pods it could be worth a go.

 

The fastest growers I've had in culture are Drosophyllum, they filled the jars in months! Very easy seeds to sterilise too, what I found to be crucial though was after sterilising in bleach and washing with water to remove part of the seed case with a sterile blade.

 

For VFT explants there is a lot of information on the flytrapcare forum. I think you can use the traps or the flower stalks but I'm not sure how important it is to collect samples at the right time of year. I've had experience with other plants where spring is the best time to collect samples as the new growth has a large amount of meristematic cells. 

 

http://www.flytrapcare.com

 

Good luck and keep us posted on your progress!

Edited by Mark Long
Link to comment
Share on other sites

Thanks guys. Drosophyllum is famous for needing scarification to break the seed casing, gibberellic acid is another way of doing so. However, it's interesting that they seem to do well in-vitro.

My attempts at in-vitro for sarracenia seed were disastrous as the seed casings seem to trap contaminants so unless you have unopened seed pods (not in my case) you are on a hiding to nothing. I've got plenty of sarra seedlings grown from seed, this is more curiosity and the mad professor in me.

The fact is, this is more for fun than anything. I have over 150 dionaeas, so quantity is not the aim, it's 'can I? '. It was the same for the nepenthes seeds, and although not an unqualified success was better than I expected.

Hey, it's got to be a laugh, hasn't it?

Les

Link to comment
Share on other sites

My attempts at in-vitro for sarracenia seed were disastrous as the seed casings seem to trap contaminants

Sarracenia seeds are not difficult to sterilize, what protocol have you tried?

Link to comment
Share on other sites

I tried a dip for a minute in a weak bleach solution, rinse in distilled water, a soak for 5 mins in a hydrogen peroxide solution, rinse again then a quick dip in PPM before putting them down on the gelrite gel. None made it past a week! The fungal bloom was rampant on all the flasks

However I had no trouble at all getting them to germinate on normal peat / sphagnum mix

A local grower has some Drosophyllum Lusitanica seeds for sale, after your comment above i couldn't resist getting some to try

Cheers

Edited by lesthegringo
Link to comment
Share on other sites

I tried a dip for a minute in a weak bleach solution, rinse in distilled water, a soak for 5 mins in a hydrogen peroxide solution

That's not enough!

1/ NaOCl 1% (available chlorine), 5-10 min

2/ Sterile water, 5 min

3/ H2O2 3%, 5-10 min

And use PPM into the medium to avoid cross contamination.

Edited by bux
  • Like 1
Link to comment
Share on other sites

People aren't saying about the culture solutions here.

The standard culture mix for CP's is one third murashige and skoog plus sugar plus agar. Not sure about the MS spelling.

Nepenthes meristem is very difficult to sterilize because Neps have a large amount fungal thingies within the plant. Seeds are easier but are not true to type.

I have heard that VFT's are very easy, they can almost be put in a blender.

Good luck

Link to comment
Share on other sites

Join the conversation

You can post now and register later. If you have an account, sign in now to post with your account.

Guest
Reply to this topic...

×   Pasted as rich text.   Paste as plain text instead

  Only 75 emoji are allowed.

×   Your link has been automatically embedded.   Display as a link instead

×   Your previous content has been restored.   Clear editor

×   You cannot paste images directly. Upload or insert images from URL.

×
×
  • Create New...