TC Lab (Tissue Culture) - Dionaea seeds

[Melle]

Helle all,

This weekend I wanna try to use some Dionaea seeds for my TC lab.

Some one have experience with this?

I have this:

http://www.ebay.com/itm/360582004583?ssPageName=STRK:MEWNX:IT&_trksid=p3984.m1497.l2649

I will put 1 seed in every test-tube.

- What is the best place to do this? Inside/outside?

- Do I need some extra light for this?

- What temperature?

- What humidity do I need?

Thanks all!

Regards,

[Ian Salter]

There is a lot to consider and I think you would do far better to look up tissue culture threads on here and other sites.

If you are using Agar with nutrient then everything will have to super sterile and I doubt those test tubes are sterile but you never know.

If you are just sowing In vitro then perhaps at least sterilise any media first, keep indoors under lights and at about 18 Deg C and the closed tube will provide full humidity.

Remember damp off too, gradually introduce seedling back into a drier atmosphere or they will wither and die.

I tried tissue culture years ago and even though I went to great lengths to keep mold spores out and bacteria they all eventually failed.

I did however grow some wonderful moulds and some funky coloured bacteria's.

Do your research and be very prepaired.

[FlytrapCare]

I remember when I first tried tissue culture of Dionaea seeds nearly 5 years ago now. As Ian stated, success with tissue culture requires a lot of time spent reading and preparing. Judging from the questions you're asking, it may be that you've not done enough reading yet.

Have you read this article?:

http://www.flytrapcare.com/tissue-culture-basics.html

Information about temperature and lighting can be found in that article. Humidity isn't a concern at all because it will always be nearly 100% in any vessel (such as your test tubes).

And there is an entire forum dedicated to tissue culture here:

http://www.flytrapcare.com/phpBB3/tissue-culture.html

If you read through threads there, you could learn a lot.

[Melle]

Thans for the answers! I will read the articles.

Hope to get smarter. But you guys say it is very difficult?

[Ian Salter]

Keeping things sterile is the hardest and most important part and requires a strict regime.

If you are wanting to do actual proper Tissue Culture then building a laminar flow cabinet with a HEPA filter to screen out all particles is essential.

You also have to spray bleach the entire inside prior to working and all tools need to be flame sterilised or sterilised in a pressure cooker and only opened in the cabinet just before use.

You also need to sterilse any tissue or seeds with mild bleach and even when you have done all this one mould spore or a slight touch of a not sterile finger and it's a failiure.

But don't let this put you off as i know someone who set his vials/jars over a steaming pan keeping the jars upside down at all times and had good results.

Study and understand first though so you know the principles and what to look out for either way it is an experiance and worth trying.

have fun and good luck.

Show us how you get on.

[Melle]

Hello,

I have read the site's.

Very instructive!

But I need many materials en stuff I don't have. (But I can buy)

The sterilization of the materials should be fine.

But I hear sometimes that Tissue Culture also has to work in this way:

Use fresh seeds, sterile test tubes and distilled water.

One or a few seeds per test tube.

18-20 hours of light per day.

And then the seeds should germinate well.

Is this a myth, or just lucky?

Melle

[Ian Salter]

fresh seed will always help, Sterile test tubes are a must or you must sterilise them and distilled water containes no impurities.

A few seeds in each will guarantee germination as a single seed that hasn't germinated is a big waste of effort.

All correct but not sure if light other than a heat source will aid germination but once they have germinated yes.

Keep us posted when you get started we are all interested.

[Melle]

Thanks.

So now go do this first:

- I will sterilise the test tubes.

- Look for FRESH Dionaea seeds. (Do you know a good place to buy?)

- Search for the best media for in the test tubes. ( vitamines and stuff) -- Tips are more than welcome!

If I am ready I will post some photo's!

Edited March 31, 2013 by Melle

[Melle]

Ingredients I need for Dionaea medium is this:

List of ingredients for preparing 1 liter of Dionaea muscipula (Venus fly trap) medium:

• 1/2 MS medium with vitamins (one half of a 1 liter packet or 2.2 grams). Most Tissue Culture Techs uses 1/3 MS for Carnivorous plants with great results.
  
• 2 tablespoons of table sugar (about 30g)
  
• 1 ml of BAP (1 mg/ml) - leave this out for seeds. It's even optional for explants. Dionaea explants will form callus without it.
  
• 1 ml of PPM
  
• Agar
  
• Distilled water
  
• Vinegar and baking soda
  

For medium I want to buy this:

2 MS, 2 Agar, 10 ml PPM, No Hormones

Two 1-liter packets of Murashige and Skoog basal medium with vitamins, two 9-gram packets of agar, 10 ml PPM

Everything I need. But ingredients list is 1/2 MS. What I buy is 2 MS. Just pick a quarter of the 2 MS, right?

[blaasjeskruid18]

Hello Melle,

maybe it is a good idea to go to the intruduction topic

good luck with your Tissue Culture!