FlytrapCare Carnivorous Plant Forums

hey guys,
I'm working on my first micropropagation experiment and atm I'm learning about Tissue culture, the science behind it and I'm also setting my own mini lab at home which I'm really excited about but being new at this I have a lot of questions and I hope you guys can help me with them

1- why is contamination such a big deal with tissue culture? normally a seed will grow in a far more contaminated environment(soil) and yet they manage to survive, is it because callus lacks the protective coatings the seed have ?
2- can I start my Tissue culture with a seed? will it have an advantage over other types of explants?
3-will using antibiotics and antifungal agents increase the success of my experiments? (also if possible, what are some common microorganisms that contaminate the Tissue culture? )
4-Can I use a Microwave to sterilize my media instead of a pressure cooker?
5-how do I use a liquid media instead of a solid one? I'm wondering about this because it seems to me that need to periodically replace the media would be much easier with a liquid one instead of a soild one, you simply drain and refill.

that's all I have for now, thank you

1. Contamination in a more concentrated growing environment would cause the very rapid growth of fungus/disease. Kind of like how they grow cultures of bacteria/fungus in labs, the medium is such that it increases the rate of growth. So if it's contaminated, mold/bacteria/fungi would go buck wild.
2. I don't know
3. If your environment is sterile, you shouldn't need either of those.
4 & 5 - best to let someone else answer these as I am not experienced in TC for plants, I have only seen it done in the lab at my college for bacterial cultures.

raedk wrote:2- can I start my Tissue culture with a seed?

Yes.

raedk wrote: will it have an advantage over other types of explants?

Yes, seeds are easier to sterilize over explants. But you can't get an exact clone of a plant if you use seed. So, for example, if you wanted to propagate B52, you'd have to use an explant. If you used B52 seeds, you wouldn't be propagating B52, but rather B52 x B52, which is genetically different than B52.

raedk wrote:4-Can I use a Microwave to sterilize my media instead of a pressure cooker?

Yes, though I believe it is more challenging to get right. I've never done it myself, so I can't comment.

raedk wrote:5-how do I use a liquid media instead of a solid one? I'm wondering about this because it seems to me that need to periodically replace the media would be much easier with a liquid one instead of a soild one, you simply drain and refill.

I've never used a liquid media, so I can't comment.

I can comment on the microwave. The concept of a microwave is different than a pressure cooker, a microwave will bombard the media with a certain low frequency of light waves and bring it to a boil but you will most likelyevaporate more moisture and end up with a more concentrated medium, furthermore, I have no idea what the effect will be on the contents of the media, you might burn it and destroy important ingredients... A pressure cooker (working on the same principle as an autoclave (big pressure cooker I use in the lab)) seals and therefor the pressure rises, and If you know your physics, this increases the boiling temperature of your media, hence less contaminants survive... The spores from fungi are very resistant to extreme envoironments, so are the spore bodies of certain bacteria. A pressure cooker raises the boiling temp above the normal 95degCelcius to 200 or above, and even at these higher temperatures the media needs to "boil at optimum temp for 20min, so take 20min to reach optimum temp and 20min to cool down, you need about an hour to kill all contaminants effectively, not sure this is a viable method in a microwave as you'd need to increase the time even more, and as I said, you arew going to dry out and burn your media... The pressure cooker is your best option...

Just to add to backer, I would be concerned with the medium boiling over in the microwave - ever tried to make oatmeal in one? Messy, and uneven cooking. Also that since microwaves vary in power, it might take longer or shorter to boil - best bet is a pressure cooker, 15 psi, and they aren't really that expensive if you don't need a huge one.

raedk wrote:hey guys,
4-Can I use a Microwave to sterilize my media instead of a pressure cooker?

Both work great one you established your media preparation methods. Microwave may take longer to get it desired level, then you repeat the same procedure.
Recently, I am only using pressure cooker. However I can say that best cultures I have observed were the ones in my microwaved jars/vessels. It should not sound that it is better than other method. Just sharing my experience about which it can be done and the results are good.