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Thread: Envenomated agar? Any one try this?

  1. #9
    OMG h8 pings MrFlyTrap2's Avatar
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    Quote Originally Posted by Pyro View Post
    AS for schedule of transferring, it depends on the plant. With those that produce a toxin it is usually obvious when it needs to be done because the media goes from clear to straw to yellow to orange to brown. The orange/brown level is where toxic effects begin to be noticed.
    VERY interesting! Thanks for sharing this. I had a culture of D. Capensis seedlings that I just let do their own thing for about half a year (or more) after taking what I wanted from the culture.

    That exact effect started happening. The media began to haze and spread as you indicated along with the color changing. I figured it was just contaminated but still it was different, the spread was -through- the agar, not mold puffs or anything growing -on- the agar. The darker stages began to kill the plants in the culture.

    I was a bit fuzzy for the need of doing sub cultures. Transferring the plant into the exact same media wasn't making a lot of sense to me. But that toxin info was new to me!

    Nate
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    N=R* fs fp ne fl fi fc L Pyro's Avatar
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    @ Swords:

    You can see the colour change in charcoal containing media but it is more difficult. I find that in charcoal media the charcoal will settle, not a huge surprise as the agar is still a "liquid". As such there is usually a "clear" zone that is about the first 1-3mm of depth. You have to view it at an oblique angle but you can see it.

    I have a few cultures that tox'd out, I'll try to photo them so you can see what it looks like.

    An alternative that may work for you is to make an expendable culture in plain media that you can watch for colour change and when it reaches the yellow stage then switch all the flasks.

    Nate,

    Never heard of capensis being one of the toxin makers, course I have never heard of anyone TCing capensis so that may be why. But in my experience no Drosera does the tox thing.

    The hazing sounds more like a bacterial swarm to me but I could be wrong. There is also a strange algae that can swim through the media as well.

    The transfer thing is to keep the plants happy. They do deplete the resources in the agar with time so you need to transfer for that reason alone. And some plants just divide like gang busters even in basic growth media (N. rajah is notorious for this, which is why Rob has 20,000 of them...) so you need to just divide and divide and divide.
    'My love was science- specifically biology and, more specifically, when placed in a common jar, which of two organisms would devour the other.'

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    OMG h8 pings MrFlyTrap2's Avatar
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    Quote Originally Posted by Pyro View Post
    Never heard of capensis being one of the toxin makers, course I have never heard of anyone TCing capensis so that may be why. But in my experience no Drosera does the tox thing.

    The hazing sounds more like a bacterial swarm to me but I could be wrong. There is also a strange algae that can swim through the media as well.

    The transfer thing is to keep the plants happy. They do deplete the resources in the agar with time so you need to transfer for that reason alone. And some plants just divide like gang busters even in basic growth media (N. rajah is notorious for this, which is why Rob has 20,000 of them...) so you need to just divide and divide and divide.
    Pyro, I'm sure it could have been a zillion different thing, and without actually examining the culture in person, or visual photos at the least I'm sure it'd be very difficult to find out what happened to it. I think that if anything you've given me some more insight to some of the 'other' reasons cultures go bad, and it seems worth a shot at least to post failed cultures for the experts out there that might be able to give me a better idea of why it crashed.

    And yes I do get snickers when I talk about culturing capes, but it's not that I want to raise a genetic army of D. capensis for a lifetime of profit... I want to learn about the process, the procedures, and tools needed in order to do TC properly. Capes are cheap (free!), seeds seem to always be viable, and I won't be heart broken if they go bad. Before I want to begin laying waste to Nepenthes seed, I want to feel a lot more comfortable with experience. 8)

    Nathan
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    rattler's Avatar
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    actually capensis seems like a good one to practice on to me.....if they dont germinate you KNOW its something you did, no second guessing weither it was the source material that was the actual problem........
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    I'm gonna be starting my TCing with droseras too (not capes specifically) but mainly cos they sell the seed on ebay by the lb. But i want t\o do all kinds of things just figure dews are easier going.

    Has anyone tried using a drosera leaf soaked in hydrogen perxiode then placed on agar to start droseras invitro that don't have seed available? I don't mind a big failure rate on such a procedure since it's only a hobby but if one piece stays clean and grows that one vial is all that's needed. I have started sundews on peat this way but not sure if it could be done clean enough to do invitro?

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    N=R* fs fp ne fl fi fc L Pyro's Avatar
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    Swords,

    I'll double check with my buddy but I think you need something stronger than H2O2 for TC sterilization. IIRC he used a bleach solution but I can not recall the %. It worked for both Drosera leaves/tubers/gemmae and Ping leaves
    'My love was science- specifically biology and, more specifically, when placed in a common jar, which of two organisms would devour the other.'

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    N=R* fs fp ne fl fi fc L Pyro's Avatar
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    Okay, sorry it took so long, had a million other things on my plate

    Here are some pics for you all

    Toxin accumulation in agar


    Charcoal agar, note how there is separation



    And to give you an idea of how different tranfer times can be...

    This flask was transfered into on 5/07


    And this one on 4/07


    So obviously different growth rates make for different frequency of transfer
    'My love was science- specifically biology and, more specifically, when placed in a common jar, which of two organisms would devour the other.'

    See You Space Cowboy

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    Neat stuff thanks for showing the pics!

    I suggested Peroxide cos I've heard bleach was sometimes too strong for seeds and merstems.

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