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Invisiblelipa


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Trametes versicolor
    #8051170 - 02/21/08 09:53 AM (15 years, 11 months ago)

Does anyone have some up close photos of trametes versicolor on agar? I would really appreciate it.


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OfflineMycoAu
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Re: Trametes versicolor [Re: lipa]
    #8060522 - 02/23/08 01:45 PM (15 years, 11 months ago)

email me @

I'm currently growing T. versicolor on agar (its just reached the edges of the plate). If you want pics, I'll send them to you.


Edited by MycoAu (02/23/08 08:58 PM)


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OfflineRogerRabbitM
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Re: Trametes versicolor [Re: MycoAu]
    #8062283 - 02/23/08 08:54 PM (15 years, 11 months ago)

Why not post them here for everyone?
RR


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OfflineMycoAu
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Re: Trametes versicolor [Re: RogerRabbit]
    #8062319 - 02/23/08 09:06 PM (15 years, 11 months ago)

Wasn't sure how to do that - looking into it. I have JPGs for upload, but the site doesn't accept that. What to do?

Lipa, if you know how to load those pics, please do that for me and send me an email to tell me how you did it. thanks. Once I know how, I'll add more to it (I've got more to post than those four I sent you)


And thanks for the suggestion (to RR). Was probably being stupid and will pay for it. I forget about stuff like that because I don't really need to CMA for anything- or at least I didn't think so. We'll see.


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OfflineRogerRabbitM
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Re: Trametes versicolor [Re: MycoAu]
    #8062418 - 02/23/08 09:40 PM (15 years, 11 months ago)

At the top of the page where it says 'Welcome MycoAu' click the link for 'pics'. Next, click the link for 'upload' and you'll get a browse-to box where you can locate the pictures on your hard drive. Make sure they're 200K or less and they should upload fine. After that, you copy and paste the html code into your post and you'll get a thumbnail that links to the larger picture.
RR


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OfflineMycoAu
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Re: Trametes versicolor [Re: RogerRabbit]
    #8066299 - 02/24/08 08:58 PM (15 years, 11 months ago)

Just testing- appears fine in preview. Let me know if you want more pics. Tthese are the two best that I have IMHO, but I do have more.




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Invisiblelipa


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Re: Trametes versicolor [Re: MycoAu]
    #8068046 - 02/25/08 09:53 AM (15 years, 11 months ago)

I am afraid mine doesn't look the same. Roger care to shed some light. His looks a little weird to me. The mycelium of my clone is very leathery cottony





I will try to open one in front of the hood tomorrow if the pics are not good enough for you. Here are the ones I have anyways.

Cloned from this fruit.



Edited by lipa (02/25/08 10:00 AM)


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OfflineRogerRabbitM
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Re: Trametes versicolor [Re: lipa]
    #8068133 - 02/25/08 10:30 AM (15 years, 11 months ago)

Agreed. MycoAu's pictures look like they're contaminated with bacteria. You can actually pour some hot agar with gentmycin sulphate right over the existing growth and allow the mycelium to work its way to the surface, scrape the mycelium to a new dish and repeat. After a few times, it should be cleaned up.
RR


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semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
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Invisiblelipa


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Re: Trametes versicle [Re: RogerRabbit]
    #8068181 - 02/25/08 10:50 AM (15 years, 11 months ago)

You know Roger I have not ever tried to use that stuff, but would like to have some to try. Where might I get that locally in a place like San Diego. Thanks for the tek too!!  I appreciate It!:cool:


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Invisiblelipa


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Re: Trametes versicolor [Re: lipa]
    #8068193 - 02/25/08 10:57 AM (15 years, 11 months ago)

MycoAu. I think also that your cultures are contaminated. If you cannot manage to clean them up. Hit me up. I am not shy. If you want a culture, pm me. I made quite a few.


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OfflineRogerRabbitM
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Re: Trametes versicolor [Re: lipa]
    #8068291 - 02/25/08 11:40 AM (15 years, 11 months ago)

I doubt you'll find antibiotic agar anywhere except for mycology supply businesses. Sporeworks.com is a sponsor here and they carry it in stock.
RR


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semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
Thomas Edison


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Invisiblelipa


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Re: Trametes versicolor [Re: RogerRabbit]
    #8068397 - 02/25/08 12:19 PM (15 years, 11 months ago)

Oh! I thought maybe that was something you might be able to get from the vet or feed store. Thanks for the reply.


Edited by lipa (02/25/08 12:19 PM)


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OfflineHotnuts
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Re: Trametes versicolor [Re: lipa]
    #8068400 - 02/25/08 12:21 PM (15 years, 11 months ago)

There's mushroom mycelium growing on the surface of that bacterial colony, so a hot agar pour should work easily on that mess MycoAu. You don't even need antibiotics. Just pour a hot layer over the whole works until the new layer has just covered the highest point of that mess. Just over the inoculation.


Edited by Hotnuts (02/25/08 12:23 PM)


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Offlinemakaveli8x8
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Re: Trametes versicolor [Re: Hotnuts]
    #8068508 - 02/25/08 01:03 PM (15 years, 11 months ago)

or you could just transfer it to a new plate and maybe the myclium will outrun the bacteria, then do one more transfer

another option is to just throw some into a grain, even when my jars are contaminated their seems to always be a place i could get a clone from as long as I don't wait to long


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Edited by makaveli8x8 (02/25/08 01:05 PM)


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OfflineHotnuts
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Re: Trametes versicolor [Re: makaveli8x8]
    #8068730 - 02/25/08 02:25 PM (15 years, 11 months ago)

You put a wedge in whole grain of what MycoAu has there, you'll end up with nothing but a mess and a waste of time.


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OfflineRogerRabbitM
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Re: Trametes versicolor [Re: Hotnuts]
    #8068876 - 02/25/08 03:07 PM (15 years, 11 months ago)

Agreed. Hot agar will often pasteurize the bacteria long enough to let the mycelium recover and get a head start. Short of putting a piece on damp cardboard, I don't know of a better way to separate the two. It's actually very common when cloning wild mushrooms. Hundreds or more species can exist together in a single culture in the wild, but when brought into artificial sterile cultivation, they fail if you don't make a series of transfers to separate them.
RR


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semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
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OfflineHotnuts
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Re: Trametes versicolor [Re: RogerRabbit]
    #8068913 - 02/25/08 03:25 PM (15 years, 11 months ago)

It is the best way to clean up cultures of bacteria. One shot with the hot agar pour and the culture's bacteria free. Much higher % than doing a transfer IMO. Thanks for turning me on to that one actually.


Damn near all of the wild specimens i've cultured have been filthy once applied to agar. One of which I couldn't even get straighted out, Sparasis crispa. Heavily eaten up with molds. :thumbdown:


Edited by Hotnuts (02/25/08 03:28 PM)


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OfflineMycoAu
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Re: Trametes versicolor [Re: Hotnuts]
    #8070227 - 02/25/08 09:14 PM (15 years, 11 months ago)

That's what I thought might be causing the greasy appearance, but I'm still new to this. It seemed odd to me that a species as quick and aggressive as T. versicolor would remain contaminated (not outrun the bacteria). And, why only contaminate the areas in which mycelium (a competitor) exists, when there is plenty of open space (for a bacterium) to exist elsewhere on the plate?

Not questioning that it is indeed contaminated, like I said, I suspected that because of the appearance.

I have placed a peice of another agar culture (also T. versicolor, tissue from the same mushroom sample as the agar culture shown in the pictures above) into a pint of popcorn. White and much more whispy appearance. No problems with contamination (no greasy spots, odd smell or anything I would normally associate with bacterial contams). I think I should take a few of those kernels and place one on each of 5-10 agar plates to make sure I have at least one master that's clean.

I might try the hot agar pour later to see if I can isolate the dirty one for practice. I'm not terribly concerned with this sample as I can find this species everywhere. However, I think the practice will help when I need to be able to recover a sample.

Thanks for the help everybody.


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Invisiblelipa


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Re: Trametes versicolor [Re: MycoAu]
    #8071889 - 02/26/08 10:14 AM (15 years, 10 months ago)

Are you running a hood. Also "Just for thought" I soak my specimens in a diluted bleach solution before I open them up and take tissue. I have had the best success so far with this method. I also belive it is very important in picking the right specimens. Sometimes it is hard to resist taking the biggest specimen for a clone or the one that has the coolest colors. But the health is of the most important factor in success ,even if you have to take the youngest, freshest specimen and hope for the best.


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OfflineMycoAu
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Re: Trametes versicolor [Re: lipa]
    #8072738 - 02/26/08 02:16 PM (15 years, 10 months ago)

yes, I have a laminar flow hood. That's not that issue- I'm positive on that one.

I use 3% H2O2 for soaks of the sample before and after opening the mushroom (and it's not the same bath, so it's not cross-contamination from that source). Maybe I should only do the pre-opening soak and leave it alone after that.


Do you dilute your bleach for that soak or just straight ~5.25%


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Invisiblelipa


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Re: Trametes versicolor [Re: MycoAu]
    #8072750 - 02/26/08 02:19 PM (15 years, 10 months ago)

4cc bleach in 1 cup of water.
I soak for 15-20 min then take it out and let it dry up in front of the hood for a bit till it's not so soggy . Then tear it open and start cutting. With the trametes I peeled the skin first.


Edited by lipa (02/26/08 02:23 PM)


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OfflineMycoAu
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Re: Trametes versicolor [Re: lipa]
    #8072792 - 02/26/08 02:30 PM (15 years, 10 months ago)

"peeled the skin first"

Is that before the soak or after soak but before cutting for cultures?


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Invisiblelipa


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Re: Trametes versicolor [Re: MycoAu]
    #8073256 - 02/26/08 04:24 PM (15 years, 10 months ago)

Before the soak.


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OfflineMycoAu
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Re: Trametes versicolor [Re: lipa]
    #8073273 - 02/26/08 04:29 PM (15 years, 10 months ago)

Are you doing that just to reduce bioburden before beginning? Do you wear gloves? If not, how do you ensure that you're not just adding bacteria from your hands during the process?

Sorry about the simple questions, but I like to have a clear and complete picture of a method before I start. It generally saves a lot of time in the long run.


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Re: Trametes versicolor [Re: MycoAu]
    #8074198 - 02/26/08 07:39 PM (15 years, 10 months ago)

Whenever you isolate polypores, always take inner tissue from the outer edge of the mushroom. The newer growth is a lot cleaner than that of the tissue closest to the stipe. Use a super sharp razer blade or exacto knife to lightly scrape tissue from the center of the outer edge. All it takes is a tiny bit of tissue from the center for cloning. That will help tremendously is the acquisition of a cleaner sample.


Edited by Hotnuts (02/26/08 07:40 PM)


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Invisiblelipa


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Re: Trametes versicolor [Re: MycoAu]
    #8076420 - 02/27/08 09:16 AM (15 years, 10 months ago)

Quote:

MycoAu said:
Are you doing that just to reduce bioburden before beginning? Do you wear gloves? If not, how do you ensure that you're not just adding bacteria from your hands during the process?

Sorry about the simple questions, but I like to have a clear and complete picture of a method before I start. It generally saves a lot of time in the long run.




Yes! I am doing that to reduce the burden and to help ensure the success of it all. I do wear gloves when I do a clone. I figure "why should I add my natural bacteria to the picture" Also keep you specimens newly exposed surface 45'd to the hood at all times while cutting.


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