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Offlinesansa

Registered: 11/17/09
Posts: 647
Last seen: 10 years, 4 months
Sterile tissue from wild shroom?
    #11711001 - 12/24/09 09:47 AM (15 years, 27 days ago)

If you tear open a wild shroom's stem lengthwise and then cut out a section of the tissue inside so you don't get any that was touching the outside, would that be pure mushroom mycelium or would you still need to go to agar?

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OfflineRogerRabbitM
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Registered: 03/26/03
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Loc: Seattle
Last seen: 1 year, 10 months
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Re: Sterile tissue from wild shroom? [Re: sansa]
    #11711013 - 12/24/09 09:51 AM (15 years, 27 days ago)

You would still need agar to clean it up.  There's no such thing as pure culture in an outdoor mushroom.  All grow in harmony with molds and bacteria, and both will be in the interior of the fruit, whether it was exposed to open air or not.  If it was a young, rapidly growing fruit, it should clone easily on agar, and should easily outrun the contaminants, letting you grab some mycelium from the leading edge of the growth to transfer away to a new dish.  Two transfers and I'll bet you can get a clean culture.
RR


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Offlinesansa

Registered: 11/17/09
Posts: 647
Last seen: 10 years, 4 months
Re: Sterile tissue from wild shroom? [Re: RogerRabbit]
    #11711704 - 12/24/09 12:28 PM (15 years, 27 days ago)

Thanks

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Invisiblecyantific
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Registered: 10/11/09
Posts: 1,323
Loc: Shakashuri Island
Re: Sterile tissue from wild shroom? [Re: sansa]
    #11712180 - 12/24/09 01:52 PM (15 years, 27 days ago)

ive cloned wild mushrooms outside in the wild using the needle pop tek a few times ... RR is right though that it is unreliable as my success rate is at about 40/60 maybe less lol ... my most successful tissue biopsies have mostly come from the center of the cap / nipple down into the stipe ... its much easier than working with the spores ... it takes many transfers of the spores to get them properly domesticated and cleaned up ... for some reason though tissue transfers perform quite well once they take ...

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InvisibleCitizen13
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Registered: 11/01/09
Posts: 1,319
Re: Sterile tissue from wild shroom? [Re: cyantific]
    #11712457 - 12/24/09 02:43 PM (15 years, 27 days ago)

Quote:

cyantific said:
ive cloned wild mushrooms outside in the wild using the needle pop tek a few times ... RR is right though that it is unreliable as my success rate is at about 40/60 maybe less lol ... my most successful tissue biopsies have mostly come from the center of the cap / nipple down into the stipe ... its much easier than working with the spores ... it takes many transfers of the spores to get them properly domesticated and cleaned up ... for some reason though tissue transfers perform quite well once they take ...




i like to take my biopsies from the bottom inch of the stem.


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Invisiblecyantific
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Registered: 10/11/09
Posts: 1,323
Loc: Shakashuri Island
Re: Sterile tissue from wild shroom? [Re: Citizen13]
    #11713575 - 12/24/09 06:26 PM (15 years, 27 days ago)

i do as well , but on a wild mushroom there is more chance of contam inside the base at ground level ...
also once you get about half an inch deep into the nipple and stipe it gets dicey again as you enter the flute of the stipe where contams may have made their way up ...

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Offlinesansa

Registered: 11/17/09
Posts: 647
Last seen: 10 years, 4 months
Re: Sterile tissue from wild shroom? [Re: cyantific]
    #11713916 - 12/24/09 07:59 PM (15 years, 27 days ago)

Quote:

cyantific said:
i do as well , but on a wild mushroom there is more chance of contam inside the base at ground level ...
also once you get about half an inch deep into the nipple and stipe it gets dicey again as you enter the flute of the stipe where contams may have made their way up ...



If I'm reading this right, you mean you want the center core of the stem that's lower than half an inch from the top of the mushroom and higher than the base of the stem? How high above the base of the stem do you suggest?

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Invisiblecyantific
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Registered: 10/11/09
Posts: 1,323
Loc: Shakashuri Island
Re: Sterile tissue from wild shroom? [Re: sansa]
    #11713944 - 12/24/09 08:06 PM (15 years, 27 days ago)

pierce the nipple/center of cap straight down from the top about half an inch , twist the syringe to dislodge the tissue , and inject into lc ...

some shrooms may be more frail than others so extra care is needed to not go past that half inch into the cap/stem and sometimes even not going even that much .... you will know if you get too far because the needle will then slip into the flute of the stipe quickly without resistance ... if that happens just twist and extract quickly and knock jar , it may or may not be okay ... even if it goes right it may or may not be ok considering its origins ...

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