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FreeSporePrints
Registered: 03/06/05
Posts: 1,139
Loc: Rome, Italy
Last seen: 3 months, 15 days
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Spores germination Petri plate VS PF cake
#8687785 - 07/27/08 10:18 AM (15 years, 7 months ago) |
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Hi, have anyone found a better chance of spores germination on PF cake when the spores don't want to germinate for a cause or another into a petri dishes?
Thanks, Fabio
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MycoAu
5thKingdomCome
Registered: 07/18/07
Posts: 1,047
Last seen: 3 years, 2 months
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Re: Spores germination Petri plate VS PF cake [Re: FreeSporePrints]
#8688259 - 07/27/08 01:04 PM (15 years, 7 months ago) |
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I've had great success germinating my L. edodes print on WBS after several complete failures for germination on agar. (With and without antibiotic, with and without pre-hydration, etc.)
hydrated spores overnight in syringe, injected into WBS and in a few days to a week- mycelium everywhere. I highly recommend this as a way to go (for starting out in strain isolation). Afterwards, use a different agar formulation for plate growth, isolation, etc.
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FooMan
Registered: 02/02/05
Posts: 8,957
Loc: Earth
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Re: Spores germination Petri plate VS PF cake [Re: FreeSporePrints]
#8693246 - 07/28/08 05:10 PM (15 years, 7 months ago) |
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I agree with the above poster. That usually happens to me when I'm working with older spore prints. Hydrating the spores before putting them on agar usually fixes the problem unless the spores are in really bad shape.
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FreeSporePrints
Registered: 03/06/05
Posts: 1,139
Loc: Rome, Italy
Last seen: 3 months, 15 days
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Re: Spores germination Petri plate VS PF cake [Re: FooMan]
#8697526 - 07/29/08 01:33 PM (15 years, 7 months ago) |
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Why does it happen for you?
Thanks, Fabio
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MycoAu
5thKingdomCome
Registered: 07/18/07
Posts: 1,047
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Re: Spores germination Petri plate VS PF cake [Re: FreeSporePrints]
#8699679 - 07/29/08 09:42 PM (15 years, 7 months ago) |
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why does "what" happen for "who"?
Sorry, but I'm apparently confused about who the question is directed at and what exactly the question itself is supposed to be asking.
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FreeSporePrints
Registered: 03/06/05
Posts: 1,139
Loc: Rome, Italy
Last seen: 3 months, 15 days
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Re: Spores germination Petri plate VS PF cake [Re: MycoAu]
#8702110 - 07/30/08 12:45 PM (15 years, 7 months ago) |
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Sorry but my English sucks.
I was asking to myself: why the spores that don't germinate on agar, sometimes they germinate on grains?
Thanks, Fabio
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MycoAu
5thKingdomCome
Registered: 07/18/07
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Re: Spores germination Petri plate VS PF cake [Re: FreeSporePrints]
#8702320 - 07/30/08 01:31 PM (15 years, 7 months ago) |
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i would imagine it has to do with the enzymes or markers (on the cell walls of the grains) being present, thereby signalling the spores to germinate and begin growth. Or it could be something simpler like a difference in pH or appropriate nutrients for that species.
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Mycelio
Stranger
Registered: 06/24/08
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Re: Spores germination Petri plate VS PF cake [Re: MycoAu]
#8706245 - 07/31/08 06:36 AM (15 years, 7 months ago) |
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Thought the same.
Fabio, I guess it is for Kuehneromyces mutabilis.
Which agar mixture did you use?
It may help if you cook the agar with wood tea. Just boil water with small wood chips for ten minutes. It will turn light brown. Then use the tea for cooking agar. Not sure if it will do the trick, but it won't hurt if the spores find molecules of their final food.
I'm having great success doing spore syringes with wood or straw tea without any other additives. I let them sit for a few days until the liquid gets clear and the spores germinate.
Carsten
Edited by Mycelio (07/31/08 06:40 AM)
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FreeSporePrints
Registered: 03/06/05
Posts: 1,139
Loc: Rome, Italy
Last seen: 3 months, 15 days
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Re: Spores germination Petri plate VS PF cake [Re: Mycelio]
#8706391 - 07/31/08 08:24 AM (15 years, 7 months ago) |
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@MycoAu: Hi Thanks for the tip!
@Carsten: Hi Carsten it was a kind of PDA..made with "fecola di patate". In Italy is sold "fecola di patate", that's potatoes boiled, dehydrated and made in powder. It's used in confectionery as thickener for creams and cakes in general..has got an high content in amid but doesn't contain gluten.
The final pH of the medium is 5.5-6.0..a good pH for germinating spores.
Thanks for the tips of the tea. But i haven't understood a thing: Do you let to germinating spores into syringes with wood tea and when you see some mycelium you inocule into petri dishes or similar?
How many days generally before the germination starts?
Thanks, Fabio
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Mycelio
Stranger
Registered: 06/24/08
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Re: Spores germination Petri plate VS PF cake [Re: FreeSporePrints]
#8706775 - 07/31/08 10:55 AM (15 years, 7 months ago) |
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Hi Fabio,
that potato powder sounds interesting. I have too look if I can find it here too.
Assuming we are speaking about K. mutabilis spores, I would expect them to prefer wood over potatoes. In petri dishes, I would try ME agar with wood tea. Unfortunately Stamets doesn't mention this species in his books and you don't find much info in the net. Two threads from here, in case you didn't see them already: http://www.shroomery.org/forums/showflat.php/Cat/0/Number/3607916 http://www.shroomery.org/forums/showflat.php/Number/3065782#3065782
The wood tea syringe is my method of choice for Agrocybe aegerita spores. Those ones germinate after two days. Then I shake well and inoculate jars with sterilized grain only using one or two drops.
Carsten
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FreeSporePrints
Registered: 03/06/05
Posts: 1,139
Loc: Rome, Italy
Last seen: 3 months, 15 days
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Re: Spores germination Petri plate VS PF cake [Re: Mycelio]
#8707395 - 07/31/08 01:18 PM (15 years, 7 months ago) |
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Hi Carsten!! If you want i can send to you some fecola di patate.
Yes it's K.mutabilis! I'll try MEA(ME is MEA?) using wood tea
Thanks for the links, the sporeprints i've are from ragadinks..but i imagine too old to germinate..however i want to try!
Thanks for the tips! Fabio
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Mycelio
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Posts: 1,636
Loc: Berlin
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Re: Spores germination Petri plate VS PF cake [Re: FreeSporePrints]
#8707566 - 07/31/08 01:43 PM (15 years, 7 months ago) |
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Thanks for the offer Fabio! I think the german 'Kartoffelstärke' (potato-starch) is the same stuff and available in supermarkets.
And yes, I meant MEA.
Good luck with the spores, if they were kept dry until now, your chances should be good.
Carsten
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umbrellamaker
Intergalactic Sociopath
Registered: 05/03/05
Posts: 573
Loc: Houston TX
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Re: Spores germination Petri plate VS PF cake [Re: Mycelio]
#8719991 - 08/03/08 02:25 PM (15 years, 7 months ago) |
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I thought I might share this experience. Agar concentration will cause old spores to not germinate. Try diluting your agar to 25%, and then try using ph 7 bottled spring water, as sometimes the tap water in my area contains bad metals in it. If you are using tap water, and have a ph meter on hand, test the ph of your agar when it has warmed to the point of pouring into dishes. I would bet a dollar that your ph is not in range due to too much malt extract, or bad tap water, or both. Either way, A dilute agar formula is always better than a stronger one when germinating spores.
I hope that helps, if not I am sorry for chiming in.
Be well! Good luck!!
(I don't practice Santeria...I ain't got no Crystal ball;) Sublime
Edited by umbrellamaker (08/03/08 05:52 PM)
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spacel0rd
anarchohippienerd
Registered: 07/21/08
Posts: 210
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Re: Spores germination Petri plate VS PF cake [Re: umbrellamaker]
#8720353 - 08/03/08 04:20 PM (15 years, 7 months ago) |
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Quote:
umbrellamaker said: I thought I might share this experience. Agar concentration will cause old spores to not germinate. [...] Either way, A dilute agar formula is always better than a stronger one when germinating spores.
thx for sharing that info. good to know. i never ran into this issue in all the zillions of posts i read here the last few months.
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FreeSporePrints
Registered: 03/06/05
Posts: 1,139
Loc: Rome, Italy
Last seen: 3 months, 15 days
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Re: Spores germination Petri plate VS PF cake [Re: umbrellamaker]
#8724925 - 08/04/08 03:07 PM (15 years, 7 months ago) |
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Quote:
umbrellamaker said: I thought I might share this experience. Agar concentration will cause old spores to not germinate. Try diluting your agar to 25%, and then try using ph 7 bottled spring water, as sometimes the tap water in my area contains bad metals in it. If you are using tap water, and have a ph meter on hand, test the ph of your agar when it has warmed to the point of pouring into dishes. I would bet a dollar that your ph is not in range due to too much malt extract, or bad tap water, or both. Either way, A dilute agar formula is always better than a stronger one when germinating spores.
I hope that helps, if not I am sorry for chiming in.
Be well! Good luck!!
(I don't practice Santeria...I ain't got no Crystal ball;) Sublime
very interesting your tips. Just yesterday i've discovered that the pH AFTER have been sterilized is not the same as BEFORE sterilizing. I would try! About the water i'll try distilled water.
Thanks a lot mate you're welcome on FreeSporePrints.org
Fabio
..and thanks also for the tip of the diluited agar.
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umbrellamaker
Intergalactic Sociopath
Registered: 05/03/05
Posts: 573
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Re: Spores germination Petri plate VS PF cake [Re: FreeSporePrints]
#8725147 - 08/04/08 03:49 PM (15 years, 7 months ago) |
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No problem. I am glad if my comments helped. Just to specify one thing though. You shouldnt be aiming for a 7ph when pouring agar. I just meant that you want your water that you ad to your extract before pressure cooking to be 7 neutral. Even a 25 percent agar mixture should keep your ph right when pouring into dishes if you started with clean neutral water.
For some people, who like to see vigorous growth from spore germination on a plate, 50% is fine too. Fact is, I have germinated many spores from full concentration on the back directions of bottle. But I always try the 25% method before I toss a print out, and more times than not, I get action from it, when I wouldn't otherwise.
Peace everyone...
Edited by umbrellamaker (08/10/08 01:35 PM)
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FreeSporePrints
Registered: 03/06/05
Posts: 1,139
Loc: Rome, Italy
Last seen: 3 months, 15 days
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Re: Spores germination Petri plate VS PF cake [Re: umbrellamaker]
#8753154 - 08/10/08 01:32 PM (15 years, 7 months ago) |
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i've created a little pictorial tutorial on how to make potato starch medium:
http://www.freesporeprints.org/smf/index.php?topic=206.msg1030#msg1030
Fabio
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