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Offlineucsdmolpath
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re: "average" agar cultivation temperature?
    #2241145 - 01/14/04 11:06 AM (12 years, 10 months ago)

hi all,

though i know all strains and species are different, etc...i was wondering if there is an ideal "average" cultivation temperature for spores on agar...

i have an incubator for low temperature bacteria and yeast (of course i would take precautions to keep my mycelium from getting contaminated)....which rests at a constant 30C (86F) which seems a bit high?  and how many days should i expect to wait at the average temperature?

also, how long (# days) is hydrogren peroxide stable for and at what ideal temperature (since peroxide degrades faster with higher temperatures and light exposure...)  i am attempting to transfer my mycelium onto peroxide plates eventually...but gotta get the spores to germinate first :smile:


thanks!


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Invisiblemicro
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Re: re: "average" agar cultivation temperature? [Re: ucsdmolpath]
    #2241153 - 01/14/04 11:14 AM (12 years, 10 months ago)

Depends on the strain, like you said. 30 should be fine, though, for most genus/species. If the ideal temp is much lower you can always just incubate them at room temp -- there's no reason you *need* to heat them. Make sure to check because, with fungi, usually there is a slow curve that goes to the best temp, but this quickly drops off much above the optimum temp, so growth is usually *really* inhibited at temperatures 5 degrees or more above the optimum temperature. This is why it's better to go lower than higher.

Usually those incubators are for cloning certain inserts, so make sure noone knows what you're doing at work.... Unless you discussed it with them beforehand....

Oh -- peroxide stays good for a little while -- it WILL break down in high heat, though. Plenty good for as long as you need to run petris (as long as you add it after cooling the media so it's warm to the touch, while stirring) but I don't know the shelf life of peroxide plates.

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Edited by micro (01/14/04 11:17 AM)


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Invisiblemicro
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Re: re: "average" agar cultivation temperature? [Re: micro]
    #2241185 - 01/14/04 11:31 AM (12 years, 10 months ago)

BTW -- to clarify what I mean -- if other people use it they'll probably get really pissed if they find out you're growing a fungus resistant to every antibiotic in the same incubator they're using to select for inserts with antibiotic resistance genes.

It may not be a problem for you, but I figured I'd throw out the warning, since I work in a lab.

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Offlineucsdmolpath
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Re: re: "average" agar cultivation temperature? [Re: micro]
    #2241219 - 01/14/04 11:57 AM (12 years, 10 months ago)

hey there,
thanks...i'll probably end up keeping the plates on my desk...and yes, i know the concerns of the lab :smile: they were all warned...they worry less about what i do, and worry more about the p-32 nucleotides that my other lovely co-worker plays with open bench, no shield... :smile:


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Invisiblemicro
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Re: re: "average" agar cultivation temperature? [Re: ucsdmolpath]
    #2241283 - 01/14/04 12:23 PM (12 years, 10 months ago)

Lol! Hopefully their kids won't have three heads.... I shouldn't talk, though -- I'm not very careful myself....

Leaving them on your desk should be fine!

Good luck!

--
Micro


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Offlineucsdmolpath
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Re: re: "average" agar cultivation temperature? [Re: micro]
    #2241354 - 01/14/04 12:41 PM (12 years, 10 months ago)

hehe thanks for your advice and encouragement (novice to mushroom cultivation here).....

insofar as my co-worker, thankfully i don't think they're having kids...(that would be a scarier than than any kind of radiation scare! :smile:

take care :smile:


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Offlinepietruk
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Re: re: "average" agar cultivation temperature? [Re: ucsdmolpath]
    #2247740 - 01/17/04 04:44 AM (12 years, 10 months ago)

Yes but is there a standard exact temp to incubate spores on agar? I just came by an incubator myself and would also appreciate some exact temp info


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Offlineucsdmolpath
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Re: re: "average" agar cultivation temperature? [Re: pietruk]
    #2249219 - 01/17/04 08:59 PM (12 years, 10 months ago)

hi there...
so after looking up information on different types of mushrooms, it seems that the ideal spore germinating temperature can vary from room temperature to 77F.  i probably wouldn't go higher than that, as it can be detrimental....

another way, i decided to try pushing the spores to grow faster so that even if temp. is off, the mycelium may have a faster start....it may or may not work, but i'll keep you updated....

--------how i've tried to push spores a bit...
1)  first i take the spores and scrape them into a clean culture tube...add a small volume(1ml or so) of liquid media (most agar media recipes minus the agar)...i'm using bacterial LB media + a little bit of glucose (sugar)...let it shake at room temperature for a few hours...add another equal volume of 2X concentration of hydrogen peroxide diluted in liquid media.

(if, as above, 1X is considered a 1:10 dilution of store-bought peroxide, which is 3%, then 1X=0.3%.  This means that a 2X should be 0.6%....in the end, about 1:5 dilution of a store bought peroxide)

2) shake again for a few hours...then add about 100?l or several drops until about 3cm in diameter puddle forms onto a normal 10cm agar plate containing 0.3% peroxide agarose.

3) grow at a temp. somewhere between room temp and 77F....i've put my plates (sealed with parafilm) on top of a mini-fridge under my lab bench...where the air is just a little bit warmer than room temp...

----------------------------------------------

In the meantime, I'm also trying some liquid innoculation into grain media jars...seems like it could spread the mycelium more evenly as opposed to pushing wedges of agar into birdseed and wood flakes...will report back later :smile:

ok...good luck...hope this helps


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Offlinesauroman1
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Re: re: "average" agar cultivation temperature? [Re: micro]
    #21021500 - 12/24/14 03:15 PM (1 year, 11 months ago)

Quote:

micro said:
Depends on the strain, like you said.  30 should be fine, though, for most genus/species.  If the ideal temp is much lower you can always just incubate them at room temp -- there's no reason you *need* to heat them.  Make sure to check because, with fungi, usually there is a slow curve that goes to the best temp, but this quickly drops off much above the optimum temp, so growth is usually *really* inhibited at temperatures 5 degrees or more above the optimum temperature.  This is why it's better to go lower than higher.
--
Micro




What happens if temperature of agar gets higher than 30C, say 34C? Can spores and mycelium die immediately at such temperature or maybe grow even faster? I'm talking about p. cubensis, cambodian strain.
Colder temperature is safer but shrooms will grow slowly. So higher temp can useful.


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OfflineBumbaclotjohnson
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Re: re: "average" agar cultivation temperature? [Re: sauroman1]
    #21023780 - 12/25/14 06:40 AM (1 year, 11 months ago)

They aren't gonna die but they won't grow fast either, 34c is about 93f I'm pretty sure death occurs in the 100s f


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OfflinetaGyo
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Re: re: "average" agar cultivation temperature? [Re: Bumbaclotjohnson]
    #21025934 - 12/25/14 10:24 PM (1 year, 11 months ago)

After 106f myc begins thermal death.


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Edited by taGyo (12/26/14 12:59 AM)


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OfflineBumbaclotjohnson
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Re: re: "average" agar cultivation temperature? [Re: taGyo]
    #21026095 - 12/25/14 11:23 PM (1 year, 11 months ago)

^ sounds about right


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Offlinesauroman1
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Re: re: "average" agar cultivation temperature? [Re: Bumbaclotjohnson]
    #21026810 - 12/26/14 03:49 AM (1 year, 11 months ago)

Quote:

Bumbaclotjohnson said:
They aren't gonna die but they won't grow fast either, 34c is about 93f I'm pretty sure death occurs in the 100s f




You mean 100f temperature? I almost have reached that temperature, detected about 36C.
How to detect death, how many days it takes for first visible mycelium to appear on agar?


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Offlinekrypto2000
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Re: re: "average" agar cultivation temperature? [Re: sauroman1]
    #21030417 - 12/27/14 02:28 AM (1 year, 11 months ago)

I'm pretty sure cubes ideal temperature is actually 86f. When growing in a jar though I've read you should not go above about 83f because they generate their own heat and thus will be a few degrees higher than that possibly, plus as stated you really don't want to go over. Due to their small size I really doubt a petri will generate a significant amount of heat so 86f ambient is probably ideal.


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Offlinesauroman1
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Re: re: "average" agar cultivation temperature? [Re: krypto2000]
    #21030777 - 12/27/14 06:34 AM (1 year, 11 months ago)

Quote:

krypto2000 said:
I'm pretty sure cubes ideal temperature is actually 86f. When growing in a jar though I've read you should not go above about 83f because they generate their own heat and thus will be a few degrees higher than that possibly, plus as stated you really don't want to go over. Due to their small size I really doubt a petri will generate a significant amount of heat so 86f ambient is probably ideal.




Thanks, with ideal temperature how long it might take for the first visible mycelium to appear?


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Offlinedrake89
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Re: re: "average" agar cultivation temperature? [Re: taGyo]
    #21032639 - 12/27/14 07:37 PM (1 year, 11 months ago)

Quote:

taGyo said:
After 106f myc begins thermal death.



I reckon tropical species could survive above these temps but I'm not sure what we're talking about species wise, here


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Offlinekrypto2000
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Re: re: "average" agar cultivation temperature? [Re: drake89]
    #21033916 - 12/28/14 03:29 AM (1 year, 11 months ago)

Not necessarily. While the ambient air may be 106f I wouldn't be surprised if the mushrooms were quite a bit lower when you consider location (eg. shade + being in or on the earth) and evaporation.


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