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M11
White Thumb

Registered: 06/14/09
Posts: 1,840
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Germination time on agar questions.
#14261411 - 04/09/11 12:09 AM (12 years, 9 months ago) |
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Hey folks,
I was reading through some old threads and RR said that fresh spores germinate within hours, even minutes on agar. He said you won't see growth for a few days. I have also read that it can take up to 2 weeks for spores to germinate on agar.
Have any of you experienced anything to support this?
Also, after I inoculated my plates with about 1-2 drops, the spore solution moved around my plate when I moved it. I was trying to inoculate 3 different spots on the petri dish.
Will this be an issue or will I simply have a wider area of germinating spores and therefore better sector selection?
Could this increase germination times?
Thanks
M
-------------------- Of course, then there's the problem of eating vermiculite. On the bright side, it makes your poop and teeth glitter. Just pretend it's christmas. -RR Those mushrooms are fine. Your friend is a pussy. -RR Outdoor Patch
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Doc_T
Random Dude




Registered: 03/06/09
Posts: 42,395
Loc: Colorado
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Re: Germination time on agar questions. [Re: M11]
#14261419 - 04/09/11 12:11 AM (12 years, 9 months ago) |
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7-14 days to see growth from spores.
-------------------- You make it all possible. Doesn't it feel good?
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m_jacobs
Man Whore
Registered: 03/11/11
Posts: 121
Loc: CA
Last seen: 10 years, 7 months
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Re: Germination time on agar questions. [Re: Doc_T]
#14261444 - 04/09/11 12:25 AM (12 years, 9 months ago) |
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I just did a single drop on agar on 4/3. Saw a BB sized ball of fuzz on 4/6. I now have a dime - nickle sized island.
**Red arrow points to were the drop landed and where the BB ball of fuzz started

Started a Agar log here.
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M11
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Registered: 06/14/09
Posts: 1,840
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Re: Germination time on agar questions. [Re: m_jacobs]
#14262575 - 04/09/11 10:52 AM (12 years, 9 months ago) |
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I inoculated mine yesterday afternoon, and I swear that I can see tiny dots of something forming already. Looks maybe like dust? I do not believe those were there when I initially inoculated. Maybe it is just clumped spores, but I shook the hell out of the syringes before using them -flicked them too.
Maybe it is just wishful thinking?
-------------------- Of course, then there's the problem of eating vermiculite. On the bright side, it makes your poop and teeth glitter. Just pretend it's christmas. -RR Those mushrooms are fine. Your friend is a pussy. -RR Outdoor Patch
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cc2
Mush

Registered: 05/15/10
Posts: 2,611
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Re: Germination time on agar questions. [Re: M11]
#14263475 - 04/09/11 03:27 PM (12 years, 9 months ago) |
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Quote:
M11 said: I inoculated mine yesterday afternoon, and I swear that I can see tiny dots of something forming already. Looks maybe like dust? I do not believe those were there when I initially inoculated. Maybe it is just clumped spores, but I shook the hell out of the syringes before using them -flicked them too.
Maybe it is just wishful thinking?
when things spread too fast on agar, it is probably molds or other contams. anyway time will tell, I think it'll be fairly easy to distinguish later healthy fuzzy mycelium from sporulating and hyphae of zygomycetes
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M11
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Registered: 06/14/09
Posts: 1,840
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Re: Germination time on agar questions. [Re: cc2]
#14264057 - 04/09/11 07:08 PM (12 years, 9 months ago) |
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I am really not concerned with contaminants. I used a flow hood, which was carefully sterilized prior to pouring and inoculating my plates. Also, I incorporated streptomycin sulfate and penicilin G into my agar mix after autoclaving, so bacterial contaminants are fucked.
I used careful aseptic technique. It is kinda hard to see through the petri dish plastic and parafilm and such.
Time will tell though!
I am so jacked about working with isolates.
-------------------- Of course, then there's the problem of eating vermiculite. On the bright side, it makes your poop and teeth glitter. Just pretend it's christmas. -RR Those mushrooms are fine. Your friend is a pussy. -RR Outdoor Patch
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Damion5050
Mush Doctor


Registered: 05/01/08
Posts: 12,493
Loc: Lost In Translation !
Last seen: 3 years, 5 days
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Re: Germination time on agar questions. [Re: M11]
#14264295 - 04/09/11 08:31 PM (12 years, 9 months ago) |
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How long did you let your dishes sit before using them ?
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M11
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Registered: 06/14/09
Posts: 1,840
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Re: Germination time on agar questions. [Re: Damion5050]
#14266443 - 04/10/11 10:46 AM (12 years, 9 months ago) |
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24 hours. Agar normally solidifies within 2-3 hours, but I had no time until the next day to inoculate. I would be hesitant to inoculate 2-3 hour solidified agar anyway.
-------------------- Of course, then there's the problem of eating vermiculite. On the bright side, it makes your poop and teeth glitter. Just pretend it's christmas. -RR Those mushrooms are fine. Your friend is a pussy. -RR Outdoor Patch
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M11
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Registered: 06/14/09
Posts: 1,840
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Re: Germination time on agar questions. [Re: M11]
#14269161 - 04/10/11 08:52 PM (12 years, 9 months ago) |
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Both my Ecuador and PE have germinated. PE germinated the fastest or is growing the fastest. Nice tiny white fuzzies. I would guess that RR's statement holds true. Spores germinate very quickly on agar, but take a few days to become visible. I would post pictures but they are tiny. Visible growth within 2 days.
I love parafilm. Great stuff to wrap agar dishes with.
-------------------- Of course, then there's the problem of eating vermiculite. On the bright side, it makes your poop and teeth glitter. Just pretend it's christmas. -RR Those mushrooms are fine. Your friend is a pussy. -RR Outdoor Patch
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Primal Call
Earth Mage



Registered: 09/05/10
Posts: 2,766
Loc: Here until here
Last seen: 4 years, 3 months
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Re: Germination time on agar questions. [Re: M11]
#14269262 - 04/10/11 09:08 PM (12 years, 9 months ago) |
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what temp range are you storing these in?
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M11
White Thumb

Registered: 06/14/09
Posts: 1,840
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Re: Germination time on agar questions. [Re: Primal Call]
#14269423 - 04/10/11 09:31 PM (12 years, 9 months ago) |
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75-80 F. I put them in a room that is currently not air conditioned. I am amazed at how quickly they germinated, let alone how quickly they became visible. I did use more spore solution than necessary.
It is definitely mycellium and not mold. I shit you not. It looks very cotton like. I bet I will be making transfers in the next couple days.
-------------------- Of course, then there's the problem of eating vermiculite. On the bright side, it makes your poop and teeth glitter. Just pretend it's christmas. -RR Those mushrooms are fine. Your friend is a pussy. -RR Outdoor Patch
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M11
White Thumb

Registered: 06/14/09
Posts: 1,840
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Re: Germination time on agar questions. [Re: M11]
#14290672 - 04/14/11 03:11 PM (12 years, 9 months ago) |
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I made my first transfers today. I selected the outermost most rhizomorphic sections of the largest colonies. I hope that was the right thing to do. The PE seem to be more rhizomorphic than the EQ at this point, although I won't see true sectors until 3-4 transfers from now.
The center of each colony, if you will, seemed to be more cotton-like than the outermost areas.
I hope to make my next set of transfers next Monday or Tuesday. I would ideally like to make 2 transfers a week, but that may not be possible.
-------------------- Of course, then there's the problem of eating vermiculite. On the bright side, it makes your poop and teeth glitter. Just pretend it's christmas. -RR Those mushrooms are fine. Your friend is a pussy. -RR Outdoor Patch
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Primal Call
Earth Mage



Registered: 09/05/10
Posts: 2,766
Loc: Here until here
Last seen: 4 years, 3 months
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Re: Germination time on agar questions. [Re: M11]
#14292022 - 04/14/11 07:12 PM (12 years, 9 months ago) |
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got any pics? I'm just learning how to clone & sector and would love some visual examples 
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M11
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Registered: 06/14/09
Posts: 1,840
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Re: Germination time on agar questions. [Re: Primal Call]
#14294896 - 04/15/11 10:26 AM (12 years, 9 months ago) |
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You can maybe see where I took some agar wedges in this photo. I think for my next transfer I need to make much smaller segments. I was just shooting for the most rhizomorphic for this initial transfer.
Those labels are just some made up ones. The second word starts with either p or e, indicating PE or EQ.
These two plates were inoculated with spores 7 days ago.
-------------------- Of course, then there's the problem of eating vermiculite. On the bright side, it makes your poop and teeth glitter. Just pretend it's christmas. -RR Those mushrooms are fine. Your friend is a pussy. -RR Outdoor Patch
Edited by M11 (04/15/11 10:38 AM)
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Primal Call
Earth Mage



Registered: 09/05/10
Posts: 2,766
Loc: Here until here
Last seen: 4 years, 3 months
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Re: Germination time on agar questions. [Re: M11]
#14298773 - 04/15/11 11:47 PM (12 years, 9 months ago) |
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nice!
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iforgot



Registered: 06/22/11
Posts: 24
Last seen: 12 years, 1 month
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Re: Germination time on agar questions. [Re: M11]
#14862454 - 08/02/11 03:15 PM (12 years, 5 months ago) |
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Hey M11 - I hope you don't mind, but I'm trying to figure out wtf I've done wrong, and I'm hoping you can help me... I purchased some spores, and made some malt extract agar plates, and inoculated them like you did yours... of course a couple places, the water moved around on the agar, but I wasn't concerned. (Don't worry, I'm a microbiologist, and prepared the plates myself, so I know there was no contamination when I got started, plus I did it in a laminar flow hood...)
Two days later, after incubating at 30 degrees C, I noticed tiny little flecks of what appeared to me to be bacterial growth - only on the spots where the spore suspension was deposited. A week and a half later, I still have the same exact thing, plus one plate with two white fungi growing on them. They look nothing like the mycelia from any of the pictures I've seen on this site.
My thought - the spores were contaminated, or they were sent in an unsterile spore solution. What do you think?
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Primal Call
Earth Mage



Registered: 09/05/10
Posts: 2,766
Loc: Here until here
Last seen: 4 years, 3 months
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Re: Germination time on agar questions. [Re: iforgot]
#14864586 - 08/02/11 10:06 PM (12 years, 5 months ago) |
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Not that you are asking me, and you have assuredly more experience than I do, but you should have probably posted a new thread with pictures for your inquiry.
You'll get a lot more attention asking the community than you will asking the few of us here in this 3.5 month old thread.
I don't mean to be a bastard, and I hope you get your agar problems addressed! 
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iforgot



Registered: 06/22/11
Posts: 24
Last seen: 12 years, 1 month
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Re: Germination time on agar questions. [Re: Primal Call]
#14866284 - 08/03/11 08:18 AM (12 years, 5 months ago) |
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No worries, Ryath - thanks for the tip! I'm still pretty new to this site, so any help or suggestions are very much appreciated...
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