27 replies to this topic

[8145]

So this is my second spore to agar experiment (first one failed). I inoculated the agar picture below a few days ago and now I am getting this funky growth. I am not sure whether is good or bad but at this stage I am just happy of growing something

Do you guys know if this plate is being colonised or contaminated. I check a few videos on youtube but neither the colonisation or contamination examples look like this... So i am kind of puzzled.

Also check up the macro pic it appears that there are some bubbles going on there wtf ?


Cheers,

Attached Files

•  Spore C1.jpg   75.5K   63 downloads
•  SporeC1_2.jpg   81.98K   77 downloads
•  bubbles.jpg   57.64K   55 downloads

[Amazonian]

Hmm, hard to say with the pics provided. It looks like contams at first, but I am thinking the brown patches are a massive amount of spores that you put onto the agar? The tufty white around the brown blotches looks like myc , but I am on an iPod and the images aren't macro? . Just wait and see huh?!.

Good luck.

[Amazonian]

Yep , looks like myc' from here.

[8145]

Hmm, hard to say with the pics provided. It looks like contams at first, but I am thinking the brown patches are a massive amount of spores that you put onto the agar? The tufty white around the brown blotches looks like myc , but I am on an iPod and the images aren't macro? . Just wait and see huh?!.

Good luck.

Yes I kind of messed it up a bit when I spread the spores - end up putting a really massive amount crumbed in a the same place. Should I transfer the fluffy bits onto anothe petri dish ?

[8145]

Yep , looks like myc' from here.

OMG THIS IS MASSIVE ! I created life

Edited by Blas, 10 July 2012 - 10:09 PM.

[jwerta]

yea looks like myc to me 2... good luck

[Amazonian]

^ lol, don't get too excited... I am not a fungi expert , and I am on an iPod, but I rekon it's myc'. The real fungi heads can confirm This before you plan the next step. (I will have a proper look from computer after)

I wouldn't be mucking with the petri dishes. Just keep an eye on them.



Edit; arrow up was an indication that my reply was to blas( not jwerta who slipped a post in whilst I was typing!)

Edited by Amazonian, 10 July 2012 - 10:16 PM.

[Therefore]

erm, dont want to be a buzzkill but it looks like contamination to me. It's to hard to tell from the pics. Dunno the last one looks like some white myc maybe.
How long was the time frame between spreading the spores and when you started to see growth.
Spores are like dust so how did you 'crumble' them? You have gill fragments in your sporeprint....?

Make a homemade inoculation loop out of an old electric guitar string (the thin E string down the bottom).
Flame sterilise the loop and google how to streak plates with the loop

The other thing is that you might want to start with a clean print from a SAB member here. Mushrroms that have been in a supermarket for weeks might be contam. I know the shrooms in my local are imported from Korea! they really cant be that fresh eh.

Start with a clean print and then if it gets contam you will know that your technique is no good.

That said i made a clean plate from a a print of a store bought button mushroom. The veil had not broken which helps.

Anyway you are getting close and progressing fast. You will succeed.

I could be wrong....it could be myc!!

[Distracted]

Section off a sector of the white rhizomophic mycelium onto another plate asap.

[LokStok]

blas, do whatever he ^ says.

the white rhizomophic mycelium he's talking about is the feathery, fractal looking stuff spreading on the outer from the satellite spots.
the more yellowish growth burgeoning from the shit-tonne of crud on there is not your friend.
you have a short window.

[8145]

blas, do whatever he ^ says.

you have a short window.

Holy crap I just got home hope is not too late... Operation myc rescue has begun

[Therefore]

Holy crap I just got home hope is not too late... Operation myc rescue has begun

^^^ shit goes down fast in mycology. Good luck!

[jwerta]

pics after please id love to find out how you went

[8145]

I section off the fluffy bits and transfer them to another plate...fingers crossed. I took some pics but you cant see much because for this emergency I had to use my left over plates which have some condensation on the lid...(first time pressure cooking agar sorry I didnt know i would be too hot and condensate silly me). So will post some pics when it gets super big and colonise the whole plate. In the mean time I can post some a pic of his healthy bro

[8145]

Yes i did cut into the agar that is why it looks funny. Also I used and scalpel.

[8145]

Make a homemade inoculation loop out of an old electric guitar string (the thin E string down the bottom).

I looked into this but I reckon the G string will be better

Also I meant crumbed as in clustered all together didn't know how many spores to put but will take your advice on board about the way to streak plates. Cheers for that.

[Therefore]

Nice work.
Yeah im new to agar and i still get condensation on my plates. Just got to wait for it to cool.

[Amazonian]

the more yellowish growth burgeoning from the shit-tonne of crud on there is not your friend.

Yes, the yellow (contam) is more visible on a computer. Looking at it from my ipod, i was thinking it might have been like color leaching from the spores?!.

Yeah im new to agar and i still get condensation on my plates. Just got to wait for it to cool.

When i have waited for the agar to cool before i pour it, it solidifies?!.

[8145]

Now this mold grew on another plate. But still I got 15 healthy ones. Is it possible to save this one ? I am afraid if I open it will contaminate my growing environment.

[Amazonian]

^ Me personally, if i had 15 healthy ones, i would just put that one out in the garden.

[8145]

^ Me personally, if i had 15 healthy ones, i would just put that one out in the garden.

No I don't want them to be eaten by mold ! I am already too attached to them

[Amazonian]

^ Lol... your checking them every five seconds...aren't you (don't worry, i do the same).

You could try to cut out the mouldy bit, but it will probably spread its spores everywhere in doing so?. Or you can do what you did earlier and transfer the healthy segment to a new plate?!. I am very slap dash with most things i do... but sometimes... i get away with it

Edited by Amazonian, 12 July 2012 - 04:36 PM.

[Distracted]

If you want to remove condensation put some hot water into a cup and place the cup onto the plate of agar, it'll remove the condensation. Stacking freshly poured hot plates can also help remove condensation. Well... it'll remove it on all except the plate on top, which you can put a cup of hot water on.


It's been a long time since I did some agar work, i didn't put enough work into it to get anything proper out of it but was surprised by the lack of contams just from PCing agar in a bottle and pouring them into plates inside a glove box then sealing the plates with micropore tape.
I'll tell you what though, people give you some funny looks when they start seeing labeled plates of agar culture randomly throughout your house.


What exactly are you trying to achieve anyway? or just having a bit of fun? :D

[SallyD]

There's no way I'd open that one in the last pic posted, it has what appears to be a fully mature Trichoderma colony in the bottom left sector.
When trichoderma matures and releases spores it can release millions of them in seconds all it takes is a bit of a jolt.

Make yourself an inoculation loop like Therefore suggested and only take spores from mushrooms that have no signs of degradation (soft mushy spots, wounds etc) that have unbroken veils. Break the veils yourself after sterilising the specimen with alcohol and make some sporeprints of known quality.

Just because you see white fluffy growth it doesn't guarantee that it is mushroom mycelium, many moulds and contaminants look white or whitish until they fruit (like that Trich in the last pic did before it matured).

I realise this post sounds a bit harsh, which is not my intention, my point is don't fuck with anything suspicious. Throw it out and use a good one.
Only mess with contaminants if it is an absolute last resort and the culture would be lost without intervention.

Edited by SallyD, 13 July 2012 - 09:35 PM.

[Bigred]

im new here but i put some rice malt , honey, water(i copied a recipe from the net and put it in the microwave in a plastic container nuked it for about 7 min i dont have a pressure cooker yet so i then steamed for half hour on high left to cool then injected spores into it in a sterile
procedure would this work and plan on if i see growth is ad h202 but i have the hydro kind and it has some
silver product in it any help would be great

Edited by bigred82, 16 July 2012 - 06:59 AM.