Jump to content
The Corroboree
TheMooseZeus

Is something wrong or am i impatient?

Recommended Posts

Hey all!

 

I on the 25th i inoculated some agar dishes with a summer fruiting mushroom.

 

After 8 days i have seen no growth (mycelial or other) on the plates.

 

The spore prints were dated 2019 and i followed pastywires tek.

 

They are being incubated at ~24o

 

Any knowledge and wisdom would be greatly appreciated!  

  • Like 1

Share this post


Link to post
Share on other sites

HI Moose,

 

Since you are incubating them i'd say something's gotta be happening. BUT this depends on the mushroom/strain/viability/etc of the spores. Search around for the strain you are trying to grow to see the average time for spore germination, i.e.some are pretty quick 2-5 days, some can take up to 4 weeks if not longer. Also technique would be another one, making sure you actually got some spores on there (I've done that a couple times). Another good idea may be to hydrate the spores, by ways of making a spore syringe and leaving it for 24hrs before going to agar. 

Just some thoughts, hope they help :)

 

Max

  • Like 2

Share this post


Link to post
Share on other sites
On 08/05/2020 at 6:21 PM, Maximt said:

HI Moose,

 

Since you are incubating them i'd say something's gotta be happening. BUT this depends on the mushroom/strain/viability/etc of the spores. Search around for the strain you are trying to grow to see the average time for spore germination, i.e.some are pretty quick 2-5 days, some can take up to 4 weeks if not longer. Also technique would be another one, making sure you actually got some spores on there (I've done that a couple times). Another good idea may be to hydrate the spores, by ways of making a spore syringe and leaving it for 24hrs before going to agar. 

Just some thoughts, hope they help :)

 

Max

Thanks so much for the reply!

 

I've used two strains so far which have reportedly quite short germination times - The remaining plates i have going have shown no contamination (The rest had contamination which i assume is a sign that it isn't a problem with the agar i'm using). I definitely have spores on the plates maybe even a few too many. Some have quite obvious smear marks. As for hydrating i've heard that can help but cant help but think that it could introduce contamination. I have also heard that because the spores are so tiny such a small volume of water is needed to hydrate the spores. An amount that could be easily achieved in the agar plate.

Although i definitely want to look into spore syringes!

 

Cheers!  

Share this post


Link to post
Share on other sites
Posted (edited)

So have you got any that germinated, even if they had contam? or do you have some empty and some with contam, and no signs of mycelium at all? If so, would come down to viability of the spores, maybe dried out? how were they stored?

Also, what agar recepie are you using, i've heard that PDYA germ rates are lower than MYA.

 

Completely agree, the liquid on the plate should be sufficient, but in some cases that produces better results that scraping to agar. Also, somewhat limits the number of spores you put down on a plate as you just need to make one drop in the center. This will disperse them more evenly rather than clumping together. Too many spores can stop/slow growth, as mycelium at this young age dont like competition, BUT thats not to say they will not germinate or that it prevents them from germinating.

 

For spore syringes, i suggest taking a syringe (wrapped in alfoil so you can wipe with alco if needed) and a shot glass filled with water (covered with alfoil), PC both for 45min. In front of your flowhood/still air box using a flame sterilized inoculation loop (remember spores dont like sharp objects), get some spores and swirl it around in the glass. Alternatively fold and rub the print to drop some spores into the glass, and use the needle to swirl them around. Then draw the water with the syringe and you are done.

 

Hope that helps.

 

Max

Edited by Maximt
added a question
  • Thanks 1

Share this post


Link to post
Share on other sites
On 11/05/2020 at 1:45 PM, Maximt said:

so have you got any that germinated, even if they had contam?

I don't think so, i had one with a vague looking white something on it but i couldnt see and ended up scrapping it because of contam. I set up a control plate which i didnt open but stupidly removed the label and lost it... oops

 

I am using PYDA, that might have something to do with it.

 

Thanks so much man, ill get round to that soon, maybe get a hold of a fresher print :D

 

Its been a few days now without checking on them so ill get back to you if i see anything

 

Cheers!

 

Share this post


Link to post
Share on other sites
On 14/05/2020 at 3:23 PM, TheMooseZeus said:

I don't think so, i had one with a vague looking white something on it but i couldnt see and ended up scrapping it because of contam.

 

That to me would indicate viability of the spores. If you've done a bunch of plates something should have germinated (except for contam ofcourse). So maybe try to hydrate via the syringe method, leave them there for 24hrs and then do a drop on a plate. And maybe try MYA, you can get malt at Dan Murphy's or brewer store for fairly cheap.

Spore prints can last a long time (i've heard 10 years) if stored properly, the biggest issue is they dry out. I don't have much experience in this area, but some key points are they should be in a zip-locked bag, and in a cool dark place (not fridge as they will dry out there as well).

 

Happy to help with what i can. Good luck.

 

Max

  • Like 1
  • Thanks 1

Share this post


Link to post
Share on other sites

Create an account or sign in to comment

You need to be a member in order to leave a comment

Create an account

Sign up for a new account in our community. It's easy!

Register a new account

Sign in

Already have an account? Sign in here.

Sign In Now

×