Petri Colonization?

Started by psilly, October 02, 2004, 11:53:28 AM

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psilly

I sterilized 22 glass petri dishes, along with a jarful of standard RBF/verm mix and a jarful of pure vermiculite.  All contents were placed in a glove box and allowed to cool.  Then I placed chunks of the RBF/verm mix into each jar, inoculated with 3 drops spore solution, sprinkled with dry vermiculite, and covered with glass petri lids.

I removed the petri dishes from my glovebox, placed them on a sterilized plastic tray, and put the tray in a black plastic trash bag, which I then set on top of a heating pad with adjustable temp settings.  After four days at least five of the dishes have signs of nice, fluffy, mycelial growth.

Once the dishes are fully colonized, which I'm guessing will take between 1 week to 10 days from inoculation, the idea is to see if I can use 1/2 the contents of each dish to inoculate a standard PF jar setup.  I'm expecting contam rates to be higher due to the transfer process, but if I can inoculate 44 jars using 5cc spore solution, I can afford to lose a few. ;)

I'll keep you informed from time to time, worst case scenario I've wasted half a syringe and a little bit of effort.

anno

You can skip the petri dish though in this case and simply use a colonized pf cake to spawn dung/straw or to inoculate grain jars.

psilly

OK, time to check back in regarding my experiment.  The petri dishes were a bad idea.  Period.  Most of them were smelly within a few days.

Back to the drawing board...

I've been playing around with cloning techniques, and I think I stumbled upon something kinda cool.  If somebody else here wants to try to duplicate my results, I'd be glad to know what you found out.

I cloned ten GT tissue samples in agar, then decided to transplant directly to fifteen sterilized PF-jars using a glove box.  Remembering a previous post to this site about using coffee filters as lids, I decided to use filters with half of the jars, and tin-foil with the other half-- just to see what would happen.

The mushroom tissue colonized aggressively in all fifteen jars, although I lost a few more jars to contamination than normal (black/brown mold).  I'd assumed that contam rates would be higher, since inoculating with agar spears just seemed like it would be a bit messier than using syringe injections.  The ones with coffee filter lids seemed to fare far worse, though, which made me question that my inoculation was solely to blame.

Here's the interesting thing.  After about two weeks in the incubator, I noticed that the jars with coffee filter lids were shriveling up... duh.  Dehydration was kicking in, and the mycelial growth all but stopped in these jars.

Still playing mad scientist, however, I kept the samples.  I thought it might prove useful to know if they would recuperate after rehydration.  So when my jars with tin foil were fully-colonized, I cased both batches in two separate plastic trays in a peat/potting soil/bone meal mix.

Guess which batch EXPLODED into full colonization of the casing material over the next week and a half?  (Keep in mind, we're talking Golden Teachers here.  Hardly the most prolific colonizer.)  The dessicated, pathertic-looking, shriveled up, barely-colonized cakes FAR OUTPACED my 'healthy' cakes when water was re-introduced.

Any thoughts?  I'm thinking I may be on to something here.

anno

Interesting. Now if you can reproduce this behavior a few more times, then you might actually be on here.