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Agar plates ...


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#1 OZZZ

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Posted 21 October 2008 - 09:42 PM

Im new to agar, and here are a few of the first plates I did.
This first one is just a scraped TX print to PDA.

Im curious as to why Im not seeing very rhizomorphic growth compared to many I see online? Did I just happen to scrape a portion of the print that didnt contain any strong substrains?

I dont really see any sections that look better then the others as far as isolation goes. Im not planning on isolating, I just wanted to try to germinate some spores on agar with a print that I didnt need for practice before I used my good prints. But if I was to continue with the plate ... I dont see anything worth selecting do you?

agar (2 of 3).jpg

agar (1 of 3).jpg

Below is a piece of GT mycelium that I saved from extinction .. haha. I got a contaminated syringe from a vendor. I love this particular strain from this vendor it has always produced fantastically for me even from MS. Unfortunately with the syringe being badly contaminated everything it touches turns to shit in a hurry. I happend to see a small whisp of mycelium in one of the jars that I grabbed with a tweezers, dunked in h202, then put on this dish after I swished some h202 on the surface of it.

It looks really fluffy and white, but not rhizomorphic. Now you can see just in the last few days it has jumped to the plate (its 15 days in). Im curious, the growth extending from the center piece looks a bit more rhizo but still quite whispy ... is it possible for this piece .. looking like it does to produce rhizo sections as it grows? In your experience? Or did I happen to just get another weak substrain on this round as well? This makes more sense that it could happen in this case since it was a small piece of mycelium I grabbed, but in the tex print situation above .. not so much I would think out of all the substrains that landed on the plate one should have been rhizo (there was a bunch of spores on the plate from the tex print scraping.)

agar (3 of 3).jpg


Edited by Sidestreet, 14 August 2015 - 09:45 AM.
Re-posted images in-line


#2 Jigalow

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Posted 21 October 2008 - 10:00 PM

How many days has your plates gone?
Opps helps if I read 15 days in


#3 OZZZ

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Posted 21 October 2008 - 10:01 PM

......... As oppossed to this! This is Quezon strain in a quart jar of coir and worm castings. I shot spores into it just like BRF and it colonized the entire jar in 14 days.

Look at the fucking rhizo's here. Ive been using it as a master for g2g's.

I think Im going to take those two rhizos clinging to the glass and put it on some plates.

Do you think my plates up above are just showing whispy growth because of the h202?

 

Attached Thumbnails

  • quezon (3 of 3).jpg
  • quezon (2 of 3).jpg
  • quezon (1 of 3).jpg

Edited by Sidestreet, 14 August 2015 - 09:45 AM.
Removed offsite image links


#4 OZZZ

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Posted 21 October 2008 - 10:02 PM

Jigalow ... Both the TX and GT plates have went 15 days. But the GT plate was a piece of mycelium so it went on h202 PDA where as the spore print went on straight PDA (obviously)

#5 Jigalow

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Posted 21 October 2008 - 10:06 PM

How is the agar, starting to dry a bit?

I'd take the marked parts and get them going on new agar.

You should be able to get more if you peel back the fluffy stuff
as it's a different layer on top of the other.

 

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Edited by Sidestreet, 14 August 2015 - 09:46 AM.
removed broken link


#6 Jigalow

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Posted 21 October 2008 - 10:13 PM

My PR's look just like your GT (Fluffy) and I used an LC.
My GT from a GT clean up is just hanging onto life...

I am on I think transfer 3 now of the PR's. It's slowly getting more Rhizo.
Just try to pic about a match head size and get it going on new agar.

Maybe some one more seasoned could help out here...


#7 OZZZ

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Posted 21 October 2008 - 10:18 PM

OK ... Ill put them on new plates I just poured some new agar tonight. Why do you think the Tex Print looks so shitty? You think the agar is dry?

I guess I expected to see areas of fluffly growth and areas of rhizo growth when doing a ms scrape to agar. Of course the idea being to isolate the rhizo growth ...

#8 Jigalow

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Posted 21 October 2008 - 10:20 PM

IMO From all the different spores and the water transfered it all over the agar.

#9 Jigalow

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Posted 21 October 2008 - 10:23 PM

The only one I have that is VERY Rhizo so far is my B+ but working on the others.....
 


Edited by Sidestreet, 14 August 2015 - 09:46 AM.


#10 sandman

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Posted 21 October 2008 - 10:50 PM

thats just how plates look from multispore swipe. Selet several areas and go from there. hen repeat a few times. Then your growth will look more rhizo. WHat you be sealin them dishes with?

#11 OZZZ

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Posted 21 October 2008 - 11:08 PM

I see .....


Those were sealed with clear packing tape because I had nothing else ... lol

But the new ones are sealed with strips of glad wrap, I saw a tek somewhere a guy uses it exclusively because it is gas permeable.

Supposedly only the glade brand not the cheap brand? But thats what Im using on the new ones since I dont have parafilm. It worked fine I only had one plate out of 9 contam and that was from taking a shiitake clump out of a half colonized/contamed clump of straw. One transfer and it was clean ... this agar stuff is making me feel good because I know my general actions and techniques are sterile as all the plates are coming out clean lol

#12 Workman

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Posted 22 October 2008 - 12:56 AM

Sandman is right. The large number of spores germinating all together in a small space inhibits rhizomorphic growth. Sometimes a really aggressive strain breaks away from the rest, but generally you don't see rhizos until after at least one transfer.

I use 2 inch cut glad wrap mini rolls to seal the edge of my petri plates.

#13 OZZZ

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Posted 22 October 2008 - 06:05 PM

Right on, good to know ... thanks again guys!

Ive transfered those few rhizos from that quart jar to a plate, and I just got done doing some portebella grocery store clones as well. I took a few sections from the GT plate and put on new plates also.

So its not the h202 that is making for more fluffy growth due to slowing it down a bit?




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