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Mex.a on agar


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#1 carpo4

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Posted 28 February 2005 - 08:54 AM

here's a recap of the grow so far:
mea plates were inoculated on the 18.1.05
with a sporeworks spore-syringe.
two (pic1 and pic2) produced sufficient growth for the first set of transfers on the 1.205 (pic3, pic4 and pic5),
and from these, a further set of transfers was made on the 8.2.05 (pic6, pic7 and pic8).
they were left to form sclerotia,
the first of which were noticed on the 23.2.05 (pic9).
currently, 6 out of the 36 plates transferred to are showing signs of slerotia formation (pic10, pic11, pic12, pic13, pic14 and pic15).
including one plate from the first set of transfers, which i had decided to hold on to (pic16).

an interesting part of the sclerotia activity
is what appears to be small globules of liquid sitting on their surface.
is this emanating from within the sclerotia itself
or due to condensation from the heat of development?
good thing/bad thing?
also, is it possible to tell from where the sclerotia are forming
as to whether a plate is composed of a single isolated substrain, e.g. pic10?

Attached Thumbnails

  • masterplate1rsz.jpg
  • masterplate2rsz.jpg
  • b1rsz.jpg
  • b4rsz.jpg
  • b11rsz.jpg
  • 3rd transfer big 1.jpg
  • 3rd transfer big 2.jpg
  • 3rd transfer small 2.jpg
  • sclerotia c25 rsz.jpg
  • sclerotia9 rsz.jpg
  • sclerotia6 rsz.jpg
  • sclerotia5 rsz.jpg
  • sclerotia3 rsz.jpg
  • sclerotia2 rsz.jpg
  • sclerotia1 rsz.jpg
  • sclerotia4 rsz.jpg
  • sclerotia7 rsz.jpg


#2 Guest_Peter Cottontail_*

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Posted 28 February 2005 - 09:09 AM

I've seen moisture on the sclerotia in a petri dish before. Most likely metabolyte. Doesn't hurt anything. I'd just let those grow out a bit more, then take the one with the most sclerotia and use it to inoculate your grain.

#3 carpo4

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Posted 28 February 2005 - 06:17 PM

thanx, rodger!
well, so far, my money's on that plate in pic10.

#4 destroy_erase_improve

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Posted 28 February 2005 - 07:19 PM

so after you pick a plate, you use some of it to spawn to jars to grow out. now do you keep that plate that was cut out of as the master. or do you cut a piece away and grow out another dish, and use that NEW dish as the master for storage?

#5 Guest_Peter Cottontail_*

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Posted 28 February 2005 - 07:57 PM

The latter. I like freshly poured agar for storage in the refrigerator.

Let's say one of his dishes has ten times as much sclerotia forming as the rest. That's a good master. Transfer a single piece of the sclerotia to a new petri dish. Allow it to grow out for a week or so until the dish is half covered with mycelium. Wrap the dish in several layers of newspaper for insulation against temperature swings, and place in refrigerator.

The original dish with all the sclerotia can be cut 8 ways and each wedge is enough for a quart jar of rye grain. For faster colonization, use one petri dish for 4 quarts of rye. Mex a does very well in grain to grain transfers, so each of those 8 jars can be expanded into ten more to give you 80. Each of those can be expanded into ten more to give you 800 quart jars within six weeks, from a single petri dish.

#6 destroy_erase_improve

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Posted 28 February 2005 - 08:15 PM

makes sense.
if your going to use a dish for four qts am i correct in assuming to cut into smaller pieces as opposed to one big chunk per jar. creating more innoc points and faster colonization. (^^^^^intrigued by sclerotia and agar work)

#7 carpo4

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Posted 10 May 2005 - 07:29 AM

time to blow the dust off of this thread
with a little update on my grow:
6 x 600ml jars of coffee-soaked rye grain
were innoculated on the 4 march
with agar wedges taken from the strongest sclerotia producer (pic10).
600ml jars were chosen over quarts,
shown here side-by-side for comparison,
due to their increased surface area-to-volume ratio,
as observations have been made previously
that sclerotia tend to form more against the glass interface.
unfortunately, these jars have been a wash out
due to the myc refusing to grow off of the wedges.
they were shaken after a week or two,
which helped spread innoculation points,
and eventually fully colonised
but even after 2 months from initial point of innoculation
there has been no visible sign of sclerotia,
apart from this single fella!
in contrast, i also decided to do a batch of 12 x 1/2pts,
syringe-injected with the same isolated substrain used in the 600ml jars,
which had been grown out in a dex jar from a cut of agar.
after only 28 days from point of innoculation
sclerotia growth is evident,
although not consistent across all the jars.
here's a comparison of the three strongest jars so far:
growth on day 28 for jar 1, jar 2 and jar 3
growth on day 34 for jar 1, jar 2 and jar 3

Attached Thumbnails

  • mex.a.3_day34.jpg
  • mex.a.2_day34.jpg
  • mex.a.1_day34.jpg
  • mex.a.3_day28.jpg
  • mex.a.2_day28.jpg
  • mex.a.1_day28.jpg
  • mex.a jar sclerotia_day60.jpg
  • mex.a dex jars.jpg
  • 900 and 600ml jars.jpg
  • sclerotia7 rsz.jpg
  • mex.a jars_day60.jpg


#8 Guest_Peter Cottontail_*

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Posted 10 May 2005 - 10:51 AM

Interesting. However, I haven't found a correlation with sclerotia growth along the edges of the glass. They seem to grow throughout the grains. With a really good sclerotia producing substrain, you'll end up with 1/2 to 2/3 jar of sclerotia from your original jar of grains.
RR

#9 Guest_novak_*

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Posted 11 May 2005 - 07:35 PM

i've been looking for agar info to read up on. anyone has some good archive threads that will explain the basic of how and why? the more info in it the better!

#10 Hippie3

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Posted 12 May 2005 - 06:05 AM

i've been looking for agar info to read up on. anyone has some good archive threads that will explain the basic of how and why? the more info in it the better!


AGAR from a to r

#11 carpo4

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Posted 12 June 2005 - 07:24 AM

i have several plates sitting in my incubator,
isolated substrains which can be seen at the beginning of this thread,
which i've been waiting on
to see if they'll fruit invitro.
well, here are a couple of solitary fellas i found today.
any suggestions as to what these plates would be good for
if anything?

Attached Thumbnails

  • c22 invitro fruit 4months.jpg
  • c10 invitro fruit 4months.jpg


#12 Guest_Peter Cottontail_*

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Posted 12 June 2005 - 07:49 AM

They're probably too old really to be of much use, in my opinion. Too many cell divisions by now so senescense would have set in. That was most likely the problem in getting the jars to colonize in April/May. If you have more spores, try again or if you're out of spores, you might cut that chunk of sclerotia into several pieces and try starting them in a fresh plate. Personally, I'd go back to spores. A really good Mex A isolate will have sclerotia forming within two to three weeks of dropping spores on the agar.
RR

#13 Hippie3

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Posted 12 June 2005 - 07:49 AM

see if you can turn that shroom into a mycellial syringe,
multiply that in liquid nute solution
and then stab some jars ?




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