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Sclerotia Concerns [ Mexicana A strain ]


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#1 goldenteacher1163

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Posted 09 July 2007 - 06:32 PM

About how long does it take before sclerotia really start to form? My foaf has got quite a lot of
experience with cubies, and decided he would like to try something else.
He shot 9 pt jars 3months ago with mex. A liquid culture ( lc is a multispore ). All jars contain 150ml rye grass seed, 75ml purified h²o, and a small knife tip of lime. The only difference in the jars is 4 of them also contain ¼ tsp of nutritional yeast just to see if it caused the mycelium to colonize faster as it has in the past when added to lc with other cubies. Anyway all the jars were fully colonized after 2 shakes and 9 days at temps around 76-78°F with almost no difference in speed.
Now is the part we found interesting all jars are still looking well except for the fact that there is almost no visible sclerotia, and the jars with the yeast have noticeably less than those without.
Is there a reason for this?
All jars seem to also have much less than he thinks they should have for 3 months worth of growth. Most only have a few small spots near the bottom of the jars and they are not that big ( I will get some pics asap ).
The jars have been kept in a black rubermaid type bin with the lid on to keep them in the dark, and the temps have been 76-79°f. One piece canning jar lids were used and they have been loosened for some air exchange.
If anyone could please give me any pointers or solutions it would be appreciated.

#2 Hippie3

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Posted 09 July 2007 - 06:44 PM

takes awhile,
check back in several more months

#3 Workman

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Posted 09 July 2007 - 06:56 PM

It does sound like less than stellar results. Pictures might help but it sounds like you are doing everything right. I usually see baby sclerotia form in about 3 weeks and 3 months they are really chunky and pretty much done.

#4 goldenteacher1163

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Posted 19 July 2007 - 08:54 PM

here are a couple of pics but they don't do it justice. better late than never
foaf was having a hard time getting good pics because of glare from jar
he is working on getting a few more to give me. Hope these help a bit, or maybe this is what the jars should look like

#5 Myc

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Posted 20 July 2007 - 03:55 PM

I am having identical results to your FOAF's with the exception that my cultures were started with hydrated spores and not LC on May 30th. I used annual rye grass seed pre-soaked, pc'd and innoculated. I use the standard canning jar lid with tyvek, and injection port which I cover with tape after innoculation, and a polyfil port in the center of the lid. One cultrue was supplemented with brewer's yeast prior to pc'ing and it is showing less vigorous growth. The plain grass seed finally germinated after around 45 days, has been shaken once and is almost fully colonized. I wonder if the method I used caused the seed to be too moist? Which would slow colonization IME.

This is much slower than the growth I demonstrated earlier. I posted a pic of P. Mexicana growing on a modified Dark Rye Flour cake recipe and was showing sclerotia formation after 30 days. The culture never produced much more than a tiny amount and I desroyed it by dissecting it for examination. I was, therefore unable to expand it further. Hence the rye grass seed experiment. Keep us posted. My FOAF would like to know how your FOAF fares.

BTW....As with all of my posts and pictures, I find it easier to speak in the first person about the results and methods of a FOAF. The reference FOAF is far too cumbersome and clutters up my sometimes wordy posts. Please take this as my blanket discaimer that none of this stuff is mine.

#6 goldenteacher1163

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Posted 23 July 2007 - 04:06 PM

foaf said jars seem to be starting to come around but they just don't seem to show the growth that was expected. He has come to the conclusion that it is a direct result of going multi spore.
This time a clone will be taken from a few of the better stones upon opening the jars. A few agar plates will also receive a deluted spore solution, and he is going to select for best stone producing strain. I guess we will see how he deos

With any luck when I see him today I will have an update on the jars at hand

#7 Myc

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Posted 23 July 2007 - 08:27 PM

Here are some progress pics. I think the thickened areas of mycelium are sclerotia being formed. Keep posting about your FOAF's results. I would like to know about cloning or final fruiting substrate should one be used. I haven't found much detailed information regarding this strain and would like to know more about it.

#8 Hippie3

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Posted 24 July 2007 - 10:08 AM

GT i'm wondering if yours are getting enough air,
why not rig up a better lid now to help,
might speed that growth up a bit.

Myc,
might yours be drying out a bit ?
i also note no growth on bottom,
you never shook those up to spread growth ?

#9 prankster239

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Posted 24 July 2007 - 10:36 AM

Here is an Mex A jar from me. It´s about 1,5 months old. The mexicana strains also from sclerotia very well on the dishes, the dishes are also 1-2 month old.

Normally it should no problem to get sclerotias, just wait longer.

#10 goldenteacher1163

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Posted 24 July 2007 - 01:50 PM

GT i'm wondering if yours are getting enough air,why not rig up a better lid now to help,
might speed that growth up a bit.


Thanks, that sounds like a good idea I will pass it along and see if it helps any.

#11 Myc

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Posted 24 July 2007 - 05:27 PM

Myc,
might yours be drying out a bit ?
i also note no growth on bottom,
you never shook those up to spread growth ?


I shook them once about 3 days ago and that sped up colonization. There is mycelium @ the bottom of the jars but it wasn't very strong at the time the pics were taken. I think the hydration is just about right. Maybe a little dry? My tek came from a book I have and suggested a 2:1 ratio of grass seed to water. There is moisture condensation on the inside of the jar which is typical of most of my cultures. Just not as much as I usually expect to see.

#12 Freaky

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Posted 24 July 2007 - 07:53 PM

Not very good luck here with my Mex A either. I just leave it and check it once a month or so.

Its been fully colonized for a couple months, no stones on it at all though. Oh well. I'll try playing with it again sometime, but not for a while as it was a big loss to take :(

Prankster, those stones look really nice. :cool:

#13 prankster239

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Posted 24 July 2007 - 10:53 PM

Its been fully colonized for a couple months, no stones on it at all though. Oh well. I'll try playing with it again sometime, but not for a while as it was a big loss to take :(

I dont understand this. The mex a always made sclerotias on nearly every substrate when i grow them.
Where did you get your spores? I hope sporeworks :greenboun
I think bags or plastic jars like my one are better for forming the sclerotia because there is more space for them to grow. I hope you understand what i mean, my english is bad. They are elastic.
Maybe your grasseeds are treated with some chemicals. Make an try with millet. In my grows they always produce very fast sclerotias on millet.

Dont give up, i cross my fingers for you :hippie:

#14 Freaky

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Posted 25 July 2007 - 07:26 PM

The rye grass seed is in myco bags. Spores from sporeworks. I think the lack of growth is from temp. fluctuations. That is my fault, not the spores or seed.

I'm wondering if the filter patch in the bags is enough for air exchange. Thinking about adding another filter or something since its colonized.

Either way, its fun to mess with the stones (in my case lack thereof) and learn :)

I'll take some pics and post them for you guys to look at here in a few days.

#15 Lazlo

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Posted 25 July 2007 - 07:39 PM

Myc's jar appears to have some bacterial problems in it.

#16 Myc

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Posted 26 July 2007 - 03:59 PM

Myc's jar appears to have some bacterial problems in it.


They seem fine to me but who knows? The spores took quite awhile to germinate but I didn't notice anything in the meantime. The growth pictured is only about 10 days worth (give or take). There certainly aren't any strange odors coming from the cultures so I guess only time will tell. I'm in no hurry.

Bacterial contamination might explain the slow germination but.....??? Wouldn't bacteria prevent germination altogether? I'll check 'em again in a couple of days and post more pics.

#17 goldenteacher1163

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Posted 01 August 2007 - 06:35 AM

Hippie foaf took your suggestion and made 4 new poly lids. The reason only 4 were made is that half of the interest in this hobby is experimentation for higher learning. This way if those 4 suddenly show a significant growth improvement then the problem is definate.
So far the growth in the 4 deos look better!! :bow: I want to thank you for the help

Some comparison pics are in the making

#18 goldenteacher1163

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Posted 01 August 2007 - 06:39 AM

one more thing if anyone out there could let us know what they do for fae while incubating jars for sclerotia production it would be great. I just wanted some ideas as to what kind of caps work and don't work or if maybe filter patch bags are better ect...... Thanks

#19 Hippie3

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Posted 01 August 2007 - 06:55 AM

Hippie FOAF took your suggestion

a wise foaf indeed

#20 truepercs

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Posted 10 August 2007 - 05:07 AM

Think its cause of the lime ...




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