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Official Copelandia/Panaeolus Thread


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#101 Microbe

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Posted 30 August 2019 - 11:38 AM

I’ve found pans do well with peptone, my recipe is 1000ml distilled water (950g)
32g dextrose (sugar)
16 g malt extract
16 g peptone
8g yeast
20 g agar
10 g potato starch

(Optional below)
.3 g ammonium citrate
.2 g thiamine
.2g magnesium phosphate
This is max Nute I’ll reduce sugars by up to 50% depending on what I’m doing

I too will still be around I’m going to put my pan tek on here for y’all.


That's interesting, quite a bit different from the "official" Pan. cyan. culture media from the CRC 'Handbook of Microbiological Media, Fourth Edition' (2010)

Dung Extract Agar
(DSMZ Medium 781)
Composition per liter:
Agar ............................................................................................15.0g
Malt extract...................................................................................5.0g
Ca(NO3)2·4H2O........................................................................0.72g
MgSO4·7H2O................................................................................0.5g
K2HPO4 .......................................................................................0.25g
Peptone .........................................................................................0.1g
Dung extract...........................................................................100.0mL
pH 6.9 ± 0.2 at 25°C
Dung Extract:
Composition per 150.0mL:
Horse dung.............................................................................. variable
Preparation of Dung Extract: Add an average sized piece of
horse dung to 150.0mL water. Gently heat and bring to boiling. Boil for
2 hr in a water bath. Filter. Use immediately.
Preparation of Medium: Add components to distilled/deionized
water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring
to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15
psi pressure–121°C. Pour into sterile Petri dishes or leave in tubes.
Use: For the cultivation and maintenance of Panaeolus cyanescens.


This media was apparently developed exclusively for Pan. cyan. and they were kind enough to use a pressure cooker setting on their autoclave, like they knew trippers were watching

Can peptone be replaced with anything OTC? My home lab has basically every inorganic ingredient to all the culture media I see but no peptone. Will whey protein work? A lifetime supply would cost me less than even the smallest quantity of any grade of microbiological peptone.
Thanks
I use brewers yeast or soy bean powder in place of peptone with comprable results. Peptone is expensive in comparison but its pharmaceutical/ lab grade. I would suspect that the proteins in peptone are more readily available to the culture also.

“It is always the simple that produces the marvelous.” —Amelia Barr
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#102 ItBeBasidia

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Posted 02 September 2019 - 11:32 PM

Got germination on pan cambo "sandose". hopefully I have better luck with these than the pan cyan Australia.a2dfe3fe0343e40ed1854639caaff5a4.jpg
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#103 peacefrog

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Posted 09 September 2019 - 12:07 PM

Amazing thread.

Sorry I’m very late to the party for this one lol.

I’m just not as active as I once was and on a mycology hiatus but y’all who know me, know I love thIs species. Keep this one alive for a while!!

Amazing pics so far.

Peace and good vibes my friends.
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#104 PanPiper

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Posted 12 September 2019 - 07:32 AM

Can anyone help positively ID the sub species?  I collected two different Panaeolus species in the wild in Cambodia last year and grew them out.  

 

I finally got the spores under a microscope to try and determine exactly what I have so I can file my prints/agar/spawn accordingly and continue to work on cultures.  Anyway some of the spores are 11um and others are 13um according to my friend who had access to the microscope.  

 

Can anyone tell from the slides which is Pan Cambo and which is Pan Cyan?  If I am not mistaken the 11um spores should be Pan Cambo and the 13um should be Pan Cyan.   Is that the distinguishing characteristic?  

 

Can Jake, Asura or any of the other pan wizards give pointers on how to ID?

 

Thanks

Panaeolus Spores 1000x 13 um
Panaeolus Spore unknown sub species
Panaeolus SpecimenB 11mm

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#105 jakeoncid419

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Posted 12 September 2019 - 06:27 PM

See
See the Copelandia key in Gerhardt 1996
However sport size alone is not going to be enough to determine beyond a shadow of a doubt what it is you could also count the basidia but even then it’s not 100% as there are anomaly’s to know for sure you need to do genetic sequencing (Send to alvalab.es for ITS sequencing)
Other than that the best you can do is measure 30 spores, then drop the largest and smallest 5 percent and look at the ranges to make a meaningful comparison. But for the most part Without comparing spore size measurements to sequence data there is no way of knowing how accurate it is. The sequencing let's you know which features to pay attention to and which are variable.
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#106 PanPiper

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Posted 13 September 2019 - 12:44 AM

Thanks Jake..I'm excited to learn more about it. Fyi, I think the first tray is pan cyan and the 2nd is pan cambo

 

 

Copelandia - tub July

 

Pan Cyan 12cm gills
 
Copelandia experimental grow

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#107 makinbones69

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Posted 13 September 2019 - 01:50 PM

That looks so awesome!
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#108 Asura

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Posted 14 September 2019 - 07:46 PM

PanPiper nice grow man. That looks great.

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#109 Foster

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Posted 20 September 2019 - 01:20 PM

So I hit about half a dozen 1/2pt jars of PC'd, whole grain, par boiled rice. (9er Tek style without the verm). They sat for 7-8 days before i got to them, so I was sure the jars were perfectly sterile and ready to go.

 

An MS syringe made from a small portion of Asura's Pan Jamaica print.  (TY Asura :) ).

 

Got 100% germination and nice growth , no contams.  Looks to be a pretty fast colonizer. I will post some pics soon as I get an opportunity.

 

So busy at the moment with real life, I haven't had time for photos, pasteurizing or much of anything else.  But I'm off to a good start and in no hurry.

 

Also got KapnDanks's Tex Orange Caps doing just as well,100% germ, clean and growing fast, with Ps, Serbica hydrating in a syr. (TY Kapn)

 

As well as ShadowLords' TC? hydrating and ready for my next round of PCing. TY Shadow old friend :)   

 

Good to be back in touch with all yall and nice to have some fresh blood at topia and fresh genetics in my grubby little hands ;)

peace n much love yall   


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#110 KapnDank

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Posted 20 September 2019 - 11:24 PM

I have two bags of Alabama pans almost finished thanks to asura.
Glad the TOC germed well. Did the serbica give you problems? I got 100% germ on both plates. Took a bit longer to germ than most but once it did it did so beautifully. Lots of strong looking sectors from the get go.

#111 Foster

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Posted 21 September 2019 - 02:58 PM

I wasnt planning on doing any agar, just straight to grain. I might save a little for some plates just in case.

I just have the spores hydrating because i wasn't sure how fresh they were and ran out of jars ready to noc.

I still have solution form both the Pan Jam and the TOC left over in the syrs i used.. And i only used a small portion of the prints to make those.

I'll be doing a pc run soon with more rice and use the Serbica, and TC then. Ill let ya know Kapn ;)



#112 Nurko65

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Posted 21 September 2019 - 04:03 PM

I had started a conversation titled Pan Cyan HELP in the Discussion Forum but it seems to have disappeared. I am not sure why. Hopefully I did not do something wrong.

Anyway, I am trying to get started on Pan Cyans, Australian strain and was having a number of issues on trying to get anything to grow. I received many useful comments and have changes my procedures going forward. Perhaps the biggest thing I learned is that starting from a MS syringe was less than desirable. I don't think molds were the issue but I had plenty of problems with what I think is bacteria which manifested itself as spots of wetness in the spawn. Anyway, I would sure appreciate some comments on the attached pictures.

The following shows some spawn that was inoculated 8/31/19. The jar on the left had a dime sized spot that appeared wet and didn't colonize. Are these ready to spawn to a fruiting substrate? Can I some how kill the bacteria spot? Another question, I've received two versions on what to do with the spawn as it is colonizing, to shake or not to shake. What are the pros and cons? Both these jars have not been shaken. (For some reason all the pictures I see are rotated 90 deg counter counter clockwise and I don't know why. If you are seeing the jars on their sides, please rotate the pictures to the right.)

Pan Cyan Spawn 83119.jpg

A couple of the many jars I tried on this project had this faint blue area on the bottom which I took as Trich however I later learned could simply be blue from bruising. Comments?

Pan Cyan Contamination 83119.jpg

Finally are three Petri dishes I inoculated on 9/17/19, 4 days ago. The top one is from a MS syringe of Pan bisporus. The growth seems stunted to me but is this normal? The bottom left dish is from a LQ I prepared directly from a MS syringe. There seems to be two layers, one growing faster that the other. If a do a transfer from this one, should I choose the outer layer or the inner one which likely contains two growths? The bottom right is a 3rd transfer from a dish. This looks the best I've seen so far. Should I do yet another transfer to improve it again or is it ready for a liquid culture of a grain substrate? The one on the lower left is 

Pan Dishes 91719.jpg
I would greatly appreciate comments on these pictures. I have mastered Cubes but these Pans seems to be a challenge.

 

 



#113 coorsmikey

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Posted 21 September 2019 - 06:26 PM

Right where you left it. You can use the "Find Content" button in your profile to look for any member's content https://mycotopia.ne...vity&mid=156581



#114 peacefrog

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Posted 22 September 2019 - 08:28 PM

I will attempt to answer your questions Nurko65.

None of those grain jar look healthy to me. I personally would not spawn them.

I have added some notes on your agar plates and attached. It’s the best I can do only going from the pictures.

Hope this helps. And good vibes!

BF30998B-C1F6-4D22-BCBC-5BD9FBBA9D24.jpeg
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#115 jakeoncid419

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Posted 23 September 2019 - 12:50 AM

Nurk
How old are the jars?
What do they smell like?
Pan myc will bruise blue on grain but those jars do look off. The colonization is not consistent.
Pan myc moves fast. Good culture should eat up a jar in 10 days
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#116 Nurko65

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Posted 23 September 2019 - 06:06 PM

I am still learning my way around the website but I am getting better, 

Thanks Peacefrog, that is about the best news I have received since starting this exercise in early June. 

All my attempts at creating spawn resulted in either no growth at all or very slow growth. The jars above were inoculated June 3, so 20 days ago. I have opened a few and the odor is not very strong at all - I suppose musty would be a good description. I have had jars smell much worst or sweet smelling.

I believe that both my Pans, cyanescens and bisporus spore syringes were contaminated with bacteria. I used the spores and a LC made from the spores to inoculate various substrates. I wasn't thinking straight but learned a lot hopefully can recover.

One question not answered was to shake or not to shake. I have heard both suggestions on the site and would appreciate the pros and cons or comments.

Thanks again for the advice and patience.


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#117 Asura

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Posted 23 September 2019 - 11:49 PM

Shaking is a great way to figure out if your culture is good. A healthy cyans culture should grow out a jar in about 10 days.

Contrary to popular belief, a healthy cyans culture will be quite vigorous and is not the delicate flower that some people

seem to believe that it is.

 

If you shake and the culture dies, you've just saved yourself a lot of time and effort. 

 

Personally, I think spores to agar then to LC is the best way to start cyans. You can put the LC to grain and do a traditional

spawn to bulk style grow or you can use the LC to inoculate sterilized substrate in bags (this is what I do).

 

For whatever reason, I have not had good luck putting agar wedges to grain. I seem to get a lot of dead jars when I do this.


Edited by Asura, 23 September 2019 - 11:50 PM.

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#118 Nurko65

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Posted 25 September 2019 - 02:31 PM

Thanks Asura. That is the path I will follow; spores to agar to LC.



#119 peacefrog

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Posted 26 September 2019 - 04:56 AM

As said above, shaking a jar will not hinder growth unless it has contamination present.

Copes/pans, whatever you choose to call them, will not be hurt by several distribution shakes IF all is healthy. I shake mine much earlier then a lot of cultivators and never see an issue unless I have contamination present.

Here is an example of when I shake for the first time with them:
51D7F278-333E-4A06-AC94-CB089D8BC3F4.jpeg

Pretty early. But you must take into consideration, that jar was my master and had very little grain in it and some manure added. If that had been a full quart or something, I would have let it grow a couple of more days before shaking. But hopefully you get the idea. Either way, the point is that shaking/distribution will not hurt a thing.

If you take a spore syringe to LC, you will have no idea what you are growing until you test it. Contamination could be either throughout or just hitchhiking along with the mycelium. Agar is your friend lol. It’s not a must, but can be a very valuable tool as a cultivator. I ALWAYS take spores to agar first, one never knows.

And it’s up to you if you want to use agar wedges verses LC/LI to inoculate grain. They have their advantages and disadvantages, but will get the job done just the same if all is clean.

#120 ElrikEriksson

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Posted 26 September 2019 - 12:37 PM

I'm curious what I can use as a casing for Pans.

I'm ready to put spores to agar, and I have enough peat left for this first grow, but I'm trying to discern if I should get more while it is still in stores. I have no other use for peat.

I've seen in old threads materials such as cactus soil are used, more recently it seems just limestoned peat is in favor. I have 30 or 40 litres of a fine peat-like store bought potting soil that I don't use for anything. Its composition is listed as peat, finely shredded tree bark, coir, compost. Will this be likely to work well or should I make a point to do a trial grow with an isolate I grew out with my last remaining peat?

On the other end of the spectrum, would composted conifer duff be worth an attempt or would that be too nutritious?

I like to make a significant portion of my food and medicine from this land and I am already in the habit of composting such materials in quantity, so the attempt would be no inconvenience.






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