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OfflineShroomasta
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!?! Info on Advanced Pinning Techniques !?!
    #629709 - 05/13/02 10:32 PM (14 years, 6 months ago)

I have a friend who is currently in the process of trying to get through a batch of jars that don't seem to want to pin. This is what he described to me as the method he used:

Firstly, rye berries were used as the substrate and placed into Ball quart jars, filling them half way. The lids were fastened upside-down and wrapped in alum foil. The jars were PC at 15psi for 45min. The grain was innoculated under a laminar flow hood and incubated at room temps (67-75) for now coming on 1 month. The jars are definitely fully colonized, however, they do not seem to want to pin.

The original plan was to use a negelct tek (Hippie3's myco-tek on PF site) on these jars, however, as invitro pinning was not initiated by incubating in light, the plan was to proceed by continuing with the neglect tek by initiated pinning in the jars now and continuing on. However, no matter what has been attempted, which has included giving it more air, keeping temp constant at 76 and aloowing for 12 hour light cycles, nothing seems to initiate them to pin.

One method of trying to use the grain was to colonize dung/straw mixture which was prepared as per the bulk neglect report (http://shroomery.org/forums/showflat.php?Cat=&Board=Forum4&Number=609532&page=0&view=collapsed&sb=5&o=&fpart=1). Dung was optained in chunks and pasteurized as said (165 at start) for 1.5 hours and then drained overnight on a rooftop. The boxes were filled on the bottom with the dung/straw and colonized with 1.5 quarts of spawn from the jars. They were cased useing an extra layer of dung. Colonization initially seemed to go well, however, 1 week in cobweb mold seem to take hold and he began to spray with a 2 Tbs baking soda to 1 quart water mixture, three times a day, as well as a 1:4 h202 mixture. This did not seem to contain the mold. Other boxes also seemed to develop other molds. Hopefully it is just the top layer, and the bottom can be recased after the infection ( tips?). However he has essentially abandoned that venue.

Now he is back to trying to work on the jars. Does anyone have any tips on getting them to pin or anything they would like to contribute about the above mentioned methods? Any ideas as to what might have gone wrong? Currently the jars seem to have over colonized are are quite fluffy and are producing some yellowish-orange'y goop. He is thinking about cold-shocking outside (~60) or in the fridge (how long should this be for?). Also, is there any way to continue the neglect tek or do these have to be birthed? Also, if he were to just birth them and place them on cafeteria trays lined with alum foil and mist them placing them in a cabinet, could this work? Ideas? Should he be thinking about using the flat cake tek and casing with verm? Any help would be greatly appreciated!! He has been wracking his brain and is really trying to figure out what he did wrong.


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InvisibleBeautifulDay
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Re: !?! Info on Advanced Pinning Techniques !?! [Re: Shroomasta]
    #630514 - 05/14/02 02:45 PM (14 years, 6 months ago)

>drained overnight on a rooftop
...interesting...

I know someone else who uses similar methods, with great success. What are the details of your terrarium? it would seem that, since you are going to the bulk neglect tek, and since none is mentioned in your post, that you have neglected to build a terarrium. Without a terrarium, there is very little chance you will get cakes to fruit, they need a very high humidity level because their bare mycellium is exposed to the raw elements and succeptable to drying out. How watertight/airtight is the 'cabinet' you refer to? You may be successful using it with perlite for humidification. Read the teks man, use the PF or MMGG recommendations. Though the bulk neglect tec appears very simple on first inspection, it is actually a full on spawning-casing tek, the beauty of it is that it allows you to forego the complex terarrium setup. Hope that helps.
Ciao
BeautifulDay


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Offlinevatoloco
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Post deleted by MOE THE MAD SCIENTIST [Re: Shroomasta]
    #630571 - 05/14/02 03:47 PM (14 years, 6 months ago)



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OfflineShroomasta
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Re: !?! Info on Advanced Pinning Techniques !?! [Re: BeautifulDay]
    #630593 - 05/14/02 04:15 PM (14 years, 6 months ago)

Thanks for the input BeautifulDay. Few questions...since you say that it needs high humidity, in teh jars, the humidity level is realtively high. At least high enough to have condensation on the sides and the lid. Is this not enough? (At least for pinning?) Also, if there is constant misting, could I birth without a terrarium? How well do you think the cakes would do without casing just sitting out on a sheet?

About the bulk neglect...what aspects of it allow the bypass of the terrarium? As in, why doesn't the humidity level matter there? WOuldn't the mycelium still need the same conditions, yet they still grow without the high humidity once cased using the bulk neglect. Also, any tips on containing and keeping mold at bay? Thanks for any suggestions you can provide.


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OfflineShroomasta
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Re: !?! Info on Advanced Pinning Techniques !?! [Re: vatoloco]
    #630597 - 05/14/02 04:16 PM (14 years, 6 months ago)

The EQ and the B+ strains have both been tried invitro and work well...so I don't htink that this is a problem. Dunno...seem to have hit a roadblock here.


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Offlinefelixhigh
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Re: !?! Info on Advanced Pinning Techniques !?! [Re: Shroomasta]
    #630984 - 05/14/02 10:44 PM (14 years, 6 months ago)

i think i've seen this before... it seems you're growing a non fruiting strain... i dont know how long you've been keeping it alive but your only chance (if there's any) is isolating the rhizomorphic sectors rather than cottony ones until a pure rhizo growth is stabilished...
anyways, good luck.


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OfflineBaby_Hitler
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Re: !?! Info on Advanced Pinning Techniques !?! [Re: Shroomasta]
    #631048 - 05/15/02 12:12 AM (14 years, 6 months ago)

Have you tried taking the tape off, and/or loosening the lids, and turning them upside down? could be an air exchange problem.



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Offlinefelix
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Re: !?! Info on Advanced Pinning Techniques !?! [Re: Shroomasta]
    #631066 - 05/15/02 12:36 AM (14 years, 6 months ago)

light, fresh air and humidity/temp.

they will not produce fruits unless the co2 content is low enough. fresh air

since you incubated at temps of 65-75, the fungus may be accustom to lower temps. try a lower, but stable temperature and keep them at that level until they fruit. OR a regular overnighter in the fridge may work.

jars of rye are not usually fit for 'cakes'. im not saying it won't happen with straight rye, but im thinking you should case those with AT LEAST a straight vermiculite layer.

plus, you say you are coming up on only 1 month. more patience.
DON'T mist cakes directly.


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Offlinefelix
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Re: !?! Info on Advanced Pinning Techniques !?! [Re: Shroomasta]
    #631072 - 05/15/02 12:42 AM (14 years, 6 months ago)

oh yeah...if you want to take them out, you should at least get a small rubbermaid and fill it with perlite/water. this way you don't have to mist. you mushrooms will LOVE a cool, dank rubbermaid.


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Real botanists laugh at HPS systems, we do however use high pressure sodium in the parking lot. - artthug


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OfflineShroomasta
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Re: !?! Info on Advanced Pinning Techniques !?! [Re: felixhigh]
    #632079 - 05/15/02 06:45 PM (14 years, 6 months ago)

Non-fruiting strains are cultured how? Was it possibly something that went wrong in the incubation phase or was it the original spores? I asked him about his jars and he says that there are ones that contain think amounts of rhizo growth and those also are not pinning. Hrm...I'll keep trying things.


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OfflineShroomasta
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Re: !?! Info on Advanced Pinning Techniques !?! [Re: felix]
    #632086 - 05/15/02 06:48 PM (14 years, 6 months ago)

I'm guessing that his problem may be the c02 content. I can't really think of anything else that seems to be wrong. It's day four after cold-shocking in the fridge for a a couple of jars and still nothing...so I guess he'll try breaking up the jars and casing them with verm or maybe coco coir and verm. Basically, his main concern is that since he lives with other ppl and they may have problems with his stuff, he can't really use large terrarium setups to get all of his stuff in it. Therefore, he hopes to do it without terrarium's (ie no real humidity control)...does this completely jeopardize the ability to get shrooms? Thanks for comments so far guys!


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Offlinefelixhigh
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Re: !?! Info on Advanced Pinning Techniques !?! [Re: Shroomasta]
    #632290 - 05/15/02 09:46 PM (14 years, 6 months ago)

taken from the mushroom cultivator:
"Of the many newly created strains intrinsic to multispore germination, some may be only capable of vegetative growth. Such mycelia can assimilate nutrients but can not form a mushroom fruitbody (the product of generative growth)."
i think this is the case (i'm assuming it was a multispore innoculation)...
it's not spores fault, the problem is that certain matches of spores can produce sometimes these vegetative mycelium... happened to me a couple times. it seems perfect, smells nice you do everything right but it just refuses to fruit...
you should read about mushroom genetics to learn more (i can barely remember this stuff)...


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OfflineShroomasta
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Re: !?! Info on Advanced Pinning Techniques !?! [Re: felixhigh]
    #632294 - 05/15/02 09:52 PM (14 years, 6 months ago)

The spores that were purchased from thehawkseye were not multispore. They were isolated strains. Some of the strains that were used inlcuded EQ, B+, Thai LB, Thai LY, and Plantasia. I had heard good things aobut hawkseye before, so I don't think it was the spores. I'll get picks up later and hopefully, you guys can help identify the problem better. Thanks.


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Offlinefelixhigh
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Re: !?! Info on Advanced Pinning Techniques !?! [Re: Shroomasta]
    #632300 - 05/15/02 10:02 PM (14 years, 6 months ago)

what? you were not able to fruit all those strains??? then you must be doing something very wrong...
but anyways, no matter if its an isolated strain or whatever, when you put a lot of spores into a substrate you're making a multispore innoculation buddy... and please, get some reading for your shrooms sake...


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OfflineShroomasta
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Re: !?! Info on Advanced Pinning Techniques !?! [Re: felixhigh]
    #632397 - 05/16/02 02:16 AM (14 years, 6 months ago)

Do be advised, that he can get them to fruit when cased, however, invitro fruiting is the question here. Is there any way to invitro fruit shrooms out of rye jars without casing? If casing is nesscesary...can it be done by just adding a moist verm layer to the tops of the rye jars and then let them sit it out? Is it possible that the verm in teh BRF cakes povides some sort of cellular scaffold for which the mycelium need in order to fruit (ie the reason why casings do produce fruits?). If he were to add verm to the rye jars and try to invitro fruit that way, do you think that might work? In essence try and turn it into a BRF cake only with rye. Thanks for your comments.


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Anonymous

Re: !?! Info on Advanced Pinning Techniques !?! [Re: Shroomasta]
    #632749 - 05/16/02 09:22 AM (14 years, 6 months ago)

The problem is CO2. They will pin eventually, but will have to consume all of the available nutrients within the rye first.

Any strain will eventually fruit invitro, some are just more CO2 tolerant, and will pin in response to temperature drop, and light exposure alone. Others require a drop in CO2 in combination with Temp, and Light stimuli. All fruitable Strains will eventually pin regardless of Light, Temp, and CO2, in response to NUTRIENT DEPRIVATION.

You are just gonna have to wait. The mycelium colonizes and feeds very slowly at the temp range you have used. It is not gonna fruit until Available nutrition has been depleted to some critical level.

I find Strains pin at a much slower rate, even when cased, if the colonization of substrate took place at lower temperatures then ideal. They do however pin in response to lower CO2, Temp. reduction, and light. They just do so slower then ideal.

In the future, you might find filter disks to be an advantage, in your scenario. Due to your inability to maintain higher Temps for colonization(Faster growth rate), resulting in faster nutrient depletion, you will be wanting your mycelium to respond to the other main trigger you have not dealt with, CO2. Filter Disks will allow for some gas exchange. They will increase your growth rate slightly, even at those temperatures, and they will allow for much needed gas exchange.


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Anonymous

Re: !?! Info on Advanced Pinning Techniques !?! [Re: ]
    #632754 - 05/16/02 09:25 AM (14 years, 6 months ago)

As colonized as they look, they are still colonizing. There is available nutrition in the rye, and they are gonna keep growing Vegetatively, unless you can get rid of the CO2.


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OfflineShroomasta
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Re: !?! Info on Advanced Pinning Techniques !?! [Re: ]
    #632842 - 05/16/02 10:22 AM (14 years, 6 months ago)

Ok, thanks a bunch. I'll have him work on the CO2 situation and hopefully that will get some things straightened out. Secondly, how rhizomorphic should the growth look normally. Many of the jars (rye) do have small fiberous looking growths all around, however, the BRF cakes seem to have much larger and longer outgrowths...this may also be a reason. I'll try and have him work on casing this stuff. Thanks again.


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Anonymous

Re: !?! Info on Advanced Pinning Techniques !?! [Re: Shroomasta]
    #633196 - 05/16/02 02:28 PM (14 years, 6 months ago)

BRF is a readily available substrate for the mycelium to colonize,EAT. The mycelium will do so at a faster rate, then Rye grain. You should have more rhizomorphs on BRF then Rye grain, in the same span of time. Eventually, if you wait, RYE GRAIN will be a more nutritoussubstrate and result in higher yields. It just takes a longer time for the mycelium to eat the ryegrain!!!

Your problem was a Temperature problem first, and all other problems are associated around the Growth rate of the mycelium and it's effects, due to this temperature problem.

In over ten years of growing cubensis, I have never had a problem with non fruiting substrains, and I rarely multispore. When multisporing, this possibility should be eliminated entirely. You have many strains growing in each jar, and one of them will be a fruiter, FOR SURE.

Rhizomorphs invitro are an indication that the jar is approaching the pinning stage. The mycelium has gathered sufficient nutrients, the available nutrition of the substrate is becoming limited, and it is ready to fruit. The quality and quantity of nutrient gathered will determine the extent to which they will fruit. Given water is not limited in any way. Rhizomorphs are feeder strands. They are designed to GATHER WATER and funnel it, with any stored nutrition, and any nutrition that is readily available in solution with the water.

BRF is a faster substrate then rye grain, this is why you have a higher frequency of rhizos in those jars as opposed to the Rye grain. It is not the quality of the substrains within those jars, but the Availability of nutrition with the substrate within those jars.

Example: in jars of sterilized manure, it is advisable to add BRF in small quantity to accelerate the colonization of that substrate. Once a sufficient network of mycelium is established, feeding on the BRF, it will readily eat the manure. The same principal works with manure tea, compost tea, etc... You are not just increasing nutritional composition, but increasing nutritional availability.




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OfflineShroomasta
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Re: !?! Info on Advanced Pinning Techniques !?! [Re: ]
    #634360 - 05/17/02 01:15 PM (14 years, 6 months ago)

Ok, thanks for the input. I'm glad to be able to post that a couple of rye jars are now pinning. Interestingly enough, two jars that my friend put aside as contaminated, have teh most pins and even fruits already. (He stopped checking on it for a while) It might be taht these jars had teh most air, as he was trying an experiment of using toliet paper as a filter, which is why he believes that they cotaminated, however, the added air exchange was a great benefit it seems. There are other rye jars that are pinning also, and these seem to be the ones that he checked more often then the rest. Therefore, the conclusion that we have drawn in addiion your other post was that CO2 was teh deciding factor. He has tried to find a good way to air out everything, and my guess is that he will go to coffee filters for now, while he waits for his filter lids. I'll keep updating on his progress as he tells me what is going on.

One question is whether it is feasible to use a large trash bag and line the bottom with moist verm, then place a tray of sorts with all the cakes in it and then close the bag to keep up humidty. As long as the bag is misted and fanned out this should work just as well as a terrarium, right? If there is anything that could be done to improve on the Poorest man's terrarium, hehe, any help would be appreciated. He has many rye cakes to deal with and not very much capitol, unfortunately. So any suggestion on how to birth or a good tek (flat cake, maybe?) would be appreciated. Thanks again.


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