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Invisibledumbfounded1600
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Registered: 07/29/07
Posts: 2,624
All Of RR's Notes (Advanced Mycology) Notes/Links I Liked * 9
    #8524923 - 06/14/08 08:59 PM (15 years, 9 months ago)

There is no talk of Psilocybin/Psilocin talk in these threads so if that's what your looking for your wasting your time.


RogerRabbit 2003 Advanced Mycology


I've been having better luck lately with lime than with bleach. If you can swing the ph of the substrate or casing to 10 or so, they mycelium will still colonize it, but the contams don't have a chance. Bleach and H2O2 will both slow down they mycelium. I think peroxide is better reserved just for cobweb mold, and bleach is great for bombing the air before sterile work by using a 1:10 mixture with water. I hope this helps. CASING PH

Another thing that will make your agar overflow is if the heat is up too high during sterilization. Anytime steam is escaping the top of your pc, it is also escaping your agar. Once you reach 15lbs, turn the heat down as low as possible to maintain that pressure. Since the pressure inside the agar bottle is also at 15psi, anytime the little weight on the top of the PC rattles and lets off steam, the agar is allowed to boil in the bottle. This causes it to 'boil over' just like a pot of sauce on the stovetop. And of course, like was said earlier, if the pc cools down to fast, the pressure in the agar bottle is higher than the pressure in the PC, and it boils over. AGAR OVERFLOWING

Don't mix lime with your grain. Lime, bleach, etc will never compensate for sloppy sterile procedure. Your jars should be pc'd then innoc'd in a sterile environment. The value of lime is for casings, compost, or straw. When i pasteurize straw, I set the 160F water to a ph of 11-13. This seems to hold the contams at bay while the mycelium colonizes the substrate. The high ph does indeed slow down the mycelium a bit, but it kicks the trich in the butt. Another good trick is to sprinkle a bit of lime right on the surface of your casing layer. This creates a high ph zone right on the top where the trich and other contaminant spores land. Hope this helps. PH/GRAINS/CASING/STRAW

The main point of the logs is to hold the CO2 in during colonization. They are pinning in 14 days as a result. I think the logs get a bit more yeild per the same amount of straw as a laundry basket, but that's not the point. The difference is faster colonization, and no misting. The Visions laundry basket tek is an excellent way to grow also, don't get me wrong. As for the PH of 12, this is what it measures right after pasteurization. As soon as the mycelia begins to rip through the substrate, the ph naturally lowers. Within two weeks it will be in the 6.5-7.5 range. The exact reason why we keep the pH so high is myc can handle it fine, but spores (of any kind) won't be able to germinate until the ph comes back down. By then the myc will be established and can hold its own. STRAW LOGS

I've never been able to get spores to germinate on antibiotic agar. I've found that if you put enough of the antibiotic in it to kill the bacteria, then it won't let the spores germinate either. I also wouldn't recommend using any antibiotics as standard procedure. You're likely to develop a super strain of bacteria that would become resistant to the antibiotic. What is recommended is to germinate the spores on regular agar, then transfer to antibiotic agar, then take another disk of antibiotic agar from a third petri dish, and place the whole disk of agar on top of the mycelium, creating a myc sandwich between two petri dish sized disks of antibioic agar. Wait for the mycelium to grow up through the top slice of agar, and transfer some myc from this point. It should have left any bacteria rods behind as it grew through the antibiotic agar. Then, wrap up the rest of the dish and throw it away. Once again, I'd only recommend this if you're sure you have a bacteria problem. Spores won't germingate on anitbiotic agar for me. Are you rehydrating them first? I wanna know what I'm doing wrong. I get anitbiotic agar from fungi.com, but have to germ on regular agar, then make a transfer. NOTHING will germ on the anitbiotic agar. Maybe that's a good thing? ANTI BOTIC AGAR

Paul Stamets has written that 86F is the ideal temperature for cubies to incubate. He was talking about substrate temperature, not air temperature. The substrate will always be warmer than the surrounding air. COLONIZATION

Incubating in a lighted room helps to speed up pinning. COLONIZING IN LIGHT

The center of the can will go anaerobic on ya and it will rot. I've found 8-12 inch diameter logs work great, but with a 14" log it's hit or miss with getting an anaerobic core. Anything over 14" has rotted and produced nothing. If you want a giant straw log, build it 10-12 inches in diameter, and a mile long. LARGEST STRAW LOG

I've tried different methods of air exchange within a log and ended up with trich growing every time. They work just fine all closed up. That's closer to the way they evolved in nature as well. STRAW LOG

I think it will work much better for you if you don't run that fan. I realize you were shooting for positive pressure to keep out contams, but the turbulence caused by the fan will have the air swirling all around in little eddy currents and they will blow contams off your hands, arms, and the interior of the box, and into your project. Just keep the air really still, lysol the interior, wear gloves, and work fast. It should work really well for ya! FAN IN GLOVEBOX

It sure seems like the salt would make it trich and bacteria resistant. After all, for thousands of years, we humans preserved our fresh meat in salt to keep it from rotting. Just gotta work out the right amount of salt to keep the bacteria from growing, while still allowing the established mycelia to grow. SALT AND MYC SPROES

Yes, grass helps. It also provides a 'straw' to pull oxygen into the substrate to aid in pin development. I've actually planted grass seed in my casing before. It's something for folks to try. GRASS IN SUBSTRATE

I've been to both the beginners and masters seminar at pauls. I'd recommend them to everyone. Expensive yes, but you'll walk away with lots of info. Paul will NOT discuss sacred mushrooms at the conferences or over the telephone. If you can 'listen between the lines' you will get all the info you need. When you hear a method described as good for "straw and dung loving mushrooms" listen very carefully. PAUL STAMETS SEMINAR

Actually, there is a procedure for crossing strains of fungi. It only takes a flow hood, and working knowlege of agar culture. Everyone has seen how sometimes with a spore print germinated on agar you have 'sectors' or zones that stay separate and don't grow together? Those are incompatable sub strains. Now, take two strains of mycelia and place them both 1/2" apart on the center of the same petri dish. If they stay separate, they are incompatable. If one simply overruns the other, it is simply a stronger strain. BUT, if the two strains grow 'together' and form what is known as a 'zipper' at the point of merging, (this will look like a third zone of mycelia) take a myc transfer from a rhizo that is LEAVING that zone, and you have created a third strain. It works, and the result will not be sterile. You may have to try a hundred times before you get a cross, but this is how it's done in the commercial mushroom industry. No gene splicing, no microscope, etc. CROSSING STRAINS OF FUNGI

I've never tried Banrot on a trich outbreak. It's used as a preventative measure. I'd hate to use ANY chemical close to harvest. Ya don't wanna be eating fungicides, no matter what brand. My rule is to throw away anything that gets the 'green'. It's not worth the risk to your other puppies to keep a trich infested project around. Just start out with twice as many casings/cakes/logs/baskets as you need, so you can afford to throw away the contaminated ones, and still have what you set out to get. KILLS TRICH DEAD

Now, if you want that massive pinset you have to remember that a log is not a casing. A log is more of an overgrown pf cake in certain ways. It needs 100& humidity to make a really good pinset. My recomendation on your next try is to leave the original plastic tubing (umbrella covers? kewl!) on until you have at least half a dozen pins or so. Don't be in a hurry to peel the plastic tubing off. (birthing) Pins will form invitro just fine, due to air exchange from the holes you punched when you built the log. Once you birth it, keep it covered loosely with plastic that has been punched with holes for air exchange. The plastic should be actually touching the log. (I noticed you have somewhat of a tent. Let the plastic fall directly on top of the logs, just lift it up and move it around a few times a day for air exchange) Fruit bodies that form can easily push the plastic out of the way. That should maintain 100% humidity under the plastic due to the moisture that is within your log. Keep trying, experiment with new things, and keep all informed! (thanks to hip for pointing me to this thread by email) UMBRELLA STRAW LOG

I mostly use mine to look to see how much bacteria is on the spores before use. I'm not sure you can see those at less than 400X. 100X gives you a nice view of the mycelia and is plenty to see the clamp connections. MICROSCOPE

We have quite a thread over at mycotopia about Banrot 40 WP. If you soak your grain or popcorn for two days in a mixture of one teaspoon per gallon of water to hydrate the grain before pc'ing, trich will not grow on that grain, but live mycelia will not be hampered. I even left a jar open for half an hour, exposed to normal room air after it had been soaked in the banrot, then sterilized. It never contaminated, yet when inoculated with live mycelia, it grew normally. Spores won't germinate on grain that has been treated with the Banrot. That includes mushroom and trich. Another big problem in mushroom cultivation is bacteria. I haven't tried onion or garlic yet, but they're both on the list. I'm also wondering if one could soak the grain in antibacterial soap before going into the pc. I've noticed most dish soap now days says it's 'antibacterial'. If it works, it's a matter of finding the amount of soap that the live mycelia can withstand, but the bacteria can not. BANROT FUNGICIDE

They aren't spraying fungicide in oregon and washington. If they tried it, ten thousand naked hippies would lay down in front of the equipment and block them. I picked lots of azures near astoria again this year, even getting directed by a park ranger who 'caught' us to where a better patch was. I don't think they're going away. Good project anyway though. Don't just spawn wood chips. In nature they grow in the beach sand right along the shoreline, nestled into the dune grasses. Most azures I've found have been within 100 yards (meters) of the shoreline. OREGON!

I tried one, and it didn't work out well. Hemp fiber doesn't hold moisture well enough. I does work VERY well however to mix hemp fiber with your straw, or any other substrate. It also seems to have an effect on potency, but more research is needed. Does anyone have access to the tryptamine content of hemp fiber? HEMP BALL SUBSTRATE

Cotton is a decent substrate. Hemp is better. Compost and manure are better than either one, from my experimements. SUBSTRATE





RogerRabbit 2004 Advanced Mycology


Growth should not slow down. If problems develop after 5-7 days, somethings wrong. COLONIZATION

My coffee experiments were done simultaneously with beer experiments. The beer helps a bit, but nothing to brag about. One thing to be absolutely sure of, let the beer sit and get flat or boil it before using. The alcohol is bad for mycelia. In my case the beer was used to hydrate grains of rye. Colonization was a bit faster, but not much. Pinning didn't seem to be affected one way or the other. BEER SUBSTRATE

Dried caps have been known to start growing again on agar. Go for it. With all due respect to those who disagree or have never done it, you can take dried mycelium and get it growing again on agar. I've done this many times with dry mushrooms. The stipe works better than the cap. This is not due to spores germinating, but due to the dried mushroom tissue rehydrating and beginning to grow again. Prepare the agar a bit on the wet side to help hydrate the tissue. Also, be ready with fresh plates to transfer into, because all the contamination on the dried fruit will also begin to grow. You'll have to transfer healthy mycelia as soon as you see it to a new petri dish. It generally takes ten days to two weeks, sometimes longer, for the dried fruit to begin to grow again. Unfortunately the contamination doesn't need that long, so you have a bit of work to do to clean up the culture, but it will work. You can even take a piece of the inside of a dried stem and get it to grow again on agar. No way a spore could be on the inside of the stem where it's protected. Dried mycelium of many species can be hydrated and coaxed to grow again. You are correct though. . .A dried cap or stem is not sclerotia. Cubies don't form sclerotia. However, a dried cubie can be rehydrated and grown out again on agar. Try it. You'll see the growth coming in many directions from the piece of stem you place in the agar. You will have no doubt that you have 'cloned' that piece if you'll try it. DRIED MUSHROOMS

An initial ph of 7.5-9 is what you want to shoot for with casings. Peat is too acidic and will favor contamination unless you lime it. PH CASING

The main problem is that to totally sterilize rye or popcorn would take 24 hours or longer in the pressure cooker, and that would leave you with a goopy mess. When we pc for an hour at 15 lbs, what we're doing is giving a 'window of opportunity' for the mycelia to colonize the substrate before the contaminants can make a comeback. You could probably do it with full, non vented canning lids, but when the customer opens the jar, it will suck in air and contaminate. One idea would be to inoculate the grains and allow them to colonize, then refrigerate. STORING

I've used coir for years as a substrate. It actually has LOTS of nutes in it. I use it mixed with verm, coffee grinds and gypsum for substrate. I'm considering switching back to peat based casings, because I'm tired of the overlay problems with coir. It has so many nutes, the mycelia wants to colonize it like a substrate, even when it's 'diluted' by 50% with verm. No, never tried coir by itself. But again, I don't use any substrate by itself. I've found that if coir is good, coffee is good, manure is good, etc. etc., if you mix them all into a cocktail, it's even better. Just an observation. COCO COIR

The main reason to soak popcorn is to germinate the heat resistant bacteria endospores, so the PC can kill them. Otherwise, many survive. Popcorn takes longer to soak up water, so I would recommend you boil for an hour or so, then load into jars and PC. My rule of thumb when using popcorn is to watch for the kernals to appear to have doubled in size. POPCORN

Guys,
This is old science. It's been done for many years in the commercial mushroom industry. It's the standard way they keep and trade isolated strains of edible mushrooms. They desiccant dry the fruitbodies, then powder them. They store the powder until ready for use, then sprinkle it on agar to regenerate the strain. It stops senescence in its tracks. Try it without soaking in peroxide first. That only weakens the mushroom myc so it doesn't take off as fast. Watch for the first signs of myc growth, and immediately transfer that to a new petri dish. Do this a time or two and you'll have a clean specimen, even from years old dried fruits. It works, trust me. DRIED MUSHROOMS

s an alternative to putting two types of spores in a jar, try inoculating two different isolated strains of one species onto the same agar wedge. Simply transfer two wedges of myc to the same plate. When the two grow together, you'll see a 'line' form between strains. Sometimes one strain will gobble up and 'eat' the other one. Sometimes, each holds its own turf and the line stays put between them, where each has its own territory on the petri dish. About one time in a hundred, along this 'line' a third sector will open up. This is by definition, a 'hybrid' produced by dikaryotic pairings from each strain. It is definately and absolutely possible, and I've done it. Next fall, when I have time to resume the hobby, I'll do it again and write a photo documented tek. Until then, anyone who has time and inclination to experiment, now knows how to go about it. HYBRIDS

Did you guys know you can take a desiccant dried mushroom several years old, and clone it onto agar? There is no need to store a petri dish for a year. Just dry a fruitbody and put it away. When you want the strain back, clone it onto agar. This is how its done in the commercial mushroom industry. In fact it was Stamets who first told me you could clone a totally dry fruit when I found a dry one that had been out in the sun for months. He was right, and it worked. DRIED MUSHROOMS

Get sticky mats, and set them up in your grow area. They're bright yellow, and attract both flies and gnats. Once they land on it, they can't get loose. They work great. I'm not talking about little pest strips, but 2' x 4' mats.
http://www.fungi.com/tools/misc.html#DFP
You’ll find sticky mats on the bottom of this page
FUNGUS GNATS

I placed one drop of solution from my syringe from workman on agar. This was then isolated into half a dozen substrains(sectors) One of these sectors began making sclerotia within a matter of days. This isolate was then used to inoculate jars. Sclerotia was forming even before full colonization. After ten days, the quart jar of rye grass seed was fully colonized and used to inoculate twenty more jars. Half rye berrries, and half rye grass seed. During the grain to grain transfer, a piece or two of the sclerotia was dumped out and injested. They were very small pieces,(less than one gram) but enough to produce slight but quite pleasant 'effects'. One thing I've found is that rye berries produce sclerotia faster and bigger than rye grass seed. This was a surprise. Another thing I've found is the sclerotia produces faster when the jars are exposed to light from day one. Conventional wisdom says to incubate sclerotia producing species in the dark. I found this not to be true. I've also found this to be a very easy species to grow and fruit. I found the rye berries work better quite by accident. Ditto for the light. It was a 400 MH on for some orchids in the same room. The jars that were shielded from the 400 produced less than half the sclerotia of the ones that got light. Next time I harvest some, I'll weigh it for ya. I don't use the scales much anymore. I just munch until I'm full...lol.

http://www.shroomery.org/forums/files/04-23/640151042-sclerotia_004a.jpg
SSS
MEXICAN STRAIN A SCELORTIA

The bleach is used as a preventative, to kill contaminant spores, not to kill established contaminants. You're looking at some two year old posts in that thread, as the process was being developed. BLEACH

I've used coffee grinds in casing material for a long time. It increases the chance of contamination, but also increases the pinset and yield. Don't forget the lime. Using weak coffee to hydrate corn or rye(or any other spawn) is a good, proven tek. Coffee has tons of nutes that fungi love. I've even used brewed coffee to hydrate my compost/manure mixtures with excellent results. COFFEE USE

A trick I've found with wild mushrooms, is to cut a small section out of the stem and put it between two pieces of moist cardboard. In a few days, mycelium will begin to grow. Take a strand of the new myc, and transfer it to antibiotic agar. The cardboard will have effectively 'filtered' most of the molds away. I take a bag of pre-cut 6" X 6" moistened cardboard when I go picking, for this purpose. One trick to outrun bacteria(it isn't necessary for fungi imperfecti) is to make an agar sandwich as you mentioned. Place your culture on antibiotic agar, and place another disk of antibiotic agar on top of it. Take the very first myc to poke through the top layer, and transfer it to a new dish. Repeat the process. After a transfer or two this way, the bacterial rods should no longer be present. After you make the transfer, look at the old dish under the microscope. If the rods are gone, the specimen is clear. If I have bacteria riding along on the myc, I transfer a very small piece to a new petri dish of antibiotic agar. Then, take a second petri dish, and transfer the entire disk of agar to the top of the transfer you just made. You now have two full sized disks of antibiotic agar, with your contaminated myc in the middle. After a few days, myc will grow through the top layer and poke out, before reaching the edges of the dish. When myc pokes out the top, I use my inoculating loop to scrape some myc without taking any of the agar. Transfer this to a new dish. Somehow by growing right through the antibiotic agar, it seems to leave the bacteria behind. Normally, to get away from molds, a couple of transfers on antibiotic agar does the trick. Bacteria can be a whole different animal, as it seems to 'hitch hike' along on the mycelium. You can't see it without a scope, but it will sure contaminate your grain if you use it to inoculate your spawn. I'm talking the bacterial 'rods' and not something obvious that you can see like wet spot. I wish I had a camera attachment for my microscope.

Question;
This is what I interpreted from Stamets in GGMM. I tried that but ran into problems holding on to the large piece. How do you transfer it from one dish to the other? I tried once but dropped it on the bench. So, I decided to use a small piece instead, it works just as well for me...

Quote, “try loosening about 1/3 of the agar from the dish with your scalpel, surface tesnion will hold it long enough to invert and align the dishes anf the agar should drop right in... “
Yes, what Prisoner above said is how I do it. Works like a charm.
I use the sandwich technique for isolation away from bacteria. For molds, simple antibiotic agar seems to do the trick. AGAR ISOLATIONS

Totally, completely dried mushroom mycelia will grow again. It's a fact and not to be argued. It happens every year in the wild. Look at oyster mushrooms. In my area, there is NO rain from about mid May until early October. The forest fire danger is very high, and all the dead wood in the forest is cracker dry. If you cut into a log, as I did last week, you will see the mushroom myc, and it's as dry as a bone and looks dead. However, come October when the rains start, there will be oyster mushrooms by the truckload popping up from nearly every fallen tree, all over the forest. It happens every year. Culture banks store dried mycelia long term as a way of preserving isolated strains of edibles. The dried mycelia grows again when re-hydrated. Those aren't spores germinating, it's mycelium. Besides, I challenge anyone to grow oyster or shiitaki mushrooms from spores. DRIED MUSHROOMS

This gives me a bit of hope. I recently received some very rare spores that are 20 years old. They've been six weeks on agar and so far. . .nothing. I won't toss the dishes, but just leave them be and hope for the best. I sure would like to see these germinate. YEAR OLD SPORE PRINTS LIABILITY

If you simmer rye grass seed, you will have a mucky mess. Be sure to use twice as much grass seed by volume as water(dilute coffee especially helps sclerotia production). Add a pinch of gypsum in each jar, and if you're using coffee, also add a small pinch of hydrated lime to each jar. Put on a solid lid, and shake the heck out of it to mix everything up. Remove the solid lid, and replace with a lid with two small 1/8" holes drilled in it. Place a filter disk on the outside of the lid, so it won't get wet. Cover with foil and pc for 90 minutes. Shake the jars while still slightly warm from the pc. This will facilitate easier breakup of the rye grass seed. When cool, inoculate. RYE GRASS SEED

Drop the spores into the water trough. Just about any cube will work in that climate. If you wish to be 'authentic' order gulf coast or treasure coast. Florida strains. REINTRODUCING MUSHROOMS IN THE WILD

The bacteria is not from the water, but from the corn itself. Simmering or even PC'ing will not kill them all. You can try a 48 hour soak to germinate the endospores, then PC, and it will help. My information comes from experience. Besides, rye grains are far superior to corn anyway. Use what is easiest for you, but my recommendation to one and all is rye grain. Without a filter, there is virtually no chance that cracked corn or any other spawn is going to make it to the fruiting stage. Peace all, and happy growing. CRACKED CORN

AM is a mychorrhizal fungi. It must grow in conjunction with a living tree. You can find it in the wild easily, but I know of nobody who has succeeded in cultivating it. AMANITA MUSCARIA

Mushroom mycelia will easily colonize a very base substrate. I use hydrated lime in pasteurization baths at a rate of 1/2 cup for ten gallons of hot water. This gives the bath a ph of 9-11. The mycelium then rips through the straw like wildfire. Get some ph test strips and test the water with coffee added. Add lime to raise the ph. Be sure to use hydrated lime, as it's water soluable. STRAW LIME

Best not to use more than 10% coffee grinds in a casing layer. The best use for coffee is as the liquid used to hydrate your grains. Mix it 50% of drinking strength and use this as the liquid. My results over the last two years show consistantly a 40 to 50 percent decrease in colonization times(of the grains), and a faster pinset once the grains have been spawned into bulk substrate. Coffee also is a big help if growing invitro. COFFEE/COFFEE CASING LAYER

Coco coir makes an awesome addition to your compost, straw, manure, etc. It's a great bulk substrate, but as I've said for well over a year now, it sucks as a casing material, too many nutes. Peat/verm is much better for casings, and as anno pointed out, don't forget to lime it to buffer the ph. COCO COIR





RogerRabbit 2005 Advanced Mycology


Think if you look at that growth under a microscope, you'll see a ton of bacterial rods attached. Try pouring hot antibiotic agar right on top of the growth. Fill up the dish. I've been having good luck cleaning up cultures lately that way. The hot antibiotic agar seems to kick the contams in the ass, but the myc survives and grows up through the layer to the top. When this happens, use your scalpel and gently 'shave' some myc off the top without getting any of the agar. Transfer a very small piece of mycelium to a new dish, and when it grows out a bit repeat the process. It takes a time or three to get a good, clean culture. Try it. AGAR

Another trick to use that helps even without antibiotic agar is to simply pour hot agar right on top of a contaminated plate. The heat seems to slow down the molds long enough for the mushroom mycelium to recover and climb up through the layer to break out on top. This usually only takes a day or two. As soon as the myc starts to appear on the top layer, 'shave' a small amount off and transfer to a new dish. Don't take any of the agar, but only scrape a bit of the myc off the top. This seems to leave the contamination behind 90+ percent of the time. Repeat the process if necessary. I've cleaned up lots of edible cultures from the wild this way. AGAR

Something I recently tried and it worked for reishi is birch plywood. Cut the plywood into 1" strips, then bang with a hammer to break up slightly. Soak for three days in water, then inoculate. You can get plywood just about anywhere. WOOD LOVERS AND EUCALYPTUS

Antibiotics aren't for green mold which is an imperfect fungi, they're anti-bacterial. True, sterile technique is foremost, but some of us are using prints that were left under the cap too long and have bacterial rods all over them. When the mushies germinate, so does the bacteria. Antibiotic agar is a godsend, imo. ANTI BIOTIC AGAR

I just use the agar from fungi.com but mix it up a bit weaker than called for in the recipe. I also add a pinch or two of gypsum to the litre of agar, and this seems to help fruiting. A bit of gypsum in jars helps invitro fruits to form also by the way. My experience with mexicana is it's all about strain isolation. Some substrains just won't make sclerotia, and others won't fruit. By growing out on agar you can find the best substrains. The chunk of sclerotia below was sliced into three pieces and each one was transferred to a new dish. These were grown out and transferred to grains as the fruiting strain. The best sclerotia producers were likewise selected and grown out on rye grain, which in my experience is superior to rye grass seed for sclerotia production. I'm not sure of the actual processes involved. Perhaps something to do with calcium aiding fruiting, but that's really just a guess. I've lately found a bit better performance all around with gypsum, and for some reason fruitbodies are more likely to form on agar when a pinch of gypsum is used. It's also very effective as you mention to keep grains from clumping up. Yes, I add the gypsum to everything prior to sterilization. For quart jars of grains or liters of agar, the very un-scientific method of using a pinch between your thumb and forefinger is the right amount. MEXICANA AGAR

Ok, I've mentioned this before, but been working on it a lot lately and this tek really works. I pick a lot of wild edibles up in the mountains and have been cloning/printing them for domestication. Often, both molds and bacteria are embedded in wild mushrooms. It's normally easier to transfer live mycelium away from molds than from bacteria, which seems to 'hitch-hike' on the myc.  If you'll mix up a batch of antibiotic agar, you can pour the hot agar directly on a petri dish with both mushroom myc and contaminants. Completely cover all the growth in the petri dish with a fresh layer of hot antibiotic agar. The hot agar will 'pasteurize' the contaminants, but the live myc will barely be set back. In a couple of days, the mycelium will grow through the layer of agar and appear on top. At this point, carefully 'shave' a bit of the myc off the top without taking any of the agar. Transfer this to a new dish and you now have a clean culture. The very dirtiest of wild specimens might require a repeat of the above, but this procedure usually gets it on the first go. Try it. ANTI-BIOTIC AGAR

My guess is yes. In fact we've seen a few grows lately where guys have deliberately mixed different strains in the same casing and had very nice results. By different, I mean such as EQ being mixed with B+ in the same substrate. The conventional wisdom in the community is the strains will 'fight' each other until one wins out. This does not appear to be the case. More experimentation is needed. SUB STRAIN CAPABILITY

A 1/10 ratio works well, so in your case I'd suggest using one entire fully colonized jar to inoculate ten more via grain to grain transfer. G2G

I'm still in love with coffee hydration. Soak your verm in it and watch all the invitro fruits pop. COFFEE

Also bear in mind, the antibiotics will only help to protect against bacteria. They will be of no use against clandosporium, aspergillus, penicillium, trichoderma, etc.,  etc. ANTI-BIOTIC AGAR

I pour about 1/4 full. Most species I grow out will fully colonize the petri dish within two weeks or so, and the lesser volume of agar will help to reduce condensation on the lid. Hyphae, try something next time you put away cultures. Pour a couple of petri dishes very shallow. Perhaps a 1/8" layer. Allow the mycelium to colonize 2/3 of the dish. Be sure to use parafilm. Place in the refrigerator and forget it for a year or so. When you pull it out, the agar and mycelium will be totally, bone dry. Take a few samples and move them to fresh agar. Within a few days, they'll be growing again. I recently took out a three year old azurescens dish that was as dry as it could possibly be. It's growing great again, while the spores from the original print it came from were no longer viable. If I can confirm this with a few more trials, I'll be throwing away all my test tubes, as they're such a pain in the ass to prepare and clean out. I would have not thought dry mycelium could recover this well/fast. AGAR

By the way, if you buy organic rye berries from a health food store, they are required by law to be fungicide free. ORGANIC RYE BERRIES

Agreed, some endospores will always survive the pressure cooking cycle. The idea with sterilization is to give us a two week window of opportunity for our mushroom mycelium to colonize the grains to the point they can take care of themselves. PRE STERILIZED GRAINS

Wheat will work fine if you can't get rye. WHEAT GRAINS

Never disturb a contaminant in a petri dish. To do so simply dumps millions of contam spores all over your good mycelium. The proper procedure is to cut a very small piece of good mycelium and transfer it to a new petri dish. Do this in a glove box and not in front of a flow hood. A flow hood will blow the contaminant spores everywhere. It takes many transfers away from the original spores to get an isolated strain. You have nothing to fear from transferring away from contamination. By the time a petri dish is grown out like that one, clamp connections have formed everywhere and the genetic information has already been exchanged throughout the entire dish. TRANSFERING AWAY FROM CONTAMINANTS

If you'll hold it up to the light and look at the dish from the back side, you'll see them easier. The middle picture has thousands of sectors. If you want to do an isolation, you need to isolate as many substrains(sectors) as you can, as most will be poor performers. It's that one isolate in a hundred or so that is going to kick ass. I'd take that top plate, and transfer 20 sectors from around the perimeter to a full sleeve of petri dishes. Take specimens to transfer that are no larger than a grain of rice. Grow out each of those twenty dishes(there will still be sectoring) and transfer each one to grains for fruiting, after saving a sample from each dish to a second sleeve of twenty dishes. Be sure to label them. When your grains fruit, you will see which is the best substrain. Go back to the petri dish that corresponds to your best fruiter, and grow that one in bulk. Be sure to save some mycelium in the refrigerator for future use. AGAR ISOLATES

I would recommend using one drop of spore solution and working very hard to keep it undisturbed right in the center of the dish. You want your spores germinating in close proximity to each other so proper clamp connections can be made, thus producing dikaryotic mycelium. I recommend MEA. AGAR


They also have them divided into two, three or four sections. For doing strain isolation, I like the four section dishes. It really saves money on petri dishes and agar, not to mention time.

http://www.cenmed.com/showDetails.asp?item=6141&strCatName=&mainCat=LS&category=565&cat=235,565&pagename=viewcart.asp ;   PETRI DISHES


If you'll place two different dikaryotic strains on petri dishes a few hundred times, you'll probably get one or two dikaryotic pairings. In the vast majority of cases, a zone of isolation will develop between the two strains and that will be the end of it. Eventually, one strain may overrun the other, but no dikaryotic pairings take place. I've been attempting this for years, and have only two success stories so far. The hybrid in this instance will emerge from the zone of isolation between the two growths on the petri dish, and will be a distinct third sector. You can test your hybrid by placing a small sample of it and the two parent cultures on a third plate. If a zone of isolation develops between all three, you know have successfully created a hybrid. Stamets teaches this tek at his master's seminar. GENETIC ISOLATING

I've used a whole petri dish to inoculate one liter of liquid culture solution. I never use more than one drop from a spore syringe right in the very center of the petri dish. You don't want a huge amount of spores germinating. That defeats the purpose and makes strain isolation that much harder. The milky water might be germinated spores or it could be bacteria. Keep a close watch. AGAR/LC/

Another good rule of thumb if your spores are fresh and viable, is less spores are better than more spores. Keep them in the very center of the dish so that they'll have the maximum amount of space to grow out in before reaching the edge of the dish. This also keeps colonies from running into each other and having to set up a 'zone of isolation' between the strains, thus sapping energy that could be better used growing. SPORES ON AGAR

Agreed. I use a bit of chicken manure in my compost mix along with cow, horse, worm castings, coir, seaweed, and a bit of verm to lighten it all up. Don't use more than about 10% of the total as chicken manure. It will help grow some giants. COMPOST

Nice work. Put a bunch of pennies around your mushroom patch. Slugs hate to crawl over copper. SLUGS

Stamets' work with biaremediation shows that of all the toxins, only heavy metals end up in the fruitbodies. Oyster mushrooms grown on diesel spill sites and PCB spills had no traces of said contaminants in them. I wouldn't worry about slug bait. TOXINS OF MUSHROOM FRUITBODIES/SPECIES

I'm going to throw something out here that contradicts the prevailing beliefs in cloning/isolation. I would like to see some of you in a position to do so, attempt to verify or disprove the following. I have noticed when cloning fruits from multispore inoculation, that more than one strain is cloned. I have seen obvious sectors on the agar from a single piece of cloned tissue. At first, I thougt I must have gotten some spores that were stuck to the stipe into the mix, but later I was very careful to only select tissue from deep inside the stem from fruits that had not yet opened their caps. My theory based on these observations is that single fruits on a cake or cased substrate from multispore inoculation, can and are sometimes made up of multiple strains(substrains in shroomy speak). Multiple strains within the same fruitbody? That is how it appears to me. Before anyone answers "impossible", do a bit of research. If there's another explanation, I'm all ears. I'm not talking about different types of mycelium such as cottony, rhizo, airy, etc., but actually separate sectors that when transferred to different petri dishes will not mix with other sectors from the same frutbody. Sometimes fact is stranger than fiction. I'd love to see a few of you myco-geeks jump on this. I know I am. Have fun. CLONING/SUB-STRAIN/ISOLATING/AGAR

It's very normal not to see the rhizomorphic growth until you make few transfers. With multispore inoculation on agar, you have hundreds if not thousands of strains growing together and all over on top of each other. Remember, any two spores that germinate and 'pair up' by forming a clamp connection is the very definition of a 'strain'. Just dig in and take a very small piece of mycelium no larger than a grain of rice. Move this to a new dish. When this dish begins to grow out, you may or may not begin to see sectors. It might take a second transfer before you get few enough strains to see the individual sectors. Take the best looking growth again and move it to a fresh petri dish. Eventually, when you hold a dish up the the light and look from the back(agar) side, you will see individual sectors like the spokes on a wheel. In the picture below, the dish on the left is the result of two transfers from the original spore swipe. Prior to the second transfer, the rhizomorphic mycelium was buried in the mass of strains and couldn't be differentiated, just like yours are. The rhizomorphic sections are now clearly showing up at ten o'clock and two o'clock. Each of those sections has four or five individual sectors that will be isolated out next time I do agar work in a day or two. The remainder of the dish will be discarded. The dish on the right is a single sector isolate, as you can see the entire dish is rhizomorphic and when held up to the light, there is no further sectoring. I fruit out each individual sector to determine the best fruiting isolate. Many isolates are mediocre fruiters, but if you want to find the one that will deliver those monster flushes time and time again, this is how it's done. I keep a small piece from each petri dish in the refrigerator until I've found the best fruiting isolate, then I get that piece out of the fridge and grow it out. I always keep a piece from the original petri dish in test tubes in the refrigerator so I can go back to that strain for years to come with no fear of senescense. It's a lot of work, but hey. . .It's a hobby right? ISOLATING
http://www.shroomery.org/forums/showflat.php/Number/4377475#4377475


One way I've found to beat those pesky contaminants is through agar pasteurization. Make up a fresh batch of agar. When it has cooled enough to pour into the dishes(+/- 120F-130F or 49C-55C), pour a layer right over the top of the dish with both healthy mycelium and the contaminants. Be sure to cover all the growth in the dish with a layer of hot agar. The mushroom mycelium, being a coherent network will usually survive the assault, but the contaminants will be neutralized for a few days. Watch the dish carefully. Usually within 48 to 60 hours, the mushroom mycelium will make its first appearance through the fresh layer of agar. Use an inoculating loop rather than your scalpel and immediately scrape a tiny bit of this mycelium off the surface of the top layer without taking any of the agar itself. This usually results in a very clean transfer of mushroom mycelium before the contaminant has had a chance to recover. CONTAMINATED AGAR PROBLEM

The way I do it is in front of my flowhood, I cut a piece of mycelium from the very center of a thick stipe. This gets 'virgin' tissue. I place this small piece of tissue on a piece of wax paper and place in a clean petri dish with desiccant under the wax paper. I then seal it up with parafilm, desiccant and all. I've pulled two year old cracker dry tissue out this way and it began growing within two weeks after placing on agar. Mix the agar a bit weak, so it's wetter than normal. Make a transfer as soon as you can see mycelium growing. CLONING FROM DRIED MUSHROOMS

Personally, I never dip in H202 when making transfers. The key is to take a very small piece of mycelium, whether you're cloning fresh tissue or dry. A piece of wet tissue half the size of a grain of rice will grow out fine, and dry tissue half that size will work. Remember, the larger the chunk of tissue you transfer, the more contaminants you transfer. You can drop a very small piece of dry or fresh tissue into distilled water and shake it up. That will wash it very well without stalling the mycelium with something toxic to it. The distilled water will also hydrate dry tissue. A second rinse in clean distilled water may be called for if the sample is especially dirty. Transfer the now clean piece of mycelium to agar under asceptic conditions with sterile tools. Always wear a surgical mask, hairnet and latex gloves. Use a glovebox if the tissue is contaminated, not a flowhood. If the tissue is not contaminated, a flow hood will give a better success rate. Watch it daily and transfer the very first mycelial growth you see to new petri dishes. Don't wait for contamination to show up. The mycelium will recover much faster if it hasn't had its ass kicked with hydrogen peroxide, and you'll usually make that first transfer from fresh cloned material within 48 to 60 hours. If dry tissue is used, usually within a week you'll see enough growth to make a transfer to a new petri dish. Save the peroxide for those cobweb outbreaks on your casing material. Good luck. AGAR TRANSFERS

A tip for cutting the price of parafilm in half is to cut it lengthwise before wrapping, so each piece can wrap two dishes. I wrap 1,000 petri dishes that way with each $20 roll of parafilm, which works out to only 2 cents per dish. PARAFILM

Interesting. Personally, I've had better yields with rye berries than with rye grass seed. Perhaps due to the higher moisture holding capacity? Have you ran side by side tests by any chance? By the way, your mex a rocks! MEXICANA SCLEROTIA

It will sector just like cubensis. Transfer each sector to a new dish to grow out. Fruit each sector separately to determine the best fruiting strain, then discard the rest. Be sure to keep all dishes in the refrigerator as soon as you've transferred from them to stop growth. Also make sure they're properly marked so you can go to the correct one when you determine the best fruiting strain. However, reishi mycelium looks and behaves like most other mushroom mycelium. There really is no way macroscopically to tell one mycelium from another with any degree of certainty.  ISOLATING REISHI

Plenty of organic material and good drainage. Essentially, just take good care of the tree and the mycorhizal fungi will take care of itself. MYCORHIZAL

The spore doesn't 'split' like a seed, but the mycelium grows from the germ pore at the pointed end of the spore. You can see a sequence of pictures of a spore germinating at the link below.
http://www.mushroomvideos.com/892340.html ; MUSHROOM SPORES

Distilled water is the correct medium for spore syringes. SPORE SYRINGES

You should sterilize the needle between jars. Alcohol is only a sanitizer, not a sterilizer. You need to flame the needle red hot to sterilize it. Also, build the syringe in a glove box or better yet in front of a flow hood. You don't want the air that's in the syringe to be unflitered, or it's likely to have bacteria or molds. NEEDLE STERILE PROCEDURE

Agreed. Coir is closer to neutral than peat, but then again, coir has just about nothing in common with peat. Coir may be fairly nutrient free as far as plants are concerned, but it has plenty of nutes that fungi love. I've spawned rye into pure coir, and had results as well or better than spawning to cow manure. I think most of us by now add gypsum to our substrates(if not we should) so the calcium is added at that time. My biggest gripe with coir is the cost. It's much more expensive than other ingredients that do as well or better job. Perhaps if one lives in an area where coconuts are produced it is cheaper, but for us city dwellers that must go to the hydro or pet store, there are other substrate ingredients that are better and cheaper. After a few years of experimenting, I've purchased the last brick of coir I intend to unless I move to a desert island in the tropics. Try it in all your manure/compost recipies. Gypsum will help with the flushes. It has calcium and sulphur, which are both essential for good fruitbody development. I use it in the coffee/water I soak the rye berries in, then use it again in the compost and manure I spawn to. I even put some in the water I pasteurize straw in, so the straw can soak up a bit. I use it at about ten percent by volume of manure. Ten cups of manure, one cup of gypsum. Give it a try. I've used it for years...just thought everybody did...guess not, lol. COCO COIR

Stamets calls it 'hybridization between dikaryons' but it is correctly a 'cross' and not a hybrid. A true hybrid would be a cross species mating. No, I don't have a picture of a third sector opening up. I'll try to get one next time. It's very rare for them to cross on a dish like that. Perhaps one in a thousand. CROSS AND HYBRID

That one hour window can be extended by making a couple of transfers instead of just one. The reason for the very limited window is because the hybridized strain, being brand new and not established will be quickly 'swallowed up' by the two parent strains which are rapidly growing.  The procedure I've developed calls for making a transfer when the two parent strains are about 1/4" apart, before the very first mycelial strands drift into the zone of inhibition. Take a small square of agar, but be sure to get the very smallest amount of each parent strain as possible. By doing this, there is not a large mass from the parents waiting to overtake any possible dikaryotization(when two binucleate mycelial cells exchange genetic material as opposed to two mononucleate cells) between the two strains that may have occured. Remember, there are millions of cells in each parent strain by the time they meet up, and only one or two cells are likely to pass genetic material between them, forming the cross. Watch very carefully over the next few days for a third segment of growth to emerge. As soon as it's noticed, remove it to a dish of it's own.(a jewlers microscope is handy for this) Once the new growth gets established, test for hybridization by placing it on a dish with two equal sized mycelial wedges from the two parents. If a three way line of inhibition forms, you now have a third, independent strain that is a cross between the two parents. Bear in mind, even using toxicated agar, this will only happen once in 50 to 100 petri dish you perform the experiment on. Get ready for a lot of frustration before you see your first success. Then, you must do it again and again until you actually get a fruiting strain. It also sucks when you're away at work during the two to three hour window you have to make the transfers. It's amazing how much closer two growths can get to each other during ten to twelve hours while you're gone to work. It's like approaching a car head on, the speed of closing is twice that of either growth. Also, it's important to know the toxin breaks down very quickly, so don't put the two growths too far apart on the agar. You only have 48 hours or so for them to grow together before the toxin degrades, so you have to have all your ducks in a row before you start.  All I could suggest is give it a try. I used crotalus atox venom which it appears they don't carry. What sucks is the stuff is five times the price of black market cocaine which makes it awfully hard to experiment with. Just be sure to add it to your agar at the last minute so the heat doesn't ruin it. CROSS STRAINING 2 DIFF STRAINS

I think it's jumping the gun to assume that a fungal cross-species hybrid would be sterile because that's what happens with mammals. Work is underway to test this theory. HYBRIDS

Caps or stems can both be used for cloning. There is growing evidence however, that mushroom fruitbodies may contain more than one set of dikaryons. This means that more than one substrain may be present in the same fruitbody. That is why clones on agar often separate into individual sectors with a zone of inhibition between them. ISOLATING GENOTYPE

You would have fruits from two different strains. You could even do it with different species. They will very rarely hybridize that way. Strains of the same species are much more compatable and dikaryotic/dikaryotic pairings are also possible, but not likely. It will happen occasionally however. I've also put different species in the same substrate before. For example, you can mix copelandia and cubensis in the same manure or straw tray and they'll fruit at the same time. I've never seen they hybridize this way however. Dikaryon to dikaryon pairings are quite common in the commercial mushroom industry. Let's say you have one strain of agaricus that has large meaty fruitbodies, and another strain that gives prolific flushes. Dikaryon-dikaryon pairings in this case would be called for in an attempt to get the best of both worlds. HYBRIDS

TMC was written over 20 years ago and a lot has changed in mycology since then. Paul no longer even holds to that "incubate in total darkness" bullshit, as his incubation rooms are under nearly 12 hours of fluorescent light per day, every day. TMC is great for a new grower to use while getting started, but that's about it. Nobody gets decent pinsets from a minute or two of light per day, and nobody has their projects ruined because a bit of light hits the jars during colonization. LIGHT AFFECTING GROWTH

I'm not quite sure what you're asking, but with multispore inoculation you'll get lots of different substrains forming, some better sclerotia producers than others. I'd transfer sclerotia or mycelium from the best producing sectors to either agar or grains to grow out. It's what I did with these plates. The best sectors were transferred, the rest were tossed out.
http://www.shroomery.org/forums/files/05-46/207873314-Mex_a_sclerotia_a.jpg
MEXICANA A

Antibiotics have an effect on bacteria, not fungi. They're sometimes added to agar to fight bacterial contaminants when culturing. Antibiotics will have little to no effect on the green molds or mushroom mycelium. ANTIBOTICS

Peat has a ph of about 4 to 5. If you were checking the ph right after adding hydrated lime, that's why it was so high. You need to add the lime, then let it sit overnight before reading ph. Also, make sure your ph meter is properly calibrated. I suggest the paper test strips rather than a ph meter. PH OF PEAT

Single sector isolates that are proved prolific fruiting strains will outperform multispore inoculation, but there are quite a few steps to get there. You'll want to do a strain isolation tek to isolate at least ten single sectors of rhizomorphic growth, then take each one to fruiting, while keeping the master petri dishes in the refrigerator. Once you determine the best fruiting strain, go back to that strain master and take it to liquid culture. It will take a minimum of three to four transfers on agar to get to single sector isolates, depending on how many spores you start out with on your original petri dish. ISOLATING DIKARYOTIC MYCELIUM ON AGAR

Keep those 'clone jars' in the refrigerator. Only take them out when you need to do a grain to grain transfer. They should last for a year or two that way, then simply do a g2g and keep one of those jars back as a strain master. CLONE JARS

I've found that a weaker nutrient mix in my agar gives faster growth. Perhaps it's because the mycelium is looking for food. Also, make sure you have gas exchange, so only wrap the dish with parafilm or some other sort of breathable tape. AGAR NUTRIENTS

Cut a piece no larger than a grain of rice when making isolation transfers. When you swipe spores onto agar, chances are the dish will grow out hundreds of substrains. You likely won't see any sectoring until after the second or third transfer for that reason. The first picture below is a dish after two or three transfers. By holding the dish up to the light, you can clearly see the sectors by looking at the dish from the back(agar) side. The dish on the left side of the second picture is after an additional transfer or two. You can see the rhizomorphic growth developing from the ten and two o'clock positions. By transfering only the rhizomorphic growth, you eventually end up with a totally homogenous, single sector rhizomorphic isolate as seen in the dish on the right of the second picture.
http://www.shroomery.org/forums/files/05-18/530157237-sectored_dish.jpg
http://www.shroomery.org/forums/files/05-27/070264929-rhizomorphic_myceliumaa.jpg
STRAIN ISOLATION

It will take at least three to four transfers to get to single sector isolates. Five transfers is not unusual. More if you get carried away and use a lot of spores. If using a syringe, use only one drop in the middle of the dish. If using an inoculating loop, take a very small swipe of spores off the print. Continue making transfers until there is no more sectoring. Fruit out each single sector isolate to determine the best fruiter. I incubate petri dishes at room temperature in a normal room. Dishes are wrapped with parafilm for protection from contaminants and gas exchange. I actually isolated some fuzzy mycelium once for a fruiting trial. It colonized slowly, then never produced even one single mushroom. When isolating strains on agar, always select fast growing, rhizomorphic mycelium to transfer. What happens with multispore inoculation is thousands of sub-strains are all growing at the same time, crowding each other and giving the visual impression of fluffy mycelium. When transfered to bulk substrates, sometimes the more aggressive rhizomorphic strains take over. By looking at the petri dish pictures in my post above, look at the one labled 0629 Poteet. That dish is after several transfers and I see five distinct substrains: 2 rhizo substrains at the 10 o'clock position, 1 rhizo strain at the 2 o'clock position, and two fluffy strains at 12 and 6 o'clock. If one were to grow out those isolated cottony or fluffy strains, results would be poor. You can see how the rhizomorphic strains are already gaining speed, and if that dish were left for another two weeks, the rhizo strains would cover the entire dish. STRAIN ISOLATION

I would recommend no more than 1/8 cup for that 15 gal bucket, whether horse manure or straw. BLOOD MEAL

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Invisibledumbfounded1600
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Registered: 07/29/07
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Re: All Of RR's Notes (Advanced Mycology) Notes/Links I Liked [Re: dumbfounded1600] * 2
    #8524930 - 06/14/08 09:01 PM (15 years, 9 months ago)

RogerRabbit 2006 Advanced Forum

They use Banrot 40WP. I ran several experiments with this fungicide a couple of years ago. It is so powerful you can soak rye grain in it, then PC, then leave the lid off the jar for 24 hours in an open room, and the grains won't contaminate. It can also be applied to casing material, and I guarantee that no trich or cobweb will grow on it.  It works by preventing spore germination, so it has to be inoculated with live mycelium, as nothing will happen if you try to inoculate with a spore syringe. After determining that the Banrot 40WP works, I stopped the experiments because I see no reason to use chemicals to replace proper clean room procedure. A properly pasteurized, NOT sterilized casing layer will have no trouble surviving two flushes, which delivers 90% of the fruits you're going to get anyway. After two flushes, I recommend tossing the tray into your outdoor garden and replacing it with a fresh tray for the most effecient use of your growroom space.  Dried Rhododendron leaves are nearly as effective if someone is really having trouble with trich or cobweb. Simply dry them, then grind up in your hands and mix with the casing material at the rate of ten percent. Pasteurize and use. Fungus spores won't germinate in the presence of Rhododendron leaves. Contaminants aren't a problem in outdoor patches. I have no idea what the active ingredient in Rhododendron leaves is, but they do prevent mold spores from germinating.  FUNGICIDE IN CASING

The mycological definition of a 'strain' is a pairing of spores to form dikaryotic mycelium. This means that every spore print can produce thousands, even millions of strains. The 'OMC definition' of strain is the region of the world where the print was collected. Such is not mycologically correct, and does not constitute a 'strain'. STRAIN

PE6 is a Tex/PE cross, and unfortunately the 'texas' shape presents itself much more than the PE. I agree with workman in another thread when he said the penis shape and albino coloration are recessive. PE6 shouldn't be considered a true PE because of this. PE 6

You won't see individual cells at 200X without very expensive lenses. Digital zoom won't get you there. I can make them out at 400X, but to clearly see the components of the individual cells, a properly prepared slide and 1000X with oil immersion is required. 200X can help identify contaminants, which is one of the most important uses for a scope in our hobby anyway unless you get into a breeding program. VIEWING MUSHROOMS UP CLOSE

You may have seen the term 'substrain' used here. It refers to the various matings of compatable mycelium that generate into sectors from the same sporeprint. Technically, anytime mycelium mates up to become dikaryotic, a strain is born. However, it's what we normally refer to in the OMC as a substrain. SUB-STRAIN

Lots of people have dunked their logs with success, others with failure. Smaller logs are definitely easier to handle when it comes to dunking. The biggest reason in my opinion for using logs for one massive flush is the increased chance of contamination from straw after a single flush, whether one uses logs or bins. It should be noted that the majority of fruits from straw will always come from the first flush. Thus, the increased chance of contamination and the resulting green spore drop must be weighed against the limited return from a second and succeeding flushes. I'm sure others will pop in with their experiences as well. A lot of it depends on the climate and the contaminant spore load in the area you grow. During the winter, your chances of success with multiple flushes will be substantially greater than during the summer when the trich spore load is much greater. I hope this helps you make an informed decision.
RR (skyypilot). Well because more than about 10% manure will screw up a strawlog. I still recommend one flush, then burying in the garden outside. Straw is so cheap, your real estate is better served by making a new log and putting the area to better use with another massive flush, rather than going to all that work for a hit or miss second flush. Of course I should say the above is just my opinion, others are free to disagree. Here's a couple of tips. Get your bale of straw at a feed store and get the huge bale for the same price as the little ones at a craft store. Stop by U-haul and get three of the medium size boxes. You can stop before you get to your apartment and break the bale down into those three U-haul boxes which won't attract attention as you carry them in. The other thing, when pasteurizing straw indoors, always put the spent water/straw chunks down the toilet, not the sink. The toilet has a 3" diameter drain, the sink only has a 1 1/4" diameter drain.  STRAW LOG

Most fungicides kill all spores, but allow mycelium to grow untouched. Thiophanate-methyl is approved by the FDA for use on mushroom crops and I ran some tests on it a few years ago. It's the active ingredient in Banrot 40WP. You can read up on it here. http://www.epa.gov/oppsrrd1/reregistration/tm/tmsummary.htm
FUNGICIDES

Plants and fungi have a long history of cooperation. Everyone should dispose of their spent cakes into their houseplants. The plants will thrive and you'll still get an occasional mushroom. The metabolytes produced by the mushrooms are plant food, and the plants roots open up paths for the mycelium to colonize. It's a win-win situation. PLANTS AND FUNGI

Peroxide breaks down very fast in the presence of organic matter. I would imagine all of the h2o2 was broken down long before the mycelium finished colonizing the top of the dish. PEROXIDE

Why would anyone pick fruits that aren't ready? Unless you're harvesting 18 wheeler loads for a commercial operation, you should have time to make several passes over each tray to only pick the fruits that are ready. Picking the entire batch of mushrooms because one or two are ready is silly. Some people believe that by picking the first few stragglers that come up you can stimulate a better pinset, while others say to leave those early birds in place until the entire flush is ready, which causes them to puke spores everywhere. I say pick individual mushrooms as they are ready. If lots of pins are going to be damaged by picking, then simply snip with sissors or a knife as close to the substrate as you can without damaging the nearby pins. PERPETUAL HARVESTS/FLUSHES

I did a grasshopper grow once. Go for it. Dry the moths very well, then rehydrate and pasteurize. I'll bet it will work. Yes, just don't let them get too soggy. BUG SUSBSTRATE

The reason fruits form around the edges of casings while boycotting the middle, is due to grower error, not the preference of the mycelium. STRAW LOGS

The idea with strain isolation is to make a dozen or so isolates, then fruit each one to find that stellar performing strain. Once you have a really good performing strain, with care you can continue to use it forever, making the time spent isolating worth the effort. STRAIN ISOLATION

A lot has been learned since the mid 80's when paul wrote TMC. Mushroom metabolytes are no longer considered to be a 'waste product', but an antibiotic response to bacteria or competitor molds. Work is underway to determine the benefits of metabolytes in other applications. Wouldn't it be sweet if we could overincubate a jar to produce harvestable metabolites, then use it to spray on a trich outbreak to squelch it? (just a theory at this point) For the time being, I use the presence of large amounts of metabolyte in a jar to be the indication of bacterial contamination. Such jars are ok for spawning to bulk, but should be avoided for grain to grain transfers. METABOLITES

All of my prints except for the redboys are nearly jet black. Spore color makes an excellent identifier for our cross-breeding experiments, and it must be admitted that both pf's brown spores and the redboy's lavender spores show a larger variation from the norm than you find by comparing EQ, PR, GT, etc. Such makes both strains significant to the community. Since both breed true and produce spores of the same color as the parents, they each truly deserve to be called 'strains', but both also show physically that they are obviously of the same species, not a new one. To be fair, Peele and I gave away the Redboy for free to the community, and it also appears that pf has given his mushroom away for free after seeing a Redboy print that hippie3 sent him. Marketing is not a factor here. Bragging rights maybe, but not marketing. SPORES

I don't consider monokaryons from one strain mating with monokaryons from another strain to be 'hybrids' because they're the same species. Of course they'll pair up and the offspring is fertile. It's similar to humans from Europe mating humans from South America or Africa. The results are not hybrids, but simply 'crosses'. A cross-species pairing would be a hybrid. SPORES/CROSSING/HYBRID

I've made many attempts at crossing azurescens with woodlovers of every species I have cultures for, including the conks such as Ganoderma, and edibles from oysters to shiitake. So far, no success. I'm sure it's just a matter of time before somebody finds the magic bullet and pulls it off. CROSSING SPECIES

If you still have problems with contaminants after using peroxide, you can also dip tissue into a ten percent bleach solution for a couple of minutes. Believe it or not, the mycelium can withstand this, but molds and bacteria can't. The bleach works really well when cloning dry tissue. Live tissue needs a week or two to recover from the bleach, but dry tissue is dormant so hardly even gets smacked. I've recovered three year old tissue. There will definitely be contamination, but you'll be able to work through it with agar. I generally try to do transfers as soon as I see mycelium without waiting to see if it contaminates or not. At other times, bacteria or mold will start to grow first, then the mushroom mycelium. In this case, all you can do is begin a series of transfers until you have a pure culture. PEROXIDE

I would consider a 'generation' to begin with a successful spore mating. In my opinion, you could expand it until the cows come home, and it's still first generation, until you fruit it to obtain spores, which could begin the second generation. Senescence is what you're thinking of when you expand the mycelium too far. GENERATION

So far, I've tried nearly all the plastic wraps, micropore medical tape and masking tape. None can compare with parafilm. You want gas exchange, but no air exchange. Micropore tape gives too much exchange and the dishes dry out. The Saran type wraps either don't allow enough gas exchange, or none at all, resulting in slow growth. Masking tape doesn't stop contaminants. I'd get parafilm. It only costs a few pennies per dish to wrap and does the job well. Try inoculating two side by side dishes with the polyethylene on one and parafilm on the other. What I found was both cultures would grow pretty evenly for the first week, then the polyethylene one would slow down, while the parafilm wrapped dish kept growing rapidly. Neither contaminated however, so the Glad does its job that way. PARAFILM

Soaking the grains first has two main benefits. First as said, any endospores that may be present will germinate so they can be 86'd by the pressure cooker. However, endospores are really more of a problem for those using popcorn, rather than other grains. People using rye or wheat rarely report bacterial contamination from endospores when they skip the soak. I soak for 24 to 36 hours after rinsing the grains very well with hot tap water. I also begin the soak with hot tap water. The heat will help to germinate organisms that may be dormant in the dry grains. After 24 hours, you'll smell a bit of fermentation, but it doesn't hurt a thing. It tells you the organisms in the grains have activated, and will be easy to kill in the PC. Perhaps one of the best reasons to soak first is to soften up the grains slowly so they can absorb moisture without cracking the kernels. If you dump dry grains into water and bring it to a boil, the grains will burst as they absorb water too fast to stretch enough to hold it all. Soaking will soften them slowly and they won't burst. Burst kernels are sticky(starchy) and much slower to colonize than intact grains. Yes, you can crumble up a piece of drywall to use the gypsum. I'd recommend getting a brand new clean piece of drywall. Perhaps visit a construction site where they're sheetrocking and ask the foreman for a few scraps. They have to haul them to the dump anyway. Add a tablespoon of finely crushed gypsum to each two gallons of soak water to keep the grains from sticking together and clumping up later. A tablespoon of gypsum in two gallons of water isn't going to dry anything out. SOAKING GRAINS ENDOSPORES

Perhaps compatible hyphae is more accurate. I think the word sex is mostly used as an analogy to make it easier for non-mycologists to understand fungi. MATING SPORES

Genetics. Look at a Pygmy from Africa next to an NBA player from Croatia, yet they're both humans. Look at a poodle next to a german shepard, etc. There's also lots of variation within each strain. Not all PE's look alike. Ditto for all the strains. GENETICS/CROSSING

Actually, bacteria will grow very well in the refrigerator, just a bit slower. That's why they stink so bad if we leave the leftovers from the chinese restaurant in there too long. Try cutting off a chunk of the sclerotia itself and transferring it to a new dish. I'll bet it takes off and grows. BACTERIA

While you're going through TMC, look up anastomosis. We can breed fungi by spores or by crossing dikaryons, either way. CROSSING

Gentamycin sulphate is the only antibiotic I know of that works well with agar. It is added BEFORE sterilization at the rate of 1/15 gram per liter. Gentamycin sulphate

Looking good. Try putting some spores on a slide with a bit of karo water so you can take pictures of them germinating. GERMINATING SPORES/MICROSCOPE

I would always recommend starting spores in the two dimensional space of a petri dish so that the germinating mycelium can be visually observed and isolated away from any contaminants that may be present, as well as to isolate the dozens of strains that emerge from a single swipe of spores onto the agar.  Later, if you wish to grow truckloads of mushrooms, move the isolates over to LC once you've proved a good fruiting strain. I see little to no benefit in starting spores in an LC, because it's very easy to transfer bacteria and molds to your grains or brf, then grow them out only to find your project contaminated and all the work you've done to that point being ruined. To answer your question on ph, spores will germinate very well in the 5.5 to 7 range. Just remember however, that molds such as trichoderma and cladosporium also prefer that range, so absolute attention to cleanroom procedures is a must for a high success rate. I am one of the biggest advocates of using coffee in growing, but due to coffee's ability to support mold growth, I'd suggest not using it in the early stages, such as during spore germination. Good luck and happy growing. STARTING WITH SPORES

Interesting stuff. I add activated charcoal to my agar when doing specialty experiments such as germinating very old spores or crossing strains. I've noticed healthier mycelium when grown on activated charcoal, but not necessarily faster or more growth. I've also noticed a decrease in bacterial contamination when charcoal is used, and bacterial colonies that do get started, seem to stall out after a couple of days, then are overran by the mycelium. I doubt it's the small amount of infrared that is suppressing the bacteria, but who knows? I add about a tablespoon (7 to 10 grams) to one liter of Agar. Be sure to powder the charcoal first. I use a bowl and pestle. It's activated charcoal. I got it at a pet store for use in aquarium filters. I never saw a recipe before using it. I just tried with a tablespoon of charcoal and it worked out ok. It makes the agar darker so the mycelium is easier to see too. CHARCOAL

Why are you trying to mix bulk substrate ingredients with your spawn ingredients? That's been tried many times as a way to save a step, but it costs more in yields than it saves by skipping a step. BRF is not a good bulk substrate, nor is manure a good spawn. MIXING BULK SUBS AND SPAWNING SUBS IN A SUBSTRATE

Do a search for protoplast fusion. Here's an article by a good friend of mine, Dr John Holliday, who is also a member here, although not still active. http://www.nwbotanicals.org/nwb/lexicon/hybridcordyceps.htm PROTOPLAST FUSION

You might get lucky with less. Put two small pieces no larger than a grain of rice from each strain 1" apart on a dish and let them grow together. A line of isolation will develop between them as you can see from the picture below on the left. If a third growth emerges from the junction of the first two, it's a cross between them as can be seen at the 12 o'clock position from the picture on the right. Bear in mind, you must start with pure strains. In other words, single sector isolates, so you'll have to do a few transfers to get to that point. It also helps your final outcome if both strains are proved awesome fruiting strains. Toxins such as rattlesnake venom can be mixed into the agar after it cools that will injure the cell walls of the mycelium enough to help the nuclei to come together. You can read about the process by searching for the thread "Return of the Redboy" here in advanced and/or read John Holidays paper on using the technique with cordyceps here: http://www.nwbotanicals.org/nwb/lexicon/hybridcordyceps.htm
http://www.shroomery.org/forums/files/05-39/791852198-zone_of_inhibition_001.jpg
http://www.shroomery.org/forums/files/06-16/576368248-cublandia_mycelium_001a.jpg

Try anything you can think of to degrade the cell walls. Read the article by John Holliday. He lists some other aides to hybridization besides snake venom. Snake venom is available commercially, but there's quite a 'clearing process' before they can ship to you. It's also several hundred dollars per gram. Massive spore dilution or streaking in a zig zag pattern on agar will help you isolate single spores to generate monokaryons, but that isn't what you're looking for here. You were wanting to breed dikaryons. You want to make numerous transfers until there is no more sectoring on the petri dish. I've been wanting to cross redboy or pf's redspore with an albino, but have not been able to find the time, so I hope you're successful.
BREEDING DIKARYONS THROUGH ANASTOMOSIS

I add about a tablespoon (7 to 10 grams) to one liter of Agar. Be sure to powder the charcoal first. I use a bowl and pestle. It's activated charcoal. I got it at a pet store for use in aquarium filters. I never saw a recipe before using it. I just tried with a tablespoon of chacoal and it worked out ok. It makes the agar darker so the mycelium is easier to see too. CHARCOAL

Other species such as agaricus will give improved fruitings with carbon. I was able to get morels to fruit by adding activated carbon to the pasteurized peat/verm mix the sclerotia were buried in. You might test it with test strips instead. Carbon is electrically conductive, so is probably throwing off any reading you'll get with the Kelway. Yes. You can fruit morels by growing out the sclerotia, then after letting the sclerotia sit for a few months, hydrate and place into peat/verm with activated carbon. Don't transfer the grains, only the sclerotia. I'm sure there's better ways to do it, but this is one. Yes. Separate the sclerotia from the substrate and allow them to dry partially. Then, rehydrate and place in various mediums until you find one that fruits well. The morels will come right out of the sclerotia, not any myc that emerges. We had one fruiting that came from dumping ashes from the charcoal grill onto the substrate, but were unable to duplicate it in the limited time we had. To the best of my knowledge, cooking charcoal is not 'activated'. USING ACTIVATED CHARCOAL/CARBON IN CASING






RogerRabbit 2007 Advanced Forum

Of course there's thousands of parents if thousands of spores germinate from multispore inoculation. Eventually, most of the substrains combine into a single organism through the process of anastomosis. If one wishes to separate out the individual strains, or substrains as they're sometimes called in order to preserve the diversity, you want to place the multispore culture on agar where you can observe the growth in the flat, two dimensional plane of the Petri dish. This process needs to begin within a day or two of germination so you can catch the individual strains before they combine with other dikaryons via anastomosis. Some strains are not compatible, so this is the reason on many substrates that were inoculated by multispore you'll see drastically different looking(and performing) mushrooms on the same flush. It's because those that are not compatible usually lay claim to their niche of real estate and fruit from there. By the time it gets to that point, my liquid slides are so thick with growth, the microscope light can't shine through them. According to stamets, and it seems to match my observation, most of the time all the substrains combine into a coherent whole. In other words, if you took a sample of mycelium from each corner of a fruiting tray and placed them on the same petri dish, they'd grow together seamlessly without forming a zone of inhibition. Still, there are times when it's fairly obvious by observation that more than one substrain is at work in the same fruiting tray. SPORE GERMINATION

Perhaps with a magnifying glass you could see this: Monokaryotic mycelium tends to look more like mold. That is, the mycelium is curly and wiggly, while dikaryons tend to be straighter. Monokaryotic mycelium also has a lot more side branching of the hypahae, but you won't see that without a scope. MONOKARYOTIC/DIKARYOTIC

The point to remember is a single swipe of spores in agar will deliver up to 100 isolates. I make the first transfers within a couple of days of visible growth on the petri dish. I'll make at least ten transfers from the perimeter, or leading edge of the mycelium. Each transfer should be no larger than a grain of rice or uncooked grain. After a few days, each of these transfers will have grown out enough that you 'might' be able to see some sectoring. a sector is a growth that is separating itself from the other growths on the plates. They will radiate out from the center like spokes on a wheel. Transfer each of these growths to a new plate. When you have completely isolated a strain, the mycelium will homogeneous like in the 10 o'clock to 3 o'clock position in the bottom section of the picture below. The other sections have similar isolates, but all need at least one more transfer to be truly homogeneous monocultures. The petri dish below represents the fourth transfer of it's mycelium line from the original swipe of spores. Other mycelial lines were being generated at the same time. Remember, you don't isolate ONE strain. You isolate ALL of the rhizomorphic strains from that one swipe of spores and fruit out each isolate separately to determine which has the qualities you're looking for. While you're fruiting them out, the original petri dish from each isolate gets labeled and goes into the refrigerator until you've determined which ones are keepers and which ones are trashers. The keepers are transferred to test tube slants as soon as possible, where they can be stored under refrigeration for years without substantial degradation. I've filmed a complete strain isolation tek for my video that is coming soon, so you'll be able to see the whole process from mixing the agar to the finished isolate in finite detail, including master slant preparation and inoculation. GETTING AN ISOLATE FROM A STRAIN

The grass above grew powdery mildew before those caps could even open, ruining the project. Plants and mushrooms do well together in nature, but unfortunately not in indoor cultivation. Any wood in a project like that would grow trichoderma long before any wood loving mushrooms could fruit. Sorry.
http://www.shroomery.org/forums/files/05-15/339833891-mex_0006a.jpg
It was rye grass seed that was planted in the casing material right after it was placed on the colonized substrate. The mold was dactylium, which is in the air everywhere. It got on the wet grass and started growing, which doomed the project.
INDOOR GRASS IN CASING/BULK SUBSTRATE

Spores from different strains are totally compatible, just as pollen from bubblegum can easily cross with purple haze. There is no challenge or or anything special about 'man made' strains. It happens every time you squirt spores into a jar. A strain is made when compatible monokaryons cross paths and exchange genetic information. It mattes little what the 'name' on the print was, or even if the monokaryons came from separate prints. Dikaryons can easily cross too via anastomosis. You need to pick up that book and do some reading. It gives a great starting point to build from and will dispel your early misunderstandings. SPORES/STRAINS

Get ready to be addicted to the miniature world for a while when it comes. There's a whole new universe you'll see and it's a facinating one. Did you get a camera too? The best microscope cameras are the ccd tube mounts, but they're a bit spendy. A cheaper alternative is the ones that replace an eyepiece, but they don't have the clarity or resolution, but still get the job done. Have fun! If you make your product in large batches, it's always best to test on grains or agar a sample from each batch. Sometimes the single drop of liquid you put on a slide will be clean, but there could still be bacteria or mold spores in the rest of it. Just a heads up. Which reminds me. Be sure to order a box of slides and cover slips. I can put a closed petri in my cheap microscope, but my better lab one doesn't have the working distance to be that far away, even if I fill the dish up. In fact, with the stage all the way down, I can't even fit a petri in under anything but the 4X objective. It's barely above a cover slip on a slide when the lenses are in focus at anything over the 10X objective. That's the only reason I don't get rid of the cheapie. A stereo microscope is not for looking at things on the cellular level. Consider it like having a super macro lense on a camera. The lense and the light source are on the same side of the object being viewed, so a stereo or a stereo zoom microscope sees the light that is reflected off the object. A zoom/stereo microscope is great for studying the gill structure of a mushroom, or looking at other objects such as jewelry or bugs. A light microscope has the light on the opposite side of what you're viewing from the lense, so it shows you the light that is penetrating the object you're viewing. With a light microscope, you can view things at the cellular level. Take a scroll through my gallery and you'll see several pictures taken through each type. I would always suggest getting the best scope you can afford. It's no fun looking through shitty lenses or using a cheap light that won't focus on the object you're studying. MICROSCOPIC ADDICTION

The best place to start is to get a copy of stamets "The Mushroom Cultivator" and read the first few chapters ten times. You can get it from fungi.com or amazon.com. There's no way you can start out without basic mycology knowledge to do something like what you're wanting. We'll be here to help with the advanced stuff that you don't get from his book. Spores from different strains are totally compatible, just as pollen from bubblegum can easily cross with purple haze. There is no challenge or or anything special about 'man made' strains. It happens every time you squirt spores into a jar. A strain is made when compatible monokaryons cross paths and exchange genetic information. It mattes little what the 'name' on the print was, or even if the monokaryons came from separate prints. Dikaryons can easily cross too via anastomosis. You need to pick up that book and do some reading. It gives a great starting point to build from and will dispel your early misunderstandings. An alternative is to isolate a couple of strains down to single sector dikaryons, and then place those on a petri and allow them to run together. If a third sector opens up and starts growing, isolate it. You can then test for the cross by placing all three on a new dish. A line of isolation should develop between all three. It sounds strange, but if you'll sip a glass of wine or a single shot of vodka, it will steady your hands for precision work. In fact, at shooting competitions, they test them for alcohol for this very reason. You'll want to duplicate your efforts several times over to increase the chances for success. Cubes have tetrapolar basidia, so hyphae from individual spores won't be compatible with all hyphae from other spores. It's the A, B, A1, B1 thing. I'm not sure paul covers that in TMC, but you might find some information on it at Tom Volks website. I know when he travels around, he gives an excellent presentation on exactly what you're trying to do. I've made numerous transfers of cottony mycelium and it stayed cottony. I've made numerous transfers of rhizomorphic mycelium and it stayed rhizomorphic. I believe there's more involved than we currently know. I have many strains that I've isolated down to single sector rhizomorphs, and these remain rhizomorphic even years later when removed from the refrigerator and transferred to new substrates. I think a lot of confusion comes from ceasing to make transfers before getting down to single sector isolates, thus the process of anastomosis is still occurring which would explain the mycelium changing to different types. I've found a difference in appearance based on the nutritive substance in the growing medium, with the strands much closer together when highly nutritive media was used, and the strands spread out more when lesser nutritive media was used, but they're still rhizomorphs when viewed under the microscope, even if they don't look so to the unaided eye. I've attempted to fruit completely cottony isolates and the results were also the same every time. Either they didn't fruit at all, or fruited poorly. Rhizomorphic isolates have almost always produced nice fruitings with only one or two exceptions out of hundreds. I go through about 500 petri dishes per month, so believe me when I say I've repeated these experiments many times over. CUBE CROSS BREADING

Most people either boil, or use cold water extractions. Who knows? We just make a broth by boiling the turkey tails for a couple of hours, then drop in a chicken and make chicken soup the normal way, using the turkey tail broth as a starting point. This method works with Reishi as well. It's the best way, in my opinion because you can then get the medicinal benefits with every meal and it tastes good too. Use the broth also for cooking pasta, rice, etc. Most myco medicinal companies grow mycelium in very large vats of LC, then do cold water extractions on the mycelium without ever fruiting it. Just what can be found online. The myco medicinal people keep it top secret. They won't even share with each other at the conferences. MEDICINAL MUSHROOM EXTRACTIONS

I've successfully cloned from dried fruits many times. You want to first cut up a few pieces of stem into about half the size of a grain of rice, and then rinse them well in sterilized distilled water. Drop the stem pieces into the distilled water, put the lid on the jar and then shake like hell. This washes the stem parts. Then, transfer the pieces to another jar of sterilized, distilled water with a filter on the lid such as you'd use for growing grains. It takes about six weeks to two months, and you'll see a bit of mycelium begin to grow from some of the pieces. Transfer these to agar. You can use a syringe to selectively suck them out of the water. Don't use any nutrients in the water at all. Just PC'd distilled water. CLONING FROM DRIED CAPS

You need to do number 3. However, a single swipe of spores on a petri dish will yield at least 50 substrains. I'm doing a series of isolations now that started with two swipes, each on its own dish. I've only been transferring the best looking mycelium, and am now up to over 30 substrains of pure rhizomorphic mycelium, and this is from just two swipes of spores onto agar. I plan to fruit out each and every one of these strains separately after placing the individually well marked dishes in the lab refrigerator. After fruiting, I will determine the best strains that I want to keep. I will then go back to the individual petri dishes that are in the fridge, allow them to warm back to room temp and start growing again, and then transfer a small piece of mycelium from each into master culture slants for long term storage. I will then be able to grow those same strains for 20 to 30 years, without having to go back to spores. A lot of work? Perhaps, but once the strains are isolated and stored in culture slants, it's a breeze after that and the results are dependable. Multispore inoculation certainly works, and it's how nature works. However, by isolating strains, you can get those killer flushes of wall to wall fruits with whatever 'other' qualities you seek. Those 'other' qualities might not come from the fastest growing or largest strain, and might not even come from the best flushing strain. That's why each strain needs to be individually tested. By the way, there is a full strain isolation tek and also a master culture slant tek on my DVD. You can see some short low res previews here: http://www.mushroomvideos.com/1809563.html MAKING A GOOD SUBSTRAIN (AGAR, ETC...)

Dikaryotic mycelium can exchange genetic material with other dikaryotic mycelium of the same, or even similar strains. This process is anastomosis. SPORES

Dipping a culture into peroxide is a horrible idea before transfers. It does harm the tissue and sets it back right at the exact time you're wanting maximum rate of growth to outrun contaminants. Mold mycelium will recover from the peroxide dunk faster than the mushroom mycelium will, so you defeat your purpose. You should transfer some of the culture to agar, so you can isolate the mycelium away from contaminants. I wouldn't even attempt to transfer non-sterile cardboard to grains. Simply work clean and sterile when you make the transfers. As soon as the mycelium starts to grow from the wedge, immediately transfer some of the new growth to new plates. Repeat(if necessary) until you have a clean culture. I've used peroxide when cloning heavily contaminated outdoor mushrooms that have been in my backpack for three days after picking, but that's a different story. It does set the tissue back significantly however. PEROXIDED DUNK BEFORE TISSUE TRANSFER

Normally, the traits in a fruit will carry over if you clone it. If you clone a mutant, you'll get harvests of mutants, provided the mutation was genetic in nature. If you clone from a cluster of large fruits, your future crops will develop clusters of large fruits. Senescence is always a problem if you try to keep a fungal cell line going for too long. It isn't necessary though. Instead of constantly cloning the clones, clone the first one and then make a few master culture slants in test tubes to store the mycelium. You can then take it out of the refrigerator and use it for years to come without degradation. Damage as the pins are forming, or for another example, damage from chemicals containing petroleum can be mistaken for mutations. I doubt there's an environmental condition you can easily create that would cause mutations. CLONING

You can mix a small amount of Borax into some sugar water or Karo and leave it in small cups for the ants to drink up. They'll take it back to the nest, and in about a week, the whole colony is wiped out. ANTS

Under FDA regulations, the tissue is autoclaved prior to loading into pills. You won't be able to start a culture unless they skipped that step. CULTURING CORDYCEPS

Very few rhizomorphic strains will be non-fruiting. I always ignore the cottony sectors anyway, so that ups the percentage. I won't even venture a guess as to percentage if you tried to fruit each sub-strain that differentiates. We're looking for more than fruiting strains though. Average number of fruits per flush, weight of fruits, quality of fruits, resistance to aborting, etc., are all factors to consider when choosing a good strain to keep in master slants. AGAR

Turning them upside down tends to force the water to drain from the agar by gravity, drying it out and ruining the mix. Colonize petri dishes in a vertical stack at room temperature. It's the temperature differential between inside and outside the dish that leads to condensation. Having them stacked vertically goes a long way to reducing the temperature differential on the lids. The condensation was caused by moisture that was forced out of the agar, so by leaving the dish right side up, the moisture will re-absorb back into it. CONDENSATION

A grain to grain transfer is done with young, rapidly growing mycelium. In no way is that the same as cloning. I would not clone a clone. It isn't necessary. If you have young cell lines, simply store them in the refrigerator for years and keep using them. You keep the original master culture slant in the refrigerator. When you need mycelium, you take a tiny piece from the culture slant and transfer it to a petri dish, then return the slant to the refrigerator, where the mycelium slowly grows to replace what you cut out. Use the petri dish(es) to start the next batch. By doing this, your master slant slows way down on cell division, thus preserving the culture without senescence. When the culture slant goes into the refrigerator, it essentially stops cell division, putting it into stasis. This way, you can take it out of the fridge, get a small piece, and then return the slant to the refrigerator. Using this system, a 20 year old culture might only have six weeks or less of growth, thus senenscence isn't a problem. CLONE A CLONE

You shouldn't have one layer of spawn, and another layer of substrate. You can mix them together, or you can make several layers of each so they can mix naturally. Once the mycelium fully colonizes the substrate, the entire mycelium becomes one organism. Tracer elements have been added to one corner of a substrate, and the fruit bodies uniformly contained that tracer element, verifying that the entire substrate feeds the flush. NUTRIENTS FROM A SUBSTRATE

Mushroom mycelium consumes air, just like we do. It also produces CO2, apparently as a byproduct, just like us. One benefit of a high CO2 level during colonization, is that less of the actual carbon in the substrate is converted to CO2 gas. In other words, if you allowed too much fresh air during colonization, more of the substrate would be 'consumed' by the time the mycelium got to the fruiting stage. A rapid and sudden decrease in the ambient CO2 levels is a pinning trigger in cultivation, so adding fresh air at the same time as full colonization and the other pinning triggers helps ensure a full, even flush. SCIENCE BEHIND GAS EXCHANGE IN COLONIZATION

It's pretty common. A single swipe of spores will generate dozens of single sector isolates, many of them rhizomorphic. It's important to grow out all of them to determine the best fruiters. Each will have it's own characteristics. AGAR

Go around the leading edge of the growth, and transfer small pieces all the way around. Take pieces about the size of a grain of rice. Repeat in a few days with the new dishes. By the second transfer, you should see clear sectoring. Transfer each sector to new dishes. SECTORING

Isolates are genetic, not substrate related. Mycelium will often turn from cottony to rhizomorphic and back again depending on substrate, moisture, temperature and perhaps other variables as well. One needs to provide the type of food the particular species he wishes to grow requires. GENETIC ISOLATES

You can put several specimens on each petri dish, whether or not you have them sectioned. I like the sections though. I won't ever go back to single dishes. It usually takes two or three transfers before rhizomorphs begin to show up. I prefer the three section dishes. You can run three experiments at once, saving time, money and space. AGAR QUESTIONS

I don't care much for either one of those. The lenses are achromatic as opposed to plan achromatic, which keeps your object in focus over the entire objective. With achromatic, you'll have a part in the center of the lens that is in focus only. In addition, budget scopes have very small objectives, so they don't gather much light. What is the main purpose of a scope for you? I use my zoom microscope most of all. It is designed to look at things from the top, using reflected light from a lamp also above the culture, rather than looking at the light that shines through your object from a lamp below. I wouldn't recommend 20X eyepeices. They block too much light, and their optics are usually poor, especially on the made in china microscopes. If you can get plan lenses, get them. The 30 mm eyepieces are great. I have a few sets of those for my scopes. I'd suggest getting one with a 100X oil emersion objective. If you want to count nuclei in a cell, you'll probably need it. You can google terms such as plan, phase contrast, dark field, anchromatic, etc. for lots of reading. Here's a page that tells a bit about phase contrast. http://www.microscopeworld.com/MSWorld/phase.aspx MICROSCOPE OBJECTIVES

Cordyceps brings a few thousand dollars a pound. They grow the mycelium, not the fruiting body. Getting a culture is the hardest part. You can't buy one at any cost that I've found. You have to find one in the wild and get a clone or spores. Here's mine. I'm using cricket agar to attempt to propagate it.
http://www.shroomery.org/forums/files/07-40/164105184-cordyceps_1a.jpg
http://www.shroomery.org/forums/files/07-40/164105197-cordyceps_2a.jpg
http://www.shroomery.org/forums/files/07-40/164105216-cordyceps_4a.jpg
Cordyceps militaris. in the picture above.
Which is exactly why I won't give mine away. I'm now several thousand dollars into DNA testing just to determine the various species I'm isolating. Cordyceps has many anamorphs, which make it very hard and expensive to cultivate. It has a 'green mold' conidial stage which further complicates matters. Those who spend their hard earned cash and time to isolate strains should be compensated. I've broken several bones climbing mountains in search of rare species, and mrs rabbit is laid up right now with a broken leg as a result of a terrible fall during a mountain top foray two weeks ago. Why should we just give our work away? Think about it. The grocery store doesn't give us free food, and Jeep didn't give us our mountain vehicle for free either. Nobody paid us to train our bodies to climb 4K to 5K vertical feet and back in one short winter day either. We've worked very hard for what we have. I would have no problem with fellow members here growing one of my strains. The problem with giving them away is one of the first ones in line will be big pharma to get a free culture they'll sell for millions of dollars in tablet form. If there's a financial benefit to be derived from my work, I intend to see that it goes to me and not to them. Alan, if you think I should give everything I work for away, be sure to tell your boss next payday that you don't want paychecks anymore, and to simply donate them to the pharmaceutical companies. You're working for the good of humanity. Bear in mind, Cordyceps militaris has many anamorphs, including 'green molds'. It's an amazing species, as I'm finding out firsthand here. Alan, if you can afford it, send your culture for DNA testing. It would be very interesting to see what comes back, and what anamorph, if any, they're selling. Actually, the first one tested morphed into a different mycelium type, which tested as Cordyceps militaris. This shows that at least one anamorph of Cordyceps is Penicillium. I'll work up a paper on this as soon as I have more info. It's a long, slow process with this shape-shifting, and apparently DNA altering organism. However, I'm busy with a new video project as well, so the Cordy cultures are in cold storage at the moment, there to remain until I have more time available. MUSHROOMS FROM DIFFERENT INDUSTRIES

Experiment. Put some bacteria on a petri dish, and then put down a line of metabolite. Watch how they stop the spread of the bacteria. Without a lab, I doubt you could make your own antibiotics, but penicillin and the other common antibiotics are made from fungi metabolite. I've also found that mushroom metabolites will kill trichoderma mycelium on a petri dish. In fact, it would probably kill off any competitor fungi, even other mushroom species. USE OF MUSHROOM METABOLITES

The last thing you want to do is allow a petri dish to fully colonize. Use the agar to isolate strains and to isolate away from contaminants. Don't forget to wrap with parafilm. PETRI DISH

Search Cordyceps on Google. You'll find a few good articles. There's little to no cultivation parameters though. I used live crickets to make the agar, and it's working OK. I keep them in the refrigerator at all times. Cordyceps mycelium will grow at just a couple of degrees above freezing. You'll also find some good articles at John Hollidays site. www.alohamedicinals.com RR QUESTION ABOUT CORDYCEPS

A sector is a strain. A sector develops when compatible hyphae join to become a single organism, sharing genetics. Strains can also join with other strains via dikaryotic pairings, known as anastomosis. An isolate is a single strain, meaning you have transfered all the sectors to their own plates.By taking tiny pieces of mycelium when I make transfers, I can often get to single sector isolates with three to four transfers. The dish in the picture on the left below is the result of two or three transfers. You can clearly see each sector by holding the dish up to the light. Tiny cuts were made in each of these sectors to move that mycelium to new dishes, and each of those became a single sector isolate. The right picture shows a dish that is still sectoring, and on the right is an isolated strain. Note there is no sectoring at all. Each and every isolate should be grown through the fruiting stage to determine the best performers. Prior to doing so, a small amount of mycelium should be carefully labeled and placed into the refrigerator on a petri dish or culture slant. This way, after fruiting, you can go back to an earlier stage in cell division for each future grow, capturing an aggressive strain.
http://www.shroomery.org/forums/files/05-18/530157237-sectored_dish.jpg
http://www.shroomery.org/forums/files/05-27/070264929-rhizomorphic_myceliumaa.jpg
TRANSFERS, ISOLATES, AND SECTORS?? PLEASE EXPLAIN

Instead of a slice, try scraping the scalpel so you 'shave' a very small piece. You should be able to get a paper thin curly from your scalpel that you can put a drop of distilled water over and then the cover slip should lay flat. There's mounting solutions if you want a permanent mount, but distilled water works great for immediate viewing. Tap water will have too much crap floating around in it. PREPARING MUSHROOM TISSUE FOR MICROSCOPE VIEWING

I've been saying layering works better and faster than mixing for years around here. It's good to see Paul notice that as well.





RogerRabbit 2008 Advanced Forum

Generally, with the exception of coffee, nutrients you add to a substrate will slow down colonization, rather than speed it up. The least nutritious substrates, such as straw, colonize fastest. If the mycelium is hungry, it will grow faster and more rhizomorphic in search of food. If you provide a very rich substrate, the mycelium will colonize it, only slower. If you want genetics for very fast growth, you'll have to isolate very fast growing sectors on agar and transfer these to your substrates. However, the fastest growing cell lines might not be the cell lines that provide the other qualities you're seeking. INCREASING TEH RATE OF COLONIZATION: WAYS?

One of these days, I'll clone a turkey tail and see how it does. It grows wild in the PNW, so we have all we could ever use. Two years ago, Mrs Rabbit went through Chemo for cancer, taking medicinal mushroom extracts, boiling turkey tail and reishi into teas every day, and eating shiitake every day. She completed chemo with 80% of her hair, while everyone else loses it all. Her doctor was amazed, but we're convinced it was the mushrooms. What properties, and which mushrooms did the most good, we have no idea, but suspect it was the cocktail of all of them. I do know the commercial growers of Coriolus grow and harvest the mycelium for the active ingredients, not the fruit bodies. TURKEY TAIL/MRS. RABBIT CANCER

You would still be at first generation after ten, or even fifty grain to grain transfers, but senescence would definitely be a factor. A clone keeps mycelium from a particular cell line growing. After about three months of rapid cell division, most mycelium will give up the ghost and performance will be poor. This is why we recommend no more than three grain to grain transfers. The reason is to make sure that once the mycelium from the 3rd G2G colonizes the bulk substrate, it still has the energy to produce a nice flush. A generation is just like with humans or other animals. If you do a grow from spores, it's first generation, no matter how long you grow it or how many transfers you make, just like you are your mom and dad's son, no matter how long you live. Once you fruit it, take the spores and germinate them, this new cell line is what would be 2nd generation, and so on. Mycelium can go for many, many generations. The PE strain of cubensis is living proof of this. It's degrading, but still viable after all these years. My PE6 and Workman's work with PE albino, etc., are efforts to infuse fresh genetics into the old strain. GENERATIONS

Peroxidated agar kills growth rates. It usually takes three to four transfers to get single sectors. Isolate all of them to determine the best performers. By the third transfer, you should see some rhizomorphic mycelium. PEROXIDE

There are salt water fungi. There is also talk of engineering them genetically to devour oil spills. Google will return several results for you. FUNGI THAT LIVE IN THE OCEAN

Dry tissue can in 'some' instances be revived, but it's certainly not something for a new grower to attempt without agar skills and a good laminar flowhood. In a glovebox, you might have success germinating spores from the dried fruits on agar. The problem is being able to separate the mushroom mycelium from the natural contaminants that will also begin growing on your petri dishes. I'm sure you can find a way to get to a state where spores can be easily obtained. The fuel would probably be less than the cost of isolating from a dried and dirty fruit purchased from someone. Good luck. CLONING FROM DRIED MYCELIA

I put a hardwood stick in each slant, and they easily last 2 to 3 years under refrigeration. If the mycelium on the agar dies, it will still revive from the wood. I use medical tongue depressors. If you don't use wood, make fresh slants every 6 to 8 months. HARDWOOD IN SLANTS

The beneficial bacteria we want to preserve in our casing layers is already in the peat. We pasteurize to kill off most of them, but the few survivors help to provide a small amount of contaminant mold protection, as well as to help stimulate pinning in some species. PEAT/PASTEURIZATION

I'd like to wring the neck of the first asshole who ever said that Lysol causes mutants. Lysol is toxic to fungi because it's mostly alcohol. However, you don't generally introduce genetic mutations in the first generation after applying a chemical-especially in a fast growing organism such as fungi. Remember, a deformed fruit is not a mutant. LYSOL/MUTANTS

The three transfers 'limit' is for grain to grain transfers where the mycelium is expanding exponentially. In a petri dish, this doesn't happen. The flat plane of the dish limits growth to the surface, greatly reducing the number of cell divisions. Also, as Lipa said, we don't fully grow out the petri dishes. We transfer when the mycelium is 1 to 2 cm in diameter. We can easily make three or four transfers in a two week period, isolating a strain or cleaning up a culture from contaminants. GENERATIONS/TRANSFERS

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Invisibledumbfounded1600
Stranger

Registered: 07/29/07
Posts: 2,624
Re: All Of RR's Notes (Advanced Mycology) Notes/Links I Liked [Re: dumbfounded1600] * 1
    #8524935 - 06/14/08 09:01 PM (15 years, 9 months ago)

Links I Liked

http://www.shroomery.org/forums/showflat.php/Number/4674364#4674364
Calcium Enriched Coco Coir

http://www.shroomery.org/forums/showflat.php/Number/5123010#5123010
Electrical Current and It's Effect On Mycelium

http://www.shroomery.org/forums/showflat.php/Number/5553299#5553299
Dikaryotic Mycelium Questions

http://compost.css.cornell.edu/calc/lignin.html
Ligning The Effect On Biodegradability

http://www.shroomery.org/forums/showflat.php/Number/2455578#2455578
A Question For The Science Nerds

http://www.shroomery.org/forums/showflat.php/Number/1893980#1893980
Straw Log Theory, Huge Fruits

http://www.shroomery.org/forums/showflat.php/Number/1880581#1880581
Salt And Mycelium/Spores

http://www.shroomery.org/forums/showflat.php/Cat/0/Number/2390329/page/0/fpart/1/vc/1
Clone From Dried Caps

http://www.shroomery.org/forums/showflat.php/Number/2548613#2548613
Fungus Gnats, Mematodes (Sheinernema Spp) and Hypoaspis

http://www.shroomery.org/forums/showflat.php/Number/2773868#2773868
Barley Straw Gives Of H202

http://www.shroomery.org/forums/showflat.php/Number/2888863#2888863
Agar Isolations

http://www.shroomery.org/forums/showflat.php/Cat/0/Number/2851542/page/0/fpart/1/vc/1
Absolutely Cannot Beat Green -- Can It Perm. Contaminate A House?

http://www.shroomery.org/forums/showflat.php/Number/3837982#3837982
Isolation Of Mycelium From Dried Mushrooms

http://www.shroomery.org/forums/showflat.php/Number/3963836#3963836
Casing Material

http://www.shroomery.org/forums/showflat.php/Number/4134022#4134022
Is This A 'Non-Sectoring' Agar Isolate?

http://www.shroomery.org/forums/showflat.php/Number/4125778#4125778
Microscope Help PLease, GRaduation Present

http://www.shroomery.org/forums/showflat.php/Number/4185523#4185523
Another Genetic Problem

http://www.shroomery.org/forums/showflat.php/Number/4345100#4345100
Sub-Strains

http://www.shroomery.org/forums/showflat.php/Number/4363162#4363162
Can You Clone From Dried Mushrooms

http://www.shroomery.org/forums/showflat.php/Number/4377475#4377475
Need Help With Strain Isolation

http://www.shroomery.org/forums/showflat.php/Number/4468631#4468631
Amorphous Diatomaceous Earth (ADE)

http://www.shroomery.org/forums/showflat.php/Cat/0/Number/4672561/page/0/fpart/1/vc/1
Calcium Enriched Coco Coir (Ca2+)

http://www.shroomery.org/forums/showflat.php/Number/4675607#4675607
The Coffee Project - Results Only

http://www.shroomery.org/forums/showflat.php/Cat/0/Number/4723693/page/0/fpart/1/vc/1
Petri Dish Pics With Following Content...

http://www.shroomery.org/forums/showflat.php/Cat/0/Number/4723693/page/0/fpart/1/vc/1
Hybrids

http://www.shroomery.org/forums/showflat.php/Number/5143032#5143032
Fungicides In Casing

http://www.shroomery.org/forums/showflat.php/Number/5340461#5340461
What Would Happen If...

http://www.shroomery.org/forums/showflat.php/Cat/0/Number/5332012/page/0/fpart/1/vc/1
New Orleans Soil And Fungi

http://www.shroomery.org/forums/showflat.php/Number/5361201#5361201
Strawlog Dunking

http://www.shroomery.org/forums/showflat.php/Number/5361223#5361223
"Hydroponic Mushroom Growing"

http://www.shroomery.org/forums/showflat.php/Number/5515002#5515002
Briefly Re-Opening Mycelia Piss Applications

http://www.shroomery.org/forums/showflat.php/Number/5518491#5518491
Spores, Usefull Identifiers Or Variable Within Species

http://www.shroomery.org/forums/showflat.php/Number/5553299#5553299
Dikaryotic Mycelium Questions

http://www.shroomery.org/forums/showflat.php/Cat/0/Number/648961/page/0/fpart/1/vc/1
Crossing Two Strains IS Possible..Here's How..

http://www.shroomery.org/forums/showflat.php/Number/5595537#5595537
Bioluminescent Mushrooms

http://www.shroomery.org/forums/showflat.php/Number/5606658#5606658
Proof That Mushrooms Are Sensitive!

http://www.shroomery.org/forums/showflat.php/Number/5651186#5651186
Strains

http://www.shroomery.org/forums/showflat.php/Cat/0/Number/5648213/page/0/fpart/1/vc/1
Mycological Career

http://www.shroomery.org/forums/showflat.php/Number/5850075#5850075
Spore Pics

http://www.shroomery.org/forums/showflat.php/Cat/0/Number/6024742/page/0/fpart/1/vc/1
Charcoal's Secret

http://www.shroomery.org/forums/showflat.php/Cat/0/Number/6029820/page/0/fpart/1/vc/1
Total Unrestricted Access To University Lab

http://www.shroomery.org/forums/showflat.php/Cat/0/Number/6144141/page/0/fpart/1/vc/1
Outdoor Cultivation: Weraroa Pouch Fungus (Updated With Microscopy Pics)

http://www.shroomery.org/forums/showflat.php/Number/6174822#6174822
Diverse Mushroom Species

http://www.shroomery.org/forums/showflat.php/Cat/0/Number/6284138/page/0/fpart/1/vc/1
Guar Gum Tek

http://www.shroomery.org/forums/showflat.php/Cat/0/Number/6029820/page/0/fpart/1/vc/1
Total Unrestricted Access to University Lab - What experiment??

http://www.shroomery.org/forums/showflat.php/Number/6972885#6972885
Capelandia Pleurocystide Apex Crystals Dissolve In KOH

http://www.shroomery.org/forums/showflat.php/Number/7335765#7335765
Manure With Extras Recipe

http://www.shroomery.org/forums/showflat.php/Number/7502181#7502181
Importiant!! Is The Cause Of cancer Common Fungus?

http://www.shroomery.org/forums/showflat.php/Cat/0/Number/7533929/an/0/page/0
(MC FORUM) Forget About This LC

http://www.shroomery.org/forums/showflat.php/Number/7674688#7674688
Mushroom For Other Industries

http://www.shroomery.org/forums/showflat.php/Number/7761069#7761069
Culturing Cordyceps On Agar

http://www.shroomery.org/forums/showflat.php/Number/7801472#7801472
Agar problem (Too Wet?)

http://www.shroomery.org/forums/showflat.php/Number/7815574#7815574
Increasing the Rate Of Colonization Ways?

http://www.shroomery.org/forums/showflat.php/Cat/0/Number/7459057/page/0/fpart/1/vc/1
Question For RR - Creating New Substrain

http://www.shroomery.org/forums/showflat.php/Number/7920810#7920810
Bioluminescent Fungi - Novel Luciferin Isolation

http://www.shroomery.org/forums/showflat.php/Cat/0/Number/7656684/page/0/fpart/1/vc/1
Growing Clean Prints In A CleanBag

http://www.shroomery.org/forums/showflat.php/Cat/0/Number/7924741/page/0/fpart/1/vc/1
Snake Venom Into Hybrids

http://www.shroomery.org/forums/showflat.php/Cat/0/Number/7940459/page/0/fpart/1/vc/1
Greensulate Insulation From Mushrooms

http://www.shroomery.org/forums/showflat.php/Cat/0/Number/7965308/page/0/fpart/1/vc/1
Quest For A Fully Automatized Climate

http://www.shroomery.org/forums/showflat.php/Number/8323042#8323042
UWATTS-SEC/CM2 NEEDED TO KILL TRICHODERMA

http://www.shroomery.org/forums/showflat.php/Number/8361944#8361944
Mycelium On Agar Picture Thread

http://www.shroomery.org/forums/showflat.php/Cat/0/Number/8410381/page/0/fpart/1/vc/1
H202 Discovery (New High Res Pics Updated)

http://www.shroomery.org/forums/showflat.php/Number/8378700#8378700
Selling Venom

http://www.shroomery.org/forums/showflat.php/Cat/0/Number/8478514/page/0/fpart/1/vc/1
Reviving Mycelium From Frozen, Dried Mushrooms

http://www.shroomery.org/forums/showflat.php/Number/8488033#8488033
Crossing Strains

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Invisibledumbfounded1600
Stranger

Registered: 07/29/07
Posts: 2,624
Re: All Of RR's Notes (Advanced Mycology) Notes/Links I Liked [Re: dumbfounded1600]
    #8531617 - 06/16/08 09:22 PM (15 years, 9 months ago)

Bookmarked

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Invisiblewisp

Registered: 04/13/08
Posts: 5,304
Re: All Of RR's Notes (Advanced Mycology) Notes/Links I Liked [Re: dumbfounded1600]
    #8532296 - 06/17/08 01:27 AM (15 years, 9 months ago)

Nice:thumbup:

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Offlinedstark
Manifesting Minds
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Re: All Of RR's Notes (Advanced Mycology) Notes/Links I Liked [Re: wisp]
    #8532662 - 06/17/08 05:30 AM (15 years, 8 months ago)

Yup! very very nice! RR rocks :rockon:


--------------------
What is a mind, if not something to be messed with? What is consciousness, if not a state to be altered?

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at Home~

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OfflineEstario
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Re: All Of RR's Notes (Advanced Mycology) Notes/Links I Liked [Re: dstark]
    #10546378 - 06/21/09 07:39 AM (14 years, 8 months ago)

Every noob should read this and the other thread with All Of RR's Notes On Mushroom Cultivation Forum . I read the book Psylocibin and now I have to unlearn most of the wrong teks that were inside the book. If you read this you will have great success. The best thing is RR not only says how things should be done the right way, but also explaines why his methods work better. You will learn a lot about mycelium, "strains", contamination, importance of pH and many, many more things.
Thank you RR for sharing your knowledge with us! :heart:


--------------------
All Of RR's Notes On Mushroom Cultivation Forum - a must read
:alert: Everything I post is completely fictitious, and for your amusement only. All the pictures I post are either photoshopped or ripped from the internet. Whenever i trade for spores it is for examining the spores under microscopes to see their characteristics. There is no reason why I would ever want to nor will I grow mushrooms containing psilocybin. :alert:

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Invisiblethe_damn


Registered: 08/26/11
Posts: 383
Loc: Hell Flag
Re: All Of RR's Notes (Advanced Mycology) Notes/Links I Liked [Re: Estario]
    #15438723 - 11/29/11 02:14 AM (12 years, 3 months ago)

Just wanted to post here because it will take me forever but I want to read every last bit of this.


--------------------
I'll be damn...

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InvisibleMr.PhilCybin
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Registered: 06/13/11
Posts: 11,642
Loc: Gnarnia
Re: All Of RR's Notes (Advanced Mycology) Notes/Links I Liked [Re: the_damn] * 1
    #15706224 - 01/23/12 09:48 AM (12 years, 1 month ago)

Definitely gonna read this.

marked for future refrence :popcorn:


--------------------
I'm stupid, Falcon91Wolvrn03 is smart.
I'm ugly, Falcon91Wolvrn03 is beautiful.
I'm a loser, Falcon91Wolvrn03 is a winner.
Someday, I hope to be like Falcon91Wolvrn03 but secretly know I never will.

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Offlineepyon
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Re: All Of RR's Notes (Advanced Mycology) Notes/Links I Liked [Re: dumbfounded1600]
    #28326894 - 05/19/23 10:22 PM (9 months, 25 days ago)

This information need to be what everyone reads before they can ask a question lmao 🤣 I have been tinkering around for a couple of years now nothing to compare to this tho I feel like trying some new stuff after reading it's been a while and I had lost the fun and challenging things about this hobby just doing the same over and over

Thank you op your a camp
Know you have Rekindled a spark that was almost out


--------------------
"I went to Magical alternate universe and all I got was vast cosmic powers"
Jason Asano.

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Offlinejohnukguy
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Re: All Of RR's Notes (Advanced Mycology) Notes/Links I Liked [Re: epyon] * 3
    #28327037 - 05/20/23 12:59 AM (9 months, 25 days ago)

You're necroing a thread that's over a decade old.


--------------------
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V: Who? Who is but the form following the function of what and what I am is a man in a mask.
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