Pine inoculation - a fungi tale

[Wires_Guy_wires]

I have a couple pinus sylvestris and a couple of them are super healthy. They have percolation issues every summer because the huge dense fungal mat that's attached to their roots.

In late summer 2021 I decided I wanted to expand these beneficial acting fungi because they seem to have a very positive effect on the trees riddled with them. Hardly any needlecast, good growth rates.

To get things going well, some changes had to be made to the receiving trees: unglazed clay containers and of course an inoculant. Something needed to carry the good fungi from one pot to another.

So I set up an inoculation station: 20 bamboo (or wood?) chopsticks were jammed into the pots with the good fungi. After a week or two, they turned white and I was fairly certain the fungus had spread its mycelium into the chopsticks. Mycelium retracts and dies whenever it runs out of resources so I knew my window wasn't very large.
Those same chopsticks were jammed into the pots with very little or next to no fungi, after a fall repot. To increase the soil humidity I slapped on a lil moss.

Fast forward a season.. I suddenly see the moss flourishing after being droopy the entire winter. Apart from the areas that were loaded with fertilizer though. I decided to have a look up close.



Fungi all around the entire circumference of the pot. It seems to have worked.

I now have a thriving culture of wires_guy_wires scots pine fungus, and the knowhow about expansion.

Sweet!
Now it's time to ramp up and tweak production, see whatever other pines can connect with it as well.

 

[19Mateo83]

The chopstick trick is pretty ingenious! Well done!

 

[Wires_Guy_wires]

The chopstick trick is pretty ingenious! Well done!

Back in ancient times, when I was still growing mushrooms that make people feel funny.. I found that all fungi need to be transferred are a good carbon source.
If you can't get a sterile laboratory, you need a medium to grow fungi and nothing else (bacteria are the worst! Yeast are even worse!). I've spent days fiddling with potato dextrose agar, liquid malt extracts, liquid cultures, slants, and basically every gelling agent and carbon source there was. Hours and hours of sterilizing, and always the same contaminations.. But being scientific doesn't always yield the best results.. I needed something practical. I've build flow hoods, sterile chambers, hundreds of methods including fire.. Didn't work either.
And then I took a huge dump and it dawned on me. I wrote down a technique that allowed people to germinate spores in a petri dish without bacterial contamination because there was no food for all the other microbes to process.
Freaking toilet paper rolls.

I was able to grow entire mushroom cultures on just toilet paper rolls. The recycled paper brown ones. And if cardboard works, so does wood.
But.. This experiment kind of fueled the old flame. I'll be starting some petri's with these cultures and see what happens. I think I'll be able to freeze them too with some tweaking.

Use this information for your own benefits! Because someone else sure will!

 

[19Mateo83]

Back in ancient times, when I was still growing mushrooms that make people feel funny.. I found that all fungi need to be transferred are a good carbon source.
If you can't get a sterile laboratory, you need a medium to grow fungi and nothing else (bacteria are the worst! Yeast are even worse!). I've spent days fiddling with potato dextrose agar, liquid malt extracts, liquid cultures, slants, and basically every gelling agent and carbon source there was. Hours and hours of sterilizing, and always the same contaminations.. But being scientific doesn't always yield the best results.. I needed something practical. I've build flow hoods, sterile chambers, hundreds of methods including fire.. Didn't work either.
And then I took a huge dump and it dawned on me. I wrote down a technique that allowed people to germinate spores in a petri dish without bacterial contamination because there was no food for all the other microbes to process.
Freaking toilet paper rolls.

I was able to grow entire mushroom cultures on just toilet paper rolls. The recycled paper brown ones. And if cardboard works, so does wood.
But.. This experiment kind of fueled the old flame. I'll be starting some petri's with these cultures and see what happens. I think I'll be able to freeze them too with some tweaking.

Use this information for your own benefits! Because someone else sure will!

Same principle of mushroom plugs which I am very familiar with. Good to know it works with beneficial fungi too

 

[HorseloverFat]

Back in ancient times, when I was still growing mushrooms that make people feel funny.. I found that all fungi need to be transferred are a good carbon source.
If you can't get a sterile laboratory, you need a medium to grow fungi and nothing else (bacteria are the worst! Yeast are even worse!). I've spent days fiddling with potato dextrose agar, liquid malt extracts, liquid cultures, slants, and basically every gelling agent and carbon source there was. Hours and hours of sterilizing, and always the same contaminations.. But being scientific doesn't always yield the best results.. I needed something practical. I've build flow hoods, sterile chambers, hundreds of methods including fire.. Didn't work either.
And then I took a huge dump and it dawned on me. I wrote down a technique that allowed people to germinate spores in a petri dish without bacterial contamination because there was no food for all the other microbes to process.
Freaking toilet paper rolls.

I was able to grow entire mushroom cultures on just toilet paper rolls. The recycled paper brown ones. And if cardboard works, so does wood.
But.. This experiment kind of fueled the old flame. I'll be starting some petri's with these cultures and see what happens. I think I'll be able to freeze them too with some tweaking.

Use this information for your own benefits! Because someone else sure will!

I found this also... whilst cultivating various fungi.. and searching for correct mediums... and sterile fruiting areas. If cultivating in controlled, clean, environment, once 'clean' is compromised.. or any 'control' is LOST, it is damn near impossible to correct the course, realistically.

I tried, too young.. and in utmost, forced secrecy...

I never had what I would call a successful harvest... it was all compromised.. and various parameters effected due to said compromise.

Buuuuut... growing in more natural situations.. like existing in your pots, outdoors. This has been.. easier.

Really neat with the chopstick, Amigo!

 

[Shibui]

Mycorrhiza seems to transfer itself throughout the pots here. New seed trays have a healthy population despite not being close to the other pines and no other transfer of soil.
I know some wood fungi are sold as dowels so I guess that's why you have tried these wood chopsticks but mycorrhiza is not a wood consuming fungi so why would they be interested in a chopstick? The presence or absence of mycorrhiza in a pot is not proof the skewers are the cause.
Chopstick in the pots won't hurt but I'd need much better controlled tests to be convinced they have been effective in transfer.

 

[Wires_Guy_wires]

Mycorrhiza seems to transfer itself throughout the pots here. New seed trays have a healthy population despite not being close to the other pines and no other transfer of soil.
I know some wood fungi are sold as dowels so I guess that's why you have tried these wood chopsticks but mycorrhiza is not a wood consuming fungi so why would they be interested in a chopstick? The presence or absence of mycorrhiza in a pot is not proof the skewers are the cause.
Chopstick in the pots won't hurt but I'd need much better controlled tests to be convinced they have been effective in transfer.

What makes you think that mycorrhizae aren't wood degrading? There are thousands of species.
They have the enzymes and the genetics to degrade lignin and cellulose, as most fungi do.
Why would oyster mushrooms be able to degrade diesel fuel? It's not really something they encounter in the wild. Yet still they do.
Those magic mushrooms degrade poo and grains, but I've grown them on paper.

The thing is, although not very scientific.. This tree has been without myc for 4 years. After the introduction of some chopsticks, the pot is filled with it in a three month timespan.
I have a couple more trees like that, same story.
It's not a controlled environment and it sure isn't fool proof. But I'll take a 19/20 'no fungi before, a lot fungi after' as an effective result.

 

[Bnana]

Most ectomycorrhizal fungi are obligate mycorrhizal and can't live as saprophytically.
Unlike oyster and magic mushroom that are saprophytic. Those are completely different organisms.
To me this is like: "What do you mean lions do not eat grass? lot's of mammals eat grass, I've raised goats and sheep and cows on grass. So surely I can do the same with lions."

How do you know that you did not introduce a saprophytic fungus?

 

[HorseloverFat]

Beneficial Sub - saprotrophic , wood eating fungus DEFINITELY exists!

In fact, Gymnopilus sp Fungus are amongst the FEW that grow here.. that Oxidation React with blues, purples, and green.



These Species of Fungi.. often 'eat' SPECIFIC deadwoods. Mostly coniferous.

 

[HorseloverFat]

But I am very interested in this conversation.

And to hear from you more experienced folks.

 

[HorseloverFat]

Because... saprophytic fungus works with and in RELATION to EctoMycorrhizal bodies in nature...

I always thought them beneficial when working in unison..

But, as I said, am curious what you more experienced have to say. I love to learn.

 

[Wires_Guy_wires]

Most ectomycorrhizal fungi are obligate mycorrhizal and can't live as saprophytically.

That's why I had to jam chopsticks into rootballs.
As you're well aware, a lot of spores never touch a rootball because sporulation happens mostly in the air, and spores are largely hydrophobic.
Just like pollen tubes, they need to grow a little to make a connection. Even in pure H2O spores can germinate and use their stored sugars to grow and search for a mate.
The further you dive into mycology and microbiology, you'll find that there is no thing as a 100% obligate. We base those characteristics on laboratory results aiming at optimal growth or division, rarely at 'survival'.
One of my favorite microbes is pseudomonas because it smells like grapes. It's a strict (obligate) aerobe but it can and will grow anaerobicly if there are nitrates around.

Fungi can and will grow if the plant dies, and they will venture into the unknown in search of new connections. It's very well possible to graft them onto new roots. Even if they don't degrade the wood, they will use its matrices for all intents and purposes.
Magic mushrooms can't degrade wood, but they will latch on to it and this allows people to freeze slanted cultures.

I do know that there is so little food in the original containers that saprophytism is out of the question: no fungus can live off of rocks for 4 years and form such a huge patch of mycelium, there just isn't enough carbon. I've grown fungi on digested rye bran or woodchips and they still had a lower biomass than what I have here.

To me it sounds like: my lion is eating vegetables and munching on plants, so it must be a cow. I've raised a lot of lions and they eat meat. So it must be a cow if it's eating anything else. We both know that that kite doesn't fly. I'm sure that if you'd dive into the genome of whatever mycorrhizal fungus, you will find the code for lignase, cellulase and other wood degrading enzymes.

There are studies out there that if a plant doesn't provide the sugars, the mycorrhizal connection becomes a hostage situation; the fungus will eat the plant for food, which is a form of saprophytism - maybe parasitism. The other way around too: if a fungus can get all it needs without the plant, it will not form a symbiotic partnership because it's a waste of resources.

 

[MaciekA]

Back in ancient times, when I was still growing mushrooms that make people feel funny.. I found that all fungi need to be transferred are a good carbon source.
If you can't get a sterile laboratory, you need a medium to grow fungi and nothing else (bacteria are the worst! Yeast are even worse!). I've spent days fiddling with potato dextrose agar, liquid malt extracts, liquid cultures, slants, and basically every gelling agent and carbon source there was. Hours and hours of sterilizing, and always the same contaminations.. But being scientific doesn't always yield the best results.. I needed something practical. I've build flow hoods, sterile chambers, hundreds of methods including fire.. Didn't work either.
And then I took a huge dump and it dawned on me. I wrote down a technique that allowed people to germinate spores in a petri dish without bacterial contamination because there was no food for all the other microbes to process.
Freaking toilet paper rolls.

I was able to grow entire mushroom cultures on just toilet paper rolls. The recycled paper brown ones. And if cardboard works, so does wood.
But.. This experiment kind of fueled the old flame. I'll be starting some petri's with these cultures and see what happens. I think I'll be able to freeze them too with some tweaking.

Use this information for your own benefits! Because someone else sure will!

Given your current and past experiences, whether anecdotal or scientific or not, I am curious to hear your thoughts about one thing that always comes up at the back of my head any time fungi are brought up in the context of pine cultivation: How does anyone ever really know if the specific species of fungus that colonizes a soil/root system is "The" one that should pair with that pine? Or a "compatible" one? or a "less compatible" one? Do the fungi that I see colonizing lodgepole pine roots in the Oregon Cascades have any kind of affinity (or disaffinity) for my JBPs? Scots? or even other pines collected from the same mountains?

Whenever inoculation (from commercial products) or cross-inoculation is discussed, I always wonder about this stuff, because I've never seen a data table that says "here are the fungi species known to work with <everything strobus> or <all Japanese species of pine> or <ponderosa , but NOT bristlecone>" -- etc.

Does it even matter, from your perspective / what you know?

 

[Wires_Guy_wires]

Man, that's a good question. I think, and I don't firmly believe because this is an evolving science with no real hardliners yet, I think there are two broad groups.

There's a group that connects with plants in general, they are not specific and they don't care which plant provides what. To do this, the fungus needs to be gentle to any and all plants, and contain the genetic code to send out friendly messages. It's like google: everyone can use its network, it's adapted to whatever language you speak. The plants and fungi evolved alongside each other and were symbiotic in the cases it was beneficial.

Then there's a group that's specific. It produces biomarkers, phenols, esters, turpenes, exudates, or whatever, that only a certain plant type recognizes. The other way around, the plant has to produce chemicals that the fungus recognizes. In a sense you can say they have to speak the same language, be from the same country, and they have to have cousins that were once friends. These co-evolved together, probably from the point where the plant branched out from its own genetic tree.

The latter however, is something curious: some species of fungi can attach to all pinacea but only pinacea, others have a very limited range within the pinacea group; some are only found on nigra and sylvestris. Part of this has to do with native ranges, habitats and so on.

The former is a good example of - once and again - showing that fungi were here first. It's probably the plants that have evolved to connect with the fungi instead of the other way around. And genetically, the land plant group has been reduced enough over extinction events that it's fairly safe to say that whatever survived, survived with a genetic basis for mycorrhizal connections, which are now common in nearly all plants.

With new stuff like nanopore sequencing and cheaper and faster mapping and recognition tools, it's getting easier to find whatever is living in the soil. In the past we had to grow these things in a laboratory, and if they didn't grow, they couldn't be classified or typed. Nowadays we can slap some dirt into a mixture, and three to twelve hours later we have 300Gb of genetic sequences (not very solid ones, but enough to divide organisms into families).

I have seen data tables, connecting fungi and plant species. But they're subject to a lot of research because everytime we think we humans have it right, the natural world surprises us.
Most store brands though, stick with the universal fungi. That's why they usually sell a herbaceous mix and a woody plant mix. But, a shovel of forest soil would contain probably a couple million times the amount of organisms. So I'm not a big fan of store bought inoculant.

 

[Firstflush]

So does anyone believe in the mycorrhizae inoculant packages? Usually they have many of the most common fungal species in the brand. I guess you would want the native mic for perfection but can pine use a general purpose mic species and still work.

Side note, the mic packages work wonders for my veggie garden. I see a difference.

 

[Wires_Guy_wires]

can pine use a general purpose mic species and still work.

Yes.
Amanita for instance is present in most mixtures and it can connect with a whole bunch of forest dwellers.
I have amanita growing in my pine soils, as well as in my juniper soils - they fruited so I could positively ID them. So it seems that it's a good one to have present.
L. bicolor is another species that generally does well with pine and spruce, and conifers in general. I found it in the backyard but I don't know if its in my pots.

Myc packages aren't bad. I mean, they contain spores and spores are like the sperm and egg of a fungus (they mate by growing their bodies into each other and combining their nuclei). But myc packages are what they are: a breakfast meal. It'll suffice, but the strength of a truly diverse community with a lot of variation can't be matched by store bought stuff.
The thing is, however, once fungi get a foothold, they start optimizing the pot environment for their own good. Which in turn provides conditions for other fungi and micro organisms to grow as a community. Myc packages are a good foundation. But in the end spores will eventually end up in there either way, unless you shower the soil with antibiotics. So is there a need for myc packages? Absolutely not. It does however, introduce a lot of species fast.
So instead of waiting for a single spore from a forest to blow over to your pot, take 14 days to germinate, connect with the roots and find a mate.. You now can skip the possibly 3 years of waiting and add them right away.

But forest or wild soil.. Dang.. It's so much richer, contains so much more organisms. It feels weird to pay 20 dollars for something I scrape from my hiking boot every time I return from a forest walk.
The mycological community is filled with hippies and free thinkers - I mean that in a positive sense. So there's a lot of resistance towards making money off of things anyone can collect for free if we'd just take the time to get out there and find some old forest growth.
Back in ye olde days there was this thing called the free spore ring, which basically was a system where you sent a letter containing a stamp and an envelope to a myco hobbyist, and you'd get some millions of spores in return for free. I don't know if the system is still in place somewhere. But it's been around since the 1990's I believe. And the people who were active then as rebellious teens, are now professional mycologists in a midlife crisis. I get that it pisses them off somewhat that some get-rich-fast cannabis dudes are drying mushroom caps and blending the spores together to make big money with something the mycologists have been doing for ages, for free. Even Paul Stamets is getting a lot of hate because of it. But he too, admits with ease that his stuff isn't magical compared to whatever can be found out there.

 

[HorseloverFat]

Yes.
Amanita for instance is present in most mixtures and it can connect with a whole bunch of forest dwellers.
I have amanita growing in my pine soils, as well as in my juniper soils - they fruited so I could positively ID them. So it seems that it's a good one to have present.
L. bicolor is another species that generally does well with pine and spruce, and conifers in general. I found it in the backyard but I don't know if its in my pots.

Myc packages aren't bad. I mean, they contain spores and spores are like the sperm and egg of a fungus (they mate by growing their bodies into each other and combining their nuclei). But myc packages are what they are: a breakfast meal. It'll suffice, but the strength of a truly diverse community with a lot of variation can't be matched by store bought stuff.
The thing is, however, once fungi get a foothold, they start optimizing the pot environment for their own good. Which in turn provides conditions for other fungi and micro organisms to grow as a community. Myc packages are a good foundation. But in the end spores will eventually end up in there either way, unless you shower the soil with antibiotics. So is there a need for myc packages? Absolutely not. It does however, introduce a lot of species fast.
So instead of waiting for a single spore from a forest to blow over to your pot, take 14 days to germinate, connect with the roots and find a mate.. You now can skip the possibly 3 years of waiting and add them right away.

But forest or wild soil.. Dang.. It's so much richer, contains so much more organisms. It feels weird to pay 20 dollars for something I scrape from my hiking boot every time I return from a forest walk.
The mycological community is filled with hippies and free thinkers - I mean that in a positive sense. So there's a lot of resistance towards making money off of things anyone can collect for free if we'd just take the time to get out there and find some old forest growth.
Back in ye olde days there was this thing called the free spore ring, which basically was a system where you sent a letter containing a stamp and an envelope to a myco hobbyist, and you'd get some millions of spores in return for free. I don't know if the system is still in place somewhere. But it's been around since the 1990's I believe. And the people who were active then as rebellious teens, are now professional mycologists in a midlife crisis. I get that it pisses them off somewhat that some get-rich-fast cannabis dudes are drying mushroom caps and blending the spores together to make big money with something the mycologists have been doing for ages, for free. Even Paul Stamets is getting a lot of hate because of it. But he too, admits with ease that his stuff isn't magical compared to whatever can be found out there.

Cool!! Fly Agarics are such a cool species. We have lots here, a little North of me.

Do you get the Golds and the Reds?

 

[MaciekA]

But forest or wild soil.. Dang.. It's so much richer, contains so much more organisms. It feels weird to pay 20 dollars for something I scrape from my hiking boot every time I return from a forest walk

If I'm hiking in a pacific northwest forest and I've got a plastic baggie with me, what's the ideal material I hunting for? Bits of decaying wood in contact with the ground?

Really appreciate this thread, it's opened my eyes a bit.

 

[Firstflush]

If I'm hiking in a pacific northwest forest and I've got a plastic baggie with me, what's the ideal material I hunting for? Bits of decaying wood in contact with the ground?

Really appreciate this thread, it's opened my eyes a bit.

Soil around the roots of species you are interested in, mostly oaks and pines. I would using your knife and grab a few tablespoons from several locations. Go a few inches down. When you get home dump it in a gallon freezer back and mix it with a chunky in ground planting mix.

“Mic“ above was supposed to read “myc”.

 

[Wires_Guy_wires]

If I'm hiking in a pacific northwest forest and I've got a plastic baggie with me, what's the ideal material I hunting for? Bits of decaying wood in contact with the ground?

Really appreciate this thread, it's opened my eyes a bit.

Dandruff, leaf litter about an inch down in the soil, some root tips you exposed. The soil on a trail is usually not ideal, but a couple feet next to a trail is perfect. Look for tree species you have, and try to find the oldest specimens and least disturbed spots.
And if you find them on the forest floor; entire full grown mushrooms. Not the ones growing on logs or wood, the ones with gills are generally better, puffbals too, the ones with pores are best avoided. Place those mushrooms on some aluminium foil and let them dry out. You'll see the spores on the foil in half an hour to an hour or so. After a day or two it's even more visible.
Porous buried rocks from healthy looking spots are good too.. You name it, if the trees look good, the soil is probably good too.