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Bolete cultivation?

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A bloke from Lismore gave me some boletes he described as Slippery Jacks yesterday. No confirm ID yet, and I'm not going to bother really, he's a lifelong forager and has been eating them from this patch for years. I'm satisfied they're edible

Have done a lot of searching, and I remember reading the other day someone has managed to cultivate bolete-type mushrooms in beds or bags without associating them with a living tree. It was a recent tek, but for the life of me I can't find it, and it could be just my bad memory

Anyone know of or has tried successful cultivation of boletes? I want to put this one on some wood pellet agar and/ or PDA + chloramphenicol for spawn, but if it's not a goer til a tek is perfected I won't take it further

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This is quite interesting

http://www.sciencedirect.com/science/article/pii/S1340354010701856

Although it is a completely different species it is still another mushroom thought to be ectomycorrhyzal but is now appearing not to be obligatory that way.

If you get some going on agar I'd be quite keen to give it a go myself.

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Wow! Thanks for that, and kudos for your google-fu

Anyone got a full copy of the article? I'd be interested to see how they got agar MS to solidify at pH4, anything below pH5 is difficult to solidify for most gelling agents. Unless they did the whole operation in liquid culture

I'll make up some agar today and if the fruit is still viable I'll give it a go :)

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agar should solidify fairly easy at pH 4 as long as you add a bit extra of the agar agar powder.

You could potentially add a gelatinous substance such as BRF to increase the solidity of the mix too. I've done that many times before with great success. Quite a wet mix sure but it is alot firmer than without the BRF

Ill try and find a full article but can't promise anything. I imagine it is somewhat guarded.

I would suggest testing the soil pH of wherever you got the bolete from. In fact try culturing some of the soil sample seperately from teh bolete. It may in fact turn out that its more of a microbial symbiotic reltionship than a mycorrhyzal relationship. It's interesting because there are a few mushrooms that do that. One prime example is the morel mushroom. And it was always thought to be myorrhyzal but is in fact not.

Best of luck to you mate. I hope that something happens. ANYTHING. even a failure with minor progress is a step in the right direction.

Cheers

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Thanks for the article Zed, looks facile

And Ace, you rock. I'll remember that soil/ agar protocol too. I don't have any soil for this bolete, but it's a very logical step to take for the next one.

I'll be trying it with some wood pellet/ chloramphenicol agar. Won't be seeking bacteria for association- at this point- that would be a few days more work to serial dilute and check that what's on the plate is actually going to be the useful association species. It's an option for further down the track- and facile, but labour intesive

agar should solidify fairly easy at pH 4 as long as you add a bit extra of the agar agar powder.

You could potentially add a gelatinous substance such as BRF to increase the solidity of the mix too. I've done that many times before with great success. Quite a wet mix sure but it is alot firmer than without the BRF

Ill try and find a full article but can't promise anything. I imagine it is somewhat guarded.

I would suggest testing the soil pH of wherever you got the bolete from. In fact try culturing some of the soil sample seperately from teh bolete. It may in fact turn out that its more of a microbial symbiotic reltionship than a mycorrhyzal relationship. It's interesting because there are a few mushrooms that do that. One prime example is the morel mushroom. And it was always thought to be myorrhyzal but is in fact not.

Best of luck to you mate. I hope that something happens. ANYTHING. even a failure with minor progress is a step in the right direction.

Cheers

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Seems pretty doubtful to me but worth trying. I got confused about how the basidiospores germinated on agar. Surely that alone would be the million dollar thing.

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Getting mycorrhyzal fungi to grow on agar is not all that hard. In fact the life cycle up until fruiting is not the most complicated thing. It is the mystery behind how the damn things fruit which makes them complicated.
I still personally believe there is chemical signalling going on. I think it would be possible to use column or gas chromatography to isolate all of the individual chemicals in the roots of plants that have fruiting mycorrhyzal fungi and also isolate the chemicals in the live fruits themselves.

Then compare that to isolates from roots of trees that dont have the fungi and isolate the chemicals in lab grown agar cultures. Compare the pair, look for the difference, analyse the different compounds and then see if they can be synthesised. If they can be apply them to an artificial growth medium and see if it initiates fruiting.

No reason it wont work. Everything is just chemical signals and the roots and fungi are obviously communicating somehow.

It is essentially the same process they initially used when isolating plant hormones.

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Got a reference for that?

my understanding is they rarely germinate in agar - in fact my understand was that they never germinate on simple agar mediums - but I must admit I've never tried.

if they do why don't sporebanks / strain libraries sell them?

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My understanding was creating a successful host innoculation was difficult - hence the reason until recently innoculated trees were so damn expensive.

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fascinating thread :D

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Just gonna chuck a heap of links. Some more credible than others.
There may be some species that are notoriously difficult but i imagine agan that it is a chemical thing ie what is or is not present in their ideal medium.

As to why sporebanks dont sell them I have no idea. Although I'm sure for research and education institutions you CAN buy them. I could be wrong there though.

http://www.ncbi.nlm.nih.gov/pubmed/12018258
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC123902/

http://link.springer.com/article/10.1385%2FABAB%3A98-100%3A1-9%3A311#page-1

http://www.oocities.org/timessquare/2933/immyco.html

http://archive.bio.ed.ac.uk/jdeacon/microbes/ectoimag.htm

http://www.sciencedirect.com/science/article/pii/S0953756209808800

http://www.sciencedirect.com/science/article/pii/S0038071704003748

http://www.researchgate.net/publication/8357337_Use_of_sucrose-agar_globule_with_root_exudates_for_mass_production_of_vesicular_arbuscular_mycorrhizal_fungi

That is a whole lot of info on growing mycorrhyzal fungi on agar, germinating spores on agar, different types of agar, different contams in agar, some techniques for innoculation of trees etc.
Really decent selection of info actually and only the tiniest tiniest bit that is available.

Id love to have a go with an amanita actually. That would be really cool.

Anyways enjoy the info there. I'm sure it brings up more questions than it answers, as always.

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Oh and another point. In all those articles it appears as though LESS nutrients in the agar is MORE conducive to germination and initial growth. Where you go from there is when it starts to get interesting and complex

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Thats a great idea.

i imagine cloning the material on a predominantly water agar with activated carbon mixed in would be a good bet too.
Let us know how it goes :)

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My understanding was creating a successful host innoculation was difficult

That's right, a lot of requirements have to be met for the mycorhizzal link to take place. First, the host tree seedling has to be very young. And I mean just germinated. Anything older than a few weeks can be very resistant to mycorhizzal links with higher fungi, and if it's collected from the forrest, the chances are it's already established a link with some mushroom species and will reject the one we want to give it. So that's the main problem, raising a small forrest indoors in sterile conditions.

Another very important thing is the pH of soil, which is extremely important to some mycorhizzal species, as well as the presence of some microelements in the soil.

I have two very good agar recipes for mycorhizzal fungi, which require a long list of inorganic compounds, salts and whatnot. One other thing you may try is to make a 'tea' of the soil that seems compatible to the species you want to cultivate, and add a bit of nutrients like malt extract.

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One other thing you may try is to make a 'tea' of the soil that seems compatible to the species you want to cultivate, and add a bit of nutrients like malt extract.

Pretty much my conclusion as well. nature doesnt work with inorganics, so i wouldnt go there.

i remember sitting in the micro lab one day looking at all the ingredients for the agar recipes and thinking we've got it all wrong....

Edited by obtuse
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I too thought it was very hard to germinate mycorrhizal spores on agar...apparently I'm not correct. It's very difficult to germinate the species that are very difficult to germinate.

The mycelium of some species will grow ok on certain agar mixes but fruiting is another matter altogether.

Tangich...what you say about a variety of mycorrhizal relationships is really interesting. I agree that there are likely lower order fungi that too make mycorrhizal relationships but one tree is able to host multiple species.

From a documentary I saw recently the fungi releases a hormone so the root lets down it's defenses (willingly) and allows the mycelium to invade its cells. It's not just a matter of the mycelium wrapping around the roots, it invades the cell walls of the roots and sets up arbuscules. The relationship is beneficial not just in a water and nutrients sense, but also in an exploratory sense. Mycelium is much faster moving and can explore a greater volume of earth. It is also quite happy to grow a number of networks and kill off those which are not successful. Tree roots don't quite operate the same way. So the mycelium will seek out nutrients and when found, the tree will then focus its root growing efforts along that path to the resource. Yes, the mycelium can transport some nutrients back to the roots but some time the roots want proprietary access.

It's really quite quirky for our little minds to try and understand.

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There is a famous quote: " plants dont have roots, they have mycorrhizae"

up to 95% of plants are able to form a symbiosis with fungi, be they ecto or endo (i.e. arbuscular vesicular) mycorrhizal. in the greater amount of cases it is essential for the growth of the plants that they initiate the relationship, up to 90% for the endo, 5% ecto. it has been going on for ages too i think as long ago as 480 million years.

the fabaceae have gone even further and develoed symbioses with nitrogen fixing bacteria, using the same (but modified) mechanism that is used to regulate the relationship between arbuscular mycorrhizae.

My own opinion is that we only just starting to plumb the depths as far as symbioses are concerned. An academic friend of mine is of the opinion that its really the fungi manipulating the plants, and not really a shared relationship. the plant gains many benefits though as the AM fungi turn on all the defensive mechanisms of the plants, which otherwise dont get turned on till after the initial infections/attacks.

What initiates the fruiting in the ectomycorrhizae? i suspect its some trigger associated with changes in sugar transfers. either the beginning of increased suger movement, or the slowing, given the nature of deciduous trees.

An additional point is that some trees swap mycorrhizal relationships. Eucalyptus sp. for example start with endomycorrhizae but get to a point where they swap to ectomycorrhizae. I would assume then that trees are able to have many mycorrhizae and are not exclusive. particularly given the dynamics of ectomycorrhizae, in that they only wrap around the tree root, and do not enter the plant cells as to endomycorrhizae.

Edited by obtuse
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I've had success in isolating *something* from a small local bolete, I was just mucking round. Still, wthout it fruiting, I have no idea if it is a bolete or just something that was along for the ride with the bolete. White fluffy stuff on a plate.

The whole wounded plant thing gives rise to a bunch of plant stress or antioxidant compounds which might work to initiate agar culture or spawn run, such as abscisic acid, salicylic acid, ascorbic acid etc. But hey, there's a PhD in that, at least one, and all I want is something yummy to eat

We're gunna have to stop thinking of humanity as being the pinnacle of life and start realising that we're just a vehicle for other life forms to play with. Once you realise you've been thoroughly used by mycelial cells and have spent a significant part of your life cycle manipulated by them to attempt to produce their desired end result, while simultaneously hosting several billion bacteria with their own agenda

I have manipulated my immediate ecosystem so it is specifically designed to produce mycelia-favourable outcomes. It's so highly specialised it has probably rendered me less capable of forming human pair-bonds to perpetuate my species. My behaviour is markedly changed as a result. I'm more nocturnal, more solitary, my focus is shifted away from social interaction with other people when compared to controls. I think I've been parasitised by our mycelial overlords.

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Obtuse and Darklight...two great posts, very different but each great in their own ways.

Obtuse, I totally agree that we are just scratching the surface and I also agree with your friend who says that fungi let plants live. I think that's very much true. Fungi colonised the land mass first and it was they who allowed plants onto their patch. They certainly do still exercise huge amounts of control over plants.

There's a local truffle farm/nursery that has some intellectual property they guard about how they get truffle to grow on agar but from there, they simply put a wedge of it amidst the roots in the pot of the tree. Very low tek at that end.

Darklight, I like to think that anyone who hunts fungi is infected by a human cordyceps. When you explain cordyceps to a lay person it's incredibly eye opening, the behaviour modification especially. I feel, like you do, that fungi modify my behaviour. They call me to the bush in my time off, I wander around, eyes down, often for hours at a time. Collecting them, then as I travel I spread their spores. I talk about them at length, informing others about them too.

I long to have a 'tree change' - just buy a 40 acre hobby farm with a creek, design and build a house and turn the whole property into a fungal forest, supporting a number of different tree and mushroom species. I'm still torn as to whether I go purely european or whether I go half native, half european.

Strangely enough, did you happen to catch the 2 part doco on SBS recently called 'Life on us'? Long story short, before we are born we are 100% human cells, as we exit the birth canal we are exposed to bacteria migrating through the vaginal wall from the large intestine, these bacteria colonise our skin and form a proctective barrier. Even through breast milk we gain gut flora and other immune defenses. Long story short, when we die, our cell count is only 10% human, 90% microbial. We are walking vectors.

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LOL. I can see me in a t-shirt that just had the word "Vector" on it. I'd prolly never have sex again, but if a chick got the joke it would prolly be the best sex ever, because biology

Maybe we should start a band called Vector, if there aren't several hundred of them already

Personally I'd go all native fungi, only because we don't know enough about them

I just hijacked my own thread. Dammit.

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I wonder if the dropping of leaves somehow signals fruiting? I did also read about how the fungus has to trick the plants roots for access to describe it crudely.

interesting thread and great links.

I did my tree change already but it appears my big eucalypt already have established passenger fungi :)

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I am certain in my early days of reading about mushrooms I came across an article that stated porcini will only fruit on trees that have themselves reached sexual maturity. I've never been able to find the article since. Nor have I found any information that supports or contradicts it.

In leaf molds there apparently are some acids produced that prompt spore germination, or at least soften the spore casing. Again, this is something I've been told and don't have evidence to support. The very few european forest I've been hunting in are utterly different to Australian forests. Incredibly thick leaf litter and no under story. They tend also to be really silent places, no noise from birds or insects even. Quite different.

Darklight...the SBS doco also talked about physical compatibility between two people. Not in a liking each other's personality sense but in an attraction that we can't describe, which may have something to do with how are biomes align or how genetically compatible we are. Sometimes we can't put our finger on why we aren't attracted to someone and it turns out it could be to bacteria on our skin not having a good time with theirs. Even stomach worms are able to alter our brain chemistry and reduce inflammation and excess immune response. So vector might not be the most appropriate name, meat puppet might be closer to the mark ;)

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