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Invisiblelorbitherize
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Spore Germination of A.Bisporus *DELETED*
    #5082682 - 12/19/05 06:43 PM (14 years, 10 months ago)

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OfflineThai_connection
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Re: Spore Germination of A.Bisporus [Re: lorbitherize]
    #5091341 - 12/21/05 10:13 PM (14 years, 10 months ago)

i've heard cloning would be the better alternative, never heard of spores even being sold thou just spawn. spore are pretty much useless.
peace
siam


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OfflineShdwstr
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Re: Spore Germination of A.Bisporus [Re: lorbitherize]
    #5092946 - 12/22/05 09:44 AM (14 years, 10 months ago)

Any success I've had, with button and Ports, was from cloned cultures.

Here is a Thread you may find helpful.

Good luck!
Shdwstr


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Invisiblelorbitherize
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Re: Spore Germination of A.Bisporus *DELETED* [Re: Shdwstr]
    #5094528 - 12/22/05 06:02 PM (14 years, 10 months ago)

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Offlinefalcon
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Re: Spore Germination of A.Bisporus [Re: lorbitherize]
    #5095155 - 12/22/05 08:45 PM (14 years, 10 months ago)

I'm starting to believe they are ( useless ).... still no growth, but just
out of curiosity - - why? Why the heck don't they germinate? Is it a
plot by the big growing companies that each and every mushroom
they turn out is completely sterile?


They are just hard to germinate. One thing I've heard that spawn makers do when they are trying to create a new strain of bisporus is put a piece of a mushroom on the agar on the same plate as the spores they are trying to germinate.

It is thought that the metabolites from the growing mushroom aid in the germination of bisporus. This is pretty tricky though, as the mushroom piece that you put on the plate will grow and if you wait to long it will grow over the whole plate.

You can also stick the mushroom to the lid of the plate, I'm not sure how they do this, maybe stab the piece into some agar that they have poured in lid.

A thing you could try is making a tea out of compost and add that to your cake instead of water. This is just an idea, I have no idea if it will work.


But if I get them to grow from a tissue culture, will the resulting
mushrooms also be infertile?


No they would be fertile, they will probably be hard to germinate.


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Invisiblelorbitherize
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Re: Spore Germination of A.Bisporus *DELETED* [Re: falcon]
    #5095974 - 12/23/05 12:06 AM (14 years, 10 months ago)

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OfflineMycena
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Re: Spore Germination of A.Bisporus [Re: lorbitherize]
    #5096959 - 12/23/05 08:03 AM (14 years, 10 months ago)

You just have to think about the role agaricus plays as a secondary decomposer

its a scavenger and a predator of other microbes.

so naturally it likes to germinate in the presence of other fungi and bacteria which it eats.
Often i find growth is slow until a plate contaminates. If you get a bad plate and leave it longer the agaricus will overgrow and eat the bacteria and moulds


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Invisiblelorbitherize
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Re: Spore Germination of A.Bisporus *DELETED* [Re: Mycena]
    #5097140 - 12/23/05 10:09 AM (14 years, 10 months ago)

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Offlinefalcon
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Re: Spore Germination of A.Bisporus [Re: lorbitherize]
    #5099740 - 12/24/05 12:00 AM (14 years, 10 months ago)

Could it be that the increased levels of CO2 resulting from a growing
piece of mycellium are partially responsible for the germination of
the spores?

I don't know.

Would there be any other substances besides this
capable of reaching from one side of a dish to the other?


There are probably a lot of things, enzymes, acids, alcohols, fungi are major chemical factories that eat the world around them.

In light of simplicity, would it be feasible to start a liquid culture
by inoculating it with a fragment of a mushroom and also with
spores at the same time?


Might work if you could keep the mycellium from the fragment separate from the mycellium from the spores. Worth looking into, sounds like a good idea.


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Invisibletahoe
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Re: Spore Germination of A.Bisporus [Re: falcon]
    #5103498 - 12/25/05 01:24 AM (14 years, 10 months ago)

your best bet is to put some button spors on one side of a petri and cube spores on the other. Or something like that. Alot of the time with bispourus the spores wont germinate until the petri, cake or grain jar contams and and the mold will cause the bisporus to germinate. So if you are handy with agar or have something that will germinate on a pf cake then inco one side of the agar or cake with one strain that will germinate adn the other side with the bispourus


--------------------
Stop experimenting half way through your first grow. Grow it to maturity, watch it, learn from it. Do this a few times then experiment with different ideas and figure out what works best for you.


Teh=The

I need to proofread


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OfflineMycena
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Re: Spore Germination of A.Bisporus [Re: lorbitherize]
    #5105913 - 12/25/05 11:51 PM (14 years, 9 months ago)

Quote:

lorbitherize said:
Does it
colonize over everything, or just small-fry stuff?




i dont think that will work

the sheer volume and high nutrient content of the grain will make it turn to a soupy mouldy sludge and thi sis no goo for the agaricus

agaricus wants a clean fresh enivironment with oxygen and mooisture

the reason it works on plates is that the mould and bacteria quickly eat the small amounst of freely soluble sugars and protein and then stop. they can dgeread agar so the substrate stays as it was
then the agaricus eats them - ALL of them . blue moulds, bacteria, yellow and white moulds etc
i havent see it go up against the green trichodermas but itll take on and consume all opportinistic 'house' moulds

i have only ever cloned agaricus, i haventrtried from spores
but i would think a good way would be to make peroxide agar
then take some cardboard squares - about the same size as blotter acid
moisten and nuke them to pasteurise
using forcpes dip these on the spores and place spore side up in the middle of the agar
the spores germinate on the cardboard and grow off and onto the agar

i think this might work cos the cardboard isnt sterile

its worked for non sterile prints of woodlovers

alternatively i would try Multispore inocualtion into jars of P/C'd straw and aged dung (field agaricus) and composted sawdust and dung ( agaricus blazei, subrufescens, augustus etc )

and just wait
i want to try all this but itll be later this summer i can can get around to it


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Offlineragadinks
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Re: Spore Germination of A.Bisporus [Re: lorbitherize]
    #5555197 - 04/25/06 05:00 PM (14 years, 6 months ago)

I had never succeeded in germinating spores from agaricus species.
But now I have found an article that says that other mycelium can
stimulate germination or some kind of alcohol.

Here is an excerpt of this article:

Quote:


MATERIALS AND METHODS
Strains and media. Strains of A. bisporus desig-
nated D-26 and 70-2 were investigated. These
strains represent, respectively, golden-white and
light-brown varieties of the cultivated mushroom.
Cultures were grown on a defined medium contain-
ing: asparagine, 1 g; glucose, 20 g; MgSO4 7 H20, 0.5
g; K2HPO4, 1 g; KH2PO4, 0.46 g; CaSO4, 0.5 g; FeCl3,
10 mg; thiamine, 120 Ag; distilled water, 1,000 ml;
and agar, 20 g. Mushrooms were obtained commer-
cially (Butler County Mushroom Farm, Inc., Wor-
thington, Pa.) or were grown in the laboratory, us-
ing cased rye grain as a substrate (39). Basidio-
spores were collected as spore prints at 20 C and
germinated either in the presence of isoamyl alcohol
or isovaleraldehyde (1 and 0.1%, respectively, added
to defined medium after autoclaving) or in the prox-
imity of a growing mycelium (23, 37, 47). Spores and
mycelia were generally inoculated onto 5 cm2 mem-
branes (DuPont PD 150 film) spread upon the sur-
face of the defined medium. Cultures were incu-
bated at 25 C, and membranes with adherent cul-
tures were removed from the medium and prepared
for microscope examination.





The interesting part is:
Basidiospores were collected as spore prints at 20 C and
germinated either in the presence of isoamyl alcohol
or isovaleraldehyde (1 and 0.1%, respectively, added
to defined medium after autoclaving) or in the prox-
imity of a growing mycelium (23, 37, 47).

I wonder what isoamyl alcohol or isovaleraldehyde is and where one can obtain it from ?

raga


P.S.:

These papers also seem to be interesting - anyone has got one of it :grin: ?

23. Lowry, R. J., and A. S. Sussman. 1968. Ultrastructural
      changes during germination of ascospores of Neuro-
      spora tetrasperma. J. Gen. Microbiol. 51:403-409.

37. Rast, D., and E. J. Stauble. 1970. On the mode of action
      of isovaleric acid in stimulating the germination of
      Agaricus bisporus spores. New Phytol. 69:557-566.

47. Vogel, F. S., and R. F. Weaver. 1972. Concerning the
      induction of dormancy in spores ofAgaricus bisporus.
      Exp. Cell Res. 75:95-104.


Edited by ragadinks (04/25/06 05:03 PM)


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Offlinethenewguy05
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Re: Spore Germination of A.Bisporus [Re: lorbitherize]
    #5581732 - 05/02/06 05:07 PM (14 years, 5 months ago)

Quote:

lorbitherize said:
I'm starting to believe they are ( useless ).... still no growth, but just
out of curiosity - - why? Why the heck don't they germinate? Is it a
plot by the big growing companies that each and every mushroom
they turn out is completely sterile? It must be, because if these were
growing in the wild, they'd be in quite a spot with no viable spores!

But if I get them to grow from a tissue culture, will the resulting
mushrooms also be infertile?


i have read about species that need an additional nutrient added to the agar to make certain species germinate.


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Offlinebluelou
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Re: Spore Germination of A.Bisporus [Re: thenewguy05]
    #5605285 - 05/08/06 04:58 PM (14 years, 5 months ago)

A haaaaaa!!


Learn something new today interesting!!!
COOL.......


--------------------
Have you tried my(black kow) pile style tek outdoors!!!!!!!!


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OfflineoO_wombat_Oo
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Re: Spore Germination of A.Bisporus [Re: bluelou]
    #5614203 - 05/10/06 08:49 PM (14 years, 5 months ago)

Lorbitherize,

Keep us updated mate, I am very interested to hear about your progess and experiments.


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Offlinehyp3rcrav3
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Re: Spore Germination of A.Bisporus [Re: oO_wombat_Oo]
    #5635754 - 05/16/06 01:52 AM (14 years, 5 months ago)

I read soemwhere there are certain metabolic gases that catylize the germination of A. bisporus. I tried and tried to get spores to grow to no avail. I finally, after several attempts, was able to clone a Portobella. Rouge spores also germinated. It was curious. I believe I was able to isolate the clone so I wouldn't have to trial and error a good stock. I put the germinated spores outside in lawn clippings this spring. I'll see what develops in the fall. They are alway clones from the store since it would be too much of a pain to repeatedly isolate high yielding, good tasting, color consistant (white or crimini) mushrooms from germinated spores. Just my two cents.


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Offlinejeffpro
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Re: Spore Germination of A.Bisporus [Re: hyp3rcrav3]
    #5687297 - 05/29/06 06:44 PM (14 years, 5 months ago)

try adding activated carbon to agar to promote germination


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OfflineFeelers
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Re: Spore Germination of A.Bisporus [Re: jeffpro]
    #5688585 - 05/29/06 11:31 PM (14 years, 4 months ago)

Why dont you make a plate of some innert fungus or bacteria - cubensis for example. Then take the mycelia and sterilize it - maybe make a syringe of it and boil for an hour. Then squirt the dead mycelial/bactarial contents onto the agaricus spores sitting on agar.

The theory being that perhaps the metabolites that make it through the sterilization process could trigger germination, but without growing on the plate. Perhaps you could even use a sterilized dirt solution - there'd be heaps of random organics in there. :laugh:


Edited by Feelers (05/30/06 12:05 AM)


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Offlinefalcon
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Re: Spore Germination of A.Bisporus [Re: Feelers]
    #8467109 - 05/31/08 01:36 PM (12 years, 4 months ago)

bump

There a couple of threads on this topic right now this one seems relevant.

and an interesting link,

http://aob.oxfordjournals.org/cgi/content/abstract/28/4/541

I found this link this winter and saw that Isovaleric acid produced by yeast has been shown to help with germination of spores. I did a search and found that Swiss cheese contains isovaleric acid.

So I made some agar and put a little chunk of cheese in with each batch of agar.

I tried to start A. bisporus, A. subrufescens and A. abruptabulbous on the agar. Didn't have any luck getting anything to germinate this way though.

Any way this abstract seems to confirm what people mentioned about using cultures or materials from other species to get bisporus to germinate, though without access to whole article it's hard to know the particulars.


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Offlineragadinks
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Re: Spore Germination of A.Bisporus [Re: falcon]
    #8516523 - 06/12/08 07:52 PM (12 years, 4 months ago)



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