Attempt at growing button mushrooms

[8145]

Hi fellow plant lovers,

I recently bought one of those portobello mushrooms kits from flower power see this post; http://www.shaman-au...showtopic=32413

Because these portobellos pins are doing really well and it is so much fun to watch these little guys grow i decided to fully get into it. I did some research, look at some YouTube videos and read that brilliant post Watertrade did a while ago; http://www.shaman-au...showtopic=22562

A few days later I sourced some materials (agar, dissection kit, petri dishes, etc.) courtesy of my girlfriend who works in a lab and I embark myself deep into this mushroom journey.

On the 15th of June I run two tests.

Test 1: I took a few tissue samples out of a buttom mushroom I got from wolies and inoculate the agar media (far out did I just write that line.. It sounded so technical for a newbie...) trust me I just learnt this lingo by watching videos and reading posts .

Test 2: I took a a couple of spore prints from different mushrooms and inoculate the agar media.

After a few days some white stuff started to grow ! Yey success !!!

But..... it also started to smell a bit and the mushroom tissue on test 1 went blackish so I am not really sure how is this going.

Does anybody have any suggestions ?

Is this going good, bad or not going at all ?

Any word of advice ?

If these are going well I am going to keep posting the progress If not I'll do more trials...

Peace.

[obtuse]

I have tried agaricus from woolies as well, but gave up because i quickly found oyster mushrooms so much easier and tastier by far.

but my most important words of advice are, sterile conditions for each stage. I work in a glove box that i can keep sterile at all times (as close to as possible), as i dont own a laminar flow hood (yet). because i dont work in a lab (i do every thing in my kitchen) I work with peroxide for my cloning, for cleaning the excised sample i wish to clone, and also within my agar (peroxide will slow down the mycelium growth, and doesnt kill everything). i've had some good success with oysters bought from woolies using this method. prints i do onto aluminium foil, that i can then store and/or easily create spore syringe in a fairly sterile space, and then dilute to create better odds for minimising contaminants.

unfortunately, your plates from what i can see are full of bacterial contamination. No go there. the smell is a bit of a giveaway i think. Also good idea to seal the plates closed, and not open them unless you are cloning, or transferring mycelium to create spawn or a bulk mix.

Problem with store bought mushrooms is they are no longer sterile, they have been exposed to contaminants at each stage, and given the amount of travel and repacking they undergo it would be a fair bit. You have to try and get a bit of flesh that is as clean as possible, from within the stem or the thick bit of the cap.

I wouldnt bother with prints till you got your cloning and sterile technique down pat, as you'll proably need to be able to isolate quickly and a few times before you get clean uncontaminated mycelium.

But congratulations, good on you for taking the first step in what is a most rewarding hobby. learning your contaminants is an essential part of the hobby, and the first handful of failures is a good, but steep learning curve.

My recommendation is to get a bag of very fresh mushroom compost that is still flushing (not sure where you are, may be hard) and try and get a clone or print from a young mushroom or three rather than from the supermarket ones.

Or try working with oysters first, they are much more vigorous, and you will proably get more success, and that rewarding feeling sooner.

Have you read any of the stamets books?

All the best with your hobby, plenty of experienced folk here to help out when you need it.

Cheers, Obtuse.

Edited June 20, 2012 by obtuse

[LokStok]

Im taking them from the inner stem and avoiding slicing them in half with a scalpel in case it takes contam from the exterior when i slice.

Tear them in half

[obtuse]

Hi Therefore,

The dilution method is simply a matter of reducing the odds of contamination. draw up 1 ml of spore solution, then draw up 9 ml of sterile water, doing this a few times will reduce the odds of contamination somewhat, by 10, 100, 1000, depending how many serial dilutions you perform, but it also reduces the number of viable spores so keep that in mind.

I use peroxide in my agar to minimise airborne contamination while i transfer cultures. Its not foolproof, but certainly works well enough for me. to 250 ml i add 2 ml, which might be a bit of overkill, im not sure.

Peroxide is only good to kill spores, it will not kill fungi. so if your cloning a mushroom that already has cobweb mould, or trich etc. then you wont be killing it. I have tried using peroxide to knock them back on contaminated plates but it doesnt work.

Peroxide may in fact inhibit growth of the mycelium you want, in favour of any contaminants, but given you've just dipped that section in peroxide, what the hell, peroxide agar isnt going to hurt to much surely. the thing is just to be quick, culture new growth to new plates before any contamination that got carried across has the opportunity to take over..

I'm no agar jedi, and i dont necesarily like peroxide, its a bit lazy in some regards, and im keen to try using selective media, when i manage to get around to it.

Sorry bit of a rushed reply. mhope that helps therefore.

Cheers, Ob.

[Sallubrious]

I established a few cultures from supermarkets a few years ago.

They're all gone now, but they were clean workable cultures.

I used Lazlo's salt agar tek and almost all succeeded on the first attempt.

http://mycotopia.net/forums/agar-strain-isolation/20398-using-non-iodized-salt-culturing-antiseptic.html

[delshanon]

I just started some Agaricus on malt extract agar with no peroxide. I shall posts the results in a few weeks with what my technique was if it works contam free (if that helps).

[NSF]

Agaricus species also aren't that found of generic agar mixes, I hear they are rather selective about what they will grow on.

Ob, what strength peroxide are you using that you use just 2ml per 250ml? From hair dressing supply type stores they sell it in various 'strengths'.

I also agree with the advice to the OP to get more sterile conditions. Even in a glove box or a hood contams are still a very real and fairly frequent issue unless you have good sterile technique.

[obtuse]

I use the 3% faulting from pharmacy / supermarket.

Again, the reason i use it, is simply because i use my kitchen. If i had a better space to do work in i would probably ditch the peroxide in favour of sterile technique.

SallyD thanks for the reminder on that salt method, i meant to try it out once and forgot about it, so will try it out soon.

Cheers, Ob.

[8145]

Bummer -

Will try some oysters this weekend

[watertrade]

\ How much peroxide to add per liter or 250ml batch of agar.

\

 

I used to add 7ml of 3% H2O2 to 1 liter agar when it is cooled to 60 degree C or below.

Edited June 22, 2012 by watertrade

[NSF]

Another little tip for myc on agar and out running contams:

Take something like a metal cigar tube, hold it with an oven mit and heat the open end with a gas torch (or stove). Then place the open end into the agar, over the tissue sample. So you burn a little circle indent into the agar. This 'moat' is another little trap for contams, where as mushroom mycelium can leap across the gap, it just gives you a little more time. Allowing the mushroom mycelium to get ahead of the contam growth.

[NSF]

Ok here's a tip I got from a recipe for agar for agarics.

100gm of good quality phase 2 compost, into 2 litres of distilled water, boil 1 hour. Let cool and then decant/filter. Bring the amount back up to 2L by adding more distilled water as necessary.

In a separate container make a paste using 30g of agar, 10g of peptone and a little of the water mixture. Continue to slowly add the water whilst stirring. PC at 121 for 30 mins (30 mins starts once the PC is at temp).

[LokStok]

^^ Erm, that's what i was doing hence 'avoiding slicing them'. Thanks though ;)

 

oops, sorry man, my poor early morning comprehension skills

[8145]

This video is YouTube. I am watching it now. Will source some more material ad do a second test this weekend. Will also post the results.

Hey Blas....see if you can get a copy of the Lets Grow Mushrooms DVD.

Just watching bits of it now and got some great tips for agar. It will give you a good overview of home mycology projects.

In the DVD he mentions washing a wild specimen in iodine to cut contams. It makes perfect sense.......iodine is used in surgery to cut down on contamination. They fucking paint that iodine shit all over people when they're doing some cutting. And best of all it doesn't shock the myc like a peroxide bath does (or bleach bath).

Also mushrooms in the pin stage have explosive growth and the myc can grow faster than the contams....giving you a head start for a clean culture.

In fact without knowing why i have just had success with the pin method on agar. Ripped a pin in half, dipped in peroxide and did 2 plates. 1 failed but 1 allowed enough growth to make a clean cut of myc