Morel and morel excited!

[TheJackal]

Have not received dual element ionizer yet, but did try innoc of rose bengal agar plate with black morel LC myc, as a test run under poor man's flow hood (see http://forums.mycotopia.net/showthread.php?t=23185 (Ionizer Reccomendations), ended up removing pre-filters to boost output cfm).

Procedure was to shut down all drafts in room, oust room, take shower, wear clean clothes and mask and gloves, wipe work area and work materials with alcohol, run flow hood, and innoc, parafilm seal, label and store agar plate at 70F.

[Bobcat]

Originally Posted by crazy1

Not to sure if a sponsor here sells it. But I get mine from xxx. That's FungiPerfecti's site.
And when you do it. Fill the test tubes first and then sterilize them, then just lean them up against something till they solidify.

Our vendor Sporeworks sells this product. They support us so please support them.

Please don't take offense at the editing of your post. [and yes, I know some info is still there, not trying to censor discussion, just the linking.]

[crazy1]

Thanx Bobcat, I will buy from workman next time. And I understand the edit. Got to support the sponsors here.
They make it available to us all.

[Bobcat]

Read about storing culture slants, but was wondering if the agar plates could just as easily be stored instead for later use.

Yeah, this will work, but probably not as well as the slants or even canning jars. There a couple of options to improve the keeping of your culture with plates. One thing I have tried with success is to pour a plate real thin, allow a pure culture to grow out to at least 50% and then do a hot pour over it. The cultures are then taped up with parafilm, placed in a ziplock and put in the fridge. When you are ready to use, remove to a warm location out of the ziplock. The culture will awaken, without oxidation harm, and grow through the second layer.

You can also use small ziplock baggies.... "crack sacks" lol. They come sterile from the manufacturer. You can pour more than you'll use in a year of them with about 200 mils of prepared agar. Put in your culture, allow to grow out a little, and put in the fridge. If done right, there won't be air in there to oxidize your culture- so it will be good for a long time.

[crazy1]

When I use Parafilm, I fold it in half, width wise, then stretch it around the plate. Seems to not snap as easily and still seals the plate up, in a nice clean fashion. No unruly edges you know.

[TheJackal]

Originally Posted by crazy1

When I use Parafilm, I fold it in half, width wise, then stretch it around the plate. Seems to not snap as easily and still seals the plate up, in a nice clean fashion. No unruly edges you know.

This would have been nice to know, mine probably looks like the red-headed step child, share a pic please! Also, if you have not already, please share your agar plate making tek, as I should be ready to try this soon!

Cheers!

Jackal

[crazy1]

Next time I pour I'll get pics for you

[TheJackal]

Originally Posted by crazy1

Next time I pour I'll get pics for you

Kewl, thanks!

[TheJackal]

Originally Posted by greysRDbest

Jackal , Ive acquired some dried Morchella Rufobrunnea if you would like to try that strain. It is an easier fruiter and supposedly from everything Ive seen thats what commercial growers are using currently. Anyone reading this thread who would like to make an honest attempt at culturing morels and would like to try working with rufobrunea - pm me, and Ill try to get you some. I only have 8 dried rufobrunnea to distribute, but Im willing to share with anyone willing to give it an honest try.

Received the 2 Morchella Rufobrunnea samples with much thanks!

Who da man? U da man!

Cheers!

Jackal

sweet dude--
Honestly im just really glad to help. Hope they work out like we hope they do.......

[Lazlo]

I'm curious of this one myself.

Sorry about the no show on the pics. I forgot, so i'll have to take some thursday night. I'm surprised on how much cleaner this culture is compared to the MEA plates. In which are eaten up with a pin type of mold.

[TheJackal]

Not sure if the rose bengal pre-poured agar was such a good choice. It seems to inhibit the growth of anything.

It's been two days and nada. How long does it take for the morel cultures to spread?

Sneezed on a couple of plates for shits and giggles to see if anything will grow on this stuff. The recipe stated "Used for isolating fungi," but it just may be a loose nut behind the wheel (yours truly).

Meanwhile, the mycobag and jar seem to be growing some ball like structures that based on the below pic from http://www.fungaljungal.org/morels/mofire.htm appear to be morel sclerotia. Will let them go for a couple more weeks to see what happens.

Cheers!

Jackal

[Lazlo]

At comfortable room temperatures a few days, but at 80 or so degrees just a couple. Once they start growing out on the agar, they colonize it quickly.

Originally Posted by TheJackal

Meanwhile, the mycobag and jar seem to be growing some ball like structures that based on the below pic from http://www.fungaljungal.org/morels/mofire.htm appear to be morel sclerotia. Will let them go for a couple more weeks to see what happens.

Cheers!

Jackal

yes!

[Lazlo]

Good grief! The DWA culture's flying! I wonder if using DWA is a good way to outrun molds in culturing all sorts of species? It appears that the mushroom hyphae are racing out to find food. Wow! The hyphae have nearly doubled in length since this morning.

[TheJackal]

Here is a morel MEA/DWA study:

http://www.rirdc.gov.au/reports/AFO/04-024.pdf

[Lazlo]

I read that a while ago. That's where I got the DWA idea.

[TheJackal]

Originally Posted by Lazlo

I read that a while ago. That's where I got the DWA idea.

Evil minds think alike!

I briefly looked at it and came to the conclusion that much brighter peeps than I and with real labs are working on this problem, so tha the best I can hope for is a few morel fruits and popping a bottle of bubbly to celebrate same.

[TheJackal]

"Prepare[d 10 cups of] the "supersoil" substrate, the standard mixture is as follows:

20 percent sand [2 cups lava sand],
30 percent soil [3 cups top soil],
50 percent organic material [5 cups] composed of [:]

80 percent small wood chips [4 cups apple wood chips],
10 percent rice hulls [1/2 cup],
5 percent soybean meal [1/4 cup],
5 percent sphagnum [1/4 cup].

Correct[ed] the ph with lime to 7.1-7.3 [using Kelway soil tester]. Mix[ed] well [in 1 gallon ziplock baggie]."

[Lazlo]

Looking good man!

Here's 2 DWA plates. The mycelium is really hard to see, but you can see how clean these 2 are compared to a MEA plate. All 3 are the initial inoculants.





Here's the MEA plate.

[TheJackal]

Originally Posted by Lazlo

Looking good man!

Here's 2 DWA plates. The mycelium is really hard to see, but you can see how clean these 2 are compared to a MEA plate. All 3 are the initial inoculants.

Here's the MEA plate.

Thanks, kewl stuff and nice agar work!

great work fellas. and thanks lazlo...to quote jackal..U DA MAN!

[TheJackal]

Originally Posted by greysRDbest

great work fellas. and thanks lazlo...to quote jackal..U DA MAN!

Thanks!

"Fill[ed] an autoclavable aluminum [20.2 cm dia x 8.0 cm] tray (i.e. c[asserole] pan)
(liberally punched with drainage holes) to a
depth of 2 inches with [supersoil] substrate.
Step 17.
Saturate[ed] substrate thoroughly with water [for 24 hrs].
Allow[ing] to drain completely. Fill[ing] a second,
identical tray with soaked [for 24 hrs], [and] drained rye grass
seed to a depth of 1/2 - 1 inch."

Will be using oven bags with 1/2 plastic tube polyfil filters for forthcoming PCing step.







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[Lazlo]

This plate is bad ass. Check this out!

This is another dry isolate. At first I thought the suspected hyphae were scratches in either the agar or on the bottom of the plate. I just took another look a minute ago and couldn't believe how the mycelium was reacting to the DWA. It's very hard to see, but there's literally one hyphal strand emerging from a small group around the inoculation, then it disappears for about an 1/2 of an inch, grows out again, disappears for another 1/2" and now is starting to split into more strands. Look closely.

[TheJackal]

Originally Posted by Lazlo

This plate is bad ass. Check this out!

This is another dry isolate. At first I thought the suspected hyphae were scratches in either the agar or on the bottom of the plate. I just took another look a minute ago and couldn't believe how the mycelium was reacting to the DWA. It's very hard to see, but there's literally one hyphal strand emerging from a small group around the inoculation, then it disappears for about an 1/2 of an inch, grows out again, disappears for another 1/2" and now is starting to split into more strands. Look closely.

Nice work, shweeeet!

[TheJackal]

"Step 18.
In a clean (use[d] 5% bleach to clean up) draftfree
area [sprayed with Oust, cleaned with alcohol]
open[ed] cooled substrate bag and mix[ed] [38.9 g]
[of sclerotia] into substrate using [alcohol wiped surgical gloves].
Reclose[ed] bag and [being] place[ed] in a cool (65-70°F) dark place for [next] 4-6 weeks.
During this period (the spawn run) the relative humidity [will] be kept at
90-100%, CO2 [uncontrolled] at 6000-9000 ppm, and
no fresh air exchanges [due to being sealed in oven bag]."

[Lazlo]

Heh, it's pretty weird how the hyphae just jump around all crazy like that huh?

Nice stuff! Where did the sclerotia come from in the first place? And why didn't you just leave it where it came from? That looks great none the less!

[TheJackal]

Originally Posted by Lazlo

Heh, it's pretty weird how the hyphae just jump around all crazy like that huh?

Nice stuff! Where did the sclerotia come from in the first place? And why didn't you just leave it where it came from? That looks great none the less!

That some wild and crazy hyphae!

Trying to answer your questions, the sclerotia were formed by spawning from sporework's syringe, to LC, and then to sterilized rye grass/soil mix, per the published procedures.

Per the published procedures, a nutrient poor substrate is innoced with sclerotia in a tray with holes at bottom and placed over an identical tray of sterilized rye grass, which acts as the nutrient source.

After the spawn run, the nutrient source tray is removed, the spawn tray is chilled (I believe simulating winter conditions), and then later is removed, soaked with water, and placed in fruiting environment (I believe simulating heavy rain followed by Spring conditions).

The morel Gods should have more input about the above theories.

Cheers!

Jackal

Just as a side note...Ive said this before and have nothing but personal observations to back it up......hard smacking rain really makes a huge flush in nature. If i get a really hard rain during morel time I know im going to see more mushrooms than usual. I do know that shiitake growers in Japan have soaked and literally beaten shittake logs with a metal rod to induce a flush and there are some people studying the effects of vibrations on morel flushes but I have no data at this point.
This is a great thread....if it gets anymore intresting ill be sitting here hitting the refresh button on my browser with a bowl of popcorn and a beer.

[TheJackal]

Originally Posted by greysRDbest

Just as a side note...Ive said this before and have nothing but personal observations to back it up......hard smacking rain really makes a huge flush in nature. If i get a really hard rain during morel time I know im going to see more mushrooms than usual. I do know that shiitake growers in Japan have soaked and literally beaten shittake logs with a metal rod to induce a flush and there are some people studying the effects of vibrations on morel flushes but I have no data at this point.
This is a great thread....if it gets anymore intresting ill be sitting here hitting the refresh button on my browser with a bowl of popcorn and a beer.

That is good to know, but I have no idea how to simulate "hard smacking rain" in my humble greenhouse.

The best I can do so far is rain version 2.0 !