P. subaeruginosa won't germinate

Boarders · 10-11-07, 00:22

Hello fellow 'topiates.

I harvested a few wild P. subaeruginosa fruits this year and created prints from these wild specimens in an attempt to see if I could colonize some woodchips for fun.

I can't seem to get these spores to germinate though

I've thrown them in LC (including sterile wort), WBS, pasteurized woodchips, sterilized woodchips and onto an agar petri; all have failed to germinate.

Any tips on how I could improve my chances? I've had spores soaking in sterile water to rehydrate prior to attempting the germination and have tried multiple spore prints. I'm getting a little frustrated here! Do they need lower temperatures for germination like they do for fruiting?

kocos · 10-11-07, 03:19

Considering they are wild prints, no matter how many precautions you take there is still a large chance of contamination. Your best bet would be streaking agar with spores, than transfering clean growth to new plates. Its strange that they arnt germinating on agar either.

Boarders · 10-12-07, 19:20

Thanks for the reply kocos. I think I'll try a few more agar plates and see what happens.

Getting OT, the other thing getting to me is a Thai spore print from FSRE contaminated with cobweb

. Hopefully I can isolate mycelium from cobweb, but the stuff grows so damn fast I don't think I'll be able to isolate

fedshtkpndrk · 10-12-07, 20:26

I would try and salvage the good myc to another plate. Try and clean it up from there

danzick · 10-12-07, 21:49

Did you try other lc media besides wort?

How long have you waited for germination?

Boarders · 10-12-07, 21:55

danzick, I've tried 4% dextrose and wort with an OG 1048 and 1024. I've used dextrose agar and malt agar, WBS and woodchips.

I've waited months for germination in some samples!

As for salvaging mycelium and cleaning it up on agar for the Thai strain, I think the cobweb will grow well before the cubensis germinates going on its growth in LC.

Boarders · 11-09-07, 16:46

I am still unsuccessful with this project, but Danzick is having a go now. I thought it would be best to get an opinion on my agar/nutrient concentrations. What I use in my malt plates is 100 mL water, 1.3g agar powder, 7g Dried Malt Extract. I have also used a mix with 12g DME. These are based on what I use for yeast culturing but am now questioning whether it's an appropriate mix for mushroom cultivation.

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10-11-07, 00:22	#1 (permalink)
Boarders Mycophiliac Join Date: Sep 2006 Posts: 42	P. subaeruginosa won't germinate Hello fellow 'topiates. I harvested a few wild P. subaeruginosa fruits this year and created prints from these wild specimens in an attempt to see if I could colonize some woodchips for fun. I can't seem to get these spores to germinate though I've thrown them in LC (including sterile wort), WBS, pasteurized woodchips, sterilized woodchips and onto an agar petri; all have failed to germinate. Any tips on how I could improve my chances? I've had spores soaking in sterile water to rehydrate prior to attempting the germination and have tried multiple spore prints. I'm getting a little frustrated here! Do they need lower temperatures for germination like they do for fruiting?

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10-11-07, 03:19	#2 (permalink)
kocos Mycotopiate Join Date: Jan 2007 Posts: 851	Considering they are wild prints, no matter how many precautions you take there is still a large chance of contamination. Your best bet would be streaking agar with spores, than transfering clean growth to new plates. Its strange that they arnt germinating on agar either.

10-12-07, 19:20	#3 (permalink)
Boarders Mycophiliac Join Date: Sep 2006 Posts: 42	Thanks for the reply kocos. I think I'll try a few more agar plates and see what happens. Getting OT, the other thing getting to me is a Thai spore print from FSRE contaminated with cobweb . Hopefully I can isolate mycelium from cobweb, but the stuff grows so damn fast I don't think I'll be able to isolate

10-12-07, 20:26	#4 (permalink)
fedshtkpndrk Pantastic Mod Join Date: Mar 2007 Posts: 1,049	I would try and salvage the good myc to another plate. Try and clean it up from there

10-12-07, 21:49	#5 (permalink)
danzick Mycophiliac Join Date: Sep 2007 Posts: 95	Did you try other lc media besides wort? How long have you waited for germination?

10-12-07, 21:55	#6 (permalink)
Boarders Mycophiliac Join Date: Sep 2006 Posts: 42	danzick, I've tried 4% dextrose and wort with an OG 1048 and 1024. I've used dextrose agar and malt agar, WBS and woodchips. I've waited months for germination in some samples! As for salvaging mycelium and cleaning it up on agar for the Thai strain, I think the cobweb will grow well before the cubensis germinates going on its growth in LC.

11-09-07, 16:46	#7 (permalink)
Boarders Mycophiliac Join Date: Sep 2006 Posts: 42	I am still unsuccessful with this project, but Danzick is having a go now. I thought it would be best to get an opinion on my agar/nutrient concentrations. What I use in my malt plates is 100 mL water, 1.3g agar powder, 7g Dried Malt Extract. I have also used a mix with 12g DME. These are based on what I use for yeast culturing but am now questioning whether it's an appropriate mix for mushroom cultivation.