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Spores on agar

- Thu, 26 Nov 2020 16:51:51 EST CwgduOPu No.147465
File: 1606427511955.png -(2947284B / 2.81MB, 1080x2520) Thumbnail displayed, click image for full size. Spores on agar
So when you put spores from prints or supposed sterile spore syringes onto agar are they supposed to look like this when healthy ?

I know there are two different types of mycelium rhizomorphic, and tomentose but my one seems too fluffy.
The one on the right colonized very fast hoping that can be used.

Also they have sat there about 4-5 weeks now and a few more I have are like the top left but probably have less mycelium on them...does that mean they are rubbish .

Any guides on how to analyze agar can only find them on how to make plates and do transfers.
press - Fri, 01 Jan 2021 09:31:58 EST m3YGjSOR No.147498 Reply
seeing as this thread is over a month old, i hope that you found the resources you needed and managed to do some clean transfers.

spore germination plates tend to very densely populated oweing to the sheer number of spores and subsequently delevoping cultures. you wont really see organized growth until youve done at least one transfer.

if you have a "clean" spore syringe sold to you by a trustworthy source, you could forego culturing on agar before inoculating lets say BRF cakes a la PF tek, wait for an attractive basidiocarp, and then take a tissue sample to culture on agar. that way youd have close to a guarantee of having a strain which has good fruiting capabilities. i sometimes take samples from petri dishes that i left laying around for long enough for them to pin in vitro.

the main motivation to germinate spores on agar is to make sure that youre not introducing a contaminated inoculant, its hard or rather impossible to estimate the exact qualities of a strain just by its phenotype on agar. a particular strain exhibiting rhizomorphic growth on agar doesnt tell you anything specific but people prefer transfering rhizomorphic samples since they are easier to differentiate from mold mycellium and its easier to identify disorganized growth due to bacterial pathogens.
when doing clean up transfers, it tends to be easiest to look for the most organized, flat, rhizomorphic areas of growth and transfer those. theres not a whole lot of point in trying to get a true isolate at that point since that would be like playing the lottery.
keep in mind that the agar composition does influence the growth patterns. the more higher a medium is in nutrional density the less likely it is that you will see strong rhizomorphic growth.

more broadly speaking:
what are you going to inoculate with your cultures?
Phineas Pockgold - Tue, 12 Jan 2021 03:26:03 EST ggQ8YrMc No.147504 Reply
not OP but I'm going to be starting to inoculate for my first time some time this month or next month, do you have any tips with using a spore syringe or just anything you wish you knew as a beginner?
press - Fri, 15 Jan 2021 16:36:13 EST ZM+0YjAU No.147511 Reply
1610746573584.jpg -(138252B / 135.01KB, 1080x608) Thumbnail displayed, click image for full size.
so thats a spore syringe from a reputable vendor or are you going to make your own from a spore print?
ive never ordered a syringe since shipping those would make me nervous, but from what ive read the syringes of reputable vendors e.g. shroomery sponsors are pretty clean.

assuming that youre going for PF tek, id suggest the following:

  • dont get some stupid air stone bubbler set up. a shotgun fruiting chamber with perlite works well. other fruiting chambers can work too for PF / BRF cakes, but ive never tried them.
  • a pressure cooker isnt necessary just yet, but i recommend it. if you plan to go beyond PF tek and just the occasional grow, try to gauge if youd like to invest the money for a bigger PC. should you be in north america a presto 16 quart or 23 quart pressure cooker might be easy to come by cheap on ebay, euro cukcs such as myself arent as lucky.
  • if you have an electric grinder, make your own brown rice flour instead of buying it. a bit coarser is better than really fine flour.
  • id use coarser vermiculite, or if you feel like complicating things a bit use fine verm for the actual cake and coarser verm for the protective dry layer on top. the finer verm absorbs water a bit better in my experience but isnt quite as good for the protective layer, since it can shift around more easily
  • absolutely use a still air box
  • do a "dry run" i.e. load up your still air box with empty jars and practise the movements, get a feel for aseptic technique. try to move smoothly and visualize steps beforehand.
  • its not strictly necessary to shower, brush your teeth, wear fresh clothes, a hair net but i like to employ that practise in order to have a ritual that gets me into a good and controled mind set. id really recommend a surgical face mask though.
  • unless youre going to store the colonizing jars in a room below 16 °C you dont have to incubate the jars. ideally youd have them in a place where they get a bit of ambient light and a bit of fresh air, just on a shelf in next to your desk works fine. you could place them on a higher shelf inside a cardboard box from which youve removed the top if you want to keep it a bit stealthier. in my experience incubation has introduced for problems than it helped.
  • dont move them too much and if you inspect them ( because lets face it, everybody will anxiously look for any sign of growth ) try not to tilt them, so that the protective verm layer doesnt get tumbled around.
  • give them enough time to form small pins while still inside the jar before "birthing" them
  • rinse the cakes under cold water when you birth them and roll them in verm. let them sit in the shotgun fruiting chamvber for a few hours before misting them, that seems to help the verm coating stick to them
  • after the first flush, let the cakes sit in the shotgun fruiting chamber for a day or two before dunking them. dont do a second roll in verm after youve dunked them
  • try to observe what ideal surface conditions look like
  • dont get a hygrometre, theyre not necessary

more broadly speaking:

  • dont be afraid to start agar work.
  • glass petris are shit. they crack, theyre slippery, you have to sterilize them anyways. gamma radiated polystyrene dishes are the way to go. ive always used ones without ventilation groves, but some people swear by them.
  • dont try liquid cultures untill youre pretty comfortable in your agar work.
  • dont be afraid to start bulk grain grows once youve done a few runs with PF or feel confident enough in your technique and judgement of surface condition. this comes with a few caveats but its a lot more time, space, and eventually cost efficient than PF.
  • dont listen to assholes on youtube. most are absolute shit ime.

and i wish i had known about unmodded monotubs and shoe boxes when i started doing bulk grains instead of drilling stupid holes into perfectly fine and inscopicous plastic totes ( not to insult monotubs, theyre great)

please excuse any typos or grammatical errors. im also a bit rusty, but am kinda proud of myself that as of recently only 1 out of 52 grain jars contaminated. suprisingly enough it wasnt even trichoderma but probably a black pin mold.
press - Fri, 15 Jan 2021 17:10:56 EST ZM+0YjAU No.147512 Reply
also wish i wouldve known how to do liquid inoculant instead of liquid culture and how easy grain2grain can be. nb.
Phineas Pockgold - Mon, 18 Jan 2021 03:01:35 EST ggQ8YrMc No.147513 Reply
yes, its a syringe from a reputable vendor, 10 mL. Thanks for the info dude, thats a pretty good run 51/52, I hope to not have any contamination for my first time. Have you had any experience with unmodified tubs yet?
press - Mon, 18 Jan 2021 16:16:02 EST 5F9Zj95X No.147514 Reply
im almost sure that there has to be a slight bacterial contamination in some jars, since some of them smelled a bit off but my sense of smell isnt what it used to be anyway. fingers crossed the 25 jars ill 'noc up wont end up turning into green dust.

no hands on experience with bigger unmodified tubs yet. im waiting until i have my method of preparing spawn bags dialed in, untill then i'll keep on working on shoeboxes.

since i had pretty good results with monos in the past, it seems likely that unmodified monos would do a bit better seeing as i tended to struggle with the dry climate around my parts.

if you get contamination try to make the most of it. their location can give you hints as to when they mightve been introduced. dont try to fruit a jar that has gone moldy and try to not spread the spores around your work area. i normally sterilize them before opening them, but i think just soaking them in water and opening while still emerged should drastically reduce the amount of spores that become airborne.

out of curiousity: what variety of cubensis did you decide to get?
Phineas Pockgold - Tue, 19 Jan 2021 19:39:16 EST ggQ8YrMc No.147517 Reply
Good luck with those jars. I have golden teachers to start off with but once I know what I'm doing I want to explore different varieties. When you put a spawn bag in a shoebox, do you wait for mycelium to colonize it more or do you go straight to fruiting conditions?
press - Wed, 17 Feb 2021 11:49:27 EST bvuDfHq3 No.147543 Reply
i dont quite understand the question.
when spawning shoe boxes i normally use a mycoquart and spawn that to 1.5 actual quarts of coir supplemented with gypsum. i then let it colonize fully with the lid snapped on before flipping the lid to initiate pinning.
ideally there wouldnt be a differentiation between colonization and fruiting conditions, but just growing conditions, but my climate is too dry.

i use spawn bags for monos, since that allows for a "one aliquot of spawn per fruiting chamber" approach, allowing me to isolate any grain spawn i mightve fucked up.

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