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OfflineManifoldPrime
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Registered: 03/16/17
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Re: Cultivation General Discussion [Re: Smooby2]
    #25575317 - 10/28/18 11:49 PM (5 years, 3 months ago)

Yeah thats a mold. you can see it making little black spores on stalks . Forgot what its called. how long ago did you inoculate the plates? Loop streak/syringe drop/swab?


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OfflineSmooby2
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Registered: 07/13/18
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Re: Cultivation General Discussion [Re: ManifoldPrime]
    #25575327 - 10/28/18 11:55 PM (5 years, 3 months ago)

Quote:

ManifoldPrime said:
Yeah thats a mold. you can see it making little black spores on stalks . Forgot what its called. how long ago did you inoculate the plates? Loop streak/syringe drop/swab?



3 days inoculation loop. All work done in SAB  I thought I was very sterile. I've got a lot to learn and the prints were given to me. Not sure how sterile they are. Any tips aporeciated


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OfflineManifoldPrime
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Re: Cultivation General Discussion [Re: Smooby2]
    #25575334 - 10/29/18 12:00 AM (5 years, 3 months ago)

yeah by 3-4 days most cube myc will be tiny little pea sized colonies. rapid growth like that is a contaminant. Start really staring/looking at whatever contams pop up, so you can learn their traits. some are easy to ID like lipstick mold, some can look just like myc if you're not experienced.


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OfflineSmooby2
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Re: Cultivation General Discussion [Re: ManifoldPrime]
    #25575337 - 10/29/18 12:02 AM (5 years, 3 months ago)

Yeah. I've lost one to green mold and now this one. The other 2 showing no growth yet. I guess I'll swipe a few more tomorrow and try to be a little more sterile.


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Invisiblestareatclouds
star eat clouds?
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Re: Cultivation General Discussion [Re: Smooby2]
    #25575486 - 10/29/18 02:06 AM (5 years, 3 months ago)

You should post a video of you inoculating your dishes.


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Offlineteladi
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Re: Cultivation General Discussion [Re: stareatclouds]
    #25575537 - 10/29/18 03:07 AM (5 years, 3 months ago)

Is someone able to get this paper?

https://www.researchgate.net/publication/327539316_Blue_light_exposure_and_nutrient_conditions_influence_the_expression_of_genes_involved_in_simultaneous_hyphal_knot_formation_in_Coprinopsis_cinerea

https://www.sciencedirect.com/science/article/pii/S0944501318301526

Sakamoto, Yuichi & Sato, Shiho & Ito, Miyuki & Ando, Yuki & Nakahori, Kiyoshi & Muraguchi, Hajime. (2018). Blue light exposure and nutrient conditions influence the expression of genes involved in simultaneous hyphal knot formation in Coprinopsis cinerea. Microbiological Research. 217. 10.1016/j.micres.2018.09.003.

I sent an email to Yuichi Sakamoto requesting a copy, but just in case I get a negative response, I was hoping someone out there has access to the journal. Thank you in advance.

Quote:


Abstract

Light and nutrients are crucial environmental factors influencing fungal sexual reproduction. Blue light induces simultaneous hyphal knot formation in Coprinopsis cinerea mycelia grown on low-glucose media but not in mycelia grown on high-glucose media. Many hyphal knots are visible in the arc near the edge of the colony one day after 15 min of blue light stimulation. These findings collectively suggest that blue light accelerates hyphal knot induction in nutrient-limited conditions. Transcriptome analysis revealed that gene expression after light exposure is divided into at least two major stages. In the first stage, genes coding for fasciclin (fas1), cyclopropane-fatty-acyl-phospholipid synthases (cfs1 and cfs2), and putative lipid exporter (nod1) are highly expressed after 1 h of light exposure in the mycelial region where the hyphal knot will be developed. These genes are upregulated by blue light and not influenced by glucose condition and mating. These results suggest that although some of the genes are critical for induction of the hyphal knots, they are not sufficient for hyphal knot development. In the second gene expression stage, genes encoding galectins (cgl1-3), farnesyl cysteine-carboxyl methyltransferases, mating pheromone-containing protein, nucleus protein (ich1), and laccase (lcc1) are specifically upregulated at 10–16 h after blue light exposure when the mycelia are cultivated on low-glucose media. These genes might be involved in the architecture of hyphal knots or signal transduction for further fruiting body development. These results contribute to the understanding of the effect of environmental factors on sexual reproduction in basidiomycetous fung





Quote:


Conclusions

Blue light can induce simultaneous hyphal knot formation, which appears in an arc in a marginal colony of C. cinerea in glucose-limited conditions. Transcriptome analysis revealed that gene expression for hyphal knot development can be divided into two main stages. In the first stage, the genes nod1, cfs1, cfs2, and fas1, which are possibly involved in hyphal knot induction in C. cinerea, were expressed 0.5–1 h after a very short exposure to blue light (at least 15 min) (Fig. 2, Fig. 5). These genes were expressed by blue light exposure regardless of the glucose content in the media and mating. Limitation of glucose content was required for further progression of hyphal knot development (Fig. 8, Fig. S5). In the second stage, galectins (cgl1-3), nucleus protein (ich1), laccase (lcc1), pheromone-containing proteins, and pheromonemodifying enzyme-encoding genes were upregulated specifically in glucose-limited conditions. These data collectively suggested that blue light accelerates hyphal knot induction in nutrient-limited conditions. The expression pattern of these genes will provide markers to evaluate the progression of hyphal knot development in knotless mutants or mutants of putative light sensor genes with disrupted genes in C. cinerea. Our observations contribute to understanding the mechanisms of induction of sexual reproduction in basidiomycetous fungi.





RR et al have indicated that direct sunlight for a bit will help initiate pinning. Blue light in sunlight could be triggering the above pathways. It's interesting to me to note that this will only occur in low nutrient environments; which most bulk substrate are. I'm gonna take a few months to build an isolate to play around with different lighting and spawn ratios.


Edited by teladi (10/29/18 03:39 AM)


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Offlineteladi
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Registered: 06/27/17
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Re: Cultivation General Discussion [Re: teladi] * 1
    #25575547 - 10/29/18 03:18 AM (5 years, 3 months ago)
Log in to view attachment

I just remembered Sci-Hub. Attached for whomever is interested.


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OfflineSmooby2
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Re: Cultivation General Discussion [Re: stareatclouds]
    #25575924 - 10/29/18 08:35 AM (5 years, 3 months ago)

Quote:

stareatclouds said:
You should post a video of you inoculating your dishes.




I did another 4 swipes last night. I will post a video next time I do it. I've got to get this figured out! I've had some very successful grows with brf to bulk, but I want to progress and learn agar and grains. I have built a SAB, wipe everything down extremely well with 70% IPA. Shower,brush my teeth, mouthwash. When I sit down at my sab I put on my gloves wash them(and all the way up to my elbows)with the IPA. Wipe down anything going inside the sab before and after they're in. Open my print, flame sterilize the loop, cool it on the agar,gather my spores and swipe and close. Careful not to hover over print or dish.


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InvisibleShroomspective
Looking into the void
I'm a teapot User Gallery


Registered: 09/26/15
Posts: 1,725
Loc: Far Far Away
Re: Cultivation General Discussion [Re: teladi]
    #25575931 - 10/29/18 08:38 AM (5 years, 3 months ago)

That's interesting, I was doing some research a couple of years back, and blue light was found to be beneficial to growth..I went as far as to buy some LED bulbs that you could change the colour output from to expose them to blue more often. My results were good, but I had no control to compare to. Below are some of the articles

"The approximately 460 to 480 nm of blue light that  is  important  to  the  production  of  the  basidoma  of  most  cultivated  mushrooms"

"UV and blue wavelengths of 370, 440, and 460 nm were the most effective"

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3627973/
http://slowacki.kielce.eu/IB/PSlusarczyk.pdf
http://www.redalyc.org/pdf/685/68528461002.pdf
https://www.researchgate.net/publication/237713813_The_Effect_of_Light_upon_Basidiocarp_Initiation_in_Psilocybe_cubensis



Edited by Shroomspective (10/29/18 08:51 AM)


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Invisiblestareatclouds
star eat clouds?
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Re: Cultivation General Discussion [Re: Smooby2]
    #25576041 - 10/29/18 09:54 AM (5 years, 3 months ago)

Quote:

Smooby2 said:
Quote:

stareatclouds said:
You should post a video of you inoculating your dishes.




I did another 4 swipes last night. I will post a video next time I do it. I've got to get this figured out! I've had some very successful grows with brf to bulk, but I want to progress and learn agar and grains. I have built a SAB, wipe everything down extremely well with 70% IPA. Shower,brush my teeth, mouthwash. When I sit down at my sab I put on my gloves wash them(and all the way up to my elbows)with the IPA. Wipe down anything going inside the sab before and after they're in. Open my print, flame sterilize the loop, cool it on the agar,gather my spores and swipe and close. Careful not to hover over print or dish.




Sounds good, but maybe next time don't use beer to sanitize.


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Offlinegizmodo
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Re: Cultivation General Discussion [Re: stareatclouds]
    #25576085 - 10/29/18 10:22 AM (5 years, 3 months ago)

Quote:

Smooby2 said:
wipe everything down extremely well with 70% IPA. Shower,brush my teeth, mouthwash. When I sit down at my sab I put on my gloves wash




:freshwtf:


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Offlinetryptkaloids
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Re: Cultivation General Discussion [Re: stareatclouds]
    #25576113 - 10/29/18 10:36 AM (5 years, 3 months ago)

Quote:

Doc9151 said:
Dude,  a case of half pint jars made up of tek style is super cheap, extremely easy and will provide you with enough mushrooms to treat your depression for months and months from a single run, bulk grows are cheap once you make the initial investment  in  a pressure canner. Bulk grows are more distribution related than personal use.

How often are you medicating with mushrooms? 1 single 5g trip is said to be beneficial for several months or taking microdoses every few days to once a week has beneficial effects. I know people using them for PTSD, migraines, depression and a few other disorders that benefit from doing it as I described above. 

It sounds like you could benefit from it,  pf tek is the fastest way to get them, you will have mushrooms in 3wks to a month, bulk can take up to 2 months if you have to start from scratch.


fuck pf tek. that's a waste of my time. no offense, but this isn't my first time around the block. pretty much any method on here I've tried other than liquids to bags. the only thing worth my time is A2G
Quote:

stareatclouds said:

Quote:

stareatclouds said:

I also don't agree with this at all. Regardless of sterile tek, your utensils will get dirty before you go to work like 99% of the time. Your sterile tek includes sanitization. I spray and wipe shit down liberally with iso and rarely smell any, so maybe you're using too much, tryp?


naw, I got airflow back to my nose and after not being able to smell for a week it's like I had super powers


Quote:

stareatclouds said:
Ive been very unstable mentally the past year or so. fruiting happens to come along right during the times I need to just check out for awhile and go adventure in order to reset.




Word. I'm sorry to hear that. I'd say the most forgiving aspect of mycology with tubs is that fruiting happens just fine without doing anything, so that shouldn't be an issue for you.

You obviously have a strong passion for mycology. Regardless of needing some huge yield for the actual fruits, you can aim for more impressive yields for your own gratification. Otherwise, what's the point of posting and reading up on shit? That's why I try and get better at shit.

You definitely possess quite a bit of knowledge on the hobby, but seems like you struggle a little with your own grows (which is totally fine, most of us know more than we practice due to the nature of the game). If you want help with any bottlenecks, feel free to PM me and maybe I can share what works for me and see if it helps you. No worries if you'd rather do your own thing, just an offer from one myco-bro to another.



I appreciate it bro

Quote:

stareatclouds said:


Oats to me are the perfect grain. run a box fan on them for an hour.





Quote:

grainbrain said:
I steam dry for a maximum five minutes & load the wheat into the jars hot & steamy.



this is why I consider wheat the best grain I've ever used.  time is money. less time I spend messing with grain the more time I can spend in the SAB or cleaning


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Offlinemushroomnate
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Re: Cultivation General Discussion [Re: stareatclouds]
    #25576580 - 10/29/18 02:42 PM (5 years, 3 months ago)

Quote:

Smooby2 said:
wipe everything down extremely well with 70% IPA.




Quote:

stareatclouds said:
Sounds good, but maybe next time don't use beer to sanitize.



:rofl2:


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Invisiblestareatclouds
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Re: Cultivation General Discussion [Re: mushroomnate]
    #25576606 - 10/29/18 02:57 PM (5 years, 3 months ago)

You don't have to watch the fan for an hour, lol. You can still clean or do agar work, brah. And I don't need a full hour, it's just what I clock off from while doing other shit. I also am doing like 40 quarts of grain at a time. Anywho, whatever works for you, of course.


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OfflineIntergalacticSpore
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Re: Cultivation General Discussion [Re: stareatclouds]
    #25576615 - 10/29/18 03:03 PM (5 years, 3 months ago)

When you shake jars, can grain touch the polyfill piece that is on the inside? Won't that put nutrients inside it, shake up the contams on the way down and create frame for contams to get in?


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OfflineFishLevelMidnight
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Re: Cultivation General Discussion [Re: IntergalacticSpore] * 1
    #25576619 - 10/29/18 03:06 PM (5 years, 3 months ago)

I've never had an issue with it


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Invisiblestareatclouds
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Re: Cultivation General Discussion [Re: IntergalacticSpore] * 1
    #25576623 - 10/29/18 03:07 PM (5 years, 3 months ago)

Your Poly should be stuffed so tightly into holes that shit doesn't fall through it. And you won't get "nutrients" on Poly from grain or anything else that would germinate on it. Polyfill is a synthetic fabric which makes it tough for stuff to grow on it. Don't let it stay wet. Even if it could, it'd theoretically only be colonized by healthy mycelium within your jar, and that wouldn't make it easier for contams to fall through the top.


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OfflineDreamReality
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Re: Cultivation General Discussion [Re: IntergalacticSpore] * 1
    #25576624 - 10/29/18 03:08 PM (5 years, 3 months ago)

Quote:

IntergalacticSpore said:
When you shake jars, can grain touch the polyfill piece that is on the inside? Won't that put nutrients inside it, shake up the contams on the way down and create frame for contams to get in?




It can bash into it violently a bunch of times. Or at least that's my experience while shaking :lol:


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OfflineSmooby2
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Re: Cultivation General Discussion [Re: mushroomnate]
    #25576721 - 10/29/18 03:58 PM (5 years, 3 months ago)

I said
Quote:

mushroomnate said:
Quote:

Smooby2 said:
wipe everything down extremely well with 70% IPA.




Quote:

stareatclouds said:
Sounds good, but maybe next time don't use beer to sanitize.



:rofl2:



I know it's sad, but I kept wondering what was wrong with using IPA(Indian pale ale) Felling a little slow today. Lol
Maybe I'll get better results with this attempt. Got some PE6 spores today and can't wait to see what it can do.


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OfflineFishLevelMidnight
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Re: Cultivation General Discussion [Re: DreamReality]
    #25576723 - 10/29/18 03:58 PM (5 years, 3 months ago)

I wonder how unstabiliEd the vendor PEU and APE are.

The PEU I posted recently looked like PF albino APE but kept the uncut look right before opening up- all leucistic and I got the caps to print...

Any dedicated peeps want to try out a print and see what they get?
Stare?


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