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Mushrooms, Mycology and Psychedelics > Advanced Mycology
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agar--isolation--senescence---lets talk turkey
    #16509911 -

im lookin into isolating a rhizomorphic growth for optimum effectiveness of my efforts---seeing as i only got 7 1/2 oz dry off my last 66qt tub with MS i need to up my game---and since a fellow cant obtain live culture of them thangs--i believe i need to make my own--im lookin at some media--particularly sabouraud dextroseagar with chloramphenicol agar and some beef extract peptone agar---i have some powder PDA on hand----i understand the phenomenon between the transfer of super nutrient rich agar to a less nutritous media---its like throwing Hugh Heffner into the ghetto---he aint gonna do well--lol---anyways i just wanna start it off right----so here's my question---if i were to miraculously isolate a nice rhizomorphic sector and ran with it for a while how long would it produce them bumper crops before the growth started to run outta juice and go into senecsence---from what i understand so far is when you're lookin at a plate with different growth patterns on it the cottony growth is senescent mycellium---anyways i look forward to an answer and some direction on how to proceed--i know yall wont let me down--lol---thanks


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Re: agar--isolation--senescence---lets talk turkey [Re: Mr. Wilson]
    #16509982 -

or am i just overly complicating a simple task(likely)!!--lol--


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Re: agar--isolation--senescence---lets talk turkey [Re: Mr. Wilson]
    #16511880 -

Assuming that you are talking about P. cubensis, just going on the general nature of this forum and its users.

I believe you are over-complicating what is essentially quite a simple process, its easy enough to optimize a culture and keep it from aging without going overboard.

I might be misinterpreting you, if i am, sorry, also there are other people with better practices out there than mine, so if they chime in, listen to them over me.

Agar ~ there are loads of tips about how to make and use it on this forum as well as the RR video, which gives quite a good demonstration of the procedure for pouring them and necessary considerations. 

Media doesn't have to be complicated, just think carefully about what the fungus wants to 'eat' and how it likes to eat it. PDA is a good start.

Now... im pretty sure this would be frowned upon by the more skilled members, but ill put this out there as well; I like to isolate as little as possible before making a fruitbody clone, on the odd chance that i accidentally isolate a 'non-fruiting' strain. With the more picky species i will start the spores on agar, but then move them into LC or grain ASAP, fruit it, clone a fruiting body and then work with that.

Im pretty sure the more skilled and organized folk prefer to make multiple series of isolates and fruit them individually, but i often dont have the time or resources to do scores of petrie dishes.

Once you've got a working culture its about taking and preserving cultures from different stages, if you find something goes awry at the 4th or nth generation of the same culture, you can always go back to a slant or syringe you made earlier and make more of that.

Anyway, cottony(Tomentose?) growth isn't really senescent mycellium as far as i know, (which isn't far btw). Senescence i believe refers in this context to a loss of vigor from age/too many transfers. I dont think you need to worry about it for the moment.

Long story short, if you're new to this, i would try to clone a vigorous multispore fruit first and then isolate that culture for aggressive growth, this should ramp up your yields a bit, other than that, you could also try other varieties if you find that the one you're using is consistently a poor performer.

Anyway, thats just my 2c, i had a spare 15 mins. =3

Also put some paragraphs in there boi, its much easier to read if you separate things a bit xD

Edited by Nobitte (07/10/12 11:47 PM)

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Re: agar--isolation--senescence---lets talk turkey [Re: Nobitte]
    #16512201 -

aye--gotcha---makes sense i guess---clone out a nice fruit and a fella would already have a few desirable traits leaning their way---im definately gonna look into this further----thanks


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Re: agar--isolation--senescence---lets talk turkey [Re: Mr. Wilson]
    #16549704 -

you could always drop a grain per dish(3 per y plate) into potato dextrose agar or ME agar in several dishes with heat sterilized tweezers and look for good growth and work from there. as far as senescence goes i had a weird experiance with that before i had knowledge of it.i did grain to grains for over a year with eq's and still got it fruiting!wasn't large amounts but it still fruited, which now i understand how crazy that is. the learning curve came with PE because it fizzled out after a couple months, and i am greatful it did because it forced me here to learn, learn, learn!


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Re: agar--isolation--senescence---lets talk turkey [Re: God Stamet]
    #16550352 -

no shit man---its that fast ehh??---they say shrooms are constantly evolving---but that's damn fast


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Re: agar--isolation--senescence---lets talk turkey [Re: Mr. Wilson]
    #16550599 -

I isolate out on PDA starting with spores (1). Locate the 2 best growth points and take a wedge from the edge creating 2 new plates. (1/1 and 1/2)
Repeat from the leading edge of 1/1 and 1/2 to create 1/1/1, 1/1/2, 1/2/1, 1/2/2.

From there I can normally spot sectioning. I take the best spot for each section creating plates 1/1/1/X, 1/1/2/X, 1/2/1/X, and 1/2/2/X Sometimes ending up with 16 plates going of isolates.

I let them grow out to 1/2 of the plate size then use a sterile syringe with distilled water in it. Open the plate under the victual flow hood and squirt 1cc of water into the plate. Scrape some of the growth loose into the water and suck it back up into the syringe. I them inject it into a LC jar and let it grow out.

Once I have all of the isolates in LC jars I will take syringes and make 1 BRF jar for each. Grow it out keeping a record of everything picked from the brf cake including wet weight and dry weight.

The best performer is the one I keep. I go back through the lab fridge and discard everything other than the selected plate and LC jar.

I then take a culture tube with MDA in it and transfer from the agar plate to the culture tube. Once it starts to grow out I put it in the fridge for longer term storage.

Now, I will grow from the LC and replenish the LC from the selected strain in the petri dish. However, the growth in the petri dish will use up the nutrients and agar within 4 months so you have to replenish with new dishes grown from samples taken from the culture tube.

So in the end I will go through about 25 perti dishes of PDA, 12 to 16 LC jars, 12 to 16 BRF cakes, and 1 culture tube.

However, when you find the right one the lbs per square foot of fruiting area goes up quite a bit :grin:


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Re: agar--isolation--senescence---lets talk turkey [Re: Cyber]
    #16554984 -

thats the answer i was lookin for---thanks


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Mushrooms, Mycology and Psychedelics > Advanced Mycology
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