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mycocurious
Mike O. Kuerias



Registered: 02/09/07
Posts: 1,265
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Very Slow Going...
#7611709 - 11/08/07 03:31 PM (16 years, 2 months ago) |
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For ease of reading, all environmental and other related variables have been listed below...
Last summer I created a master-slant from a single-sector isolate of a B+ strain fruited from a multispore. Back in middle of July of this year I used that slant to inoculate several agar dishes, and over the course of two transfers isolated what was the thickest, most rhizomorphic sector of that growth. I grew it out onto several 3-sector partitioned petri dishes and by the end of August had the wedges colonized to the point of being able to use them to inoculate rye grains as a spawn medium.
This is where it all gets interesting...ly slow going. While I don't have a large number of wedge-to-grain inoculations under my belt, especially with cubensis, this still seemed an abnormally long time for incubation. I didn't have 25% colonization for the first shake until the 10th day, with the remaining "shakes" over the next ten days as colonization picked up significantly. At that point I figured it was just a slow starter as the growth exhibited in the jars was just as rhizomorphic as I seen on the inoculate wedges used and it seemed to be picking up speed as would normally be expected with each subsequent shake.
The whole point of the experiment was to document the differences between a standard recipe of amended coco-coir as a substrate compared to a homemade compost from an aerobic composting project. So I prepared a single tray of each as well as several trays of various blends of both types of substrates. All "blended" trays were lost to fermentation but the pure compost and pure coir survived...reasons still unknown. I'm not going to rule out the possibility that the substrates were over-hydrated beyond field capacity but that's not a mistake I often make and I check it before and after the steam-pasteurization.
So up to this point I wrote off the initial delay in the spawn medium as genetics and being a slow-starter and even began to believe that my substrates probably were too wet when spawned, however when I added the casing layer it took another 12 days before I could spot the first indication of a hypheal knot forming under 20x magnification...I have no experience with that at all. Normally my casing layers only takes about 5 days tops until I can spot a knot or two.
At that point it's exposed to the fruiting chamber under the environmental conditions listed below...and nine days of keeping the casing layer at the very-near saturation point per Hyphea's Pinning Strategy and those few knots finally became pins visible without magnification. As with everything else, the maturing pins took almost another two weeks to fully mature. In terms of health and quality, the fruits are not weak bodied or otherwise show any sign of weak-genetics but if you consider the poor first pinset and the 72 day long period it took from inoculation to flush...it is curious.
------------------ Or it is simply that my previous grows with this species and strain have been faster and this is my first "typical" timeline? Still it makes no sense, I was under the impression that rhizomorphic growth tended to be faster colonizers... ------------------
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Timelines: Colonization Of Rye Spawn: 20 days Colonization Of Bulk Substrates: 18 days Colonization Of Casing Layer: 12 days (measured until hypheal knots could be seen under 20x magnification) Days Spent Inducing Pinning: 9 days (until those first hypheal knots begin to show as pins) From Pin To Mature Fruit: 12 days
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Material Variables: Spawn Medium: Rye Grains @ 70% capacity per quart sized jar. Inoculate: Malt-Extract Agar Wedge containing single-sector isolate. Spawn Ratio: 4:1 substrate to spawn (6 quarts substrate to 1.5 quarts spawn) Incubation: 73-76(F) for both spawn and substrate incubation periods. Bulk Tray: Dollar store 12q dishpans with several even coats of a black vinyl dye on it's exterior to prevent excessive side-pinning. Compost Substrate: 6 quarts of homemade compost amended 5% gypsum by volume. Coir Substrate: 6 quarts of coco-coir. Amendments: (by volume) 30% used coffee grounds, 5% gypsum, 15% vermiculite. Casing Layer:5/8" Layer of Jiffy Mix Seed-Starting Mixture (a standard peat/verm mix) pH balanced to 7.2 using pickling lime and steam-pasteurized.
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Fruiting Chamber Environmentals... Air Exchanges: Entire chamber completely cycles it's air every five minutes using a constant-operation blower fan @ 25CFM through the humidifier. Lighting: 12/12 cycling using three 6500K (daylight color) fluorescent bulbs. Temperature: Controlled cycles between 70(F) in the early morning rising to 78(F) in the late evening corresponding with natural circadian cycles. Humidity: Constant at 92-99% with the low point being during the three "hotest" hours in the day.
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Don't mistake my tone for a "matter-of-fact" attitude. I'm just presenting what I believe to be correct, until I'm corrected... - How Myco-Curious Prepares Coir & Compost Substrates - How Myco-Curious Builds A Bulk Humidifier - How Myco-Curious Builds An Automated Greenhouse ------------------------------------ figgusfiddus said: Keep in mind that inoculating or whatever in front of a flow hood won't help your bad substrate, your bad inoculant, your bad sterile procedure, etc. etc. etc. It's not a +3 flowhood of magic, it's just a tool.
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c0_hush
Stranger



Registered: 07/16/06
Posts: 417
Last seen: 12 years, 4 months
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have you tried incubating at a higher temp?
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mycocurious
Mike O. Kuerias



Registered: 02/09/07
Posts: 1,265
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Re: Very Slow Going... [Re: c0_hush]
#7611986 - 11/08/07 04:46 PM (16 years, 2 months ago) |
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lol, yes, I've incubated cubensis several times in the 78-80(F) range and colonization times were roughly a day or three sooner. Keep in mind however, that most colonization times for my rye grains are 13 days...not 20.
It's fascinating in it's own right but I'm interested to see if Monstermitch, RoadKill or RogerRabbit have any input on the topics, especially since it wasn't a multispore - which is my typical answer for these type questions.
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Don't mistake my tone for a "matter-of-fact" attitude. I'm just presenting what I believe to be correct, until I'm corrected... - How Myco-Curious Prepares Coir & Compost Substrates - How Myco-Curious Builds A Bulk Humidifier - How Myco-Curious Builds An Automated Greenhouse ------------------------------------ figgusfiddus said: Keep in mind that inoculating or whatever in front of a flow hood won't help your bad substrate, your bad inoculant, your bad sterile procedure, etc. etc. etc. It's not a +3 flowhood of magic, it's just a tool.
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