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Workman
1999 Spore War Veteran
Registered: 03/01/01
Posts: 3,601
Loc: Oregon, USA
Last seen: 6 hours, 6 minutes
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PF Albino and Penis Envy Cross Breeding (updated 6-5-06 with F3 picture) 9
#5194890 - 01/18/06 12:42 PM (18 years, 2 months ago) |
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This experiment is ongoing.
PF Albino crossed with Penis Envy = F1 generation. No albino or Penis shaped caps expected at this stage. The next step (in progress) is to self (normal multispore culture) the F1 spores to see if any recessive traits can be revealed. The F2 generation is where the real action is.
Interestingly, the F1 culture seems to be showing hybrid vigor with very aggressive mycelium growth and fast mushroom growth.
+ = mature
Spores from F1 generation selfed to produce F2 generation.
+ =
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And now a little bonus
PF albino crossed with Costa Rican cubensis. I actually did this experiment first so its a little farther along.
+ =
Backcross the F1 generation spores with the PF Albino monospore culture
+=
Although backcrossing did reveal some albino mushrooms, the culture overall is sluggish and prone to aborts. ----------------------------------------------------------------------------
Method:
First I germinated a single PF Albino spore on malt agar to produce a monokaryotic culture. Dikaryotic cultures form clamp connections that are easily visible when magnified and this culture lacked these distictive clamps.
I am pretty lazy and I didn't want to spend time isolating more single spores from other cubes and then do several controlled crosses and hope for the best. I was also aware of a mechanism where a monokaryotic mycelium can become a dikaryotic mycelium by nuclear migration and not just at the new growth at the point of mycelial contact. Essentially the entire monokaryotic culture becomes dikaryotic by replicating and moving nuclei in a sort of a chain reaction through the already existing mycelial network. Sooooooo.......... here is what I did.
I put the monospore PF albino culture into a jar of sterile grain (grass seed in this case) and let it colonize almost completely. Every grain was covered with mycelium but it would need a few more days to get really dense. Then I made a fresh solution (adding just dry spores also worked) of normally pigmented cubensis spores (Penis Envy or Costa Rican), injected that into the nearly colonized grain and then shook the jar to mix. The idea is to deny any uncolonized substrate to the injected cubensis spores for colonization if it combined (mated) with other cubensis spores. In theory, the cubensis spores will germinate and some will combine with other nearby spores due to clumping, but most will combine with the PF Albino mycelium. Nuclear migration should then transform the entire jar of monokaryotic Albino mycelium into several strains of dikaryotic Albino X (Penis Envy or Costa Rican) Cubensis mycelium. There shouldn't be any room left for any significant amount of pure Penis Envy or Costa Rican dikaryotic mycelium to grow and later form mushrooms.
Edited by Workman (06/05/06 10:26 AM)
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Lana
Head Banana
Registered: 10/27/99
Posts: 3,109
Loc: www.MycoSupply.com
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Re: PF Albino and Penis Envy Cross Breeding [Re: Workman]
#5194899 - 01/18/06 12:45 PM (18 years, 2 months ago) |
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Nice work...
Lana
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ohmatic
searcher
Registered: 02/28/04
Posts: 6,742
Loc: europe
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Re: PF Albino and Penis Envy Cross Breeding [Re: Lana]
#5194927 - 01/18/06 12:55 PM (18 years, 2 months ago) |
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how did you do that ? peace ohm
-------------------- MONOTUB tek HEATBOMB tek RIP #cultivation! ....can't associate? well FUCK U !
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Trunkspace
Televangelist
Registered: 07/11/05
Posts: 87
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Re: PF Albino and Penis Envy Cross Breeding [Re: ohmatic]
#5194962 - 01/18/06 01:08 PM (18 years, 2 months ago) |
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Wow.. very nice work, it's too bad the albino/PE seemingly lost both traits. Perhaps later generations could show both phenotypes. Interesting that the albino trait emerged with the Costa Ricans though. Fascinating, Workman.
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waixingren
Registered: 03/14/05
Posts: 2,644
Loc: SW Florida
Last seen: 2 months, 7 days
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Re: PF Albino and Penis Envy Cross Breeding [Re: Workman]
#5195039 - 01/18/06 01:33 PM (18 years, 2 months ago) |
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great.
--waixingren
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ohmatic
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Registered: 02/28/04
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Re: PF Albino and Penis Envy Cross Breeding [Re: waixingren]
#5195059 - 01/18/06 01:37 PM (18 years, 2 months ago) |
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hmm, why didnt you try to mix monocaryotic cultures of PE and PF ? that's atleast what i would have tried if i had the equipment to do monocaryotic cultures. peace ohm
-------------------- MONOTUB tek HEATBOMB tek RIP #cultivation! ....can't associate? well FUCK U !
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Workman
1999 Spore War Veteran
Registered: 03/01/01
Posts: 3,601
Loc: Oregon, USA
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Re: PF Albino and Penis Envy Cross Breeding [Re: ohmatic]
#5195108 - 01/18/06 01:50 PM (18 years, 2 months ago) |
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Currently I only have the one monospore culture (PF Albino). They are a bit of a pain in the ass to generate and I was looking for a down and dirty short cut to get some F1 material. I am doing a multispore self cross with the F1 (PE x Albino) but I am considering putting the extra effort into generating several F1 lines for controlled crosses. The ultimate goal of the PE x Albino work is to generate an Albino PE.
I was suprised at how normal the F1 PE x Albino mushrooms look. Since both strains are mutated in some manner you would expect at least some problems with crossing them. Apparently Penis shaped caps and Albinism are both recessive traits. I expect the F2 generation to be filled with monsters, but we shall see.
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Edited by Workman (01/18/06 02:13 PM)
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Trunkspace
Televangelist
Registered: 07/11/05
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Re: PF Albino and Penis Envy Cross Breeding [Re: Workman]
#5195212 - 01/18/06 02:20 PM (18 years, 2 months ago) |
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Keep us updated, I can't wait to se the F2 PE/Albino generation.
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Pinback
Stranger
Registered: 07/20/02
Posts: 836
Loc: Europe
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Re: PF Albino and Penis Envy Cross Breeding [Re: Workman]
#5195256 - 01/18/06 02:40 PM (18 years, 2 months ago) |
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Very interesting. For how long did you incubate the jars after you injected the spores?
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Shdwstr
FSRCanada
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Re: PF Albino and Penis Envy Cross Breeding [Re: Workman]
#5195315 - 01/18/06 02:59 PM (18 years, 2 months ago) |
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Next step is to cross a SRA and PE and get a 3 foot tall Penis... Now that's something to ENVY
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Workman
1999 Spore War Veteran
Registered: 03/01/01
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Loc: Oregon, USA
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Re: PF Albino and Penis Envy Cross Breeding [Re: Pinback]
#5195334 - 01/18/06 03:05 PM (18 years, 2 months ago) |
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Quote:
Very interesting. For how long did you incubate the jars after you injected the spores?
About a week, but the spores were really fresh. I'd probably go at least 10 days with older spores that might take longer to germinate. Nuclear migration may continue after placing in a fruiting tray, but I didn't see that as a problem.
-------------------- Research funded by the patrons of The Spore Works Exotic Spore Supply My Instagram Reinvesting 25% of Sales Towards Basic Research and Species Identification
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armedia
Dikaryon
Registered: 11/03/01
Posts: 169
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Re: PF Albino and Penis Envy Cross Breeding [Re: Workman]
#5196150 - 01/18/06 06:44 PM (18 years, 2 months ago) |
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Workman -
I'm sure many here would like to hear your method of single spore isolations. Please share your secrets, if you are willing.
- A
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blackout
Registered: 07/16/00
Posts: 5,266
Last seen: 4 months, 22 days
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Re: PF Albino and Penis Envy Cross Breeding [Re: Shdwstr]
#5197860 - 01/19/06 03:26 AM (18 years, 2 months ago) |
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Quote:
Shdwstr said: Next step is to cross a SRA and PE and get a 3 foot tall Penis... Now that's something to ENVY
Too right! It is good to see 2 such distinct strains so the results are really obvious. Many were doubting Rogers crossings since it is hard to see the difference, hopefully this will sway the doubters and encourage more experiments.
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ohmatic
searcher
Registered: 02/28/04
Posts: 6,742
Loc: europe
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Re: PF Albino and Penis Envy Cross Breeding [Re: armedia]
#5197862 - 01/19/06 03:30 AM (18 years, 2 months ago) |
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Quote:
armedia said: Workman -
I'm sure many here would like to hear your method of single spore isolations. Please share your secrets, if you are willing.
- A
hes got a lab to work with, and that is what you need in order to create monocaryotic cultures, a lab.
no way to do it at home - therefor there are no secrets. peace ohm
-------------------- MONOTUB tek HEATBOMB tek RIP #cultivation! ....can't associate? well FUCK U !
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armedia
Dikaryon
Registered: 11/03/01
Posts: 169
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Re: PF Albino and Penis Envy Cross Breeding [Re: ohmatic]
#5197879 - 01/19/06 04:04 AM (18 years, 2 months ago) |
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Ohm -
I was being ironic when I used the term "secret". But I disagree that single spore isolations can't be done at home. I think a fair number here have "labs" of one kind of another in their homes, or elsewhere. The method may require some fancy equipment, but it is not that complicated.
The traditional method involves doing serial dilutions of spore suspensions on to agar, observing individual spores or colonies on the plate through a high power microscope, and transferring true monokaryons to a clean plate. I was merely curious to know the specifics of Workman's method.
Peace A
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blackout
Registered: 07/16/00
Posts: 5,266
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Re: PF Albino and Penis Envy Cross Breeding [Re: ohmatic]
#5197904 - 01/19/06 04:56 AM (18 years, 2 months ago) |
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Quote:
ohmatic said: no way to do it at home - therefor there are no secrets.
I thought it was possible too, I think there is a small bit in TMC about it. Wasnt it selecting strands that show no splitting/branching?
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RogerRabbit
Bans for Pleasure
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Loc: Seattle
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Re: PF Albino and Penis Envy Cross Breeding [Re: blackout]
#5197952 - 01/19/06 05:53 AM (18 years, 2 months ago) |
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Nice stuff workman. The PE6 displays much the same results. It's a cross of texan and PE, but most fruits look normal, while occasionally a flush comes out that looks like pure PE. RR
-------------------- Download Let's Grow Mushrooms semper in excretia sumus solim profundum variat "I've never had a failed experiment. I've only discovered 10,000 methods which do not work." Thomas Edison
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blackout
Registered: 07/16/00
Posts: 5,266
Last seen: 4 months, 22 days
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Re: PF Albino and Penis Envy Cross Breeding [Re: RogerRabbit]
#5197983 - 01/19/06 06:35 AM (18 years, 2 months ago) |
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Roger, if you clone a PE looking shroom from a PE6 grow, do they all end up looking like PE. Also do you notice the difference in effect that Ryche Hawk mentions with normal PE, i.e. do the PE6 shrooms have the same wavy high, the normal ones and/or the PE looking ones.
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Cyano
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Re: PF Albino and Penis Envy Cross Breeding [Re: blackout]
#5198629 - 01/19/06 11:36 AM (18 years, 2 months ago) |
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If a Cubensis is a Cubensis, why do different strains refuse to mate with each other? Does the fact that one needs to perform these complicated procedures to cross two strains not prove that not all Cubensis strains are alike?
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armedia
Dikaryon
Registered: 11/03/01
Posts: 169
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Single spore isolates [Re: blackout]
#5198671 - 01/19/06 11:46 AM (18 years, 2 months ago) |
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It depends upon whether you isolate before or after spore germination. You can visualize individual spores in the scope immediately upon dilution and place the on new plates, or wait until colonies germinate, look for the absence of clamps, and transfer those colonies (which is what Workman apparently did).
You can also work out the appropriate number of dilutions from a stock spore solution that will give you so many spores per drop.
Here's one good method: http://ejournal.sinica.edu.tw/bbas/content/1997/1/bot381-06.pdf
- A
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