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Serani
Stranger
Registered: 03/05/06
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Last seen: 18 years, 21 days
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Too far-fetched?
#5564488 - 04/27/06 08:41 PM (18 years, 26 days ago) |
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I had an idea that I wanted to run: a cross between a non-psilocybin/cin containing mushroom and one with;
For instance a mix of P. Cubensis and Pleurotus ostreatus
And why? Well P. Cubensis of course contatining the psycoactive ingrediant and Pleurotus ostreatus because it grows so well, so fast (being easier to cultivate than even P. Cubensis) and that it is so resilliant to many bacteria.
This isn't the best example to portray my thoughts, but maybe to produce a psycoatctive strand that was bioluminescent? Any of which just to expand the grounds.
Is an of this feasible in any sense?
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shroomydan
exshroomerite


Registered: 07/04/04
Posts: 4,126
Loc: In the woods
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Re: Too far-fetched? [Re: Serani]
#5564704 - 04/27/06 09:37 PM (18 years, 26 days ago) |
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Quote:
a psycoatctive strand that was bioluminescent
That would be cool!
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whatever123
Whatever I did, I'm sorry


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Re: Too far-fetched? [Re: shroomydan]
#5566319 - 04/28/06 09:01 AM (18 years, 25 days ago) |
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Yea, I have wanted to see more plants(never thought of bioluminescent fungi, except for the Jack-O-Lantern) like that be genetically altered and have a bioluminescent bacteria/gene put into them. Glow in the dark buds, GiD flowers....use your imagination!
Good luck on the cross-breed. I'd like to see the results of it, because even if it does not work as planned, it is always good to experiment!
-------------------- Koala Koolio said: there should be a 3 month waiting period between registration and posting.
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Serani
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Registered: 03/05/06
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Yea I am setting up my new laboratory this weekened-ish and I am hoping to do some work on it, I'll be sure to post logs with pictures of course; just wanted to see if anyone has tampered with the thought at all.
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RogerRabbit
Bans for Pleasure


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Re: Too far-fetched? [Re: Serani]
#5567080 - 04/28/06 01:51 PM (18 years, 25 days ago) |
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I've made many attempts at crossing azurescens with woodlovers of every species I have cultures for, including the conks such as Ganoderma, and edibles from oysters to shiitake. So far, no success. I'm sure it's just a matter of time before somebody finds the magic bullet and pulls it off. RR
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Serani
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Yes, what methods or such are you working with in your attempts?
Maybe provide a partial check-list of some things I am thinking of that you've already done away with.
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Zen Peddler


Registered: 06/18/01
Posts: 6,379
Loc: orbit
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Re: Too far-fetched? [Re: Serani]
#5583746 - 05/02/06 10:35 PM (18 years, 20 days ago) |
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Arent the properties that make mushrooms glow actually toxic?
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Feelers
Anti-Myth-Rhythm-Rock-Shocker


Registered: 06/18/02
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Are you guys just crossing? I have come into a wee bit of money - and I'm looking to be a freelance "biologist inventer". I have a current project with salvia divinorum thats in the planning stages, but I also want to move onto fungi too.
I believe I might be able to do some protoplast fusions when I get my act together (talked to my lecturer he said it was feasable). Perhaps edibles mixed with edibles, or actives mixed with edibles(as suggested). However one thing that should be looked at is that the fusions have a better success rate the closer they are related in the first place. Anyone know much about this process and common problems?
I was thinking a azure/cubensis cross would be cool, or perhaps a panaleous cubensis cross. So many possibilities. After this year I'll have freedom to try this crap.
Or how about a fungi/plant hybrid - cacto-cubensis. I love this shit!!! When GM comes into public acceptance theres gonna be some amazing things.
Ok so first up what are some very closely related edibles to the psilocybes?
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Zen Peddler


Registered: 06/18/01
Posts: 6,379
Loc: orbit
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Re: Too far-fetched? [Re: Feelers]
#5584029 - 05/02/06 11:17 PM (18 years, 20 days ago) |
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The problem is that mushrooms are particularly resistant to cross-breeding.
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fastfred
Old Hand



Registered: 05/17/04
Posts: 6,899
Loc: Dark side of the moon
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Re: Too far-fetched? [Re: Feelers]
#5584282 - 05/03/06 12:19 AM (18 years, 20 days ago) |
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Feelers said: I believe I might be able to do some protoplast fusions when I get my act together (talked to my lecturer he said it was feasible).
Wow, I've sort of dreaded hearing "protoplast fusion" mentioned here. It means I'll have some competition to be the first to do it successfully...
Protoplast fusion is the only way I can see any possibility of doing the crosses suggested. There is really no other way to cross different species. It's a very powerful tool and there are reports of successful inter-species, inter-genera, and even (gasp) inter-phylum crosses. I haven't heard of any inter-kingdom grosses though, so the fungi-plant is probably not possible.
Quote:
Anyone know much about this process and common problems?
Well... I could discourse for several pages, but rather I'll suggest a book: Fungal Protoplast: A Biotechnological Tool by: D. Lalithakumari
It's a must read if you're interested in the subject. The author's english isn't the best, but he's compiled many, many studies and charted and graphed them and discusses general principles, enzyme concentrations, times, temperatures, media, etc.
The long and short of it is that it's a simple idea with relatively simple procedures, but also quite difficult. If you plan to just bang your head and brute force it, you won't get very far. There is a lot of thought, planing, and strategy involved. I'd be happy to offer any advice you might need and/or collaborate.
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When GM comes into public acceptance theres gonna be some amazing things.
GM has defacto acceptance despite what some ignorant wackos would have you think. The people out there protesting are just too behind the times to realize that a lot of what they are already eating is GM. Most of the processed food sold today is GM. I've heard the figure of 80% (of processed food) thrown out there quite a bit. Stuff like ketchup, corn syrup, corn meal, etc. has had a lot of GM crops in it for quite some time.
But regardless, one upshot of protoplast fusion is that the regenerants aren't considered to be GMOs!
Quote:
Ok so first up what are some very closely related edibles to the psilocybes?
Stropharia rugoso-annulata (King stropharia, Garden Giant) comes to mind.
-------------------- It drinks the alcohol and abstains from the weed or else it gets the hose again. -Chemy The difference between the substances doesn't matter. This is a war on consciousness, on our right to the very essence of what we are. With no control over that, we have no need to speak of freedom or a free society. -fireseed "If we are going to have a war on marijuana, the least we can do is pull the sick and the dying off the battlefield." -Neal Levine (MPP) I find the whole "my drug should be legal but yours should be illegal" mindset disgusting and hypocritical. It's what George Bush and company do when they drink a cocktail and debate the best way to imprison marijuana users. -Diploid
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Feelers
Anti-Myth-Rhythm-Rock-Shocker


Registered: 06/18/02
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Re: Too far-fetched? [Re: fastfred]
#5584337 - 05/03/06 12:50 AM (18 years, 20 days ago) |
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You can have inter kingdom fusions! (I think animal and fungi have been joined) obviously its not gonna be very stable - and not much use.
Yeah the reason I'm looking at it is because its not considered GM - (there's effectivly a ban on GM here), and it's such a powerful tool. The enzymes involved apparantly cost in the hundreds of dollars, so not too bad.
I know very little specifics on the subject, other than a basic overveiw. I do know its not as complicated as many would think. Looking for the "clamping" etc might take a while and some experience. I read a good account of a successful fungi fusion - but I cant find what it was.
I have some crazy idea's I wanna market. (this is more of a career plan)
Actually I have a class next semmester and I could probably use the uni equipment to do a fusion for a project. Hmmmm
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fastfred
Old Hand



Registered: 05/17/04
Posts: 6,899
Loc: Dark side of the moon
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Re: Too far-fetched? [Re: Feelers]
#5584505 - 05/03/06 04:42 AM (18 years, 20 days ago) |
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You can get 5 units of chitinase for $40.00 from Sigma. Your mileage may vary, but if you go for 48 hours instead of 2 you will get around 50 units worth out of it. You can look up how many mg of chitin that will digest on Sigma's website.
-FF
-------------------- It drinks the alcohol and abstains from the weed or else it gets the hose again. -Chemy The difference between the substances doesn't matter. This is a war on consciousness, on our right to the very essence of what we are. With no control over that, we have no need to speak of freedom or a free society. -fireseed "If we are going to have a war on marijuana, the least we can do is pull the sick and the dying off the battlefield." -Neal Levine (MPP) I find the whole "my drug should be legal but yours should be illegal" mindset disgusting and hypocritical. It's what George Bush and company do when they drink a cocktail and debate the best way to imprison marijuana users. -Diploid
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thenewguy05
The Mushroom Man


Registered: 02/11/05
Posts: 2,123
Loc: My Underground Layer
Last seen: 2 years, 9 months
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Quote:
RogerRabbit said: I've made many attempts at crossing azurescens with woodlovers of every species I have cultures for, including the conks such as Ganoderma, and edibles from oysters to shiitake. So far, no success. I'm sure it's just a matter of time before somebody finds the magic bullet and pulls it off. RR
are azures a 2 clamp or a 4???
i've always thought it would be nice if you could cross the azures with some dung lover to get the intenceity of the azures without haveing to mess with wood. not that wood is a pain in the ass it's just that i have so much dung already. to get wood I have to take a special trip up north.
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Workman
1999 Spore War Veteran


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Re: Too far-fetched? [Re: fastfred]
#5585652 - 05/03/06 12:42 PM (18 years, 20 days ago) |
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Oh man, now the race is on. Sporeworks.com is providing me with materials for protoplast fusion experiments as well.
-------------------- 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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thenewguy05
The Mushroom Man


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Re: Too far-fetched? [Re: Workman]
#5585666 - 05/03/06 12:45 PM (18 years, 20 days ago) |
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NICE!!!
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fastfred
Old Hand



Registered: 05/17/04
Posts: 6,899
Loc: Dark side of the moon
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Re: Too far-fetched? [Re: Workman]
#5586722 - 05/03/06 05:09 PM (18 years, 20 days ago) |
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Quote:
Workman said: Oh man, now the race is on. Sporeworks.com is providing me with materials for protoplast fusion experiments as well.
A race indeed! I don't really have the international resources to pursue it legally, so I'm only working on oysters right now. I probably won't have any fusion products for about a year. I'm working on creating auxotrophs through induced mutation right now. I'd be happy to collaborate with you though. I have the book I recommended earlier in the thread if you need any specific info or data.
One of the first steps that need to be sorted out is how to select fusion products. You can use auxotrophic mutants, drug resistance/antibiotic markers, fungicide resistance markers, inactivated protoplasts/dead donors, colony morphology, and use of irreversible biochemical markers. Those are the most useful methods mentioned in the book. Colony morphology isn't that useful as a primary selection method. The auxotroph method seems most useful to me. Fusion partners are selected from mutants auxotrophic for different amino acids, that way when you generate the protoplasts and fuse them only the fusants will survive to regenerate cell walls. Nutritional complementation is what it's called.
Anyway, that's my two cents. Good luck hunting for auxotrophs. It's tedious work, but there are ways to speed it up tremendously.
-FF
-------------------- It drinks the alcohol and abstains from the weed or else it gets the hose again. -Chemy The difference between the substances doesn't matter. This is a war on consciousness, on our right to the very essence of what we are. With no control over that, we have no need to speak of freedom or a free society. -fireseed "If we are going to have a war on marijuana, the least we can do is pull the sick and the dying off the battlefield." -Neal Levine (MPP) I find the whole "my drug should be legal but yours should be illegal" mindset disgusting and hypocritical. It's what George Bush and company do when they drink a cocktail and debate the best way to imprison marijuana users. -Diploid
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Feelers
Anti-Myth-Rhythm-Rock-Shocker


Registered: 06/18/02
Posts: 1,806
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Re: Too far-fetched? [Re: fastfred]
#5586916 - 05/03/06 05:56 PM (18 years, 20 days ago) |
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The paper I read about it looked for myceial clamping - it stated that this is not necessarily the most efficient way - I think they found 4 clamps out of about 500 possible clapms that fit the description - and they cultured them. 2 were sucessful.
Interesting there was a big difference between the fusonts(is that the word for them?) , with both having varying traits of both species.
How do you find mutants that cant process certain aminos? I think I would try resistance to antibiotics - at least its easy to find the ones that survive.
Something you guys might be interested in - the NZ government gives grants out for specific questions that businesses have - for example if I were to say - Can you make a shitakki/button mushroom fusant using the following method - the government will pay up to $2000 for it to be done at the university.
It's designed to help out small busines, but is only open to graduates. I think its actually easily to get approved - I was asking my leceturer about my salvia experiment - and apparantly that could get funding(he said he could approve it)!
Now how cool is that? Obviously it means the active variets are off limits - but theres plenty of other things out there.
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Workman
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Re: Too far-fetched? [Re: fastfred]
#5592033 - 05/04/06 10:32 PM (18 years, 18 days ago) |
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I was thinking more along the lines of monokaryotic mycelium of two different species, that are incapable of fruiting, fused together. Since the point is to produce mushrooms, any fusants that fruited should be competent hybrids. I have no plans to complicate the procedure more than necessary with auxotrophic mutants and selective media.
-------------------- 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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Feelers
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Re: Too far-fetched? [Re: Workman]
#5592120 - 05/04/06 11:06 PM (18 years, 18 days ago) |
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I thought monokaryons could fruit? (sorry I'm a bit behind - any good online texts that go into a bit of overall detail on fungi?)
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fastfred
Old Hand



Registered: 05/17/04
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Loc: Dark side of the moon
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Re: Too far-fetched? [Re: Feelers]
#5593099 - 05/05/06 07:32 AM (18 years, 18 days ago) |
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That might work Workman. It would also eliminate the problem of fusants that can't fruit. On the other hand you'd have the problem of non-fused mycelium out competing viable fusants before they get a chance to fruit. Fusants usually aren't very vigorous and would likely never gain enough territory to produce fruits.
Feelers asked about how to find auxotrophs... The often used and tedious way is to induce mutation, plate the cells on standard media, replica plate them to minimal media, observe the ones that don't grow, then go back to your standard media and pick out those colonies.
Here is the procedure I've devised based on newer methods. Mutate, grow in liquid media, blenderize until you have 1-3 cell size pieces, allow media a few hours to recover, filter to remove larger pieces, filter and wash again with a finer filter to remove all cells, transfer those cells to minimal media, incubate for a day or two, filter to remove any cells that were able to grow in the minimal media, collect filtrate and transfer to standard media, then repeat the procedure or plate on standard media, then test colonies to find auxotrophs.
That may sound like a lot of work, but it's really only a couple of flirtations and media transfers. In this way you can screen thousands or even tens of thousands of colonies at once with minimal work. You have to look at where to invest your limited resources. You can spend more time in design and preparation that will save you work later, or you can jump right in and then spend all your time later trying to screen thousands of potential fusants later. These are just my ideas, the only real way to figure out the best procedure is to jump right in and find out the problems in your procedure. Then you can redesign and try again.
Good idea about getting grants. If you can get access to advanced labs through the program then the actual costs wouldn't be very much. Most of the costs are in the equipment. Paying researchers would be the largest cost if you could get access to the equipment. Maybe the program uses undergraduate researchers, if so $2000 could probably produce plenty of fusants.
Monokaryons don't fruit that I know of. There would be no point since they can't undergo meiosis since they don't have a full set of chromosomes.
What would be your goal with fusing salvia? I don't really know much about salvia.
-FF
-------------------- It drinks the alcohol and abstains from the weed or else it gets the hose again. -Chemy The difference between the substances doesn't matter. This is a war on consciousness, on our right to the very essence of what we are. With no control over that, we have no need to speak of freedom or a free society. -fireseed "If we are going to have a war on marijuana, the least we can do is pull the sick and the dying off the battlefield." -Neal Levine (MPP) I find the whole "my drug should be legal but yours should be illegal" mindset disgusting and hypocritical. It's what George Bush and company do when they drink a cocktail and debate the best way to imprison marijuana users. -Diploid
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Feelers
Anti-Myth-Rhythm-Rock-Shocker


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Re: Too far-fetched? [Re: fastfred]
#5594600 - 05/05/06 03:05 PM (18 years, 18 days ago) |
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I'm making a polyploid of salvia - it will hopefully reinstall its fertility (my lecturer thinks this will happen too) and it will be grow bigger, faster and more potent ect.
The thing is you cant use cholchine - as there arent many seeds around and if you can get them it'd cost thousands.
But as for this mushroom fusants, tell me what "crosses" would people like to see? After some suggestions I'll email my lecturer and see if I can do a fusion for my mycology project.  I'm thinking since all mycelium looks pretty much the same - it probably wouldnt matter if an "active" made it into the mix. haha.
Say, what happens if you fuse dikaryons? Do mushrooms function ok/better as polyploids?
Ok so what are some closely related species of edible easy to grow mushrooms to the actives?
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BlimeyGrimey
Collector of Spores



Registered: 08/24/05
Posts: 3,799
Loc: Puget Sound
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Re: Too far-fetched? [Re: Feelers]
#5600019 - 05/07/06 02:30 AM (18 years, 16 days ago) |
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there are some Psilocybe species that ARENT active, and remember you could also try an active-Conocybe/non-active-conocybe mix etc. Breeding within the same genus seems like it will make the task easier to me.
here's a cross i'd like to see.
Gymnopilus ventricosus x Gymnopilus spectabilis
spectabilis is active and ventricosus are huge. seems like a good combo to me.
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Feelers
Anti-Myth-Rhythm-Rock-Shocker


Registered: 06/18/02
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Can you get a monokayotic culture from dikaryotic mycellia? I'm asking because say I dont have access to spores how can I get a monokaryon from a clonal culture. Is this possible? I'm geussing it isnt?
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Workman
1999 Spore War Veteran


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Re: Too far-fetched? [Re: Feelers]
#5604306 - 05/08/06 09:18 AM (18 years, 15 days ago) |
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It can happen spontanously, but I am not certain if there is a method to induce it. I haven't really looked into that in detail since the isolation of single spores is pretty straight forward.
-------------------- 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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Feelers
Anti-Myth-Rhythm-Rock-Shocker


Registered: 06/18/02
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Re: Too far-fetched? [Re: Workman]
#5605975 - 05/08/06 06:02 PM (18 years, 15 days ago) |
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Well I have emailed my lecturer if I can do a protoplast fusion, so I'm just waiting for a response. The cool thing is he'll be real stoked someone wants to do such an interesting project, so he'll probably help me a lot.
Crazy guy my lecturer - he's in a ZZ TOP live video climbing the wall smashed on acid.  Now he's a respectable looking man with glasses and white hair. Definately my favourite lecturer. I might point him in the shroomerys direction.
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fastfred
Old Hand



Registered: 05/17/04
Posts: 6,899
Loc: Dark side of the moon
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Re: Too far-fetched? [Re: Feelers]
#5631173 - 05/15/06 02:15 AM (18 years, 8 days ago) |
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You asked about haploidization... Haploidization can be caused by many factors. Errors in cell division, stressful environment, lack of nutrients, chemicals, etc.. The standard way to induce it is by using benomyl.
Here is a quote... "...colonies can be induced to undergo haploidization by treatment with parafluorophenylalinine, of benomyl, UV or X-rays. (Stipiczki and Ferenczy, 1977; Peberdy et al., 1977; Sarachek et al., 1981)."
Sorry for not providing a complete citation, if you're really interested I can look it up for you.
-FF
-------------------- It drinks the alcohol and abstains from the weed or else it gets the hose again. -Chemy The difference between the substances doesn't matter. This is a war on consciousness, on our right to the very essence of what we are. With no control over that, we have no need to speak of freedom or a free society. -fireseed "If we are going to have a war on marijuana, the least we can do is pull the sick and the dying off the battlefield." -Neal Levine (MPP) I find the whole "my drug should be legal but yours should be illegal" mindset disgusting and hypocritical. It's what George Bush and company do when they drink a cocktail and debate the best way to imprison marijuana users. -Diploid
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