Alright Hr you probably dont know me but i want to tell you now that im silly. I am all over the place at times and i am not ready to actual ready to attempt hybridization. I want to get 2 different monocultures each with very obvious phenotypes.
I am aware that hybridization can result and although both parents genotypes are present it is possible that the phenotype may resemble that of a single parent but it is also possible that they can resemble both, which i would hope for. I believe this is a result of the weaker genetics conceding to the more dominant just as humans do. 1 of my kids looks exactly like me while 3 resemble both of us and 1 exactly their mother.
I know making babies isnt hybridization but i use that as a reference for genotypes and phenotypes.
I believe the inly true way to tell if you have successfully mated to monokaryons is to view under a scope and look at the clamp connections. I have the appropriate stains and scope to observe this i think.
Now we could wait and see how it fruits, and if it clearly has the phenotypes of both parents, then you can rule that a success but if they show the phenotype of 1 parent, then you cant exactly rule that a failure without looking at clamp connections.
Im probably going to work in this order, and i will try to shut up after this so we can limit this thread to actual demonstration and practical applications, that is unless everyone is cool with reading through all this.
Anyways here is the order i will be working;
Between all the said-below i will be practicing and performing techniques for monospore germination and isolating a monokaryon and growing it out. As i stated in the other thread monokaryons, at least with cubesis, are very easy to differentiate with the naked eye. I could even take it to another step and do a extremely small microgrow to make sure it doesnt fruit. I have no intentions nof breeding these and they will be released into wild.
I. I will establish 2 mono cultures of the same substrain and each with their own distinguished phenotypes.
II. I will print each one and then probably do a streak and serial dilution method or which ever i was repeatedly successful at during my practice techniques.
III. I will then attempt to breed them. This isnt hybridization and is more along the lines of selective breeding which is the same process but with a much higher success rate then hybridization. On the contrary to what i said-above, a scope should not be needed at least with cubes because it is obvious when you have dikaryons growing across the plate but it will be a best practice for me to observe under a scope the clamp connections to as it will help with my non existent microscopy skills!
IV. Fruit and observe the phenotype(s)
V. Repeat steps I-IV but this time using 2 separate strains to be determined.
VI. Take a monokaryon and induce double haploid production using Colchicine. If this works then it is instant monoculture, there is difference of opinion on rather or not it will sporulate. I think it will but even if it doesnt, and for production purposes, i work with only living cultures anyway. Couple that with the cryoprotectant LC im going to attempt, and im certain it will work, who the hell needs spores? ;)
VII. Im skeptical on being able to cross species and im not sure its possible, but i would like to attempt crossing a species within the same genus such as Psilocybe cubensis with Psilocybe cyanescens. I know wood lover and dung lover, but there is also enzyme adding and all kinds of other stuff that is going to be advanced on this already advanced subject.
Happy hybridization to all of you and best of luck! Lets create some monsters!
Edited by Microbe77, 16 November 2016 - 12:28 AM.