Is a clone from an MS grow an isolate/monoculture?

Discussion in 'MUSHROOM CULTIVATION' started by teamfdn, Apr 11, 2021.

  1. teamfdn

    teamfdn New Member

    Joined:
    Apr 11, 2021
    Messages:
    2
    Gender:
    Male
    I recently got into agar. Last weekend, I cloned a nice looking fruit by putting a tissue sample onto a plate. Everything looks great so far. Is this considered an isolate/monoculture already, or would I still need to do a series of transfers just like I would if I were starting from spores?

    20210411_005937.jpg
     
  2. MadChatter

    MadChatter Member

    Joined:
    Apr 8, 2021
    Messages:
    48
    Gender:
    Male
    Usually people clean up that culture and make a couple transfers, I’m not sure if they transfer in order to get rid of contaminants or not, because it would seem that a fruit will only have one set of dikariotic mycelium genetics, but you can test if you have a mono or not:
    Take two transfersfrom your clone plate and put them both on a clean plate side-to-side. If they separate when they meet up(sector), then it’s not a mono, if they merge, it’s a mono.
    Hope this helps, sorry I can’t answer your question.
    :cube1:
     
  3. Goatrider

    Goatrider Member

    Joined:
    Apr 3, 2021
    Messages:
    46
    Gender:
    Male
    Location:
    Germany
    You can see lots of sectors on that plate.
    It could be up to hundreds of transfers to get a true monoculture.
    Not to mention you may have a non-fruiter then.
    So you`ll always have to transfer, test, transfer, test...
    I like your way. Putting a clone on agar, maybe a few transfers to narrow sectors a bit.
    Personally i don`t aim to get a monoculture, but if you wanna spend some time...:wink2:
     
  4. teamfdn

    teamfdn New Member

    Joined:
    Apr 11, 2021
    Messages:
    2
    Gender:
    Male
    Interesting. I had definitely put myself under the impression that each fruit was an individual culture.

    I'll do a couple transfers to make sure it's nice and clean, then expand it a little and inoculate some jars. Does this seem like a logical course of action?
     
  5. Goatrider

    Goatrider Member

    Joined:
    Apr 3, 2021
    Messages:
    46
    Gender:
    Male
    Location:
    Germany
    That sectors are many different strains from multispore, each with different genetics.
    They will fight for their territory, but also rub their shoulders.

    Your plan sounds good.
    I prefer to do a little test grow after taking a clone to see if i get what i want.
    For that i've got some small boxes half the size of a shoebox.

    :rockon1:
     
  6. AlexLPD

    AlexLPD New Member

    Joined:
    Sep 25, 2020
    Messages:
    23
    Gender:
    Male
    Interesting response, I make a clone from one big cluster (550gr) and cut tree pieces from a single stem, and put them in a single petri to clone them, until today the tree have a small gap between them on the plate.
    Thanks.
     
  7. Moby

    Moby Member

    Joined:
    Feb 27, 2021
    Messages:
    34
    Gender:
    Male
    Location:
    Europe
    just recently I have been told that clones are single strains.
    The info came from a breeder with connections to someone who tests things in his university.

    An additional info because I aswell brought up what appears to be sectors was, that mutations within the mycelium happen all the time and some other more complicated things.

    You dont need hundreds of transfers from a MS plate to get a monoculture or isolate ;)
    after 30 you can find single strains, probably even earlier if you use a very little amount of spores to inoc the first plate
     
    the_chosen_one likes this.
  8. AlexLPD

    AlexLPD New Member

    Joined:
    Sep 25, 2020
    Messages:
    23
    Gender:
    Male
    Well thats interesting, I recently make my own first rizomorphic agar plate, its from a strain I have been pasing in to agar for quite awile, more for he need to keep thing up, to the urge need to make rizo, still... a nice way to clean the micellium, I will look in to that and make one or two more before say I can make monocultures.

    Hope this will happens any time soon.

    Kind Regards.
    -Alex.
     
  9. the_chosen_one

    the_chosen_one there are no answers.. only choices Moderator Mushroom Doctor

    Joined:
    Jan 13, 2012
    Messages:
    984
    Gender:
    Male
    Location:
    1984
    Nailed it. :hi5:

    Recessive genes and mutation can wreak some havoc for the person craving consistency.
     
    Moby likes this.
  10. Moby

    Moby Member

    Joined:
    Feb 27, 2021
    Messages:
    34
    Gender:
    Male
    Location:
    Europe
    I'm always willing to learn but not sure where to find stuff like that and mostly I get no answers when I ask people, do you have any educational source for me about things in this area ?
     
    the_chosen_one likes this.
  11. the_chosen_one

    the_chosen_one there are no answers.. only choices Moderator Mushroom Doctor

    Joined:
    Jan 13, 2012
    Messages:
    984
    Gender:
    Male
    Location:
    1984
    Welcome to the edge of the Rabbit Hole...

    https://phys.org/news/2018-04-paternal-maternal-dna-fungi.html

    Many types of mushroom have two different nuclei in their cells, one from the 'father' and another from the 'mother." Researchers at the universities of Delft, Utrecht and Wageningen have discovered that the genes from the parental DNAs are expressed at different times in mushroom development. "This means that when genes involved in mushroom formation are identified, we first need to find out whether the paternal or maternal nucleus is active," says TU Delft doctoral candidate Thies Gehrmann. The research results were published in the journal PNAS on 11 April 2018.

    Fungi, such as mushrooms, play an important role in our ecosystem. In nature, they recycle dead plants and animals. As humans, we not only eat fungi, but also use them in making food, such as bread and beer, and as bioreactors in the manufacture of drugs and other substances. They also play a direct role in human health since they can cause infections. In order to take full advantage of mushrooms (and fungi as a whole) and prevent their undesirable effects, a better understanding of fungi is essential. However, mushrooms are highly complex organisms, as are their genetic processes.

    In this study, the researchers from TU Delft, Utrecht and Wageningen investigated gene expression in mushrooms, both in a model mushroom and in the common, edible mushroom. They studied the extraordinary phenomenon discovered by TU Delft doctoral candidate Thies Gehrmann. "Many fungi have two different nuclei in their cells, each with different genetic material. A mushroom inherits DNA from both parents, but this is not mixed in a single nucleus as in humans. We have now seen that genes on both of the parental DNAs are expressed at different times in the development process – something that was not known before."

    The impact of this new discovery is that, from now on, it will first be necessary to find out whether the paternal or maternal nucleus is active when studying mushrooms, for example in the quest for genes involved in mushroom formation. This new understanding of the molecular mechanisms in mushroom DNA can be used to breed new strains that can improve the cultivation of edible mushrooms, such as the common mushroom.

    Much of the article in PNAS originates from the dissertation by Thies Gehrmann, who has obtained his Ph.D. on Friday 6 April at TU Delft. "For my doctoral research, I developed and applied methods of bioinformatic analysis in order to understand variations within and between mushroom-forming fungi. The phenomenon we are now describing in PNAS is an example of that. In addition, I also demonstrate that another process, known as alternative splicing, causes some of the variations. This phenomenon is difficult to study in fungi and so it was generally believed that no splicing took place."

    Thousands of cases

    Alternative splicing is a process in cells that enables a single gene to produce different proteins, each with its own function. Abnormal alternative splicing and mutations in the products of alternative splicing have been linked to cancer, autism and serious development disorders, both in mice and in humans. Despite these serious consequences for mammals, there has been very little research into alternative splicing in fungi. Thies Gehrmann has now demonstrated that thousands of cases of alternative splicing occur in the fungus S. commune.

    Here's the full article..

    https://www.pnas.org/content/115/17/4429
     
    Last edited: Apr 17, 2021
    Moby and lennon like this.
  12. Moby

    Moby Member

    Joined:
    Feb 27, 2021
    Messages:
    34
    Gender:
    Male
    Location:
    Europe
    Thank you!

    Was absent for a moment, pretty wrapped up into sourcing of spores, got a solid collection of new stuff and a microscope that I'll work into in the next weeks.

    I got an old Falbino print from 2012 from Mrs Hipp3 a while back, gave it to someone else to revive it hopefully but kept three swabs of it, will try to get it back running in the closer future too.

    Freaky joined us here btw
     
    the_chosen_one likes this.
  13. the_chosen_one

    the_chosen_one there are no answers.. only choices Moderator Mushroom Doctor

    Joined:
    Jan 13, 2012
    Messages:
    984
    Gender:
    Male
    Location:
    1984
    You're quite welcome. Wish we had that info 15-20 years ago. Could have saved a lot of arguing lol.
    Thanks for the hook up too! Just sent her a message.

    Yeah, I fired Falbino back up last year. She still does her thing. Even got a couple of Chimera on this one. I really need to get back on that project.
    Falbino Chimera.jpg
     
    Moby likes this.
  14. Moby

    Moby Member

    Joined:
    Feb 27, 2021
    Messages:
    34
    Gender:
    Male
    Location:
    Europe
    Luckily more and more people invest time into studies that at least contain bits of information buried under irrelevant stuff, at least some info tho.
    With the legalizations they gotta do more tests it will go forward in a faster tempo now.

    Here a picture of Falbino that Nicked posted on topia, he got spores from Wharfrat.

    post-160797-0-32638600-1610112014.jpg

    Guess Wharfrat has stabilized it.


    I really like the diversity in your casing.
    Is it a f2 or f3 ?

    PF Albino btw is very rare now, I dont know anyone who has it (except Nicked but dont really know him), make sure to keep it going ;)

    Cheers