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CreonAntigone
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Plant cell culture
#27698184 - 03/16/22 08:17 PM (2 years, 2 months ago) |
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Maybe 'advanced botany' as opposed to mycology, but -
I see that many labs grow plant cells in culture media. How do they go about this? What media could be used? What kind of nutrients might a plant need that could be added to culture media?
I have read about the growth of moss in culture. But moss is a special case as it has spores. How would I go about culturing, say, a tomato plant? Could I go from a tomato leaf to a culture plate of tomato cells?
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MrBlueshrooms



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There is a book I read many years ago called plants from test tubes that describes this. Generally you don't culture cells, but explants. Cells can be cultured though by using physical methods to get the cells (like a blender) or chemicals or enzymes. Media is more complicated but sterile technique is the same. To get tissue into agar several washes and rinses are used. I think diluted bleach and water iso and dish soap did the trick.
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hazyhorse
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i saw an old thread about this recently. it sounds cool, but what is the point of doing this? i just didn't understand the practical purpose of it & no one really mentioned WHY they were talking about it. can you grow more plants from it or something?
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pablokabute
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Re: Plant cell culture [Re: hazyhorse]
#27698432 - 03/17/22 03:26 AM (2 years, 2 months ago) |
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Quote:
hazyhorse said: i saw an old thread about this recently. it sounds cool, but what is the point of doing this? i just didn't understand the practical purpose of it & no one really mentioned WHY they were talking about it. can you grow more plants from it or something?
In my country, Most, if not all, banana plant cultures for farming bananas are from tissue culture and most of the time are affordable(but when you need, say, a thousand plants or more, then it quickly adds up) so my guess is,depending on what specific plant youre trying to grow, it might be practical.
Yes you could probably grow way more plants vs traditional propagation methods.
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Fermented Mushrooms!! --- https://www.shroomery.org/forums/showflat.php/Number/23378638/fpart/1/vc/1 'The second seal: βAll CONTAMINATED things and events are unsatisfactory.β' "I envy you. You North Americans are very lucky. You are fighting the most important fight of all - you live in THE HEART OF THE BEAST." --Anonymous Guerilla, or is he..
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hazyhorse
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thatβs super interesting!! i didnβt realize it was such a viable propagation method. iβll have to look more into it, the concept is pretty cool
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pablokabute
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Re: Plant cell culture [Re: hazyhorse]
#27699288 - 03/17/22 06:47 PM (2 years, 2 months ago) |
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Quote:
hazyhorse said: thatβs super interesting!! i didnβt realize it was such a viable propagation method. iβll have to look more into it, the concept is pretty cool
You can search for 'micropropagation'.
Id also like to learn to do it someday.
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Fermented Mushrooms!! --- https://www.shroomery.org/forums/showflat.php/Number/23378638/fpart/1/vc/1 'The second seal: βAll CONTAMINATED things and events are unsatisfactory.β' "I envy you. You North Americans are very lucky. You are fighting the most important fight of all - you live in THE HEART OF THE BEAST." --Anonymous Guerilla, or is he..
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clueless
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I've been doing tissue culture work for a few years now. Murashige and Skoog media is most commonly used, but orchids and walnuts have their own formulas. As for coming from a stem or leaf, that's absolutely possible.
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CreonAntigone
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Quote:
hazyhorse said: i saw an old thread about this recently. it sounds cool, but what is the point of doing this? i just didn't understand the practical purpose of it & no one really mentioned WHY they were talking about it. can you grow more plants from it or something?
One of my intentions is to explore it for mycorrhizal or parasitic fungal species that need plants to live. Some species only grow with plants and won't germinate on MEA. So the idea would be 'co-culturing', both fungi and plant cells.
Example: there is a fungus that lives on the leaves of morning glory plants and makes alkaloids. It was tested on sterile media and it didn't grow. One might be able to produce alkaloids though in a dish where you grow out morning glory leaf cells.
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SingularFusion


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I have this in my list of things to do sooner rather than later
The myriad applications for high value plants are pretty spectacular
Consider if you grow cannabis and have a couple dozen mother plants you keep as a genetic library, well that is using resources and grow space and obviously if you could keep most of that in cold storage and wake it as needed, that would be quite a saving of resources and of the daily effort to keep that all going in live plant form
Not to mention that now if you want backups kept to ensure the clone is never lost to law enforcement or sudden death, a friendly grower buddy is not needed anymore, anyone with a fridge can hold your cuts essentially. And with most folks not having the necessary skills to wake that up off a plate, the chance of theft is minimal, whereas taking a cutting from a live plant any idiot can do
Orchids are also interesting but that is a whole other discussion I don't know shit about... yet
Guess these techniques might be able to be applied to cacti as well, I dunno, hoping to learn more soon. I have seen there is added complexity to the plate media from the small amount of research I already did, I guess maybe some premixed media you buy is the way here, even though I would always make my own agar for mycology
good stuff fellas
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pablokabute
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Quote:
Nef said: I have this in my list of things to do sooner rather than later
The myriad applications for high value plants are pretty spectacular
Consider if you grow cannabis and have a couple dozen mother plants you keep as a genetic library, well that is using resources and grow space and obviously if you could keep most of that in cold storage and wake it as needed, that would be quite a saving of resources and of the daily effort to keep that all going in live plant form
Not to mention that now if you want backups kept to ensure the clone is never lost to law enforcement or sudden death, a friendly grower buddy is not needed anymore, anyone with a fridge can hold your cuts essentially. And with most folks not having the necessary skills to wake that up off a plate, the chance of theft is minimal, whereas taking a cutting from a live plant any idiot can do
Orchids are also interesting but that is a whole other discussion I don't know shit about... yet
Guess these techniques might be able to be applied to cacti as well, I dunno, hoping to learn more soon. I have seen there is added complexity to the plate media from the small amount of research I already did, I guess maybe some premixed media you buy is the way here, even though I would always make my own agar for mycology
good stuff fellas 
Thanks for all the info, this is the kind of info Im trying to absorb from this site! Cheers.
--------------------
Fermented Mushrooms!! --- https://www.shroomery.org/forums/showflat.php/Number/23378638/fpart/1/vc/1 'The second seal: βAll CONTAMINATED things and events are unsatisfactory.β' "I envy you. You North Americans are very lucky. You are fighting the most important fight of all - you live in THE HEART OF THE BEAST." --Anonymous Guerilla, or is he..
Edited by pablokabute (03/17/22 11:44 PM)
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CreonAntigone
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Quote:
clueless said: I've been doing tissue culture work for a few years now. Murashige and Skoog media is most commonly used, but orchids and walnuts have their own formulas. As for coming from a stem or leaf, that's absolutely possible.
Quote:
Nef said: I have seen there is added complexity to the plate media from the small amount of research I already did, I guess maybe some premixed media you buy is the way here, even though I would always make my own agar for mycology
Yeah I'm sure these special medias provide the best results... but I would highly doubt if there wasn't something that could be made in a home pretty easily. This site seems to have taught me that a lot of complex cultivation tools have a DIY equivalent. I guess it needs testing.
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7Suns
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Hey guys I was actually just about to do a write up on a home made version of the murashige and skoog or modified fries medium that I concocted from published science articles
Iβm deep into the study of ecto/mycorrhizal species and this is something Iβve been trying to mimic in house so Iβll share the link as soon as I finish the write up.... everything can be bought locally except ph paper
Also if youβre going this route I also recommend a quick look at my post about ph effects on agar solidification
https://www.shroomery.org/forums/showflat.php/Number/27696637
Also hereβs my home made antibiotic agar recipe if you are starting from a wild specimen itβs very helpful since itβs more then likely heavily contaminated with bacteria especially chanterelles that can have bacteria incorporated internally
https://www.shroomery.org/forums/showflat.php/Number/27576801
-------------------- Following the lost souls as they transcend the seven dimensions, through playful dancing, like wandering shadows in the heat of a mirage 7suns grain sac hack 7suns straw log hack 7suns official OTC antibiotic agar recipe 7suns DIY filter bags 7suns cheap AF flowhood 7suns cold weather fruit chamber
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7Suns
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Re: Plant cell culture [Re: 7Suns]
#27700171 - 03/18/22 12:59 PM (2 years, 2 months ago) |
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Also there a Facebook page I believe itβs called the βcactus laboratoryβ Where the guy uses this medium for rooting cactus cuttings, really fascinating work
-------------------- Following the lost souls as they transcend the seven dimensions, through playful dancing, like wandering shadows in the heat of a mirage 7suns grain sac hack 7suns straw log hack 7suns official OTC antibiotic agar recipe 7suns DIY filter bags 7suns cheap AF flowhood 7suns cold weather fruit chamber
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hazyhorse
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Re: Plant cell culture [Re: 7Suns]
#27700290 - 03/18/22 02:45 PM (2 years, 2 months ago) |
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fuck yeah, this is really cool. glad i found this thread! some home DIY methods for getting into this kind of work would be really awesome. the example of keeping a library of mother plants alone would be so practical & worth it. iβm keeping one as a βbonsaiβ mother right now but fuck, she grows like 6β in a week lol. plus itβs so easy to under water once they get root bound, not to mention nutrients etc. the cacti applications are also quite fascinating. would love to go down this rabbit hole more
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nerdintheshell



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For media murashige and skoog agar seems to be a common one. The ingredient list is pretty varied and may not be as easy to make at home as other mediums like pda or mea.
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pablokabute
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Quote:
nerdintheshell said: For media murashige and skoog agar seems to be a common one. The ingredient list is pretty varied and may not be as easy to make at home as other mediums like pda or mea.
Which reminds me of overcomplicated Cordyceps liquid culture recipes and yet its progressing to somewhat more accessible ingredients now.
Maybe if more people catches on to doing plant cell cultures, easier, more accesible recipes will be opened up?
--------------------
Fermented Mushrooms!! --- https://www.shroomery.org/forums/showflat.php/Number/23378638/fpart/1/vc/1 'The second seal: βAll CONTAMINATED things and events are unsatisfactory.β' "I envy you. You North Americans are very lucky. You are fighting the most important fight of all - you live in THE HEART OF THE BEAST." --Anonymous Guerilla, or is he..
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7Suns
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Did you guys not read my comment?
I have a simple home recipe I created as a substitute for this recipe
-------------------- Following the lost souls as they transcend the seven dimensions, through playful dancing, like wandering shadows in the heat of a mirage 7suns grain sac hack 7suns straw log hack 7suns official OTC antibiotic agar recipe 7suns DIY filter bags 7suns cheap AF flowhood 7suns cold weather fruit chamber
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pablokabute
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Re: Plant cell culture [Re: 7Suns]
#27703188 - 03/21/22 07:54 AM (2 years, 1 month ago) |
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Quote:
7Suns said: Did you guys not read my comment?
I have a simple home recipe I created as a substitute for this recipe
I only saw low pH agar and antibiotic agar recipes, is that what youre referring to?
--------------------
Fermented Mushrooms!! --- https://www.shroomery.org/forums/showflat.php/Number/23378638/fpart/1/vc/1 'The second seal: βAll CONTAMINATED things and events are unsatisfactory.β' "I envy you. You North Americans are very lucky. You are fighting the most important fight of all - you live in THE HEART OF THE BEAST." --Anonymous Guerilla, or is he..
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7Suns
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Re: Plant cell culture [Re: 7Suns]
#27703203 - 03/21/22 08:07 AM (2 years, 1 month ago) |
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Quote:
7Suns said: Hey guys I was actually just about to do a write up on a home made version of the murashige and skoog or modified fries medium that I concocted from published science articles
Iβm deep into the study of ecto/mycorrhizal species and this is something Iβve been trying to mimic in house so Iβll share the link as soon as I finish the write up.... everything can be bought locally except ph paper
This βοΈ
-------------------- Following the lost souls as they transcend the seven dimensions, through playful dancing, like wandering shadows in the heat of a mirage 7suns grain sac hack 7suns straw log hack 7suns official OTC antibiotic agar recipe 7suns DIY filter bags 7suns cheap AF flowhood 7suns cold weather fruit chamber
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SingularFusion


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Re: Plant cell culture [Re: 7Suns]
#27703242 - 03/21/22 08:49 AM (2 years, 1 month ago) |
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look forward to hearing your take on it 7suns
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SharaVabdas



Registered: 03/23/22
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Re: Plant cell culture *DELETED* [Re: clueless]
#27723093 - 04/06/22 10:38 AM (2 years, 1 month ago) |
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Post deleted by SharaVabdas
Reason for deletion: Shroomery is full of bigots and I'm leaving.
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7Suns
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Iβm going to do a write up on my media recipe and post the link here, been busy but itβs in the works just need a little more time, maybe this weekend after the farmers market Iβll finish it if I sell out before 12 I should have enough time afterwards
-------------------- Following the lost souls as they transcend the seven dimensions, through playful dancing, like wandering shadows in the heat of a mirage 7suns grain sac hack 7suns straw log hack 7suns official OTC antibiotic agar recipe 7suns DIY filter bags 7suns cheap AF flowhood 7suns cold weather fruit chamber
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SharaVabdas



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Re: Plant cell culture *DELETED* [Re: 7Suns]
#27723125 - 04/06/22 11:06 AM (2 years, 1 month ago) |
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Post deleted by SharaVabdas
Reason for deletion: Shroomery is full of bigots and I'm leaving.
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bambus
Amphibian furniture maker



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I can't wait to hear more about the home made recipe for this. @7suns, What do you do with your chanterelle cultures? I've tried to clone them unsuccessfully, but wasn't sure what to do with it after I got it on a plate anyways.
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CreonAntigone
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Re: Plant cell culture [Re: bambus]
#27730938 - 04/12/22 12:25 AM (2 years, 1 month ago) |
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Quote:
bambus said: wasn't sure what to do with it after I got it on a plate anyways.
I suggest 'in vitro mycorhizal snythesis' - illustrated here -

Isolate the fungal culture, propagate as LC to increase the volume. Then mix the pure culture with a mycorhizal partner invitro -
so for example, chanterelles are partners with birch and oak. So soak oak acorns in chanterelle liquid culture, and similarly you can put rooted oak cuttings in a jar of chanterelle culture. Wait for the roots to mycorhize indicated by the new white growth. Then plant in a greenhouse or just right outside.
It is the same principle behind growing pure culture, except it is two instead of one - dual culture, the trees and the chanterelles together.
It may take several attempts for the cuttings to survive. What you can do is start some saplings in a greenhouse that are chaterelle-partnered, then plant a set of new plants inoculated with a different liquid culture (ie genetics derived from a different fruit body). It seems in my experience this strain-strain competition increases the growth of the mycorhizal species.
And if the first set of cuttings doesn't work, you might that next season they do. I have been trying to mycorhize quaking aspen, the first attempt all cuttings died, this second attempt every single cutting rooted, lived and grew. It is a learning process.
Working with old trees helps - you can inoculate them with quite a few strains and they'll survive and be fine. Yes old trees can be inoculated, they just take a stronger innoculant owing to their maturity - one can just mass-produce a liquid culture of the mycorhizael sp. and pour it on a tree, which is a very simple method of inoculation but nonetheless, effective and sustainable.
This should establish oak trees that are partners with chanterelle. But it still would take a long time, potentially years, before there is any fruiting. It is still worth doing. There's that saying about 'the best time to plant a tree'.
Edited by CreonAntigone (04/12/22 12:26 AM)
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bambus
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Wow! Thanks for the detailed response. I have a great chanterelle spot, and a bunch of oaks on my land, so I'll have to give this a try at some point.
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CreonAntigone
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Re: Plant cell culture [Re: bambus]
#27732746 - 04/13/22 11:54 AM (2 years, 1 month ago) |
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Quote:
bambus said: Wow! Thanks for the detailed response. I have a great chanterelle spot, and a bunch of oaks on my land, so I'll have to give this a try at some point.
So the way it works is, if there is a chanterelle spot, those trees are chanterelle trees - the way the partnership works is that fruiting happens at the partnership's strongest points.
So, what trees are next to your chanterelle spot? Take cuttings or samples of those trees, as they already carry chanterelle culture in them and already are in partnership.
Then you can plant these natural chanterelle trees next to ones you synthesized invitro (as in my last post) - plant them every other tree. This would create strain-strain competition.
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bambus
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They're Oak trees, not sure what kind though. I'm pondering the idea of collecting some acorns from that spot, sprouting them and trying to inoculate them immediately as they sprout. If they're then planted in a nutrient poor soil, maybe they'll be more inclined to bond through necessity? The native soil is heavy clay.
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CreonAntigone
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Re: Plant cell culture [Re: bambus]
#27734912 - 04/14/22 07:37 PM (2 years, 1 month ago) |
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Quote:
bambus said: They're Oak trees, not sure what kind though. I'm pondering the idea of collecting some acorns from that spot, sprouting them and trying to inoculate them immediately as they sprout. If they're then planted in a nutrient poor soil, maybe they'll be more inclined to bond through necessity? The native soil is heavy clay.
I do think a mycorrhizal association can make up for deficits in the soil.
Mycorrhizal species seem to prefer loam which is about in the middle when it comes to soil types. But clearly there are plenty of natural chanterelles that come up in your spot, so I'm going to guess that your soil is fine for chanterelles.
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