Cloning Plants By Tissue Culture

OregonMeds said:

That smart lighting controller seems gimmicky. Is it just me?
Why push ballasts 24/7 when you can buy 10 more cheap (or not so cheap) 1kw ballasts and timers for less than that thing.

Neat idea, but seems as practical as tissue culture is over cloning in most cases.

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no its not just you , its an idea stolen from a standard weed nerd an turned into an "invention" that cost probably way too much. we have been makin "flip flop" controllers for years around here. (with just a few cheap parts)

in real life the controller is way cheaper then a ballast so it helps if you dont wanna buy another one.





i love the tissue thing , it wont be efficient in my opinion , but its something to play with. i love science.


i have a leaf in my fish tank that developed roots and i dont know what to do with it ? its a underwater plant for the tanks. will that leaf grow another plant , if not on its own , can i help it ?




thanks soil

your roots will grow fine, given you've got good 02 saturation, and a stable system. ot may continue to grow many different ways under water. you wont be able to manipulate it though, with fish. It should be cool to watch though.
tissue culture isnt't practical for most, you're right, with the ease of cloning. but one fine example of practicality is availability. imagine instead of seeds you could purchase verified pure rootballs ready to sprout up in a couple days, and the advantage to the grower to have fine genetics delivered overnite, with golf ball size roots, before the thing has even sprouted. It's an incredible jump start that allows me to keep a full time perpetual grow successful, with many more pants flowering than without. This system is already in use by most greenhouse/flower suppliers right now, and they will tell you how very efficient and practical it is, and they dont even have legal plant limits to consider like we do

peace

pharmacoping said:

your roots will grow fine, given you've got good 02 saturation, and a stable system. ot may continue to grow many different ways under water. you wont be able to manipulate it though, with fish. It should be cool to watch though.
tissue culture isnt't practical for most, you're right, with the ease of cloning. but one fine example of practicality is availability. imagine instead of seeds you could purchase verified pure rootballs ready to sprout up in a couple days, and the advantage to the grower to have fine genetics delivered overnite, with golf ball size roots, before the thing has even sprouted. It's an incredible jump start that allows me to keep a full time perpetual grow successful, with many more pants flowering than without. This system is already in use by most greenhouse/flower suppliers right now, and they will tell you how very efficient and practical it is, and they dont even have legal plant limits to consider like we do

peace

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i was wrong ...... with little to no knowledge it would be in-efficient. if done right it could be way more efficient.




soil

I love honest people. thank you I've heard if an honest person who is mistaken is given the truth, one of two things happen, either they will no longer be misunderstood, or no longer an honest person. once a shelf is filled with "unusable roots" not using plant counts,of your favorite herd you continue the dividing/rooting, while selecting (biweekly for me) the "unusable roots" of the strain you want to run, allow it to shoot up, now taking up plant counts, and put in the veg room to cycle.exactly 45 days later they go to flower,for roughly 60 days, and then to harvest.(weekly here) it could take a month or two to get a stocked shelf of these roots, but after that, control is yours. I dont manipulate any more than we all do when we use "rooting gel" to start clones. same hormones too, in a sealed jar, little light, agar,sugar, until I need their service. frankensteins are not my goal, and are destroyed each time. I have used colchi**** in vitro, and did produce a fine(techy name) plant with twice the normal sets of chromosomes, pictured here. PlushBerry, from TGA Subcool, attitude seeds.

On the original note, I have one of those flip switches PowerBox, nice, works, if your ballasts are identical brand, and no issues with hot fire bulbs exist. digital ballast come with an internal fuse that can, and does trip occasionally(high temps, short,expired bulb,???) and these cause it to happen also, in my experience, on one product, used only by me, brand new. I used it some time ago to light two flower rooms at opposite times(12/12 to 12/12) and it did work mostly, but one discovered lighting failure is unnacceptable, so it sits on my shelf 'o toys. not sorry for the purchase, but was a waste of time/thought. If you get another ballast/timer, you won't regret it. plus, I think these digital ballasts like a break daily, as opposed to running 24/7, imo.

peace

You know I never considered cloning a plant from the roots up...

My understanding of the law and its definition of a plant was: containing both root and foliage.. Having dealt with conventional cloning methods exclusively for so long that I condensed my personal definition to "has roots" and I think that has gotten in the way of me thinking about cloning plants this way..

I like your idea about mailing a root ball that could be induced to sprout by the recipient.. Many times I've wanted to share clones with someone across the country and there didn't not seem to be a good way to do it without breaking the law.. However in my understanding a cutting (assuming it is either/only root or foliage) should not be illegal to ship?

How long could you keep one of those root balls dormant and stable for shipping before they start becoming less viable? Could you priority ship them (instead of overnight)?

mailing the foliage is illegal. even seeds are illegal here. however, an invitro callus or rootball is not recognizable by any means. cept dna. if you could regulate sugars and protect the rootball from attacks, theoritically you could raise roots in your system. a piece of a live root can be told to sprout up, or keep growing roots/dividing. In vitro culture allows all to be sterile, with some luck of course. as the culture matures it can fight off many attacks, and you can re rinse/transplant again, if needed. I have callus over two years old, and also rootballs the same age(older,maybe?). I dont push it, but pretty sure they survive in the fridge for weeks, and in a fair temp package for days and days. packaging can be manipulated easily also, heated/etc. as long as they are not dried up, are transferred every couple months or so and not moldy/dead, their viability is limitless, I believe. been sharing this way for a long time. some top seed breeders are approachable, and they tissue culture. Instead of years to an F1, these guys can do it in months, in vitro. some will gladly share callus for test grows, instead of seeds, because they are pure, and sterile, and exact. with a punnets square they can purify a strain quickly, its amazing what they're doing. cannabis breeders bible explains much of it.

I thought as long as a plant were not complete it was basically seeds/stems, the dry weight of a cutting could never be considered more than personal use, and even thats a bit of a stretch (the law like everything else is subject to compromises as long as you have a reasonable position to argue). A decent part of standing up against the law is, well.... standing up against it... Help set reasonable legal precedents (isn't that more or less how laws get changed, when you don't have the money to successfully lobby).

I should probably apologize in advance, I'm sure I have not covered all the suggested reading material at this point..

"in-vitro callus" Would mean something like cultured scar tissue? If so is it at all specific to where the plant is scared to start with (I was under the impression that "growing tips" were most manipulable)? Are there specific ways to obtain scaring or can I do something like "burn" a plant with say, peroxide?

Could I use say.... the remnants left over after a harvest (roots and main stem)? They tend to want to live, there must be an incredible amount of life/energy left to them (can that be put to good use?)...

What if the culture were cross contaminated with the cells from another closely related plant, say its seed sister or clone sister (the mess of roots you might get from a hydro table after harvest for instance)?

very cool.....thanks

seeds, stems..you are correct, but we are taliking about shipping, not posession. ship one leaf and you have broken multiple federal laws. a stem stuck into rockwool becomes a plant count, with or without roots attached/formed.

callus is not a scar tissue, caused by knicking or damage to a plant, they are a grouping of disorganized undeveloped cells proliferating. when you tissue culture mj you'll see explants, callus, mold, roots, leaves, and some mutants. callus wil look like a puffy chunk of cauliflower. This is the basis fo rstorage of my genetics, cutting off of them is very successful. meristem or tip culture is the easiest, but these plants are totipotent and any piece will work, albeit with different patterns and success with each. The tips are the best for beginning as they are most likely without virus, and have the most hormones within.

you will notice some swelling in tissue on a stalk. this area is full of mutated genes/virus' that the plant has overcome. Taking material from this area can produce the frankensteins you might be looking for.

one cell wont rub off into another, in a tray or in vitro, otherwise we could breed via grafting root stock.... seedless breeding can take place invitro,(embryogenesis?) but I dont do this, however seed companies do.
the bacteria for these types of manipulations are available online, and are responsible for some wild creations in the plant word.

those roots are usable, in vitro and even out(regeneration). If you leave a couple leaves on the stalk after harvest you can put it back into veg and regenerate the plant with success, if desired, but you know that I think. I have rooted leaves, but it proves to be a much longer process to bring it to harvest. I use new shoots, trimmed of all foliage, scraped, and sterilized in bleach/alcohol before beginning.
many are hung up on moldy failures. I get some, sure, but 100% of the time its brought in with the plant piece, not dirty equip, or non sterile equip. I know this because you can easily see it under a scope, ////eyelash,dirt, etc. when I mix agar I've never seen mold form, unless there was a plant in it. maybe I'm an awesome sterilizer, or just fol
low directions without fear of the unknown.

(grafting hint..no agar used. wax, elmers glue, rooting hormone. easiest way is to fill a hollow new stem with a scraped stem of a new lateral tip.)

Guile said:

I thought as long as a plant were not complete it was basically seeds/stems, the dry weight of a cutting could never be considered more than personal use, and even thats a bit of a stretch (the law like everything else is subject to compromises as long as you have a reasonable position to argue). A decent part of standing up against the law is, well.... standing up against it... Help set reasonable legal precedents (isn't that more or less how laws get changed, when you don't have the money to successfully lobby).

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well seeds can be sold for the purpose of helping someone do something illeagal. -----conspiracy /federal
the stems contain a horrible schedule 1 narcotic. THC ------ can be charged as possession and/or dealing / can be ugly, but the judge would have to be a dick to give out a lot of time for that.

this is dead on.

Guile said:

A decent part of standing up against the law is, well.... standing up against it...

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now our lawyers argument is gonna be "what the fuck , your honor , sticks an marbles! i demand this garbage be thrown out"...... thats a decent lawyer. your everyday cheap lawyer is gonna get the dealing dropped if you sign guilty to the possession......... and now i got a felony for weed over stems and seeds , nothing more. and then automatically my next joint was a felony just because i already had a felony for pot.
so two drug felony's for seeds an stems.

bottom line.... keep a high paid lawyer around when fucking with pot and charges will stay minimum.

i dont really think they can be too rough with this plant matter shit because of loopholes. plus like pharma said , they would have to run a DNA test just to see what it is.
its all illeagal but consequences are determined by who you know and where your at.








if your going to ship stuff (legally of course) just make sure its airtight and it will make it to its location. if it dont , the box busted open an the postman is running out to get a 400 right now.




soil

pharmacoping said:

I love honest people. thank you I've heard if an honest person who is mistaken is given the truth, one of two things happen, either they will no longer be misunderstood, or no longer an honest person.

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well facts are facts , i cant argue with facts.

pharmacoping said:

On the original note, I have one of those flip switches PowerBox, nice, works, if your ballasts are identical brand, and no issues with hot fire bulbs exist. digital ballast come with an internal fuse that can, and does trip occasionally(high temps, short,expired bulb,???) and these cause it to happen also, in my experience, on one product, used only by me, brand new. I used it some time ago to light two flower rooms at opposite times(12/12 to 12/12) and it did work mostly, but one discovered lighting failure is unnacceptable, so it sits on my shelf 'o toys. not sorry for the purchase, but was a waste of time/thought. If you get another ballast/timer, you won't regret it. plus, I think these digital ballasts like a break daily, as opposed to running 24/7, imo.

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i dont like digital ballast for that reason. more money and more problems. ive got magnetic ballasts from the 80's that still work just fine.
lighting failure is not an option for me or you.
the flip flop switches we build dont really have the option of fucking up , all they do is switch contacts. so as long as the timer is working correctly then there is no probs. the magnetic ballast dont need a break.

now with that being said i would still recommend buying the extra ballast and timer , specially if your using digital ballast. the 12 hour break will help the ballast to last forever plus you wont fuck up 2 rooms if one single ballast goes out.



soil

I almost miss the humm of my old ballasts !! really. you're right, they just hardly ever fail, and usually gave an audible warning while dying, enough to fix it for 30 bucks!. the digis do cut out for various reasons, failures suck, but several more are sparked at the same time, so tomorrow, fixed. your ballasts are a better deal too. I still like the idea of switching for heating, (i do with timers now) but instead of a 2000 watt baseboard heater cranking, 2- 1k lights are lit, and the heat is recirc to the opposite rooms. it works great.

some bud porn..cheers !

p

pharmacoping said:

I almost miss the humm of my old ballasts !! really. you're right, they just hardly ever fail, and usually gave an audible warning while dying, enough to fix it for 30 bucks!. the digis do cut out for various reasons, failures suck, but several more are sparked at the same time, so tomorrow, fixed. your ballasts are a better deal too. I still like the idea of switching for heating, (i do with timers now) but instead of a 2000 watt baseboard heater cranking, 2- 1k lights are lit, and the heat is recirc to the opposite rooms. it works great.

some bud porn..cheers !

p

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ah... so what your sayin , is when room #2 is sleeping , room #1 is used to heat #2? thats a beautiful idea. i never even considered that.
i dont use two bloom rooms , but the more i think about it , my cheap ass is considering the savings.

hmmm a 1500w heater for 12 hours a day or a divider in the room. easy decision.


is that pink an green bud the same ?


good knowledgeable posts p !




soil

this is waaaay too much good information before coffee...... subbed up and will read properly later...... <3

california orange bud(real hashy favor, good stone) and Plushberry(top 10 medicators)
(seperated by a light trap curtain) one side vegging, other in flower currently. veg is 18hrs on, all nite, the 6 off is during high noon. flower room is lit 8-8 during the day. each has its own heater(elec baseboard,digi thermo, hardwired240), air is circed full time between the two, including c02, 8-8. very efficient for me. all the lights are 240volt, even the t-5's(Badass brand) and the 8 light controller is hardwired in, so timing is a breeze. still costs several hundred dollars monthly in electricity for ac,dehumid,heat,lites,pumps, etc. but worth it.

peace

you would almost have to make your own micropropagation media for cannabis. that can be a science in itself.

you are correct Bonkleesha, but the base mediums readily available work well. It's the rest of the mix that took forever to figure out. The kits sold and info available multiplies some flowers well, but if multiplying was needed, cloning would be the key. Different formulas are needed for each stage of tc. first one is callus, then dividing, then rooting, then sprouting...each requiring a special set of circumstances inside their womb. I"m in the tissue culture tent daily, transfers, inspections, disposals, etc. I'm 46 in 2 days, and remember being in the first marijuana growroom when I was 4yrs old. my parents had several, before you could buy all the toys and right nutes/lites online. they had lots and lots of hps streetlamps and some strange experiments, but a pretty successful garden biz for lots of years. was hooked then, nothings changed, cept technology, legality, and the price paid for quality herb !

Sorry to interrupt the continuity of the conversation.

So the callus is not a response to plant damage?

While looking around at information on callus culture One of the impressions I got was that you are probably working with predominantly "undifferentiated" cells.. "Parenchyma" (living cells from the core of roots/stems)..
"Meristematic" (found in zones of the plant where growth can take place). To form the callus

Is there an easy way to isolate these cells? If left with the "differentiated" cells will they have an influence on callus formation?

Is damage to the differentiated cells related to undifferentiated cell formation? (or just consequential to obtaining it?) Are we manipulating part of a repair response (or just completely manipulation the situation)?

What I was after with my foreign plant material question was more aimed on weather of not it could be mixed without causing the callus material to attempt to isolate/reject it? Would the entire experiment fail because everything is rejecting each other? Would things sort themselves out by creating 2 genetically different callus formations?

Could you use UV lighting (or an ozone generator) in your incubation aria to keep molds/bacteria at bay while still allowing the callus formation? Both can be relatively "harsh" but if callus is a response to damage it almost seems that it could work in the favor of a crude experimenter.

no apology necessary.
callus " a proliferating mass of disorganized, mostly undifferentiated or undeveloped cells" (Plant From Test Tubes).
this mass can resemble a chunk of cauliflower partially submerged in medium. I use this stage to multiply and divide, as well as to store genetic moms. I cut off off these and induce rooting, for genetic storage, although more resources are necessary than storing callus culture, or to be transferred to a "shooting" vessel to begin the plant stage.

Osmosis will not transfer dna from one plant to another, without certain circumstances. electricity, polypropyene glycol,bacteria/virus,mechanical dna injection, radioactivity, uv light,radiation all are being used currently(NOT BY ME) to do that. although worm learning comes to mind.....
hormones are available already cut for ease of use. most are unnecessary for keeping culture alive and flourishing for experiments, storage. if you can imagine a grain of salt dissolved in 5 gallons of water, and 1/2ml drop will make another gallon of hormoned water, and a drop of this makes a handy litre of medium mix..not scientifically accurate, but shows how dilute these are at the end. uv light would most likely cause genetic disruptions, failures/freaks. and ozone is unknown in vitro to me. I stay clean, works, mostly. I

my incubation area is a grow tent no longer needed. vents taped shut, no air movement. lysol everytime in and out, wash,clean clothes. minimal peaking. tent was sprayed with alcohol in the beginning. your issues will not be the area, or your sterile medium. It will be in transfer from one jar to another, and most will come from your plant material. there is a fine balance between sterilizing live (or dead) plant material, and disrupting the dna and ending its life. when they die, they can mold. strong established plants mold seldom, even when exposed to air. I no longer use a positive hepa air in the transfer chamber either, so there....I believe this caused more issue than not. 1 year without, and I have way better results, maybe it was failing in design, not sure, but not necessary here. best frankenstein experiments come from the first knot on your stalk of a mature plant. this is where a virus attacked, changed dna,restabalized and the plant continued/survived. this area is full of strange cells, ready to be cultured.

I apologise for hijacking this thread.
please visit my thread for great updates https://www.rollitup.org/grow-journ...-5000perpetual-grow-extracts.html#post6994167
thanks
pc

Sorry again if its messed up the continuity of things. I have been editing my post for a while not realizing you had responded to it in the meanwhile..

I keep seeing things that look more like what I would call "micro-propagation" than cell culture.. Maybe My impression of what "cell culture" might be is wrong... If you start with a seed or seedling it seems that you are only stretching the seed or clone..

I somehow figured we were looking to "clone" a mature plant using parts that would not otherwise be (growing tips or other conventional clone/propagation material). More or less stems and/or roots..
I was also under the impression that we were exploiting a survival/repair response from the plant to achieve this..

An ultra simplistic perspective would be if you were to be able to say quarter a plants stem length wise to expose the Parenchyma, (assuming its undifferentiated cells inside) then influence it to spring up a column of miniature clones (or atleast callus formations that could be used to make them)? Otherwise extract the undifferentiated cells and provoke them to make the callus that can be manipulated in that direction.
If the Parenchyma of cannabis contains the undifferentiated cells you are after, could the differentiated cells be dissolved or mechanically stripped away so that it were predominantly the undifferentiated cells being incubated? Would this provoke more successful callus formations?