Clone from leaf
Ok, lets shed those thousand page threads on this subject filled with, ahem, dubious information.
Theoretically it should be possible to clone herb in a different way. Possibly from a fan leaf. No guarantees, but it *should* be possible.
So here it is, Im throwing down the science gauntlet. Ill supply any information I can about tissue culture/cloning/hormone usage, and some of you guys actually put it to practice. Anyone willing? (Id do it myself, but I dont have a grow going on, dont have the space for this at the moment, and just got a new job)
Here is what youll need, and then what youll need to do with it:
Plant tissue of varying ages. We really need to try using a lot of different types of material, and of different ages. If the goal is ONLY to be able to take a mature fan leaf and clone from that, then use that but keep in mind the most regenerable material is the newest, youngest tissues.
Media to hold it in. You can do it with soil, you can try and do it all aseptically (without germs) in agar, you could use sphagnum moss, etc etc. a way to supply a rather high humidity will be necessary, a humidity dome, a plastic bag with a single small hole, etc etc. we cant let the material dry out, it has to stay moist. Misting and humidity control will be the small nuances which will largely determine the success of this venture.
Hormones: You will need a cytokinin and auxin source. Techhead has been cool enough to look up some links already, (cytoninins: Nitrozyme - Organic Fertilizer, Hydroponics, Diatomaceous Earth, Hydrogen Peroxide, Agricultural Fertilizers, Gardening Products but I imagine there is more looking to be done. The best scenario would be powdered hormones so you can control the exact amount to be used. If that is not possible youll need to tweak the levels a bit, experiment it will require more material, more room, and some decent bookkeeping on your part.
Thiamine (thiamine HCL): serves to keep the leaf healthy and green longer after excision from plant. Once again, not sure where to get this Id imagine you could find it in pill form somewhere. I only used 0.4 mg/l, so well assume a little goes a long way. You could get by without this I think but have a higher chance of success with it.
Nutrients: Im not sure if nutrients will be needed, or if the healthy leaf tissue will simply be cannibalized to support the new growth. But you guys know how to grow, so just use your judgement. I would say dont feed for the first week or so at least. After that perhaps ¼ strength organic nutes.
How to do it: (Ill try and make this succinct as possible, if youd like more of the reasons and rationale behind the procedure please ask, Im just trying to avoid a marathon post)
1)soak the media with the shooting hormone balance, and the thiamine.
Rooting: Auxin
Shooting: High cytokinin/Low auxin
Note that I cannot give exact amounts, because I dont know what concentration your hormone sources will be, nor the amount of water youll use. If you can manage to find that stuff out, well figure out what amount of hormone material you need per liter of water. But be systematic about it keep track of what youve done, and make various mixtures using various amounts we need to dial in the amounts used.
It is up to you whether you wish to attempt to make shoots or roots first. I suggest trying both at the same time in different pots, seeing what happens. This is biology, were almost sure to be surprised by something.
To mix hormones: Mix it up all in one batch of water. Including the thiamine. Ph the water at about 6-6.7 but it would be well worth it to try a range from 5.8-6.7
2)collect your tissue:
Use the newest fan leaves you can. Also try some smaller leaves from the top of the plant, newest tissue is best. If you happen to have some still living cotyledons, try those they are often considered the best for this purpose.
Cut the petiole off near the stem/branch, then make a cut or two about halfway through the petiole. This is all up for interpretation, wound the thing how you see fit. We want opened cells that can uptake our hormones.
Also make wounds on other parts of the leaf. We dont know if this will be successful, but it wont hurt. Make some wounds near the edges, near the midvein, in other interior parts of the leaf. Id say one wound per inch? Figure it out.
3)place the tissue on the media.
Pay attention to this part it is important. We have to find a way to have it in the media where the cut cells are able to uptake hormones. That means you might want to put it down in the soil a bit, but not so much that it will stay super wet. Maybe pile media around the wounded parts? Maybe just place it on top of the media but have a mist containing the hormones that you can keep the leaf moist with? Up for interpretation.
Supply a high humidity environment, a dome saran wrap, etc. dont let it get moldy, air it out when needed, just dont let it get dried out, that quickly kills new shoots and roots.
4)fresh media:
when you have achieved either rooting or shooting, transfer that segment of the leaf to the other media, soaked with the correct hormones for what you want next. No need to take the whole leaf, cutting off the segment with the new growth will be fine. (as in, you have shoots coming off the leaf, so you cut about an inch of the leaf off around the new growth and transfer that chunk to the new media. if the shoot is large enough you could probably separate it from the original leaf entirely and make it root.)
5)environment:
LOW light, high humidity, around 78-80 degrees and I think a heating mat might help.
6)you:
Patient, observant.
Good luck! Lets hear any questions or ideas! I've surely left something out anyways.
Ok, lets shed those thousand page threads on this subject filled with, ahem, dubious information.
Theoretically it should be possible to clone herb in a different way. Possibly from a fan leaf. No guarantees, but it *should* be possible.
So here it is, Im throwing down the science gauntlet. Ill supply any information I can about tissue culture/cloning/hormone usage, and some of you guys actually put it to practice. Anyone willing? (Id do it myself, but I dont have a grow going on, dont have the space for this at the moment, and just got a new job)
Here is what youll need, and then what youll need to do with it:
Plant tissue of varying ages. We really need to try using a lot of different types of material, and of different ages. If the goal is ONLY to be able to take a mature fan leaf and clone from that, then use that but keep in mind the most regenerable material is the newest, youngest tissues.
Media to hold it in. You can do it with soil, you can try and do it all aseptically (without germs) in agar, you could use sphagnum moss, etc etc. a way to supply a rather high humidity will be necessary, a humidity dome, a plastic bag with a single small hole, etc etc. we cant let the material dry out, it has to stay moist. Misting and humidity control will be the small nuances which will largely determine the success of this venture.
Hormones: You will need a cytokinin and auxin source. Techhead has been cool enough to look up some links already, (cytoninins: Nitrozyme - Organic Fertilizer, Hydroponics, Diatomaceous Earth, Hydrogen Peroxide, Agricultural Fertilizers, Gardening Products but I imagine there is more looking to be done. The best scenario would be powdered hormones so you can control the exact amount to be used. If that is not possible youll need to tweak the levels a bit, experiment it will require more material, more room, and some decent bookkeeping on your part.
Thiamine (thiamine HCL): serves to keep the leaf healthy and green longer after excision from plant. Once again, not sure where to get this Id imagine you could find it in pill form somewhere. I only used 0.4 mg/l, so well assume a little goes a long way. You could get by without this I think but have a higher chance of success with it.
Nutrients: Im not sure if nutrients will be needed, or if the healthy leaf tissue will simply be cannibalized to support the new growth. But you guys know how to grow, so just use your judgement. I would say dont feed for the first week or so at least. After that perhaps ¼ strength organic nutes.
How to do it: (Ill try and make this succinct as possible, if youd like more of the reasons and rationale behind the procedure please ask, Im just trying to avoid a marathon post)
1)soak the media with the shooting hormone balance, and the thiamine.
Rooting: Auxin
Shooting: High cytokinin/Low auxin
Note that I cannot give exact amounts, because I dont know what concentration your hormone sources will be, nor the amount of water youll use. If you can manage to find that stuff out, well figure out what amount of hormone material you need per liter of water. But be systematic about it keep track of what youve done, and make various mixtures using various amounts we need to dial in the amounts used.
It is up to you whether you wish to attempt to make shoots or roots first. I suggest trying both at the same time in different pots, seeing what happens. This is biology, were almost sure to be surprised by something.
To mix hormones: Mix it up all in one batch of water. Including the thiamine. Ph the water at about 6-6.7 but it would be well worth it to try a range from 5.8-6.7
2)collect your tissue:
Use the newest fan leaves you can. Also try some smaller leaves from the top of the plant, newest tissue is best. If you happen to have some still living cotyledons, try those they are often considered the best for this purpose.
Cut the petiole off near the stem/branch, then make a cut or two about halfway through the petiole. This is all up for interpretation, wound the thing how you see fit. We want opened cells that can uptake our hormones.
Also make wounds on other parts of the leaf. We dont know if this will be successful, but it wont hurt. Make some wounds near the edges, near the midvein, in other interior parts of the leaf. Id say one wound per inch? Figure it out.
3)place the tissue on the media.
Pay attention to this part it is important. We have to find a way to have it in the media where the cut cells are able to uptake hormones. That means you might want to put it down in the soil a bit, but not so much that it will stay super wet. Maybe pile media around the wounded parts? Maybe just place it on top of the media but have a mist containing the hormones that you can keep the leaf moist with? Up for interpretation.
Supply a high humidity environment, a dome saran wrap, etc. dont let it get moldy, air it out when needed, just dont let it get dried out, that quickly kills new shoots and roots.
4)fresh media:
when you have achieved either rooting or shooting, transfer that segment of the leaf to the other media, soaked with the correct hormones for what you want next. No need to take the whole leaf, cutting off the segment with the new growth will be fine. (as in, you have shoots coming off the leaf, so you cut about an inch of the leaf off around the new growth and transfer that chunk to the new media. if the shoot is large enough you could probably separate it from the original leaf entirely and make it root.)
5)environment:
LOW light, high humidity, around 78-80 degrees and I think a heating mat might help.
6)you:
Patient, observant.
Good luck! Lets hear any questions or ideas! I've surely left something out anyways.
