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Cannabis grafting
- Thread starter BobbyIronsights
- Start date
Darpa
Member
Here is an update on post 337 - 338 (scion desinfection)
The treatment of the contaminated plant material with insecticidal soap and bleach seem to have work perfectly. Clones are healthy. However, the mother plant is still infected with aphnid, even after weeks of treatment at a frequency of 3x a Week.
Here is my new batch of futur rootstock! This time I'll perform grafting on freashly rooted clones. I think It will have many advantage….
Cheer
Darpa
The treatment of the contaminated plant material with insecticidal soap and bleach seem to have work perfectly. Clones are healthy. However, the mother plant is still infected with aphnid, even after weeks of treatment at a frequency of 3x a Week.
Here is my new batch of futur rootstock! This time I'll perform grafting on freashly rooted clones. I think It will have many advantage….
Cheer
Darpa
Darpa
Member
Hi folk,
And since I've to deal with heavy stress, I need to change my mind with some science experiment. I hope some of you appreciate. It's a way for me to clear my mind from shit….
I checked on the fridge scion and they all look perfectly fine for grafting. So, in orther to get Something to think about, I deceided to perform severals additionnel cuts since my desinfection protocol seem to be working just fine.
Bigger plant material with more auxilary buds site….
Same desinfection protocol (3x insecticidal soap - 3 x bleach 10% - 3 clean water wash)
Contaminated plant material - that are special and that I Don't want to lose….
Scion preparation:
First wash : insecticidal soap
Bleach wash 10%
Next on next post....
And since I've to deal with heavy stress, I need to change my mind with some science experiment. I hope some of you appreciate. It's a way for me to clear my mind from shit….
I checked on the fridge scion and they all look perfectly fine for grafting. So, in orther to get Something to think about, I deceided to perform severals additionnel cuts since my desinfection protocol seem to be working just fine.
Bigger plant material with more auxilary buds site….
Same desinfection protocol (3x insecticidal soap - 3 x bleach 10% - 3 clean water wash)
Contaminated plant material - that are special and that I Don't want to lose….
Scion preparation:
First wash : insecticidal soap
Bleach wash 10%
Next on next post....
Last edited:
Darpa
Member
Hi IC friends,
The clones you see in post 344 are going to be use as root stock material for the scions cold storage experiment and this time I'll use freshly cut scions as quality control. This will be micro chip grafting of auxilary bud site, and I'll combine with longitudinal insection graft of auxilary buds.
The root stock will be freshly rooted clone, minimal size, on which all leaves will be removes. I'll perform a first cut through the steam, and the base of the clone for the longitudinal graft (2 auxilary bud graft), and then several side chips cuts for the insertion of different single auxilary bud.
Sorry for the poor quality of the following picture, but I guest that I could help. The yellow part are the micro scions, and the red dot are the auxilary bud site from which a new strain will grow on the main root stock…
To evaluate the succes of the cold preservation scion experiment, I'll perform the same protocol on different cold storage scion starting at one month of fridge preservation, and I'll compare the survival rate to freshly cut scions…
Cheer
Darpa
The clones you see in post 344 are going to be use as root stock material for the scions cold storage experiment and this time I'll use freshly cut scions as quality control. This will be micro chip grafting of auxilary bud site, and I'll combine with longitudinal insection graft of auxilary buds.
The root stock will be freshly rooted clone, minimal size, on which all leaves will be removes. I'll perform a first cut through the steam, and the base of the clone for the longitudinal graft (2 auxilary bud graft), and then several side chips cuts for the insertion of different single auxilary bud.
Sorry for the poor quality of the following picture, but I guest that I could help. The yellow part are the micro scions, and the red dot are the auxilary bud site from which a new strain will grow on the main root stock…
To evaluate the succes of the cold preservation scion experiment, I'll perform the same protocol on different cold storage scion starting at one month of fridge preservation, and I'll compare the survival rate to freshly cut scions…
Cheer
Darpa
Love&Peace
Member
This is incredible research!
Darpa
Member
Hi folk,
Update of post 339: cold scion preservation for grafting purpose.
In this experiment, I used scion stored in the fridge for 25 day and grafted them on unrooted clones.
Cold scion preservation doesn't seem to be a problem. The limiting factor in this test will be the use of unrooted clone for this experiment.
As soon as I removed them from the humidity dome, I have limited time to graft the cold stored scions before the rootstock steem start to show tumescence problem... (Saggy.. falling down)
But after all, we will see how it turn out...
Unrooted clones:
Removing some of the rootstock plant material:
On the left is the 25 days old cold storage scion perfectly healthy... On the right, it's the plant material I removed from the rootstoock. I wanted to use this material for QC but I'll wait for QAQC when I'll be working with rooted rootstock
Update of post 339: cold scion preservation for grafting purpose.
In this experiment, I used scion stored in the fridge for 25 day and grafted them on unrooted clones.
Cold scion preservation doesn't seem to be a problem. The limiting factor in this test will be the use of unrooted clone for this experiment.
As soon as I removed them from the humidity dome, I have limited time to graft the cold stored scions before the rootstock steem start to show tumescence problem... (Saggy.. falling down)
But after all, we will see how it turn out...
Unrooted clones:
Removing some of the rootstock plant material:
On the left is the 25 days old cold storage scion perfectly healthy... On the right, it's the plant material I removed from the rootstoock. I wanted to use this material for QC but I'll wait for QAQC when I'll be working with rooted rootstock
Darpa
Member
Then I performed auxillary tip and side grafting:
This time I use Teflon tape instead of saran wrap...
2 to 3 graft of cold stored scion were performed on each of the 4 unrooted rootstock material and then I place them separatly in mason jar with a saran wrap lip on them.
As I mentionned, this is just a pre-test for long time cold storage scions for grafting purpose on newly rooted clone as rootstock material...
Cheer
Darpa
This time I use Teflon tape instead of saran wrap...
2 to 3 graft of cold stored scion were performed on each of the 4 unrooted rootstock material and then I place them separatly in mason jar with a saran wrap lip on them.
As I mentionned, this is just a pre-test for long time cold storage scions for grafting purpose on newly rooted clone as rootstock material...
Cheer
Darpa
Darpa
Member
Hi IC Friends,
Since I'm convinced that the 25 day cold storage graft on unrooted clone material will be a succes, I deceided to push the experiment a little bit further.
Here are the unrooted grafted clones on which 25 day old fridge scions were grafter... All of the grafted scion look perfectly healthy except maybe one on the 4th clone on the right…
NOW!!! It's time to test Cryoprotective Agents (CPA) for the scion freezing storage part of the experiment.
The use of CPA is essential to avoid Intracellular ice crystal formation (IIF) within the scion plant tissue... that would causes several problems to preservation in a freezing environment. I won't discuss the damage process here but if you are interested, there is several scientific publication on this subject.
I wanted to test the formulation of an known Plant Vitrification Solution (PVS2), which consists of 30% (w/v) glycerol, 15% (w/v) ethylene glycol and 15% (w/v) DMSO. I would have change the ethylene glycol by the non toxic Propylene glycol (PEG)
Unfortunately, I when shoping today and I didn't find food grade PEG, so I'll perform a test with only glycerol at different concentrations at this time. In order to follow the glycerol absorption by the plant material, I'll use a red dye as a tracker.
However, it is already known that exogenous glycerol can be absorbed by plant cells, and this can be further phosphorylated to G3P by glycerol kinase (GK) in the cytosol, thereby increasing endogenous G3P levels and enhancing pathogen resistance.(Chanda et al., 2011; Zhang et al., 2015).
Here is a tab of glycerol freezing point vs concentration:
Next to follow in next post
Since I'm convinced that the 25 day cold storage graft on unrooted clone material will be a succes, I deceided to push the experiment a little bit further.
Here are the unrooted grafted clones on which 25 day old fridge scions were grafter... All of the grafted scion look perfectly healthy except maybe one on the 4th clone on the right…
NOW!!! It's time to test Cryoprotective Agents (CPA) for the scion freezing storage part of the experiment.
The use of CPA is essential to avoid Intracellular ice crystal formation (IIF) within the scion plant tissue... that would causes several problems to preservation in a freezing environment. I won't discuss the damage process here but if you are interested, there is several scientific publication on this subject.
I wanted to test the formulation of an known Plant Vitrification Solution (PVS2), which consists of 30% (w/v) glycerol, 15% (w/v) ethylene glycol and 15% (w/v) DMSO. I would have change the ethylene glycol by the non toxic Propylene glycol (PEG)
Unfortunately, I when shoping today and I didn't find food grade PEG, so I'll perform a test with only glycerol at different concentrations at this time. In order to follow the glycerol absorption by the plant material, I'll use a red dye as a tracker.
However, it is already known that exogenous glycerol can be absorbed by plant cells, and this can be further phosphorylated to G3P by glycerol kinase (GK) in the cytosol, thereby increasing endogenous G3P levels and enhancing pathogen resistance.(Chanda et al., 2011; Zhang et al., 2015).
Here is a tab of glycerol freezing point vs concentration:
Next to follow in next post
Last edited:
Darpa
Member
In this experiment, I'll test glycerol concentration of 30% (w/v) and 50% (w/v).
Plant material explants will undergo a sterilisation process as described in post 337 (insecticidal soap 3x, rince 3x, bleach 3x, rince 3x, dry)
Plant material selection:
Photo
Then, they will be expose to the CPA liquid medium for absorption of glycerol. (1 or 2 day at 4C) Then they will be wrapped as in post 340 and stored in the freezer for about 30 days.
The dye used is a food grade red dye (allura red) in corn syrup, glycerol and water (it also contain trace of sulfite, modified corn starch, agar gum, citric acid, potasium sorbate and sodium citrate)
I'll do the same with the PVS2 solution in a near futur.
Let's see if we can store plant material in the freezer for several weeks and then use them as scions for grafting purpose or as a clone....
Cheer!
Darpa
Plant material explants will undergo a sterilisation process as described in post 337 (insecticidal soap 3x, rince 3x, bleach 3x, rince 3x, dry)
Plant material selection:
Photo
Then, they will be expose to the CPA liquid medium for absorption of glycerol. (1 or 2 day at 4C) Then they will be wrapped as in post 340 and stored in the freezer for about 30 days.
The dye used is a food grade red dye (allura red) in corn syrup, glycerol and water (it also contain trace of sulfite, modified corn starch, agar gum, citric acid, potasium sorbate and sodium citrate)
I'll do the same with the PVS2 solution in a near futur.
Let's see if we can store plant material in the freezer for several weeks and then use them as scions for grafting purpose or as a clone....
Cheer!
Darpa
JustSumTomatoes
Indicas make dreams happen
Awesome work Darpa, nice to see some solid lab work and testing. Respect 
Darpa
Member
Hi IC Friends,
I deceided to test a different preservation method for the sub-zero cold storage of plant material.
This time sterilized scions were directly immerged in the CPA solution (30% glycerol) and were stored right away in the freezer without any acclimatation in the fridge….
We will see if this could be an alternate method for the cold storage of plant material for futur grafting experiment…
Darpa
I deceided to test a different preservation method for the sub-zero cold storage of plant material.
This time sterilized scions were directly immerged in the CPA solution (30% glycerol) and were stored right away in the freezer without any acclimatation in the fridge….
We will see if this could be an alternate method for the cold storage of plant material for futur grafting experiment…
Darpa
Darpa
Member
Awesome work Darpa, nice to see some solid lab work and testing. Respect
Thanks JustSumTomatoes! I'm glad that you like the experiments!
Cheer!
Darpa
