Clones, Mother plants, How to "Clean" cuttings!

[acespicoli]

Cuttings & Propagation

Clones and more can be discussed here.

www.icmag.com

In Progress... Feel free to share your failures and success

Plant hormone - Wikipedia

en.wikipedia.org

Phenotypic plasticity influences the success of clonal propagation in industrial pharmaceutical Cannabis sativa - PMC

The burgeoning cannabis market requires evidence-based science such that farmers can quickly and efficiently generate new plants. In part, horticultural operations are limited by the success of cloning procedures. Here, we measured the role of ...

www.ncbi.nlm.nih.gov

Propagation of Cannabis for Clinical Research: An Approach Towards a Modern Herbal Medicinal Products Development - PMC

Cannabis has been reported to contain over 560 different compounds, out of which 120 are cannabinoids. Among the cannabinoids, Δ9-tetrahydrocannabinol and cannabidiol are the two major compounds with very different pharmacological profile and a ...

www.ncbi.nlm.nih.gov

Clone References

Clone References Work in progress Adams, A. N. (2015). Elimination of viruses from the Hop (Humulus lupulus) by heat therapy and meristem culture. J. Hortic. Sci. 50, 152–160. doi: 10.1080/00221589.1975.11514616 CrossRef Full Text | Google Scholar Braemer, R., and Paris, M. (1987). Biotransfor...

docs.google.com

 

[acespicoli]

Why Aloe Vera is an Amazing Rooting Agent!

Aloe is a perfect all-natural way for you to get your clone on. Aloe is packed with enzymes, amino acids and secondary metabolites.

"Remember! Aloe has Salicylic acid. Salicylic acid is involved in local and systemic plant defense responses against pathogens. It plays a role during stresses such as drought, chilling, heavy metal toxicity, heat, and osmotic stress. SA can volatilize and warn neighboring plants of attack." -

Additionally – Aloe Vera has a rich history of assiting growers with cloning as is demonstrated by this quote from the same source above.

Aloe Vera: contains Salicylic acid is a plant compound, which has been used as a rooting agent for over 120 years in the nursery industry. This is the compound that's found in Willow trees, which you might have run across in posts on rooting a cutting. This plant material is also very high in Saponins (30,000 ppm) and this adds another level of benefits."

In order to clone with Aloe you’ll essentially follow the same steps EXCEPT for a few more steps:

Prepare an Aloe-Water solution. You can use a half-inch chunk of leaf and mix it with some distilled water. Don’t worry about the PH.

Soak the cuttings in the solution for about 12 hours prior to transplanting

Some growers even take the cutting and stick it directly into the aloe leaf. Make sure that it’s covered with the “goop” and then go ahead and plant it.

 

[acespicoli]

Katsu on Cloning
1. Take your cutting from a healthy looking branch that isn't very "woody" - green and flexible cuts seem to do the best.
2. Use a STERILE razor to make a diagonal cut in the stem to remove it from the plant, lightly shave the lower inch of the stem of the outermost layer, then use a rooting powder or gel like clonex.
3. This is the biggie - whatever medium you are using, make sure it is BARELY damp with distilled or RO water. If your medium is too wet it will take forever to root and your stem may rot before that happens.
4. Dome your cuts to keep humidity in. If they seem to wilt, spray the inside TOP of the dome (not the cuts) to raise humidity.
5. After 3 days, start cracking your dome until they wilt, then cover again.
6. Only give your medium enough water to stay slightly damp - never wet. Squeeze out your plugs if they get too wet.
7. Adding Azos (as directed) to the distilled/RO water you use to moisten your plugs before you stick your cuts in will speed up rooting.
8. Your goal is to keep the cuts BARELY alive so as to trigger the plants survival mechanism and throw off some roots. If they are too wet they get lazy.

Peace, KB

INFO FROM KATSU

 

[acespicoli]

Figure 5. Illustration of the experimental setup. A mother plant from each C. sativa cultivar (left) provided 12 cuttings, which were separated into three groups (middle) according to their respective immersion treatment: A4 inoculation; IBA exposure; or H2O treatment. Lastly, in the (right) part, there is the aeroponic system and cutting representation. Cuttings were immersed for 20 h in their respective treatment and maintained in aeroponic growth units for 25 days.

Figure 3. Visualization of different root/shoot ratios in C. sativa cuttings. “Hindu Kush” cuttings from each treatment during harvest and processing of repetition 2. (a) Cutting treated with R. rhizogenes A4. (b) Cutting treated with IBA. (c) Cutting treated with H2O with shoot lengths measuring 22.6, 11.5, and 20.5 cm, respectively. Bar ≤ 2 cm.

Conclusions​

Rooting performance was best in IBA-treated cuttings with increased root length, root dry weight, and root/shoot dry weight ratio in cuttings compared to those treated with R. rhizogenes or water. However, IBA inhibited shoot growth, which could be a problem later during commercial production establishment. Superior shoot performance was obtained in R. rhizogenes (A4) and H2O-treated cuttings, thus showing advantages for the later establishment of commercial production of cannabis. These results are preliminary as a deeper analysis could be obtained on WinRhizo. A variation in cultivar susceptibility toward R. rhizogenes was noted in 38% of inoculated cuttings that were successfully infected. The aeroponic growth unit was successful in generating rooted cutting in all tested treatments; therefore, it is an alternative to cannabis producers.
Plants 2023, 12(11), 2216; https://doi.org/10.3390/plants12112216

 

[Ca++]

I'm not sure about that last article. We need to know how consistent the grower is, before we can see if their treatments did anything. To judge consistency, the experiament was run twice. On the second run, everything rooted. Treated or just wet, is was 100%. However, on the first run, some were 0%. Exactly the same experiment, done to prove consistency, actually shows us there is an absolute worst case scenario. If we remove the two odd results, The A4 treatment is comparable to no treatment. It all becomes so close, as to become meaningless. When there is such low confidence, due to the water inconsistencies. Then we turn to the pictures of the roots, and the water looks best to me. Though standing in RO for 24h just seems crazy. There should be a test of not messing about at all.

 

[acespicoli]

I'm not sure about that last article. We need to know how consistent the grower is, before we can see if their treatments did anything. To judge consistency, the experiament was run twice. On the second run, everything rooted. Treated or just wet, is was 100%. However, on the first run, some were 0%. Exactly the same experiment, done to prove consistency, actually shows us there is an absolute worst case scenario. If we remove the two odd results, The A4 treatment is comparable to no treatment. It all becomes so close, as to become meaningless. When there is such low confidence, due to the water inconsistencies. Then we turn to the pictures of the roots, and the water looks best to me. Though standing in RO for 24h just seems crazy. There should be a test of not messing about at all.

Plain (sterile) water could be an ideal environment. It works well with IPA
Im looking for best data, you know the interesting part of the above studies to me ?
and need to evaluate your questions/statements against the paper on a re-read.

But the thing of problem is contamination bacteria, fungus, and yeast entering into the phylum
My method currently, sterilize blade with alcohol, cut directly to sterile distilled water
We thought (most growers) the failure as a air bubble... but what is in the air ?

problem is contamination bacteria, fungus, and yeast entering into the phylum
maybe why gel is good
or plant extracts because they contain things that fight off the prior mentioned contaminants

 

[acespicoli]

Had made a fogponic a aeroponic even used to just use a humidity dome over cut in perlite and vermiculite
Its not very difficult to do just that I even stopped messing with it all for awhile so im updating
whats the best new latest greatest and why?

Meristem - Wikipedia

en.wikipedia.org

Primary meristems​

[edit]
Apical meristems give rise to the primary plant body and are responsible for primary growth, or an increase in length or height.[2][3] Apical meristems may differentiate into three kinds of primary meristem:

• Protoderm: lies around the outside of the stem and develops into the epidermis.
• Procambium: lies just inside of the protoderm and develops into primary xylem and primary phloem. It also produces the vascular cambium, and cork cambium (secondary meristems). The cork cambium further differentiates into the phelloderm (to the inside) and the phellem, or cork (to the outside). All three of these layers (cork cambium, phellem, and phelloderm) constitute the periderm. In roots, the procambium can also give rise to the pericycle, which produces lateral roots in eudicots.[4]
• Ground meristem: Composed of parenchyma, collenchyma and sclerenchyma cells[4] that develop into the cortex and the pith.

10x microscope image of root tip with meristem

1. quiescent center
2. calyptrogen (live rootcap cells)
3. rootcap
4. sloughed off dead rootcap cells
5. procambium

DEEPER UNDERSTANDING OF HOW ROOTS FORM FROM STEMS?
ALSO A MICROSCOPIC CUT OF A MERISTEM GROWING TIP IS HOW TO GET HLVD FREE

 

[acespicoli]

Callus (cell biology) - Wikipedia

en.wikipedia.org

Morphology​

Specific auxin-to-cytokinin ratios in plant tissue culture medium give rise to an unorganized growing and dividing mass of callus cells. Callus cultures are often broadly classified as being either compact or friable. Compact calli are typically green and sturdy, while friable calli are white to creamy yellow in color, fall apart easily, and can be used to generate cell suspension cultures and somatic embryos. In maize, these two callus types are designated as type I (compact) and type II (friable).[18] Callus can directly undergo direct organogenesis and/or embryogenesis where the cells will form an entirely new plant.


THIS IS THE IDEA BEHIND MANY CLONING TREATMENTS ADDITIVES
Specific auxin-to-cytokinin ratios in plant tissue culture medium give rise to an unorganized growing and dividing mass of callus cells.

Indole-3-butyric acid - Wikipedia

en.wikipedia.org

 

[Ca++]

Baby Bio (brand) did their 'roota' gel, which was basically a fungicide. 0.0894% w/w 1-naphthylacetic acid. It's disappearance from the market was a disaster for me, and while I then found one last bottle, I have a bleak future. The usual clonex is nothing like as good, but does gain some usefulness if I go from clonex to a powder, giving a combination that's maybe 75% of the Roota's success rate. I have been trialing a few products, and combinations. My method for cuts is sticking them in a substrate though, and I'm likely to make some sort of aero system soon. I have a lot of learning to do..

The Roota probably suits me, as I don't do all that slightly damp effort. I do soaking wet, and leave them alone. What is done in 14 days gets to live, but I give the process 21 days, for unforeseen issues. I think they ran their test 24 days. For me, a cut that took 24 days, didn't make the grade.
I used to get the excess water out, when using rockwool and a bit of a wrist flick, and 4 finger pinch that wouldn't miss-shape them. It was no different, and after many talks with friends, it seems the extra visits required to maintain a lower water content, was a disturbance the plants don't like. I dare say in a room of cuts, this might be mute, but with a dome to lift, they would rather I didn't. It's that first critical week really. Leaving them alone has the best rates.

 

[Dime]

KOG taking clones,old school

 

[acespicoli]

Agrobacterium rhizogenes as a root agent​

The propagation of plants through stem cuttings is commonly used in the commercial production of ornamental plants, medicinal plants, and timber trees. However, some commercially important tree species exhibit a low root formation rate. As far back as 1930, the induction of hairy roots was observed in nursery apple trees (Riker et al., 1930), and it was subsequently determined that the root induction was caused by A. rhizogenes (Hildebrand, 1934). Since then, A. rhizogenes has been used to enhance root formation in plants that are difficult to propagate through stem cuttings (Zavattieri et al., 2016).

The use of A. rhizogenes as a rooting inducer for difficult-to-root plants has been found to be remarkably effective and suitable for a wide range of plants. The determination of the root-promoting ability of A. rhizogenes is contingent upon the combination of bacterial strains and plant genotypes. Within woody plant species, there exists variation in rooting efficiency. Additionally, the specific strain of A. rhizogenes employed is a critical factor. For instance, in the case of the challenging-to-root woody species, “Golden Delicious” apple, the induction of rooting rates ranges from 0% to 20%, depending on the strain of A. rhizogenes utilized. Notably, strains A4 and 232 successfully induce adventitious root formation, whereas strains 178 X A4T and R1000 are unable to do so (Pateña et al., 1988). Cuttings from mature jujube trees (Ziziphus jujuba Mill.) exhibit a significant challenge in rooting. However, the application of A. rhizogenes, specifically strain TR105, resulted in the highest root formation percentage (65%), which was twice as high as that of the uninoculated cuttings (32.5%). Conversely, strain A4 did not show any significant difference in root formation (Mochammad et al., 1996). Interestingly, in the case of Corylus avellana, the rooting rate reached 100% when inoculated with a combination of A. rhizogenes (A7 + 22) (Bassil et al., 1991). In addition to bacterial strains, the success of inoculation and production of hairy roots is significantly influenced by plant genotypes and states, as evidenced by multiple studies. For instance, in hazelnut, the stimulation of rooting of cuttings was found to be influenced by both the cultivar and the date of cutting collection (Bassil et al., 1991). Similar results have also been observed in other research groups (Magnussen et al., 1994; Mihaljevic et al., 1996; Sarmast et al., 2019). The infectivity and adventitious root production of A. rhizogenes in host plant tissue are contingent upon the compatibility between A. rhizogenes and host plants, the responsiveness of the plant tissues to the T-DNA, the production of phytohormones, and the juvenile state of the host tissues.

Auxin is a well-known root inducer, and exogenously applied auxins have been shown to accelerate the rooting process in cuttings of a wide variety of plant species. When auxin and A. rhizogenes are combined, they exhibit varying effects on branch rooting, including synergy, antagonism, or no effect. Previous studies have observed a synergistic action between IBA and A. rhizogenes in inducing rooting in radiata pine (Li and Leung, 2003) and walnut (Caboni et al., 1996). Conversely, an antagonistic action between IAA and A. rhizogenes has been observed in inducing rooting in Pinus monticola (McAfee et al., 1993). The combined effect of A. rhizogenes to stimulate rooting is contingent upon the specific species and genotypes of micro propagated fruit trees. In all tested genotypes, root formation was observed following infection with A. rhizogenes. Three distinct responses were observed: genotypes that rooted without the presence of auxins showed a decrease in rooting percentage when auxin and infection were combined; genotypes that rooted only with auxin exhibited either no effect or a synergistic effect between auxins and infection; genotypes that rooted solely with A. rhizogenes displayed either no effect or an antagonistic effect between auxins and infection (Carmine et al., 1998). According to Zarei et al. (2020), the induction of rooting in Picea abies, a species known for its reluctance in rooting, cannot be achieved solely through the use of A. rhizogenes. However, when A. rhizogenes is combined with auxin, successful rooting can be achieved (Zarei et al., 2020).

A histological investigation revealed that the development of auxin-induced roots differs from that of A. rhizogenes. In the presence of NAA, adventitious roots are generated endogenously, originating within the vascular tissues of the stem. Conversely, adventitious roots formed in response to A. rhizogenes infection exhibit both endogenous and exogenous growth patterns. In the process of endogenous root formation, calli are generated within the cortex, leading to the subsequent formation of tracheid nests, which results in a bulge in the stem. Additionally, exogenous callus, formed at the base of shoot, also gives rise to tracheid nests. Consequently, roots form from both of these callus structures (Ellen and Juvenal, 1993). The rooting process in walnut was found to be influenced by the combined action of IBA and A. rhizogenes, as well as the antagonistic effect of IAA and A. rhizogenes. Notably, a significant reduction in the roots containing bacteria was observed when A. rhizogenes was combined with either IAA or IBA (Falasca et al., 2000).

It is imperative to conduct strain screening to optimize the rooting rate for each specific plant species. The utilization of multiple strains of A. rhizogenes offers a captivating approach to enhance rooting. The modulation of plant sensitivity to auxin by A. rhizogenes is believed to contribute to the promotion of rooting (Shen et al., 1988; Spanò et al., 1988; Petersen et al., 1989; Christophe et al., 1991). This modulation is thought to occur through variations in endogenous auxin levels and auxin sensitivities across different plant species (Carmine et al., 1998).

Front Plant Sci. 2023; 14: 1196561.
Published online 2023 Nov 14. doi: 10.3389/fpls.2023.1196561
PMCID: PMC10682722
PMID: 38034586

Agrobacterium rhizogenes:​

paving the road to research and breeding for woody plants​

Baby Bio (brand) did their 'roota' gel, which was basically a fungicide. 0.0894% w/w 1-naphthylacetic acid. It's disappearance from the market was a disaster for me, and while I then found one last bottle, I have a bleak future. The usual clonex is nothing like as good, but does gain some usefulness if I go from clonex to a powder, giving a combination that's maybe 75% of the Roota's success rate. I have been trialing a few products, and combinations. My method for cuts is sticking them in a substrate though, and I'm likely to make some sort of aero system soon. I have a lot of learning to do..

The Roota probably suits me, as I don't do all that slightly damp effort. I do soaking wet, and leave them alone. What is done in 14 days gets to live, but I give the process 21 days, for unforeseen issues. I think they ran their test 24 days. For me, a cut that took 24 days, didn't make the grade.
I used to get the excess water out, when using rockwool and a bit of a wrist flick, and 4 finger pinch that wouldn't miss-shape them. It was no different, and after many talks with friends, it seems the extra visits required to maintain a lower water content, was a disturbance the plants don't like. I dare say in a room of cuts, this might be mute, but with a dome to lift, they would rather I didn't. It's that first critical week really. Leaving them alone has the best rates.

It's that first critical week really. true,
basically a fungicide. 0.0894% w/w 1-naphthylacetic acid
should get a product label and try to mix one from off the shelf chemicals

KOG taking clones,old school

Good old school series im gonna have to spend some time on that channel NIMBIN TELEVISION
Enjoying that video thanks for sharing it here

 

[acespicoli]

Some Science in your face!​

You see plants contain certain substances that help them form new growth and save off bacteria, infection and fungi. The mighty willow just happens to be loaded with these substances which is why you can basically stick a freshly trimmed branch into the ground and it will grow into a new tree in short order.
Salicylic and Indolebutyric acids more specifically. They really help to speed up the rooting process.
The chemical structures of Salicylic and Indolebutyric acids
“But, them there chemicals are all trapped in the tree man” you say? I can dig it, but there is a simple way to leech these acids out simply by soaking the clippings!

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The Witches Brew:​

It doesn’t get any easier than this folks! All you need is a willow tree, or access to one. Weeping willow contains the highest levels of these chemicals by the by. So let’s get started making some homemade rooting hormone!
Here’s a detail of my willow tree. Gracefully sweeping my lovely little babbling brook. I like to trim off the tips that dangle in the water for a cleaner look which gives me exactly what I need for my willow water!
You want to fresh green new growth. The freshest and greenest you can get. That’s where all the magic hides.

You’ll only need a handful, so don’t get to greedy. In this tutorial, we’ll be making a Ball jar’s worth so adjust your needs accordingly if you want to make a large batch. The ratio of willow to water is 1:2.

Clip off the tips of the branches and then strip off all the leaves and put them into the compost bin. Then take those thin little shoots and cut them into smaller one inch segments.

Fill up your jar one third of the way with the willow and then top it off with boiling water.

Pop the lid on and let it sit for at least 24 hours. For a stronger batch, you can put the jar in the sun and let it steep for a few days.

When it’s done to your liking, strain out the willow and store the water in a cool dark place. In a cupboard it will last for about two weeks, in the fridge, it’ll last about a month. Super simple isn’t it?!

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How to use the Willow Water:​

This will be a little different than your usual overpriced rooting hormone.

• For cuttings that can be rooted in water you can use a 50/50 ratio of willow water to regular water and leave it on a North-facing window sill to keep algae growth down. This will speed up the rooting process. You can then plant the clipping once roots have been established.
• For hardwood cuttings or plants that are a bit more difficult to propagate, you’ll want to soak the cutting in full strength willow water for several hours so that the nutrients can be taken up into the cutting and then plant it in well draining soil.
• for new plantings of young plants, use full strength willow water for the first few waterings to help give a boost to aid in the plant becoming established in its new home. After that, use regular water.
• If an established plant undergoes stress or damage, use willow water to help give it that little extra boost.

There you have it folks! Simple, fast and 100% free and natural. It doesn’t get any better than that! And seeing as willows are all over the place, it should be relatively easy to find one that doesn’t mind a little trimming. I’d love to hear some success stories if you give this a try, so keep me posted on your experiments with mad science…er, I mean botany.

SEPTEMBER 5, 2015 BY JAMES GIELOW

Salicylic acid - Wikipedia

en.wikipedia.org

Indole-3-butyric acid - Wikipedia

en.wikipedia.org

So yeah we already discussed IBA but not a natural off the tree source

 

[acespicoli]

CLONES!!! 7-14 Days! 99% Success Rate!

Hi everyone! Been a long time since I posted up a tutorial.. I hope you enjoy it.. I tried to make this as clear as possible so for you experienced people sorry if it seems a bit redundant... i wanted even the newest grower to be able to use this method. I told some people that were having...

www.icmag.com

Knight of the BlackSvn​

• Jul 12, 2017
• Add bookmark
• #22

Nice tutorial. This works even on a MUCH smaller scale. I use the shot glass version of the red solo cup, hole 1/3 way up, and filled with perlite. My cuts are only 2" tall and I use a small seed propagating tray and dome. It works great!

This is the way we always did it ... way back
(like that he used the mini red solo shot cups have some of those collecting dust :think)
Then it fit in a 1020 tray with a regular low top hmmm
Looks like LED tape?
With a seedling heat map @ 80F

You know im on topic off topic, lets hit on the cleaning cuts part as well?
Fine sticky found on forum search @ ICmag someone made it a sticky for good reason

 

[acespicoli]

Hop Latent Viroid (HLVd) & Pathogen Diagnostics: A Comprehensive Overview - Cannabis Industry Journal

Hop Latent Viroid (HLVd) threatens cannabis crops and results in reduced yield, quality and profit. However, the lack of education and research about this pathogen poses a bigger threat to the cannabis community. To safeguard the productivity and quality of cannabis crops we must learn accurate...

cannabisindustryjournal.com

 

[acespicoli]

Page_Serena_202104_MSc.pdf

drive.google.com

https://atrium.lib.uoguelph.ca/server/api/core/bitstreams/be7dc9eb-0422-4b56-8f42-8f97f1b5310e/content

 

[acespicoli]

DeBacco University

DeBacco University: Cultivating Students to Grow Beyond Their Teachers DeBacco University is designed to help the spread of education on a variety of topics. There is a focus on the sciences with current playlist including Biology, Agriculture, Forensic Science and the Horticulture of...

www.youtube.com

@Dime got lost on youtube
found some more videos, nice break from white papers

 

[acespicoli]

 

[foomar]

If you start with the best possible material from a lush and vigorously growing plant in veg then i see little difference in quality and speed of rooting with hormones/aloe or anything else , compared to tapwater.

Root zone temp seems to be the most important factor , more so than humidity.

 

[CharlesU Farley]

DeBacco University

DeBacco University: Cultivating Students to Grow Beyond Their Teachers DeBacco University is designed to help the spread of education on a variety of topics. There is a focus on the sciences with current playlist including Biology, Agriculture, Forensic Science and the Horticulture of...

www.youtube.com

@Dime got lost on youtube
found some more videos, nice break from white papers

I've watched quite a few of his videos in the past, as I remember they were basically pretty sound. He's just kind of a boring speaker and like all YouTube videos, it just takes too long for me to get the information I want. I would rather _read_.

I used to go to a site that would convert voice to text for YouTube videos but it didn't add punctuation. That made it almost impossible to read at a fast rate, so it was almost as bad as sitting there watching the video.

 

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Marijuana Botany: An Advanced Study: ​

The Propagation and Breeding of Distinctive Cannabis​

Robert Connell Clarke​