Bavarian Buds

fermented fruit juice cannabis

FFJ - Fermented Fruit Jiuce - The terp booster from Korean Natural Farming

Why do I need Fermented Fruit Juice?

The name says it all: FFJ is the sieved juice from homofermentative converted fruits which can be produced with the help of fermentation without the formation of alcohols. Depending on the time of harvest, these contain different proportions of relevant nutrients. (Ju-young Cho 1992)

In particular, these are the molecules that have not yet been completely broken down into their basic building materials (NO3-), such as amino acids and polysaccharides. These can be absorbed directly by the plant and save it, so to speak, binding energy, since it no longer has to synthesize it itself.

The homofermentative destruction (see LAB article for a detailed explanation) is particularly important here, as we only want to receive lactate and acetate in the first steps of lactic acid fermentation, as alcohol would kill the responsible microbes.

Which fruit can I use?

The most important thing in the production of FFJ is to use locally homegrown or at least Demeter / organically grown fruits, whereby the latter will perform significantly worse. That's because for the planned fermentation we need a large number of microbes, especially Lactobacillus, which we get more by adding LAB. Natural yeasts and other microbes help in this process.

Once we have fulfilled this point, we can concentrate on the fruit. Nutrient analyzes can be used here, as these substances are then released into the liquid through micronization (decomposition of the plant tissue during fermentation).

High concentrations of potassium make apples, for example, a well-suited candidate for our FFJ. But it doesn't just have to be the obvious fruits; vegetables like the pumpkin also achieve excellent results. In principle, all kinds of fruit can be used as long as they contain one of the 17 elements that can be used by plants.

 

Figure 1: Composition of mineral content of different apple varieties (Henríquez et al. 2010)

Immature or overripe - which is better in which phase?

When choosing the fruit, special attention must be paid to the degree of ripeness. This determines the timestamp of use in the growth cycle of cannabis in flowering phase. The flowering phase can generally be divided into three phases.

The first is the formation of inflorescences, paired with temporary, rapidly increasing elongation growth. Here the plant needs the highest amount of phosphorus in the flowering phase in order to produce enough inflorescences. Unripe fruits are the best choice for our FFJ, as they contain a high amount of oxalic acid. This organic acid is even better suited for dissolving phosphorus than sulfuric acid.. (Mendes et al. 2020)

Figure 2: High oxalic acid content in unripe bananas (Heather Wyman und Palmer 1964)

Overripe, but not yet bad fruit is excellently suited for use as an addition to irrigation water from the main phase, also known as "bulk weeks", ie "rapid biomass growth". These have the highest levels of organic acids, especially malic and citric acid.

Figure 3: Organic acid content in ripe banana (Heather Wyman und Palmer 1964)

These acids have already shown in several studies to significantly increase the dry weight of plants in particular in a concentration of 100mg/L (Talebi et al. 2014)

If you look at Figure 1, you will find that ripe bananas contain, for example, approx. 600mg / ml (6.2meq / 100g) of magic acid. This means that even small concentrations of FFJ are sufficient in the application.

But that was not all: Malic acid also strengthens the symbiosis with benign rhizobacteria such as B.subtilis FB17, which increases endogenous pathogen tolerance. These acids are normally produced in a complex manner by the plant itself. This also saves plant energy as ATP, which can be used for other processes. (Rudrappa et al. 2008)

How is FFJ made?

We've already covered the main part, but you need a few things first before you can start. Here is the list of ingredients you need:

  • The fruit (unripe or overripe)
  • A large mason jar (preferably two liters)
  • Cane sugar (for every gram of fruit, add one gram of sugar)
  • A rubber band
  • A cover, like a paper towel, that allows gas to be exchanged.
  • A bowl to mix
  • A scale

First cut the fruit into thumb-sized pieces and weigh them into the bowl. Now add the equivalent amount of sugar and a shot (2-4ml) of LAB.

Then mix it vigorously so that everything is moistened with sugar. You pour the mixture into the glass until it is too full. The edges / places with sugar on the outside of the glass can be cleaned with vinegar, otherwise ants or vermin could be interesting in the glass. You complete this with the paper towel and the rubber. Then it should be placed in a warm, dark place.

After 3-5 days, the solid mass should have lifted from the liquid. Simply sieve and saturate the liquid again with a little sugar.

How is FFJ used and applied?

Our FFJ is ready now. The leftovers can either be incorporated as head fertilization (fertilizer on substrate), but pay attention to the amount, otherwise you can catch unwanted guests.

You can also use the liquid normally in the maintenance spray (FPJ + brown rice vinegar + OHN), i.e. with rice vinegar and OHN. The concentration remains at 1: 500 as with FPJ. Read more about OHN in our first Korean Natural farming article.

Foliar fertilization should only be used if there are still small or non-existent blooms, as otherwise there is a risk of mold. As a rule of thumb, flowering week 2-3 is the last opportunity to apply FFJ to leaves.

Bibliography

Heather Wyman; Palmer, James K. (1964): Organic Acids in the Ripening Banana Fruit. In: Plant Physiology 39 (4), S. 630–633. Online verfügbar unter http://www.jstor.org/stable/4260273.

Henríquez, Carolina; Almonacid, Sergio; Chiffelle, Italo; Valenzuela, Tania; Araya, Manuel; Cabezas, Lorena et al. (2010): Determination of Antioxidant Capacity, Total Phenolic Content and Mineral Composition of Different Fruit Tissue of Five Apple Cultivars Grown in Chile. In: Chilean J. Agric. Res. 70 (4), S. 523–536. DOI: 10.4067/S0718-58392010000400001.

Ju-young Cho (1992): Cho`s Natural farming: Recipes and Instructions for use. Japan: Modern Agriculture.

Mendes, Gilberto de Oliveira; Murta, Hiunes Mansur; Valadares, Rafael Vasconcelos; Da Silveira, Wendel Batista; Da Silva, Ivo Ribeiro; Costa, Maurício Dutra (2020): Oxalic acid is more efficient than sulfuric acid for rock phosphate solubilization. In: Minerals Engineering 155, S. 106458. DOI: 10.1016/j.mineng.2020.106458.

Rudrappa, Thimmaraju; Czymmek, Kirk J.; Paré, Paul W.; Bais, Harsh P. (2008): Root-secreted malic acid recruits beneficial soil bacteria. In: Plant Physiol 148 (3), S. 1547–1556. DOI: 10.1104/pp.108.127613.

Talebi, Majid; Hadavi, Ebrahim; Jaafari, Nima (2014): Foliar Sprays of Citric Acid and Malic Acid Modify Growth, Flowering, and Root to Shoot Ratio of Gazania (Gazania rigens L.): A Comparative Analysis by ANOVA and Structural Equations Modeling. In: Advances in Agriculture 2014, S. 1–6. DOI: 10.1155/2014/147278.


Fermented plant juice Knef cannabis

FPJ (Fermented plant juice) - the KNF terp booster for Cannabis

1. What is FPJ and why do I need it?

Many people make little to no use of their green waste or weeds. The highest of feelings is the compost pile, which for most is a slow rotting compost. This requires long waiting periods. A thermophilic compost is faster, but also destroys important enzymes and amino acids through temperature exposure.

To preserve these essential plant nutrients, an enzymatic or fermentative conversion is excellent. This gentle, cold process is not only fast (less than two weeks) but also space-saving and odorless.

The amino acids obtained in this way, such as glutamine, can be used directly by the plant and do not first have to be synthesized from nitrate with an energetic (ATP) effort. This feeds the plant and provides it with energy-rich food.

1: Assimilation of amonia

Enzymes such as glucanases and amylases help to convert the contained secondary nutrients, like polysaccharides and amino acids, into usable substances.

Furthermore, a broad spectrum of probiotic microbes is present in the finished FPJ. These colonize the plant surface and the soil. In doing so, they displace pathogens and promote beneficial nutrient converters such as PSB (phosphorus solubilizing bacteria).

2. What plants are suitable for FPJ?

First we need to collect plants or plant in the garden for specific use. Care should be taken that the plant is very watery, otherwise complete fermentation will not occur.

Examples would be fresh, pathogen-free cuttings of tomatoes or dandelions.

Both are particularly suitable due to their nutrient accumulation in the tissue. Other plants are Beinweil, hemp leaves (only large, thick awnings) or flowers of heavy eating plants like zucchini, eggplant or bananas

If you want to make it more specific, you can just look online to see what nutrients are included. However, the water content and other categories must of course be respected there.

An important point with collected plants is the time of collection. Here you should go out early in the morning to be able to harvest the plants with morning dew. This will give you the greatest density/diversity of microbes. (Source 2)

3. How FPJ is made

The production is very simple and cheap. For 0.5L FPJ you need the following utensils:

  • 1x 1L jar
  • Approx. 500-800G brown unrefined sugar (can also be made with normal sugar)
    • Note: Unrefined sugar still contains its molasses content
    • Note: Approx. weight because you need the same amount of sugar as plant matter
  • Enough plant matter to fill your ¾ glass jar.
  • Mixing bowl
  • 1-2 tablespoons of LAB serum (speeds up the process, but is not essential).
  • Paper towel or a tissue that breathes
  • Rubber band
  • Dark storage space

Now that you have collected your material, you will need to chop it into thumb-sized pieces. It is best to shred the plant tissue directly in your bowl so that no juice is lost. This is essential for complete fermentation. Tearing provides a larger surface area for the sugar to attach to. Now we add the same amount of sugar as plant material and mix vigorously. It sucks the water out of the tissue and provides an excellent food source for our microbes. Then we add a shot of LAB, this stimulates the degradation processes by starting the homofermentative Lactic acid fermentation. Pour the resulting pulp into our jar until it is ¾ full and press it down evenly so that there is no air left in the mass. Make sure you really get everything out of the bowl, otherwise you will lose valuable juice.

On top of the pressed down parts of the plant, we add another 1-2cm layer of sugar to seal it and prevent the growth of unwanted fungi.

Tie the paper towel or a coffee filter (if the jar is small enough) with the rubber over the opening of the jar to allow gas exchange to continue. After all, CO2 is produced during fermentation. The rims must be clean, otherwise flies will be attracted. The best and most microbe-friendly cleaning you get with vinegar. This should be in the repertoire of every Korean Natural Farmer.

4. How is FPJ applied?

Like most Korean Natural Farming remedies, FPJ should be given in combination with others. A table on this can be found in the first part of the article series.

FPJ is mostly given as a "Maintenace Spray". This includes FPJ (1:500), OHN (1:1000) and BRV (1:500).

FPJ acts as the nutritional part here, OHN as pathogen prevention and terpene stimulant (Source 3). The vinegar contains many important amino acids and buffers the PH on the leaf surface. It thus ensures the rapid conversion of the remaining sugar on the leaf.

Therefore, BRV is indispensable in the administration of FPJ, otherwise pests will be attracted.

5. At what stage of my cannabis plant do I give FPJ?

Maintenace spray can be given from the end of the seedling phase until the second week of the flowering phase. The application is done by foliar fertilization. It is important not to use an atomizer but a normal pump sprayer, otherwise the microbes will burst due to the pressure and the small openings.

Spraying should not be done during full light or high temperature, otherwise there is no possibility of transpiration. This leads to overheating of the plant and corresponding reduction of photosynthetic performance.

It is also possible to apply with the help of watering, but not as efficient as via foliar fertilization. The concentrations remain the same.

6. The importance of local raw materials

Now we have gone through the topic FPJ, which can be meanly called Vegbooster. We will deal with the flower counterpart in the next article.

As a small conclusion, I would like to emphasize again the importance of local ingredients. The plant material must come from your area or at least your climate zone, otherwise the microbes are not optimally adapted. This will cause you to lose tremendous potential. Bought plants are also often contaminated with pesticides, which destroys the whole concept of KNFs.

7. Quellen

1. Spektrum.de; „Ammoniakassimilation“, (https://www.spektrum.de/lexikon/biochemie/ammoniakassimilation/338)

2. „Fermented Plant Juice for Cannabis“; (https://rollinggreenacres.com/fermented-plant-juice-for-cannabis/)

3. The_worlds_last_hope; (https://www.instagram.com/the_worlds_last_hope/)


Lactic acid bateria

LAB - lactic acid bacteria I The bouncer of Korean Natural Farming (KNF)

What are lactic acid bacteria actually?

The term lactic acid bacteria is the German translation for Lactic Acid Bacteria. This English term is used internationally in the Korean Natural Farming scene. Lactic acid bacteria can be isolated via fermentation processes and used for garden applications. We will go into more detail about the exact application possibilities in the next chapter, but first let's look at the structure and taxonomy of the microbes.

LAB are a large group of bacteria and cannot be narrowed down to a single species. They have been used for centuries by many different cultures for fermentation and preservation of foods such as sauerkraut or kimchi. These bacteria survive even the low/acidic PH produced by fermentation, which also allows them to survive our digestive system when consumed. These bacteria are very good for our intestines and help with constipation and keep the microorganism milieu in balance.

LAB in the Gut and its effects
(Source 3)

As if that wasn't enough, LABs are also responsible for the production of yogurt and cheese, as they form the basis for separating the rennet from the liquid. However, in today's high-tech production, native rennet strains are no longer used but specially grown cultures. This is because different population compositions also produce the different types of cheese.

What are LAB used for in Korean Natural Farming?

Now we have heard a lot about commercial use, but how does it benefit us now in gardening? Since there are three different purposes, we will make a small separate chapter for each.

Use as a single preparation

Now that we have isolated our LAB serum, we can use it as is. The only thing to keep in mind is to dilute it with water, otherwise it can give off a rather strong odor after application. The rate for this is about 1:1000 with pure, antibiotic-free water. Unfortunately, an exact figure cannot be given here, as each LAB serum is colonized to a different extent.

It can then be sprayed directly onto the soil around the base of the plant. This prevents colonization with harmful bacteria and speeds up the conversion of organic fertilizers. A special advantage that LABs offer is the conversion of sugars to hexanoates (salt of caproic acid = hexanoic acid shown in source 4), which are precursors and building blocks for terpenes. This means that the plant does not have to produce these substances itself, which is energy-intensive, but can use them directly. This saves energy, which can be used again for the production of sugars and other metabolic processes.

Biosynthesis of Cannabinoids and Terpenes
(Source 4)

The next and important point is the use in the phytosanitary field (pest prevention). Again the LABs can shine by being extremely strong and competitive against other microorganisms. As already described in the production, after a certain incubation/propagation time they can take over almost the entire area by claiming the available food (sugar, especially lactose) for themselves.

Not only against other bacteria but also against fungi such as powdery mildew they can successfully assert themselves. This is how the oldschool trick of the milk/water mixture against the uninvited guest came about. However, this is now much more efficient and faster with the help of our LAB serum.

The concentration for this also remains at 1:1000, since the LABs can multiply very quickly, if enough food is available. The whole thing should be applied as a foliar spray by spraying the plant with an atomizer, but take care not to go below a particle size of 5µm, otherwise this will kill the bacteria.

Lactic acid bateria
(Source 5)

The treatment is extremely effective in a protective (preventive) way, but its effectiveness decreases with the severity of the infestation, so it is only partially curative (plant is infested but does not show symptoms yet) and poorly eradicative (strong visible infestation). In case of eradicative planned action, a synthetic fungicide should be resorted to, if it is really absolutely necessary to save this cultivar as in the case of phenotypic selection.

Another point to consider with lactic acid bacteria is their ripening delaying effect, which can give undesirable results especially in the last 2-3 weeks of the flowering phase. From week 3 at the latest week 5 (depending on the duration of cultivar flowering of course) the addition should be stopped, otherwise neither the calyx hairs will turn brown nor the trichomes will mature properly.

Use as compost starter

Now we come to one of the most important point of LAB application: Its role in the initial processes of any compost.

First, we should look at the basic process of lactic acid fermentation, which is an anaerobic (without oxygen) catabolic (degrading) process. Here, glucose is converted to pyruvate and this is converted to lactate (salt of lactic acid) with the help of a coenzyme. This lactate is the starting material for the following decomposition processes, which finally leads to our beloved compost.

Depiction of Lactic acid fermentation and subsequent degradation of Pyruvat

 

This process can be further divided into homo- and heterofermentative processes, after which the respective bacterial strains are also differentiated.

The first process is the "pure form", which means that this degradation is pure, i.e. apart from lactate no other by-products are formed (apart from the "used" coenzyme). Although lactate is also acidic, it is much gentler for microbes than acetic acid, thanks to its lower pKA value. Because with a low pKA value, the acid or salt of an acid can interact poorly with the microbial cells.

Heterofermentative fermentation, on the other hand, as you can probably imagine, produces other substances such as acetic acid and ethanol, which are highly toxic to our compost. This can lead to a lowering of the PH levels and the subsequent tilting of the population. You can recognize this at home by a strong smelling compost, which looks rather slimy and structureless than decomposed.

Homo- vs Heterofermentative Metabolism
(Source 6)

The starting point of these microbial conversion processes in compost is almost always started by two different strains of LAB.

The first is "Pediococcus acidilactici" (homofermentative), which accounts for more than half of the bacterial strains grown, this inhibits the synthesis of acetic acid, which in turn is extremely toxic to most auxiliary bacteria.

The second one is "Weissella paramesenteroides" (heterofermentative), which is the counterpart of the above mentioned specimen. This one produces exactly large amounts of acetic acid and slows down the composting process accordingly. This synthesis drastically decreases the PH in the entire colonized area and thus prevents the start of the decomposition process.

Accordingly, one should pay attention to the ratio in which it is applied. In a study of the "the science of total Environment" Magazines a ratio of 10^1,5 was found as optimum.

In this optimal ratio, the lactate blocks the PH reduction by the acetic acid and thus allows the colonization of the rotting raw material with fungi, which can degrade complex organic compounds such as lignin or chitin. The resulting degradation products again form reactants (starting materials) for terpene synthesis and activate systemic (plant internal) pest defense processes such as the formation of R-proteins.

If these fermentation processes are now finished after approx. two days, Paecilomyces species begin with the colonization of the material, the degradation of organic acids (PH increases -> aerobic bacteria settle from approx. Ph=6.5). Thus, the microbial aerobic degradation process can be started.

 

Combination with other KNF products

In the last part of the application in the horticultural field, the synergetic effect with the other KNF preparations must of course be mentioned. Mostly it is used right from the beginning as part of the Seed-Soak Solution (SSS), an organic solution that optimally prepares the seed for germination.

As mentioned before, LAB can repel competing bacteria or fungi (e.g. Pythium) lurking on the small, weak seedling. The exact mixture of the SSS will be discussed in a separate chapter, but roughly speaking, it is a combination of BRV (brown rice vinegar), FPJ (fermented plant juice), OHN (oriental herbal medicine) and our LAB.

But what are the other preparations for? Good question, because these also bring massive benefits to your sustainable garden. The rice vinegar buffers the PH range around your seeds into the right range so that the formation of anaerobic metabolites (alcohol etc) is suppressed.

The fermented plant sap gives a wide range of beneficial raw materials such as yeast species/hormones/enzymes and nutrients. This little lunch package helps the seed to establish itself quickly and strongly so that it quickly enters the "safe" phase where it can no longer be destroyed by just a small fungus.

Finally, the OHN is added, which together with the LABs plays the bouncer against fungi. The combination of alcohols (very very low concentration, which damages pathogens, but cannot harm our seed) and the active ingredients of ginger, angelica, licorice, garlic and cinnamon makes it almost impossible for pathogens to damage the seedling.

 

Another important application is the combination with IMO (indigenous microorganisms). Here LAB forms one of the starting materials in the collection of IMO also again with the protective aspect. Because we want strong, composting fungi and not fast-living bacteria.

But also in the preparation of a new soil mixture it can be added together with IMO or Liquid IMO to create a good base and stimulate the nutrient cycle.

Inputs: Tasks: Mixing ratio: Seedling: Vegetative state Flower I (Week 1-4) Flower II (Week 5-8) Flower III (Week 9-10)
OHN Medicine 1:1000 4ml 4ml 4ml 4ml 4ml
BRV Catalysor 1:500 8ml 4ml 8ml 8ml
FPJ Food Veg 1:500 8ml 8ml 8ml
FFJ Food Flower 1:500 8ml 8ml
LAB Starter 1:1000 4ml 4ml
FAA Fuel 1:1000 4ml 4ml
SW Minerals 1:30 120ml 130ml 150ml
WCP Bone builder 1:800 5ml 5ml

How can I grow the LAB myself?

We have now heard about its many uses, but how do we put it into practice?
First, we need some materials:

  • rice
  • A lactose source (milk/milk powder/isolate)
  • Clean water
  • A large glass jar
  • A breathable cover (paper (uncoated)/silk cloth/fabric)
  • Rubber band/thread
  1. First, we soak the rice in water for 48std to filter out the starch. The rice should be completely covered and stirred 4-5 times.
  2. The water is filled into the previously washed jar to about 2/3, covered with the help of the cloth and tightened with the rubber band. This prevents pests from creeping in. It is especially important that the jar does not contain any residues of detergents/vinegar cleaners or similar agents.
  3. Now you can incubate the jar at room temperature out of reach of sunlight for 3-5days. The time varies depending on the temperature and richness of the environment in lactobacilli.
  4. After the waiting time, a semi-solid film should have formed on the surface. Do not worry it is not mold Lactic Acid Bacteria Züchtungsbeginn                (Source 1)
  5. Now you can remove the lid and pour off the water, but be sure to remove the semi-solid part that floats on top.
  6. Then the lactose source, in our case milk, can be mixed with the opaque water in a 10:1 ratio. However, the final glass should still not be more than 2/3 full, otherwise it will overflow.
  7. The jar can now be sealed again as before in a breathable manner and stored. The mixture should now not be shaken during the fermentation phase.
  8. After another 4-6 days the mass should have separated as shown in the picture below. Again, the temperature is crucial, however, 25 ° C should not be exceeded, otherwise it can go bad.
    .              (Source 1)
  9. The yellow liquid is now our LAB serum, which we strain and take great care not to mix it with the solid again.
  10. The mixture should have a slightly sweet smell and if it starts to stink (sour milk), pour it away.
  11. If the serum is to last longer than 3-4 days it must be stored half-open (CO2 is formed) in the refrigerator. For very long storage (3 weeks +) it should be mixed with equal weight of brown sugar so that there is no unbound liquid left. This puts the LAB into a kind of cryo-sleep also called dormancy, where they become active again as soon as enough water is available again.

Can I still use the curd/lab?

The real rennet or curd, which has settled on top, can be removed and eaten. Especially for animals like dogs, pigs and chickens it is very nutritious and helps the digestion by its probiotic effect.
For human consumption, however, it should first be made into cheese, for this there is a super
tutorial video by Chris Trump

 

Lactobacilli and the human intestinal flora

As already mentioned in the introduction, lactic acid bacteria contribute significantly to intestinal health. Here they keep the balance between acidifying and alkaline acting bacteria. Because by fighting for nutrients, they can outcompete smaller, faster-digesting bacteria. This is good for us, because when bacteria convert nutrients, mostly H+ ions are released, which can lead to a rapid drop in Phs (PH = negative decadic logarithm of H+ concentration). This causes an unbalanced intestinal flora, resulting in digestive problems and even ineffective utilization/absorption of nutrients.

Unpasteurized dairy products contain a lot of lactobacilli, but can also contain pathogens so we would personally advise against it unless you have a microscope and can really differentiate the species.

A good option would be tiny doses of LAB serum (too much can easily lead to unpleasant side effects like diarrhea or abdominal pain). The best alternative in our opinion is to process the "curd" into cheese and eat it. But even here you should pay close attention to the quality, because the curd can sometimes tip over due to the non-sterile production, if it is too warm. This has a similar effect as fermented milk on one so it is not recommended.

But there are many good tutorials on cheese making and ripening with separate microbial strains, so you can easily make your own dream cheese.

 

 

Sources

 

  1. "Natural Farming: Lactic Acid Bacteria"; David M. Ikeda1 , Eric Weinert, Jr.1 , Kim C.S. Chang1 , Joseph M. McGinn1 , Sherri A. Miller1 Cheyanne Keliihoomalu2 , and Michael W. DuPonte2 1 Cho Global Natural Farming Hawai‘i, Hilo, HI
  2. "Lactic acid bacteria modulate organic acid production during early stages of food waste composting";Quyen Ngoc Minh Tran 1, Hiroshi Mimoto 1, Mitsuhiko Koyama 1, Kiyohiko Nakasaki 2Q
  3. "Lactic Acid Bacteria and Bifidobacteria with Potential to Design Natural Biofunctional Health-Promoting Dairy Foods"; Daniel M. Linares1,2, Carolina Gómez1, Erica Renes3, José M. Fresno3, María E. Tornadijo3, R. P. Ross2 and Catherine Stanton1,2*
  4. "Complete biosynthesis of cannabinoids and their unnatural analogues in yeast"; Veronica Benites;
  5. "The value of Lactic Acid Bacteria in Bees’s stomachs and honey for human medicine"; Mark;
  6. "Biotechnological valorization of agro industrial and household wastes for lactic acid production";Juliana Romo-Buchelly, María Rodríguez-Torres, Fernando Orozco-Sánchez;


Korean Natural Farming I The future of sustainable agriculture

Why should I use KNF?

Our environment is not doing well, whether it is due to climate change, overuse of pesticides or over-exploitation of fertilizers.
One major influence is the large monocultures that make up a big part of the crops grown annually around the world. In the graph below, we see that corn accounts for almost a third, the majority of which is used for animal feed.

Vergleich der Erntegutmengen nach Kultur

(Illustration 1)
 

This intensive land use has resulted in an enormous removal of phosphorus from the soil. This, coupled with a steadily increasing world population, poses a challenge to conventional agriculture which will not have enough mineral phosphate fertilizer to meet global demand by about 2050. Below we have included a small comparison between phosphorus supplies and its consumption.

Darstellung des zukünftigen Phosphatmangels auf der Erde

(Illustration 3)
 

The occurring shortage of phosphorus also has the effect that there is less yield per plant. This again increases the demand for agricultural land, which is decimated by the increasing sealing of areas. Now the question is: How do we combat this trend?
One promising option is to combine conventional methods in a mitigated form with a sustainable, regenerative farming style such as Korean Natural Farming. Especially by using indigenous adapted microorganisms and the very effective composting/recycling of organic waste, we can counteract this trend.
It should be clear, however, that on a large, world-supplying scale, not only natural farming methods can be used, since one must ensure a basic security or basic yield with fertilizer salts.

Vergleich Mais mit/ohne Phosphormangel

(Illustration 2)
 

However, we as cannabis growers and "small scale producers" can fully work with these methods and thus reduce our Co2 footprint. This is because our beloved indoor growing method in particular is unfortunately very resource intensive.
While this is usually essential for legality reasons, KNF and Living Soil methods can save quite a bit.
How to do this and still keep your yield or even increase it, we will take a closer look at in the next articles.

What exactly is Korean natural farming?

The origin of this concept can be found in, as the name suggests, Korea. South Korea to be exact. Here the inventor Cho Han-Kyu, also called Master Cho, thought about how to achieve good results in horticulture/agriculture with old techniques, implemented in a modern way. Through this, a holistic concept was created, which owes its extraordinary results to the interaction between indigenous microorganisms and fermentation processes.

Master Cho Portrait

Master Cho (Source 1)
 

Special attention was also paid to cost minimization. By using locally available inputs and waste recycling, KNF is one of the cheapest methods to achieve good results.
Master Cho got the basic ideas behind local inputs from his studies in Japan, where he spent several years before that with highly respected horticulturists such as Yasushi Oinoue.
Back in South Korea, he combined this with the "old" techniques of the Koreans, who had already done some research in this area through kimchi and other fermentation products.

This resulted in the concept that is now trending among sustainable farmers worldwide.
It works so well that Master Cho has already been arrested due to pressure from agricultural companies in South Korea and has been imprisoned for a short time. But his teachings were still spread around the world and eventually he was released again.

How do we make use of the full potential?

As described in the previous chapter, KNF works by combining different preparations that perform different tasks.
This division makes it possible to create the perfect mixture at each stage of growth. But what is absolutely necessary, what is optional and how do you obtain your preparations?

This and much more will be described in more detail in the coming articles. This article will give you the basics of Korean Natural Farming so that you can choose which mixtures to apply at which time.

The Basics

We have already said several times that the interaction between the living organisms and parts of the soil plays the most important role in KNF, but why not just take bottles from fertilizer manufacturers, which promise the highest possible yield with their mixture?

Although these can also give good results, the labor and environmental aspect is crucial here.
To get the same yield and quality standards on a hydroponic system as on a Living Soil, you have to grow the same clone more often to meet and not exceed the respective nutrient requirements.
This is because mineral nutrients actually mean nothing other than that they are already present in their charged (ionized) form. Nitrogen, for example, is present in fertilizers as nitrate (NO3-  for annual plants) or ammonium (NH4+ for perennial plants). Through the charge, the nutrients are more or less "pressed" into the roots, as substances are absorbed here via charge gradients.
Many compare this procedure with force feeding, because the plant has no chance to reject excess nutrients and thus the famous "fertilizer burns" occur.

Aufnahme Nährstoffe durch Wurzel

 
(Illustration 4)
 

In so-called organic cultivation, on the other hand, one uses the symbiosis between plant and microorganisms/fungi. It is true that the term "organic" is difficult to define, since rock flour is also used here to supply minerals. These are partially ionized and therefore immediately available.
However, the majority, just like other inputs, is mineralized only gradually. How fast this works depends on the mineralization rate, which in turn depends on many factors.
I will break this down in detail in the Living Soil article, but here I have written down a small list of influencing factors.

  • PH-value of the soil (6-7 is best)
  • Temperature (20-25°C is optimal )
  • Microorganism composition
  • Microorganism quantity
  • Soil moisture
  • Soil structure

All these factors can be positively influenced by adding KNF product. These either directly promote the microbial population, provide nourishment or displace pathogens (harmful organisms). The soil structure is also positively influenced, as the bacteria release a kind of slime that cements the soil particles together, thus creating better water-holding properties and nutrient storage.
These microorganisms can be added exogenously (from outside) to push mineralization in a certain direction. However, this is not an instant solution; the goal should always be a balanced population of different species.

In fact, if you have a complete biotope, the plant controls the amount of certain bacteria through its root exudates. Root exudates are sugar compounds on which microbes can feed excellently. The plant exchanges them for nutrients, which in turn promotes the particular microbial species that provides the desired nutrient.
A small example: The plant wants more potassium, then it releases a specific exudate matrix, which is detected by the potassium-releasing bacteria and these are then stimulated to exchange as shown by the illustration.

Symbiose Pflanze/Bodenorganismen

 
(Illustration 5)
 

Furthermore, VOCs (volatile organic compounds) and organic acids are released for pathogen defense and release of nutrients.

The importance of local Inputs

Why do I talk about local inputs all the time? I know I sound like a broken record, but this factor has a serious impact on the success of KNF products.

We are taking advantage of the full range of organisms that are perfectly adapted to our location. In other words we use natural selection to find out which microbes are best suited for our spot.
We also protect our native biotope, which would otherwise be displaced by sometimes invasive species. While this may be partially intentional it breaks down the natural homeostasis (balance) and can lead to undesirable side effects.
Another point is that the plant can develop to its maximum potential by controlling its own nutrient supply. This refers not only to yield but also to the quality of the material such as terpene content and trichome number or density.
This does not mean that locally there are only good varieties. You should follow the recipes exactly and pay attention to identifying characteristics like color and odor. How to determine these exactly I will show you at the respective products.

Gesammelte IMO Stämme

 
Collected indigenous Microorganisms  (Illustration 6)
 

If you are interested in the philosophy and further insights into the development and application of KNF, you should read the books of Master Cho himself. Some very good Youtube videos are also available from KNF greats such as Chris Trump or PureKNFDrake.

The Life Cycle of Plants

Now we have learned that with different inputs you promote different populations which again have an impact on the control of the plant. Now let's take a look at the life cycle of a cannabis plant and what is needed in which phases to be able to compose the different recipes.

Seedling/Clone

In this phase we don't need strong nutrient products yet, but we just need to prepare the soil and the seedling for the coming growth phase. For this we provide good microbes and prevent the expansion of the population of pathogens.

Vegetative Phase

In this phase the foundation for a successful harvest is made and we have to feed the plant accordingly. Products with very high nitrogen or amino acid content should be given together with the basic supply. This is where the root system and a stable branch/leaf system establishes itself. That is why we should apply both soil and foliar fertilizers.

Flower initiation

We now have a strong and thick growing plant and want to move into the flowering phase as quickly as possible. This should not only be fast but also corresponding with the yield potential.
To create the right basis for a high yield, the plant needs the basic supply and especially a boost of calcium and phosphorus in this phase.

Main Flower Stage/generative Stage

As the name suggests, this is the phase in which most weight is gained and quality can be significantly influenced.
Special attention should be paid to the supply of minerals and potassium/phosphorus.
It is also important to mention that we work here only with soil application, because we do not want to have residues of the agents or a risk of mold (Botrytis cinerea).

Ripening

We enter this phase in the last 1-2 weeks before harvest. Now we want to induce maximum terpene formation and maturation of the trichome heads.
For this we use similar inputs as in the main phase

A brief overview of KNF products

Now you are probably wondering, what should I use in each phase? The answer will probably be superfluous after this chapter, because I will now explain to you the main components of the KNF regime.
Here, however, it is not possible to distinguish by a strict NPK specification as with conventional preparations, because the products are so much more than pure fertilizer salts. In order not to go beyond the scope of this short overview, I will deal with production and exact modes of operation in the respective articles on the specific products.
For this reason, KNF tends to speak of tasks that the product performs. For OHN, for example, the term medicinal component is used because this strengthens general plant health.

Structure: Abbreviation = original name written out = translation = effect.

OHN = Oriental Herbal Medicine = orient. Herbal Medicine = medicine -> general plant health is strengthened by stimulating the immune systems
BRV = Brown Rice Vinegar = catalyst -> without this the PH partly fluctuates and some other inputs can not be implemented
FPJ = Fermented Plant Juice = food -> nutrients broken down/available from green plant material through fermentation + carbohydrates for
microorganisms
FFJ = Fermented Fruit Juice = food -> same principle as FPJ except that here fruiting parts are taken and therefore in the flowering phase
Application
LAB = Lactic-acid Bacteria = Lactobacteria = support -> these very strong microorganisms displace pathogens, fight Botrytis and accelerate
composting
FAA = Fish Amino-acids = fish amino acids = fuel -> this preparation gives especially in Veg really gas by the immense N-content and the completely preserved
amino acids the soil life is stimulated so strongly that the soil temperature rises and the plant makes large
growth leaps in a short time
IMO = Indigenious Microorganisms = backbone -> IMO makes up to 80% of the KNF performance and is therefore the most important ingredient, since it
brings the basic stock of soil life without which nothing works. Here there are some
gradations, but we will only go into this in the designated chapter

WSCP = Water-soluble Calciumphosphat = Bone-soluble Calciumphosphate -> Hereby we support the plant in the build-up of flower buds through
additional calcium and phosphate. This allows more nutrients to pass through the
ER (endoplasmic reticulum) and thus a better supply can be guaranteed.
supply can be guaranteed
SW = Sea Water (water + 5% salt) = Mineral Complex -> This rather simple preparation consists of only two inputs, but has a strong influence. Salt is extremely
full of nutrients and should therefore be used sparingly.

How do I combine these Inputs?

Now the question is what fits best in which stage. We have already outlined it in the description of the stages, but we still have an exact list for you here. This is the compilation as master Cho personally created it for 4L of water

Inputs Aufgabe Mixtureratio Seedling Vegetative Flower- Initiation Main Phase Flower Ripening
OHN Medicine 1:1000 4ml 4ml 4ml 4ml 4ml
BRV Catalyst 1:500 8ml 4ml 8ml 8ml  
FPJ Food 1:500 8ml 8ml 8ml    
FFJ Food 1:500       8ml 8ml
LAB Supporter 1:1000 4ml 4ml      
FAA Fuel 1:1000 4ml 4ml      
SW Minerals 1:30 120ml     130ml 150ml
WCP Bonebuilder 1:800 5ml   5ml    
               
(Source 2)

From this you can already deduce it: The standard administration in each phase consists of the so-called "Maintenance Spray", which includes OHN, BRV and FPJ except in the ripening phase, here you replace the FPJ with FFJ from ripe fruit. From there you can see what the plant needs or what would still be beneficial and can then add it on top of the Maintenance Spray (MS).
For example, if there is a calcium deficiency at the beginning of flowering, you add WCP to the MS and use it as a spray.
Now we only need the recipes and seasonal tips, then we have also worked through the chapter KNF.
One tip in advance, store a lot of brown sugar and by that I really mean a lot. You will need it for the next recipes.

If you like our articles and want to stay informed about competitions, events and our passion for writing: we are also available on Facebook, Linkedin and Instagram.

Sources

  1. „Korean Natural Farming: Master Cho Biography“; Nico Hill for Gardenculturemagazin; 06.05.2019; (https://gardenculturemagazine.com/korean-natural-farming-master-cho-biography/)
  2. „Cho’s Natural Farming: Recipes and Instructions for Use“; Cho Han-Kyu

Illustrations

  1. Pie Chart; VOX; (https://www.vox.com/a/explain-food-america)
  2. Maize P Mangel; Mary; (https://www.pinterest.nz/pin/38632509278445644/?autologin=true)
  3. Improving Plant Phosphorus (P) Acquisition by Phosphate-Solubilizing Bacteria; (https://www.researchgate.net/publication/318963768_Improving_Plant_Phosphorus_P_Acquisition_by_Phosphate-Solubilizing_Bacteria)
  4. Root Nutrient Foraging; R. Giehl, N. v. Wiren; (http://www.plantphysiol.org/content/166/2/509)
  5. „A Return to the Wild: Root Exudates and Food Security“; C.Preece, J.Penuelas; (https://www.cell.com/trends/plant-science/fulltext/S1360-1385(19)30247-X)
  6. „KNF and IMO“; Nico Hill; ( https://gardenculturemagazine.com/korean-natural-farming-and-indigenous-microorganisms/ )

Pineapple-Express-Ice-O-Lator-Even-Stone-Ice-Water-Hash

Ice water hash - a complete guide from harvest to dab

The amazing title picture was taken by the one and only Even Stone. Follow him on Instagram!

What is water-hash?

As you probably already expected from the heading for this method of separating trichomes from cannabis flower, we use a combination of ice and water. This mechanical procedure uses the force of the water vortex to brush of the oleoresin containing trichomeheads from yours buds. The ice in ice-water-hash making is only used to cool the whole mix down in order to make the stalks of the trichomes brittle so that they break of more easily. But the exact steps you need to take and much more will be discussed in the following article.

Why choose water-hash?

When you hear about ice-water-hash most times it sounds pretty labourintensive and it is, but the ratio of quality per worktime is the highest of the different solventless methods. Dry-ice-sifting for example oxidises the trichomes so fast that they burst and leaks out the desired compounds like the really volatile monoterpenes (limonene). And not only the trichomes burst, the plant material gets also really brittle, really fast so the contamination rate also increases rapidly. This results in a drastic loss of quality and worth, because even when pressed this can only achieve low prices on the market.

Dryice-Hash with a lot of Contaminants
Badly contaminated, green dry ice sift from GrowWeedEasy

Then why not use drysift, there's no dry ice used and no enhanced oxidation. Thats completely true, but as we're going for most quality per time drysifting also loses in that category for it's time intensive cleaning process of the material to get to 90% purity. Don't get me wrong I love drysifting and such fire as this fullmelt drysift from CubanGrower x The Cuban Hash Queen (Pheno by TheVillage/Photographed by Erik Nugshots/Bred by Symbiotic-genetics)makes it worth it.

Cuban Grower Drysift 99% Cleanliness

But to be profitable with that kind of premium product is really difficult and the market is not that big. So in order to sell at a lower price, but still nearly the same high quality as with drysift only ice-water-hash remains an option. "Why not just blast it all"?

Good question, and yes you could do that, but the price for bho currently is not as high as decent hash rosin. Additionally the startingcost is really high if you want to do it right. If you need deeper info on hydrocarbonextraction and it's specifics, visit Murphy Murris Instagram account

And for home users I wouldn't recommend it either if you don't have a proper, safe setup and are quite experienced with hydrocarbon handling. Hash on the other hand doesn't need a C1D1 room or other security measurements than a rubber mat on the slippery floor.

So if you want to make fullmelt the fastest way possible use ice-water-hash to separate your resin like the italien hashmaker slite23 from Barcelona with this beautiful Sundae Driver 149/90u Fullmelt (Pheno by Alorganics). Now we will look a bit further into the mechanics behind hash-making in order to build a good knowledge base, that will let you achieve resin like this with a bit of practice.

Fullmelt Hash by Slite23

The principles of Hash-making

Terminology

In the first part of this chapter we will look at some basic terminology that we need to describe the process precisely

  • Micron: This term refers to the opening space in the bubble bags. It tells you how many micrometers the wholes in the screen are wide. These mostly get abbreviated as "u" or ". To put that into perspective, a human hair is 75 micrometers thick and our eyes can only see down to 40 microns. Thats why it's important to buy a scope and look at the holes in the screen to see if they are all open and not clogged with nylon or bad stitching. Always wash your bag properly before using them. For washing the first time after buying new bags, I would recommend to make a 10% alcohol/water solution and rinse the heavily. But be careful not to rip the seams.
  • Terpenes: These pure hydrocarbons are a part of the famous entourage-effect firstly mentioned by Dr. Ethan Russo in his paper "Taming THC: potential cannabis synergy and phytocannabinoid-terpenoid entourage effects" (see source 1) They induce a synergistic effect in combination with Cannabinoids in your brain, basically pushing the high in a certain direction. Like Limonene for example, that on its own already causes a sort of high if you breath it in at a certain concentration. Similar to that strains with high linalool make you relaxed and the same for other strains with a lot of terpenes. There are also other components like thiols and alcohol esters in Skunk e.g but they aren't as well researched as terpenes At last there are also terpinoids, which are similar to terpenes in effect and structure, but the have additional chemical groups. But because of their similarities we mostly count them to the terpene content.
Different Terpenes Cannabis
(Source 12)

  • Fullmelt: We already used this term in this article and some of you probably wondered what that means. It stands for the quality of the hash in regards of melt % tage. Hash that melts like water in your banger and nearly leaves no residue is the best of the best in hash. At room temperature it sometimes already melts into a oil like mass of trichomes. More info on what differentiates the melt further and how you classify it will be discussed in the chapter Rating-Systems
  • Contaminants: All particles that aren't trichome heads be it hairs, skin, dust or plant particles are considered contaminants. More on how to prevent them and proper hygiene in the coldroom in the designated chapter
  • Cultivars: Every plant that is derived from sexual reproduction is, like humans, different even from his siblings. This term refers to one unique genotype with a specific phenotype that gets expressed. So for example just because some Cake clones yield not all of them will and even if you have a selected cut it might not dump a lot of weight if grown under completely different conditions like those that it was selected in. That's why it's good to ask the clone seller how it was selected or select your own cultivars
  • Flowrate: Like in coffee brewing you can control some parameters like speed of water drainage and with that force that pushes through the screens. Using that you can safe time by not having to spray and rinse that much. But that needs the perfect speed and that takes practice as you have to be careful not to blast the water through as that could drag contaminants through the higher micron screen and also degrade your trichomes by removing more terpenes than necessary. But not enough pressure prevents the contaminants from getting flushed through
  • RO-Water: Growers already know the term, but for everyone else: it refers to water filtered through a reverse osmosis filter. This device pulls out all the minerals and other compounds that could affect the taste. For optimal results you should make your own RO-Ice cubes (Protip: Insulate your washing vessel to save ice and prep time)
Reverse Osmosis vs Osmosis Principle
Reverse Osmosis vs Osmosis Principle (Source 11)

 

The Separation Process

How exactly can you isolate the trichomes without plucking them off one by one? For that to work we need Physics. Trichomes are secretion-organs with bulbous, oleo-resin filled heads on them. The oleo-resin is trapped by a layer of lipids that releases terpenes and other compounds slowly over the day on the plant to fight of pest. It also shields the flower from UV-Damage so to increase your trichome-coverage you should incorporate UV in your lighting mixture.

As we know from chemistry class, fatty substances don't mix well with water as they're non polar. This phenomenon is the reason why trichomes stay intact while beeing washed most of the time. If your strain as a to thin lipidlayer the trichomes smear in the bag and you won't catch most of them

Thevillage Trichome Close-up Shot
Trichome Close-up shot by theVillage

That's why some cultivars won't produce hash yield numbers that justify washing the material. Even if it has a lot of good trichomes you have to separate them completely in order to catch all. Therefore they have to be evenly brittle, but not brittle.

Thats where water comes into play. This natural "solvent" stays, even with ice, at 0-5°C for the whole washing-process to insure maximum Quality and efficiency. Now that we have the Trichomes ready for action we will start the stirring process. If you're hand-washing try to get a small water vortex as the current of the water strips the flower from the Trichome heads.

If you picked a good cultivar the water should turn slightly golden/purple (depends on the strain) and you should start to see the heads drop to the bottom of the wash container. If you think you got most of them, you can stop agitating and let the trichomes settle down. After that you skim the water on top that doesn't contain resin and either filter it or throw it away. Let about 1-2/3s in the wash vessel and strain them through the Bubblebags.

How to collect the Resin from Water

But in which order do I have to put the bags in? We go from big to small openings but how many bags you have depends on your bag-set. Different companies also use different mesh sizes but most of them are in the same region so we want to give you a small List of bags that there are.

  • 220μ = Filter-/Workbag: most of what's catched here is dust and plant material. This is to clean up most of the contaminants before catching anything. If you buy one that is big enough to fit into your washing vessel, you can wash in it and pull out most of the plant material even before emptying into the filter vessel.
  • 180μ = Second Filter: Most bag-sets don't have them but they're a good option if you want to add another filtration process to your regimen. Most of the heads catched here also don't have a lot of value
  • 160μ = Rosin-/Foodgrade: This bag collection is worth it, especially when you grow strains with really large trichomes like a GMO. You can even find some partial melt in here, but I would recommend to turn this into rosin or edibles/RSO as most of the time it is still a bit dirty.
  • 120μ = great Quality Resin: This is where the real fun begins. In this range you start to see real fullmelt or at least 4 Star quality, that is either worth keeping as hash or if it's doesn't melt fully to press it and sell it for premium prices
  • 90μ = exceptional Melt: The melt coming out of this bag is the true Creme de la Creme and goes over the counter for astronomical prices up to 130$/g. The fullmelt nearly leaves no residue on the banger and the taste is amazing. As said before, all non melt result should be processed to rosin or oldschool hash (Frenchy Cannoli-style)
  • 70μ = is used more like a substitute for the 90μ bag. To test which one you like better, you should test wash and compare how the cultivar performs with the different bags. Most intact, ripe Trichomes are not smaller than 70μ so this will be the last bag with exceptional results.
  • 45μ- 25μ = Foodgrade: This bag catches some debris, but mostly unripe or already dried up trichomes so it can be used similar to the 160μ
  • "Full-Spectrum": This term is a tricky one, because a real full-spectrum Oil refers to it containing the whole range of cannabinoids and terpenes. But most Water-Hash fullspec is a 45μ through a 160μ or similar micron sizes. This leaves out a lot of other compounds so technically it's not a fullspectrum oil in my opinion. But as with everything I teach, I would be really happy to discuss these topics in the comments. Real fullspec is only achievable with solvent-extraction as it can extract everything contained in the flowers.

 

BubbleBags by Mila Jansen
BubbleBags by Mila Jansen

High-End Bags from Iceextract
High-End Bags from Iceextract

What do I do with the wet Hash now?

Now that you catched all your trichomes, you want to make them smokable by drying them really thoroughly to prevent it from molding. This can happen in the dry stage aswell so be fast with the process. There are two main options for drying Ice-water-hash. The first and cheapest option is air drying it by freezing it immediately after collection and spreading it out really really fine after that. How to do this and the exact steps will be mentioned in the drying chapter.

The most used method today is the use of a lyophilisation ofen also called freeze dryer. This neat piece of equipment uses certain physical phenomenons to quickly remove the moisture from the trichomemass. It also is the most terpene and colour preserving technic for drying. For commercial ops this device is essential for production speed and success as a failed air drying attempt could cost you a lot of money.

How to prepare yourself before setting up a hash lab

Now most of you probably would want to immediately wash their whole crop, but before doing that, we need to set up a proper lab and everything that goes with it. For starter we need the most essential item to be of the highest quality possible. Of course I'm talking about the starting-material as it is our base for good hash. That's why the mantra for most hash makers is: "Fire in, Fire out" To give you an idea of what I mean with that, we will look at our material in depth and disect what makes good Hash starting-material

The different aspects of good startingmaterial

Live vs cured Flower

The first choice with have to make after harvest is whether we want to make "live" or "cured" hash . These terms refer to the drying process of our material. Live Hash is taking the fresh flowers and directly freeze them for washing. That way, you can conserve the most terpenes especially those that flair of easily like monoterpenes. This brings more sharp, intense flavor that many people nowadays really enjoy. also the colour gets "better" more easily with this method as nearly no degradation happened. What we mean with better and which role colour plays in hash, can be looked up in the designated chapter With this method it's vital to have an extremely clean freeze, if possible you should have a separate one only for fresh frozen flower. But if you only wash once every 2-3 Months this isn't really worth it, so you can use tupperware to protect your buds from the other freezer content.


Cured Hash Rosin
Cured rosin by Mammoth Melts from Rhode Island (Source 8)

Often the whole plant gets frozen for this purpose so you will sometimes see the term WPFF on your bought hash. This stands for "Whole plant, fresh frozen"

Live Rosin High-End Example
Sour Diesel Live Rosin

Cured Resin on the other hand has a more "rounded" flavor as the highly volatile Monoterpenes have flaired of or turned into another terpene profile. Most of this is still in research, but you definitely taste a difference. This rosin is bit harder to get as light in colour as the live version, because the dry time has to be perfect. But as we will see in the further chapters, colour doesn't always say much about quality.

How to visually rate Material

Now that you picked your methode of choice, we can start to analyse if our product will be cut out for making hash. As mentioned before the trichomeheads and their size play a mayor role in it.

Trichome detailed description
Trichomaufbau Schema Quelle: Hightimes

First of all lets look at this depiction of a trichome. This shape is what you're looking for in the perfect trichome category. It has a large bulbous head with a really thin "neck". This is what we want as it will make it easier for us to break them off and separate from the plant material. What we don't want are secretory glands without heads, their "hairs" may look frosty af on a plant, but if you wash them, you will be really disappointed. A typical strain with these attribute is purple punch. It looks fantastic but washes really bad most of the time. Another unfavourable trait for hash making are varieties with really long trichome stalks, small heads and a thick waxy cuticle as they won't contain a lot of active compounds. These strains are also valuable, but just not for water hash.

Don't get me wrong, if you select a fire cut, it can dump, but you would have to go through a lot of cultivars to find one. I recommend 20-50 regular seeds depending on the breeder. You can do smaller hunts, but the probability of finding a keeper cultivar that satisfies both quality and quantity aspects will be quite low. 

 

 

 

 

 

 

 

 

The importance of hash specific breeders

This is where the breeders come in. If you want to select a hash-cut but don't have the time to sift through hundrets of plants, you should pick a breeder that selects for hash. That will give you a lot higher starting chance of finding a good specimen as the breeder has already pre selected the genetic material. There are lot of different great breeders and most probably I will forget to name some and I'm sorry if I left someone out. But to give you a direction, here are some breeder recommendation from my side: Karma genetics, Oni seedco, Bloomseed co, Dying breed seeds, Archive Genetics, Truecannabliss and Cannarado to name just a few.

Some strains especially are good for hash-making as they have a high rate of successful cultivars. Names like GMO, Sour diesel crosses or the famous Gorilla glue are well known for their ability to dump wpff yields upwards of 5%. Even most crosses with there lineages will give you your desired results so they're a good starting point for your hash-journey. For companies I would also recommend to implement a really heavy yielding strain at all times in their rotation as a safety net, if some new selections perform worse than expected.

Now that we have picked a good strain, we can start growing and watching it. When the heads are nearly finished you can scope it with a jewelers lupe or a electronic microscope. This should be done extremely thoroughly because if you don't do it and do a test wash, you potentially waste money and thats not good when you're starting a company. To give you a comparison chart we depicted one picture from a study by Dr. Potter

Trichome Types in Cannabis

Figure D depicts the best usable trichomes as we don't need excessive force for separating these from the stalk. Trichomes like in figure C aren't the optimal, sure they have a large head and contain more oleo-resin than the ones in figure B, but they require so much force to get separated that there also is a lot of potential for contaminants to get into the product. All these criteria are only the beginning of a long r&d (research and development) journey that should look at terpene-make-up, terpene Content, rosin yield, stability, colour etc

The distribution of Trichome size

Another important performance indicator is how much a strain dumps in each quality area. A cultivar that puts nearly all its resin out as fullmelt is much more valuable than one that produces 70% edible grade hash. As with every property of hash that we look at, we need to do our tests here as well.

For this purpose you should use a register/excel sheet to get an overview of your data. This data is a treasure chest of usefull information about your process, that can give you the opportunity to optimize every little detail. This would be a example of how to analyse a strain:

  • Terpene make up
  • Terpene content
  • Yield WPFF vs Cured
  • Market value
  • How long is the overturn time (harvest-harvest cycle)
  • Trichome Size
Trichomesize distribution Alchimia
Trichome Size Distribution Visualised by Alchimia

How to test your cultivar for yield beforehand

To test and find the perfect cultivar is quite a challenge so I highly recommend keeping a cut from said plant. Not only for hash making, but for sharing /selling the clone. The high end concentrate marktet is really hot right now and new, good clones go for a lot of money. So this would be the perfect example for horizontal expansion of your production line.

But to get such a cut, we have to select and before selecting we should do some tests. The first is a cannabinoid and terpene test which you need regardless as a commercial cultivator. If you buy flowers you should ask for their Coa so that you can compare your yield to the given data and check if you catched all the trichomes. This is also a great opportunity to fine tune your normal washing processes. So if you don't get the same % in yield that is given in the testresults you should go through your process step-by-step in order to find the mistake. If you don't find anything, it could be the strain. Look at how the resin handles the water and if it smears the bags. Most times this is a large opportunity for product loss.

If thats not the problem, try using the leftover material for edibles/rso, that way you can use every little bit of the material and minimize loses.

COA Hash Rosin Cannabis

Another test is the mason jar shake. This simple method has a bit more to do with experience, but is also usable for beginners. You simply take 10-15 grams of your desired flower, put it in a mason jar together with Ice and water. After letting it sit for 2-3minutes you can swirl it around a bit and then you should see golden trichomes dyeing the water golden. Let it sit for another 2 minutes without stirring and look at the bottom of the jar. There should be a small layer of trichomes if you have a fitting cultivar.

Cultivation Techniques for better Hash

Swabianextracts King Sherb Cannabis Flower
King Sherb ( OGKB V.2.1 x Blue Sherbert ) grown by SwabianExtracts

The best way to get great hash yielding flowers is to grow them yourself, so that you can learn the subtleties that go into growing the best hash flowers. One sure way to increase resin content is adding UV-lighting to your grow as we mentioned before. This lets the flower build up its defences against this high energy radiation. Not directly for resin content but for terps, most hash cultivators use organic living soil system to grow as this lets the plant fully express its genetic potential. This debate is highly discussed in the hash community so I don't want to get too deep into it. But it has to be said, that salt based fertilizer can also achieve great hash if they're perfectly tuned in for the cultivar. A general booster for terps and secondary metabolites is a healthy amount of sulfur in our regimen. Organic growers can use gypsum or epsom salt for this purpose

The necessary Equipment for Washing

Now that we know our basics and how to choose material, we need to look at our setup. To configurate it the best way possible we need to choose between the two kinds of washing. A pros and cons list should be sufficient to give you an idea of what you want to choose

Hand vs Machine Washing

Pros washingmaschine

  • Multiple maschines can be operated by one employe
  • Less manual labour
  • simple SOPs can be made for unexperienced operators
  • "easy" for beginners

Cons washingmaschine

  • Lots of work to keep perfectly clean
  • High initial investment cost
  • cheap maschines need a draintube change in the beginning
  • often not very customizable

Pros Handwash

  • Easy to clean vessels
  • Fully customizable washing program
  • Really gentle on the material
  • Smaller initial investment
  • Large batches of 10-15k grams possible

Cons Handwash

  • hard physical work
  • Not automated
  • Experience is needed to get perfect results

The next big decision is whether to air dry or freeze dry. But it would blow up this chapter so we will look at this in the specific chapter.

A short Equipmentlist

Now for a better overview we collected a list of small and big items every wash room needs in order to function properly.

  • Bubblebag-Set (+ Workbag)
  • RO-Water
  • Ice-cubes (Ro-cubes if possible)
  • Washing Device (Maschine vs Vessel)
  • Stiring device for handwash (stainless steel paddle)
  • Drainvessel for filtering the hashwater (should be big enough to mount the bags in it)
  • Thermometer for water temps
  • Extremly clean work enviroment
  • AC for cooling the room
  • Cleaningagent (isopropanol 70% works best)
  • Waterhose with RO-water to spray down the bags
  • Cold Spoon for collection
  • Dryingdevice (Freeze Dryer or Microplane/Sifter)

How do I prepare my tools?

The most important factor is cleanliness, I know I sound like a broken record, but as everything gets concentrated with hash making, the dirt also gets concentrated. Thats why the phrase: "cleanliness is next to godliness" fits perfectly for hash making. You should start with your wash room and wipe everything down with a 50/50 alcohol/water mix. Because if you only clean your tools and put them back into the room, they get contaminated aswell. Even the ceiling and the walls should be cleaned as dust can settle there.

Next we need to take a look at our tools. They also should get wiped down and put in a cold place as the resin won't stick as much then. Your Bags should also get washed with a very mild alcohol/water mix and then rinsed with RO-water. After that you can put them in the drainvessel over night in order to dry them.

Bubblebag Sizes from 45 Micron to 220micron

If you make your own ice you can make it the night before with RO. The best shape in my opinion is half round about the size of your thumb. This minimizes the surface and thus lets your ice stay longer like this.

How to keep proper hygiene in the coldroom

We explained how to clean your room + tools, now we go to the steps you can take yourself. The biggest vector of contamination is the human, so in order to mitigate this risk we wear the corresponding PPE (personal protection equipment) Lab coats are a standard in most hashlabs but a full-body tyvec-suit would be more appropriate for this usage as it contains the dirt on your trousers as well.

Tyvek Suit for clean Hash Production

As you can see in the picture a mask and glasses are also a good thing to contain bacteria in your breath and skin pieces. But thats more important if you're in the medical sector. For home use you can get away with a suit, a hairnet and proper nitril gloves. These are really practical as your hands would get sticky really really quick without them

Nitrilgloves for clean Hash Production

A coldroom buildout

Now that we protected ourself and the hash from contaminants we need to build a coldroom that makes our job as glorified janitors easier.

The best working surfaces

The first thing we should direct our attention to are the surfaces. Every desk, floor and wall should be wipeable as they collect dust. The best option would be to use stainless steel everywhere because it can be safely sterilized heavily without showing any degradation. Another reason is the faster cooling off the room because of it being metal. But such coldrooms are really expensive and only for professional operators For the homeuser, I would advice to lay out paintersfoil on the floor and walls. Looks like out of the movie american psycho, but trust me it is worth it. Also it is quickly removed if some uninvited guests want to look into your flat

Painters Foil for Contaminationprevention
Painters Foil

I would also clean the ceiling with a broom or a mob and some water if possible before washing. If you want to be really thorough, you could put in an air filter. You can use a normal carbonfilter similar to the ones in growtents.

How a professionally built shell for your coldroom

https://youtu.be/zBCXAs3xFVY

Optimal climate control

For washing and separating we need constant temperatures around 0-10°C to get the best results. Otherwise the hash will get greasy and really bad to collect. This is quite difficult if you don't have well insulated place to wash. That's why we recommend using cellars or insulated sheds for home users. If your growroom is clean and sufficient you can use it as well, but every room needs a strong AC to wash in the summer months. But how do we know which one to buy? This depends on your budget and coldroom-size. Most Acs are measured in BTU/H or Watt/H which can bei converted into each other. 1000BTU/H are equivalent to about 300 watts/h and how many of those you need can be looked up in the chart we made

Space Watts
Up to 30m2 2350
30-37,5m2 2500
37,5-42,5m2 3000
42,5-50m2 3500
50-60m2 4000

But not everyone can afford the best Ac so we're going to look at some models for each price class

Small to medium budget

In this category we have the well known mini splits, which most of you will have in their grow room. This modell is really efficient and in my opinion the best choice for small to mid sized ops as you can get down to freezing temperatures in a medium insulated place because it doesn't use air, but a refrigerant. They're powerefficient and don't take up to much space, but it needs a second device on the outside of your building which has to be connected via copper wiring. That and the cleaningafford are the only negativ points about this system. You can get them as cheap as 800$, but you have to count in the installationcost if you're not doing it yourself. But DIY is only recommend if you're a professional or have done it before as improper installation could be a fire hazard. 

Central AC Units
Central Large-Scale AC-Unit

These ACs are really powerful and normally used to cool entire houses. Thats why they can be quite expensive and use lots of power. If you buy one of these and use them all the time, you should think about buying a solar panel as this would pay for itself pretty quickly. This model is mostly used when already preinstalled in a large grow or a housingcomplex You will need a licensed electrician for this installation as this is really complicated and easily done wrong, which again poses a fire risk

How to setup a coldroom for good workflow

Now that we set up our basic shell for the coldroom we need to bring in our equipment. If we want proper workflow for e.g. commercial use (most hash/time) we have to think strategically on how to setup everything in an order where you don't block others from doing their part. So in order to give you an idea what one could look like, we depicted a example in here

If you don't understand all the terms, no problem, we will now take a look at the step by step tutorial. This should clear up most of the confusion

A step-by-step approach to washing Ice-water-Hash

So we have our room prepared, material sourced and ourselves clothed accordingly. Let's beginn the best part, the wash It's advisable to really check if you have everything you need in the room as opening it again could introduce more contaminants in your room Have the icecubes stored in the freezer aswell so that you can directly take more if you need it

Our material should either be frozen or dried and ready. First we prepare the material by breaking down larger nugs in thumb sized pieces in order to get the optimal surface area while not making them so small that they get grinded up between the ice. Be really gentle with the buds as every handling "destroys" trichomes and with that, reduces your yield.

Gastown Cannabis Flowers Mimosa
Mimosa Bred by SymbioticGenetics Grown by Gastown-Collective

When this step is finished, we continue to loading up the wash vessel. First put in the workbag, then a thick layer of ice to cool right off and keep the material from being scrubbed against the bottom of the vessel as we're trying to not damage the buds too much. Now put in a layer of material in (a bit thinner) and then a thin layer of ice again. Repeat this till all your material is in

After this, it's time to get it soaked. Use a gentle setting on your water hose and distribute it evenly over the mix until everything is floating. You want the right ratio of ice/water in order to keep the mix cold but not grind the material between ice cubes. Again, the water vortex is whats separating the trichomes from the flower. The best indicator is listening when you stir it up. Maybe do a testrun beforehand without material to get the mix right. There shouldn't be any crushing noises instead only a light collision sound of single ice cubes hitting the side of your vessel. For a example of this sound and maybe one of the greatest videotutorials out there visit Frenchy Cannolis Youtube channel for his course on hash

Prewash Bucket with Cannabis&Ice
Pre-wash soaking material by KENNWALL

Let the material sit for 5-7min for WPFF and 10-12min for dried flower in the water to resoak. We do this to let the flower get more flexible so that it doesn't release to many contaminants when agitated.

Then we beginn with the stirring process. Remember to be gentle to the material, but stirr it thoroughly to get everything nicely mixed. You can start with clockwise stirring or a canoeing like move. Continue this for 5-10minutes depending on how golden the water is and how many washes you want to do. If you want to get everything out in one wash and don't care too much about getting the cleanest of the clean fullmelt you can go long 10-15min. This method is quite time saving if you only go for rosin. If you want to separate every grade of hash you go for 5-10min depending on the strain/stiring strength.

We stop stirring and let the trichomes settle for 2-3 min again so that we can skim of water from the top. We do this as it would be really heavy to empty out the vessel in the drainvessel if it's completely filled. If you're running a maschine you only set the timer and come back to open the drainvalve on the bottom to release the water into the drainvessel. I would advise you to take your sprayer/hose and spray down the drained leftovers again as a lot of hash gets stuck between cubes/material if you only drain it from the bottom. Now that we have our bags "filled" we spray down every bag down like this to get contaminants and contaminants pushed through. We continue this with every bags because trichomes that are the right size don't fall through if they're in the right bag (micron = trichome-diameter) and the contaminants fall through to the last bags if you did it right. How to that properly we will watch a video of eldaggy of him spraying down his hash

https://www.instagram.com/p/B-P_mojBYwE/?utm_source=ig_web_copy_link

Afterwards we can scoop the hash from the bags with our spoon that we put in the freezer earlier. It's advisable to cool 4-5 spoons as they warm up pretty quick again. Now depending on our dry method we put the hash on our trays and spread it evenly to not get any water pockets. If we freeze dry we can put it aside/directly in the dryer. If we air dry we need to squeeze out most of the liquid with a 25micron screen, cover the hash and put it in the freezer in order to microplane/sieve it later on. We can repeat these steps until no more hash comes out or we don't like the work/quality ratio anymore and use the rest for blasting/alc extraktion

How to dry wet Hash

As we can't smoke wet hash we have to dry it now. And as said before we need to decide between air/freeze dry as a home hash maker. For commercial washing you can't use airdry as the risk of productloss and the dry time will make it unprofitable. I mean you can do it, but in order to make enough money of that you need brandawarenes.

Whats a Freeze Dryer

This lovely device is also called a lyophilization ofen and dries your hash in a short time. It does this by freezing it down to about -40°C and then slightly heating the trays with the hash, causing the remaining moisture to directly sublimate into a gas. This effect is encouraged by a vacuum that you pull on the drying chamber. This leaves the highest possible content of terpenes in your hash and prevents it from oxidizing which would darken your hash aswell. For info on how exactly this works and how a freeze dryer looks like we will watch a short video from a big drying manufacturer

https://www.youtube.com/watch?v=eE993Vs7j8s

For the tech heads under you, we have a second option to buying a prebuilt unit. There is the possibility of building your own freeze dryer. This will safe you a lot of money, but beware, it won't give you the same results that a commercial, specialized unit will give you. Here's a short video tutorial

https://www.youtube.com/watch?v=obhYSf1ogoo

How do I choose and use a Freeze Dryer

Let's assume you want to buy one of these babies but don't know where to look. We would recommend different models depending on your capacity needs and budget. First we have the harvest right models, these are what most people use in the industry as they beginn at 2500€ and a capacity of 800gs of Hash each day of usage. The only problem is the customer service and life time of these maschine as they require intensive care especially with an Oil pump. The second brand are labconco dryers. They get used in the pharma space and are the Ferrari under freeze dryers. These units are built for a lifetime, but also really expensive as they begin at around 15000€. We would recommend them only for commercial users as they won't make sense for the home user.

But then there is another decision to make. Do I go for the cheaper oil vacuum pump or do I invest in an oilless version

This mostly depends on your budget as I would advise you to always go with the oilless version as this is way less work. You would have to regularly check and change your oil and if it malfunctions you get oil all over your hash. But the oilless version also costs an extra 2000-3000€ depending on the manufacturer.

 

This brings us to our next point: how do I use my freeze dryer? We already told you about the pump maintenance, but this is not the only thing you have to check before using your dryer. Before each cycle you should check the drain valves. Let out all the drainwater and the close it again tightly, because if you forget that you won't be able to produce a full vacuum. Now we clean the inside of the dryer and remove any leftover ice (if you haven't defrosted). We connect the pump to the dryer and inspect the doors if they fit snuggly on the sealing part that guarantees the ability to pull a vacuum. If we prepared our trays right and they look like this we can load them into our FD and start the process. This will take about 24-36h depending on how thick your hash layer is

Full Water-Hash Tray for the Freeze Dryer
Full tray of wet hash by Kennnwall

To check if it's dry, take a card and move the hash around a bit. If it behaves like sand and has no clumps left it is dry and ready for smoking or further processing

Drying Hash without a freeze dryer

This method is mostly used for home made hash for it's cheaper tools. As said in the washing guide you should immediately freeze your hash after collecting until it's a hard brick. This gives you the opportunity to separate the individual trichomes better in order to get more surface area for water to evaporate. You need a temperature controlled room with about 30-40% humidity at all times! The room should also be clean to insure no mold spores get into your hash and multiply.

Traditional Hash Air Drying Racks
Drying Room from Aqua Tech Labs

Now you take baking paper sheets and lay out your drying area. You can use baking trays in an open room setup or put it in a clean (!) pizza Box as the cardboard will pull out even more moisture. Then take your frozen hash bricks and grate them either with a microplane or a siever finely over the paper. Be careful to really spread it out good as clumps may lead to inconsistent drying. Wait approximately 1-2 weeks and check regularly for dryness.

How do I calculate my yield from flower to Hash

Now that we weighted out dry hash and have a g number we can simply take the % Value e.g. 5g Hash out of 100gs Flower would equal a yield of 5% These values differ greatly between WPFF and cured Hash as you have to subtract the water weight from the fresh frozen flower. The fresh flowers are about 75-80% water so 5% is a high yield compared to cured hash yield where 20-25% would be a good yield. Good fresh frozen yields are between 3-8% depending on the strain, 8% being the top of the top. Most times these numbers are only achieved with selected cuts asa lot of strains yield <1% For cured Hash a good range is between 15-30% return, but this also requieres a hash specific cut

More Hash-Education

If you want to learn more about Hashtech and Hash culture visit my buddy over at TheHashishInn and listen to his Podcast

What does colour say about my Water-Hash

This is a very controversial discussion in the water-hash space and I hope to not step on anyones to toes, but this needs to be said. Colour doesn't affect the quality of Hash! Most use this phrase but forget to say that, while this is true, there can be bad dark hash. Sometimes it's not a sun ripened or cured resin, but just contaminants. This is a important factor to consider when buying Hash. You have to look really close and do the ultimate test: how does it smoke! No real hash connoisseur cares about the colour when the flavor, smoothness and high is right and so should you. Some people go as far as harvesting way to early in order to get the whitest hash, while it only leads to a speedy, weak high and nearly no flavor

"Dark" or better said amber hash can come from strong UV-Radiation, CBN-Content or long air exposure which oxidizes the compounds in hash

How is Hash rated?

Most of you will have heard about 6 star hash or lower quality, but exactly is that measured? Tbh the star-rating system is used quite liberally so a lot of "six-star" hash is worse or not even close to being fullmelt. To give you a quick overview of how it should be used and a measurable alternative shall be given here

The star rating system

1-2 Star

This grade is mostly used for edibles/RSO or further cleaning processes as it's from low quality. You can identify it by it's green/dark colour that stems from the high content of contaminants. You will typically find this in the 180u or 25u bags

3-4 Star

We use this grade of water-hash for edibles on the worse end and on the better end of the quality spectrum we press it to rosin as this will still yield a proper, terpy oleo-resin. Typically found in the 150u Bags and characterized by it's light colour but missing meltyness

5-6 Star

This premium product will be processed to high-end-rosin on the lower quality spectrum and the best of the best will be left as it is if you have a market for melt. If not, you can also press it and sell it for the maximum rosin price, but beware I will call it a sacrilege to press such high quality melt haha True fullmelt shouldn't leave residue in your banger. A tiny bit is normal as the trichome heads still have their waxy cuticle and this "burns"

The gravimetric measurement, a good alternativ?

As you might have noticed the star-rating leaves a lot of room for interpretation and therefore is unprecise. Fletcher from Archive Genetics wanted to adress this issue and bring in a measurable test. As he was the one to bring freeze dryers into the hash industry, he has a lot of credibility in this regard.

His method is to use an enail, take a 15micron metal screen and weight it before. Now you put exactly 0,1g of your Hash on the screen and put all that on the enail until no oil is left. Then take the screen and weigh it again. Now subtract the previously taken screen weight and there you have your residueweight. This also can be converted into a %melt value which should be used to categorize the Hash Visit his Instagram for fire flower and melts

 

 
 
 
 
 
Sieh dir diesen Beitrag auf Instagram an
 
 
 
 
 
 
 
 
 

 

Let’s end the star system for water hash, solvent less, ice wax, and move onto a system that accurately grades how “MELTY” your hash really is. just simple gravimetric analysis and it’s relatively accurate for the end consumer’s needs. Tag your favorite hash maker in this post! @trichomeheavyextracts1 @k3nnnwall @cannabisreverend @kush4breakfast @710labs @3rd_gen_familyfarm @resin_ranchers.pdx @hashcamp @icedoutextracts @beezlegardens @theccc420 @dannyflavors @shane_omac831 @pureoptions @jungleboys @jungleboysfullmelts @humboldtorganiccollective @michigan_made_melts @icextract_equipment2.0 @cubangrower @tharealogkushman @eldaggy @turtle_trees @megaraw_melts @voodoomelts @solventlessismore @puaextractions @fullflavaextracts @golden_trichome_extracts @dankczar_icewax @connected.california @alienlabs @returnofthealien @ogkushbreath @therealogkushstory

Ein Beitrag geteilt von Archive Seed Bank (@archiveseedbank) am Apr 19, 2020 um 6:09 PDT

What can I do with my finished product?

This depends on your needs, but most people dab/press it or use it for high end edibles. If you process it to rosin you can also make carts with it or fill preroll "donuts" with it as this is quite popular right now The leftover washed flower can be freeze dried and be blasted/alc washed to get every cent out of it. If you already separated all trichs you can use it as mulch for your garden aswell. I hope you learned something new and had fun doing it. If you have any tips/tricks or corrections you want to be included, feel free to leave a comment or shoot me an email as I want to improve the quality of my hash making aswell and you never stop learning Happy dabbing everyone PS: Sorry for my english, I'm not a native speaker :D

Disclaimer: This tutorial is intended for legal use in legal states only. We do not condone any illegal activity

Sources

  1. Dr. Ethan B Russo; "Taming THC: potential cannabis synergy and phytocannabinoid-terpenoid entourage effects"; (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3165946/)
  2. Analytical 360; „Bubble Gum Hash Rosin“ 16.06.2016 http://archive.analytical360.com/m/concentrates/565901I
  3. Iceextract; ICExtract Bag Singles and Kits 5Gal V3 https://icextract.com/icextract-bag-singles-and-kits-5gal-v3/
  4. Pollonator/Mila Jansen; MEDIUM ICE-O-LATOR® 7 BAG SET; https://pollinator.nl/product/medium-ice-o-lator-7-bag-set/
  5. GoPurePressure; Bubble Now 220 Micron Work Bag https://gopurepressure.com/collections/bubble-bags-machines-bubblehash/products/bubble-hash-bubble-now-220-micron-work-bag
  6. homeairguides.com; How To Choose An Air Conditioner (Step-By-Step Guide On Picking The Right AC Unit; https://homeairguides.com/air-cooling/how-to-choose-an-air-conditioner-guide-on-picking-the-right-ac-unit/
  7. Deborah Hucht; Kühlleistung berechnen: Welche Klimaanlage eignet sich für welche Raumgröße?; Kühlleistung berechnen: Welche Klimaanlage eignet sich für welche Raumgröße?
  8. Ab Hanna; "Hash Rosin 101: Lessons from Experienced Solventless Extractors"; https://hightimes.com/guides/hash-rosin-101-lessons-from-experienced-solventless-extractors/
  9. PurePressure; Andrew Ward; "How Bubble Hash is Rated (1* to 6*)"; https://gopurepressure.com/blogs/rosin-education/how-bubble-hash-is-rated-1-to-6
  10. Titelbild: CubanGrower/https://www.instagram.com/cubangrower/ / https://www.instagram.com/_thevillage/
  11. "HOW DO REVERSE OSMOSIS SYSTEMS WORK?"; (https://kraaiwelldrilling.com/how-do-reverse-osmosis-systems-work/)
  12. "Different Types of Terpenes in Cannabis"; (https://tetrahorizon.com/2020/07/10/different-types-of-terpenes-in-cannabis/), TheProfessor

plant tissue culture photo 3 tissues in magenta box

Plant Tissue Culture - the future of Cannabis clone production

featured image by the one and only @kandidkush

1. Introduction

In recent years, automation and standardization of the crop production process are becoming increasingly important. Especially in the production of cannabis, it is crucial to have uniform and disease-free planting material in order to reduce any plant protection measures to a minimum and to ensure a final product with consistent quality. For this reason, more and more growers decide to source their seedlings from in-vitro propagated or tissue cultured material. But what does In-Vitro production and tissue culture even mean? And how is it done with cannabis? That is what we want to explain in this Article.

In Vitro is Latin and means “in glass” or “in the glass” therefor when according to plant production, it is a technique that uses glass containers such as petri dishes and test tubes as a controlled artificial environment for the propagation of plantlets. This contrasts with in-vivo ("within the living") and in-situ ("on site") production, which are commonly used in horticulture.

Within in-vitro production there are various methods and ways to propagate the desired type of explant. Tissue culture and micropropagation are two terms that you will most likely come across while looking closer into that topic. But what exactly is the difference between those?

The main difference between micropropagation and tissue culture is that micropropagation is the production of a large number of plants from a small amount of plant material, whereas tissue culture is the first step of micropropagation, where plant cells are grown in an artificial medium to develop them into a large number of plantlets. In addition, micropropagation requires tissue culture for the propagation of plantlets.

As the global market advances to a greater reliance on plants for active ingredients, delivery of consistent and pathogen free plant material is crucial. Anyways, in the production of medicinal or recreational cannabis, the use of these techniques is not yet widespread, as cuttings can also be produced via "normal" vegetative propagation but, the higher the degree of automation and the producers' demands for cleanliness become, the more this method comes into focus. In addition to the possibility of producing clean plants, in-vitro culture is also suitable for space-efficient growing, improved yields due to vigorous plants and for saving production costs.

However, the in vitro cultivation of plants is not only suitable for propagation but is also of great interest for research and especially for breeding. For example, pathogens can be eliminated from plants, mutations can be created and sterile backups of rare or hard-to-retain plants can be secured. There are many ways to utilize tissue culture and other biotechnological methods and there are even more to be discovered.

2. Basic Terms for In-Vitro production

2.1 Medium

2.1.1. What's Agar Agar?

Some of you might know it as a plant derived gelatin substitute and that's nearly the same purpose we will use it for. The thickener is based on a galactose containing extract taken from red-purple marine algae. This brings the typical nutrient solution to the right consistency that supports vigorous growth, but gets easily penetrated by the roots to insure the best development possible. In order to generate the perfect Agar mixture we have to add it in a ratio (depending on the source) 1-2:100. This leads to a nearly complete solid medium. If you want to cultivate in anaerobe/liquid conditions, you can simply half the concentration.

Comparison Gelatine vs Agar Agar
Comparison Gelatine vs Agar Agar (Source 7)

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But why not just use gelatin instead?

That's a good question, but there's a simple answer to it. It's edible to bacteria and most other microbes so you would have to refill every plate after a short time. The nutrient dosing and validity of the experiment undertaken would be in danger as you would have to add the gelatin to the feeding chart of the microbes that are cultured.

It comes in different forms, but the most common are flakes, that need to be mixed with water und constant heat application until it's fully homogenized. These flakes are easy to store and quite cheap to get. If you have the necessary budget, there are pre-mixed/pre-sterilised versions, mostly bottled up, that can be used instantly without waiting for the hot agar mix to cool down. The cooling process can take up to 30min per batch before use.
Not only the distribution form differs but also the additives. Many agar mixes have added ingredients for certain purposes as this is used for all types of cell cultures from human cells to the infectious disease causing Escherichia coli. 

The potato-dextrose agar for example is the most used form for fungi especially for varieties like Botrytis cinerea also known as the common grey mold. Another one is blood agar, which as the name already says, contains animal blood in order to study special microbes. For tissue culture the most common type is the standard galactose agar in combination with a nutrient matrix and a hormone, depending on the phase.

Different kinds of agar media for cell/tissu culture
(Source 6)

The standard nutrient matrix in nearly all cell culture work is the Murashige-Skoog-Medium, which contains simple amino acids, nutrient salts and photosynthesis products (saccharose) as the plant can't photosynthesis in most cases. Inositol is also added in order to mitigate plant stress, strengthen cell walls and phosphate storage.

2.2 Hormones for plant tissue culture - is that even safe?

Many myths rank around plant hormones, which are mostly "feared" by the Cannabis community as it only has found real application in large scale agriculture. They get mixed up and thrown in the same bucket as GMO and Glyphosate. Not that those would be bad substances but the social stigma around them is strong.
The hormones on the other side are nearly identically to their natural/plant counterparts, which control nearly all the physiological responses of the plant to outside influences like the movement of the sun or the alignment of the plant to the gravity

2.2.1 Auxin

The first of these phytohormones (hormones in plants) is responsible for a multitude of responses to environmental stresses. Most auxin derivates in the plant and made by humans are based on the Indol-3- acetic acid. This compound is mostly synthesized in the apical region refering to the main growingtip of the plant. From there on it gets transported via the phloem and cell-to-cell by the PIN 1-9 transporterprotein.

The basic effects include wound response, apical dominance (the main branch is the tallest one), photo-/gravitropisms and it plays a important role in fruit development. But lets dive a bit deeper in the interaction of Auxin and the use cases of it.
The first practical application would be induction and promotion of root growth. This is the most used purpose and nearly all commercial rooting gels contain it. But not only in Cannabis, but also the floral industry uses it predominantly in their rooting SOP.
We also use this by giving our explants (plants in in-vitro conditions) a certain amount of it in order to achieve the highest rooting rate possible.

But as with everything in life, there is a limit to whats good for the plant. Excessive application of this substance can result in growth inhibition and plant death. Weedkillers like 2-4-D are based on the synthetic counterparts of auxin like indole-3-butyric acid. But not only damaging substances also "good" ones like common rooting gels are based on synthetic auxin.
"Natural" Auxin can be found in variety of plants shoots especially the ones of the willow tree.

Willow Tree
(Source 5)

2.2.2 Cytokinin

The next hormone in our repertoire is the counterpart to the previously mentioned Auxin. This compound focusses on the lateral growth of both roots and shoots. To be more exact the ratio of Cytokinin:Auxin is the driving factor in the morphology of the plant. They not only contradict each other, but also work together when influencing cells. When only Auxin is applied cells elongate and become big, but won't expand or differentiate. Same takes place when the ratio is 1:1. This is used for Callus multiplication or expansion.
Complementary to Auxin it is synthesized in the roots and travels to the shoots via Symplast and Apoplast. Production of this chemical is regulated by two types of response regulators. One being the B-Type and the other being the A-Type. Both are regarded as transcription factors as the either activate or stop the production of Cytokinin via influencing the transcription of the corresponding gene loci.

Directions of hormone flow in the plant
(Source 1)

Using this knowledge we can manage the morphology of the plant into growing more squat thus increasing internodal stacking and use of space.
But not only the space can be used more efficiently also the time saved by breaking seed dormancy can save time when germinating seeds for a phenohunt. The dormancy is induced by abscisic acid and by increasing metabolic activity the cytokinin decreases the level of abscisic acid in the seed. It's commonly used in plants that have a hard time germinating.
The next effect might be especially interesting for the photographers and florist under you. Cytokinins can delay senescence in other words the decay of the plants. This is achieved by increasing synthesis and slowing decay of certain proteins. Also nutrients are drawn into the treated area from nearby tissue. It's suspected that an enzyme is responsible for these actions, but no scientific consensus has been reached yet.
When using Cytokinins you have to differentiate between the plants own, adenine based, Cytokinins like Kinetin and Zeatin. Others have been found outside of plants, but they are based on Phenylurea. These exceed the effectivness of the Cytokinin in certain plants. Examples for this variety are TDZ and Diphenylurea, which are widely applied in agriculture

2.2.3 Gibberellic Acid

One of the most interesting phytohormones is the counterpart to abscisic acid. Exogenous GA3 (short version) is also responsible for breaking dormancy where it activates indigenous synthesis of more GA3, which adds to dormancy breaking factors. These are hypothesised to be a combination of growth promoting hormones (GA3, Cytokinin, Auxin etc) and reduced nutrient storage in the endosperm. Enzymes, primarily α-amylase, lead to the processing of sugars and other storage units. Another restricting factor is the Cuticula strength of the seed.
The application on an adult plant leads to a big stretch in all shoots. This is principle is also used in plant tissue culture and for virus cleansing. But this has to be done with the utmost focus on the concentration of GA3 as a high amount leads to different sex expression. This mechanic is commonly used in feminized seed breeding as it "reverses" a female plant into producing pollen with female Genome. In even bigger concentration it can lead to the complete sterilization of the specimen. Most fruit growers like in the citrus industry use the mechanic to suppress undesirable seed development

That said, when a trained horticulturist with a strict, thought through regime applies GA3 in flower, it can increase inflorescence mass and trichome development. For homegrowers and people that don't have access to these chemicals, Kelp/Seaweedextract is a perfect allrounder as it contains a variety of growth hormones aswell as Macro-/Micronutrients.

The compound was discovered by japanese scientist while researching the Gibberella fujikuroi fungi. Cytokinin is a secondary metabolite to the pathogenic pathway of the fungi on rice plants.

2.3 Basic Tools for Tissue Culture Propagation

2.3.1 Laminar Flow Hood

It's one of the most important, if not the most important part of a Culturelab. The Laminar Flow Hood (LFH) insures that all steps are done in a steril environment. There is a differentiation between normal Flow hoods and special models for working with microbiological organisms. The first doesn't filter the incoming and outgoing air, leaving the worker exposed to aerosols and microorganisms.

That's why we use the second model in most cases. This LFH got a specialized HEPA-Filter before the air enters the Hood. After getting in it can be distributed in two ways. Vertical flow hoods and horizontal flow hoods can be differentiated easily as word explains itself

Laminar flow hood or cabinet
(Source 4)

All work including the plant have to be done inside to keep out potential contaminants. It's therefore of vital importance to wipe down all areas inside with ethanol (70%) and clean the outside of culture vessels aswell when introduced in the steril environment. Gloves and hair-protectants are a must, if mass-propagation is planed. At home you can easily get away with disinfecting your hands and arms up to the elbow. Another concern is your breath as it often contains bread-mold spores and a variety of microorganisms that thrive on tissue culture medium.
If a infection takes place and you have no copy/other option, you can add antibiotics to the medium, but be very careful. Most higher plants are more resistant to these compounds than the microorganisms, but the optimal balance is very hard to reach. Another factor is the difference between every plant genome/phenotype thus a cultivar-specific test would be necessary before adding antibiotics.

2.3.2 Autoclave

Speaking of sterilisation, we will subsequently be lead to autoclavs. These laboratory certified pressure cookers are an essential tool for cleaning glassware and used culture vessels. They come in different sizes and forms, which you can choose specific for your application.
Despite the different shapes, the mechanism is the same. The chamber is evacuated via a Vacuum pump in order to simplify the sterilisation progress. It gets filled with hot steam and thus put under a high pressure to increase the cleaning efficiency. After 15-20min the optimal cleansing effect is achieved, which means that the content can be removed. Be careful not to touch it with your bare hands as it's still really hot.
You should also size the autoclave big enough for the operation, but not too big for your batchsizes as this will lead to unnecessary cost for energy.

Temperature/Pressure correlation when using an Autoclave
(Source 3)

2.3.3 Magnetic Stirrer

This device is rather simple and doesn't need much explanation, but it's still vitally important that we talk about it. You probably wonder why and we will will explain it shortly. Not the stirrer per se needs explanation, but rather how to use it and what for. It's the base of a good tissue culture lab as it's used for mixing and homogenizing of the medium components that we discussed earlier.
Each component has a specific boiling/inactivation point that you have to consider when mixing.
The goal is to find the right compromise between solubilityspeed and retention of effectiveness. Some compounds like hormones derivates have to be added after the mixing + autoclaving of the medium as they can't even withstand the autoclaving procedure.
For these substances you have to consider other ways of sterilising them such as syringe filters or specific solvents.

Magnetic Stirrer with integrated heatplate and ring stand holder
(Source 2)

It enables the production of large amounts of media as it's fully adjustable in rotationspeed and temperature, if you have a heated version. Be careful to pick a appropriable sized stir bar as this determines the homogeny of your mixed substance.  But even with a large stir bar the stirrer has it's limitations when the mixing materials are too viscous. If that's the case, we would advise a mechanical alternative as they can plow through it better. Keeping that in mind, you have to pour your hot, mixed medium out of the vessel before it cools down as it gets really hard to remove the bar after.

2.3.3 Consumables

This chapter is for items that are important, but didn't get a separate chapter as it would go beyond the scope of this article.
Firstly we have your PPE like gloves, masks, Labcoats and hair nets. These are especially important, if you have a large team and thus many vectors of contaminants/pests. But keep in mind that PPE doesn't eliminate the necessity of disinfecting the whole work environment before and after you do explants in it.
Another vital point are the culture vessels. These are the new homes for your little clones and need to have a large enough volume to accommodate them. This refers especially to the amount of medium that a plant needs in its stage of growth. There are mayor differences from cultivar to cultivar so the best way to go about it, is to  experiment with it and keep tight journals in order to create SOPs. This takes a lot of time, energy and labour so be mindful of that, when trying to establish a plant tissue culture production.

Our last important item is the scalpel.  You can find many different forms of it for specific applications, but we recommend that you go for a more environmentally friendly version by using a scalpel holder with exchangeable blade. Using this will decrease your waste of material and money as the amount of cut plant material in an TC production puts a great stress on the blade thus making it dull. The alternatives would be to use whole single use scalpels, but the produce large quantities of plastic that we don't want or high end versions. The later stays sharp for longer, but not long enough to make up for the huge price difference.

2.4 Standard terms for plant meristem tissue culture

In order to understand the basic mechanics of saving genetic material and growing callus cultures we have to take a look at the structure of a growing plant tip. This is the location we're using to explant a meristem from. But what is a meristem?

The term comes from the ancient greek word merisein, which means divide. It was used by the swiss scientist Carl Wilhelm von Nägeli to describe the undifferentiated, multiplying Cells that made up the growing tips of the plant. The apical meristem is the highest growing tip in the plant and sits on top. The counterpart of this region is the basal meristem, which sits of the tip of the roots as you might have guessed.

By Jon Houseman and Matthew Ford (licensed under the Creative Commons Attribution-Share Alike 4.0 International) Photomicrograph of a Coleus stem tip. A=Procambium, B=Ground meristem, C=Leaf gap, D=Trichome, E=Apical meristem, F=Developing leaf primordia, G=Leaf Primordium, H=Axillary bud, I=Developing vascular tissue. Scale=0.2mm.
By Jon Houseman and Matthew Ford (licensed under the Creative Commons Attribution-Share Alike 4.0 International)
Photomicrograph of a Coleus stem tip. A=Procambium, B=Ground meristem, C=Leaf gap, D=Trichome, E=Apical meristem, F=Developing leaf primordia, G=Leaf Primordium, H=Axillary bud, I=Developing vascular tissue. Scale=0.2mm.

The plant builds new meristemcells in the top middle parts as the lower parts differentiate into the predetermined functional parts. Some go to be Parchenym cells, some are going to be part of the vascular system of the plant. Knowing this, we can extract these cells before they're differentiated in order to get totipotent cells (cells that can be any part of the plant). These are full of potential and are kept in a cryofreezer to be saved for later usage, as backup or as basis for mass cell production in a bioreactor.

3. Basic mechanics of mass micropropagation

To get an idea of how micropropagation could benefit your growing operation we're gonna draw a little plan of how it could look like.
First of all we need a small motherstock from which we can take our microcuttings. These are about 2,5cm/1 inch long and at best, taken from a meristematic region.
Then we plant it in a multiplication medium under sterile conditions in order to avoid contamination and the subsequent demise of our plant matter. After 2-3 weeks we should see 2-4 new shoots coming from our tissue, which we can also cut up into separate tissue samples. The we put them in individual containers with the same medium mix again and repeat the previously mentioned cycle. Even thought there will be loses due to contamination, the replication-rate is extremely high due to each new tissue replicates itself 2-4 times. If we calculate this through, we can see that with minimal space use, an enormous plantcount can be achieved in a short period of time.
After we finished the replication cycle, we can move onto the rooting phase. This uses a special medium containing auxin derivates, which induce the production of roots on our previously made shoots. After 2-4 weeks again, we have to inspect the plants accordingly and if they're sufficiently developed, we beginn the hardening phase.
For this part of the productioncycle the plants get translated in the medium of choice that will be used later on in production. Most growers will use rockwool as it has excellent water holding capacities, but also coco or small soil pots can be used. Light plays a huge role now and has to be increased in order to acclimate the plants to the later vegetative conditions.
The humidity also has to go from the comfy 80-90% RH in the boxes down to the values of your vegroom.
This was a quick overview of the process and as you could see, there are many obstacles along the way, but in the end it's not that hard to achieve at home. For this exact purpose we will compare DIY kits that can be bought online with a home made microlab.

4. What are ready-made home kits?

There are some suppliers on the internet who offer hobby kits for micropropagation. Even if these kits are suitable for experimenting and for first experiences, they cannot replace a professional laboratory, because similar to mushroom cultivation, a sterile working method and environment must be guaranteed to avoid failures. In addition to all the materials and equipment, professional application requires trained personnel and constant quality control of the work process.

5. Future vision

As cannabis production becomes more and more automated and the costs of labor, electricity and rent continue to rise, we at research gardens see in-vitro production as a key advantage for companies that want to remain sustainable in a rapidly evolving industry. Due to the prospect of a long-term reduction in production costs as well as a simultaneous increase in product quality your company will get an advantage against your competitors. Particularly in pharmaceutical operations, a consistent quality of the plants and ingredients is indispensable. For the recreational market, however, this is just as important since similar requirements exist here as well.

Interested? Then let us start working out a concept for an in-vitro laboratory in your company today! Send us an e-mail to info@research-gardens.com

Sources:

 

  1. "Communication by Plant Growth Regulators in Roots and Shoots of Horticultural Crops" by Anish Mallad; Jacqueline K. Burns; August 2007; HortScience: a publication of the American Society for Horticultural Science 42(5) (https://www.researchgate.net/publication/237379591_Communication_by_Plant_Growth_Regulators_in_Roots_and_Shoots_of_Horticultural_Crops/figures?lo=1)
  2. "Hot Plate with Magnetic Stirring: 6.7"x6.7" SS Plate Max.300C - SH3"; MTI Cooperation; (https://www.mtixtl.com/hotplateswithmagnetcstirring67x67stainlesssteelsurfaceupto300c-eq-sh-3.aspx)
  3. "Sterilization cycle phases for a steam sterilizer"; rsd-engineering.com; (https://www.rsd-engineering.com/steam-sterilization-cycles)
  4. "Laminar flow hood/cabinet- definition, parts, principle, types, uses";  
  5. "Baum HD"; Dreamliner (https://wall.alphacoders.com/big.php?i=612687&lang=German)
  6. "CULTURE MEDIA AND ITS TYPES USED UN MICROBIOLOGY LAB."; milan; (http://biologyofmicroorganisms.blogspot.com/2010/09/culture-media-and-its-types-used-un.html)
  7. "[Tuesday Scoop] Puzzlement Between Agar-Agar And Gelatin"; Namrataa Mahalley; (https://bulbandkey.com/blog/tuesday-scoop/puzzlement-between-agar-agar-and-gelatin/)