WO2019186568A1 - Physical means and methods for affecting cannabis plants - Google Patents

Abstract

The present invention discloses a method for increasing at least one parameter of at least one cannabis-derived compound to a range being at least 5% over normal in a non-treated plant; characterized by step of emitting a non-thermal plasma modified field (PMF) on a cannabis plant or a portion thereof; said parameter is selected from a group consisting chemical parameters, wherein the chemical parameter is selected from a group consisting of amount and concentration, weight percent and molar percent of said cannabis-derived compound in the plants or portions thereof. The present invention further discloses a method for producing at least one de novo cannabis-derived compounds by changing at least one parameter of a defined PMF treatment aimed at emitting PMF on a cannabis plant or a portion of it, said parameters are selected from a group consisting of the number of PMF emitting events, number of pauses in-between PMF emitting events, number of pulses used of the PMF emitting events, and any combination thereof.

Description

PHYSICAL MEANS AND METHODS FOR AFFECTING CANNABIS PLANTS

FIELD OF THE INVENTION

The invention generally pertains to modified non-thermal plasma application. More specifically, the invention relates to a non-thermal plasma modified field (PMF) generating system and methods for inducing effects on cannabis plants and portions thereof.

BACKGROUND OF THE INVENTION

Non-thermal plasma (NTP) systems have been emerging as useful tools for various clinical applications. Plasma is known to catalyze biochemical activities when applied on tissue and is able to regulate cellular processes such as proliferation, differentiation, and apoptosis. This, in part, is due to the reactive oxygen and nitrogen species (ROS and RNS) generated by application of non- thermal plasma. Most of the non- thermal plasma research has been performed in vitro or ex vivo, which has led to investigation of potential applications such as disinfection of surfaces, promotion of hemostasis, enhancement of tissue regeneration, acceleration of wound healing, and for anti cancer therapy. However, there has not been an extensive characterization of non-thermal plasma in in vivo plants.

The influence of non-thermal plasma treatment on seed germination has recently been studied, see e.g., I. Filatova et al. The effect of non-thermal plasma treatment of seeds of some grain and legumes, on their sowing quality and productivity, Rom. Journ. Phys. Vol. 56, 139-143, 2011 ; S. Bozena et al, Influence of non-thermal plasma treatment on wheat and oat germination, IEEE Transactions on plasma science, vol. 38, 2010, both are incorporated herein as references. These studies describe an apparatus comprising a vacuum chamber, a rotary pump and a microwave resonator. The exposure time to the non-thermal plasma treatment was for a period of several minutes to more than 40 min. Furthermore, the effects reported by these studies mainly relate to the seed coat surface and seed coat sterilization.

US Patent 8,725,248 to Gutsol et al., incorporated herein as a reference, discloses that non-thermal plasma help establish mechanical connection between tissue parts through several possible mechanisms including non-thermal plasma-chemical modification of bio-polymers on the surfaces of tissue and formation of fiber material during blood coagulation. A barrier insulator or semiconductor is placed between the electrode and tissue resulting in limiting the current through non-thermal plasma and through tissue to minimize tissue heating. The disclosed non-thermal plasma treatment can be employed to promote coagulation of blood, sterilization, disinfection, re connection of tissue, and treatment of tissue disorders without causing significant thermal tissue damage. US Patent 8,896,211 by current inventors, which is incorporated herein as a reference, discloses a system for generating modified non-thermal plasma. However, it does not teach how to achieve a biological effect on living plants. US Patent 9,295,280 to Jacofsky, et al., incorporated herein as a reference, discloses a non-thermal plasma device for exerting an in vivo non-thermal plasma effect by killing or reducing a microbiological pathogen, or denaturing a protein in food.

Cannabis ( Cannabis sativa L.) is an annual herbaceous plant. It is a dioecious plant. In many countries, cannabis is cultivated as a narcotic substance or a source of narcotic substances like hashish and hashish oil. Now a day cannabis is cultivated on the large areas with the mild and tropical climate for the cannabis oil and fibre (United Nations on Drugs and Crimes 2009). The cmde drug can be obtained from leaves, flowers, seeds and stem of cannabis. The female plant yield more drug than the male. It can be smoked in cigarettes or pipes and can be snuffed or added to food. The chemical composition of cannabis varies with the type, age and part (flower, root, leaf, fiber, etc.) of cannabis plant as well as with the type of preparation; see Zerihun, Agalu et al. “Levels of Selected Metals in Leaves of cannabis Sativa L. Cultivated in Ethiopia.” Springer Plus 4 (2015): 359. PMC. Web. 19 Mar. 2018 which is incorporated herein as a reference.

Cannabis contains a unique class of compounds known as the cannabinoids. In total, more than hundred cannabinoids in some eleven subclasses have been characterized in cannabis and are concentrated in the glandular trichomes of the female inflorescences and other cannabinoids classes include cannabigerols (CBG), cannabichromenes (CBC), and cannabinols (CBN). The cannabinoids occur primarily in acid form, with neutral cannabinoids formed during drying, storage, and decarboxylation during smoking. D9-THC, the main psychoactive cannabinoid, can be over 20% by weight in specially bred cannabis strains. CBD, known for its anti-inflammatory activity and antagonism of D9-THC-induced anxiety, can range from below 0.5% up to 6.5% by weight.

Pharmacologically, the principal psychoactive constituent is D9-tetrahydrocannabinol (THC). The amount of THC in conjunction with selected additional cannabinoid compounds (cannabidiol/CBD, cannabinol/CBN), determines the strength or potency of the cannabis product. While D9-tetrahydrocannabinol is responsible for psychoactive properties of cannabis some of the other components modulate its activity. Besides D⁹-THC, there are also non-psychoactive cannabinoids with several medicinal functions, such as cannabidiol (CBD), cannabichromene (CBC), and cannabigerol (CBG), along with other non-cannabinoid constituents belonging to diverse classes of natural products. Today, more than 560 constituents have been identified in cannabis; see Wilcox, M., et al. "Improving Sample Preparation Methods For cannabis-infused Edible and Topical Products." Planta Medica 82.05 (2016): PA30 which is incorporated herein as a reference.

Medicinal cannabis is an invaluable adjunct therapy for pain relief, nausea, anorexia, and mood modification in cancer patients and is available as cookies or cakes, as sublingual drops, as a vaporized mist, or for smoking. However, as with every herb, various microorganisms are carried on its leaves and flowers which when inhaled could expose the user, in particular immunocompromised patients, to the risk of opportunistic lung infections, primarily from inhaled molds; see Ruchlemer, R., Amit-Kohn, M., Raveh, D., and Hanus, L. (2015). Inhaled medicinal cannabis and the immunocompromised patient. Support. Care Cancer 23, 819-822 which is incorporated herein as a reference. There is a variety of contaminants on cannabis plants: pesticides, heavy metals, microbes etc.

Pesticides: For most of the 20th century, the majority of marijuana produced in the United Sates was grown outdoors. With more aggressive law enforcement, marijuana agriculture moved indoors. Although this provided the benefit of year-round cultivation, it also required the use of agricultural chemicals, typically synthetic fertilizers and pesticides.

Heavy metals: Cannabis has been shown to be especially effective in absorbing metals such as cadmium and copper from contaminated soils. Making matters worse, cannabis is also intentionally contaminated with metals to increase the market weight. In 2008, 150 people in Germany developed lead poisoning as the result of using adulterated cannabis.

Microbial Indoor growth results in increased susceptibility of Cannabis to contamination by microbes such as fungi, bacteria, and plant viruses. Growing and drying also increase the risk of microbial contamination. Penicillium species are the predominant microbe contamination in marijuana grown indoor. Cannabis has even been shown to be contaminated with human pathogens such as hepatitis A, hepatitis B, and salmonella. Chronic pulmonary aspergillosis has been found in immunocompromised individuals using medicinal marijuana; see Joseph Pizzomo, N.“What Should We Tell Our Patients About Marijuana (Cannabis indica and Cannabis sativa)?” IntegrMed (Encinitas). Dec; 15(6): 8-12. (2016), which is incorporated herein as a reference.

Hence for example, when harmful microbes or fungal spores are inhaled during e.g., vaporizing or smoking, they may directly enter the bloodstream and cause opportunistic infections. Such contamination risks are not merely hypothetical: cases of chronic pulmonary aspergillosis associated with smoking unsafe cannabis are well established in the scientific literature. To minimize contamination risks to patients, following European or US Pharmacopoeia standards for inhaled preparations, certain specific pathogens must be completely absent, i.e., Staphylococcus aureus, Pseudomonas aeruginosa, and any bile-tolerant Gram-negative bacteria such as E. coli (EP, 2015; USP, 2015). Furthermore, the absence of fungal mycotoxins must be confirmed by additional quality control testing.

Table 1 shows a list of the toxic contaminants that have been found in in both medical and recreational marijuana. Research has shown that cannabis extracts (see Solvents section below) contain considerable amounts of pesticides. Reduction of microbes can be achieved by various treatments, as listed in Table 2. The optimal choice of decontamination depends on the nature of the product to be treated. For herbal materials such as cannabis, the only currently viable option for treatment is the use of ionizing radiation. Any of the other decontamination treatments would either affect chemical content or texture (i.e., heat, chemicals, pressure, steam); or would not penetrate beyond the surface of the dense cannabis flowers (i.e., UV-light).

Table 1 Toxic agricultural chemicals found in cultivated cannabis; see Joseph Pizzomo, N.“What Should We Tell Our Patients About Marijuana (Cannabis indica and Cannabis sativa)?” Integr Med ( Encinitas ). Dec; 15(6): 8-12. (2016), which is incorporated herein as a reference

Table 2 List of current main methods available for decontamination or sterilization of (food) products; see Hazekamp, Amo. "Evaluating the effects of gamma-irradiation for decontamination of medicinal cannabis." Frontiers in pharmacology 7 (2016): 108 which is incorporated herein as a reference.

Because medicinal cannabis is often used by chronically ill patients affected by a weakened immune system, pharmaceutical regulations in countries such as The Netherlands and Canada specify that these products must adhere to strict safety standards regarding microbial contamination. Decontamination of medicinal (herbal) cannabis is a necessity, as it has yet not been possible to grow cannabis plants under sufficiently sterile conditions to keep contamination levels below the required safety limits. Even if this were feasible, the multiple steps involved in harvesting, drying, processing and packaging cannabis buds would make it extremely hard to maintain near-sterile conditions throughout the entire production procedure. As a result, medicinal cannabis in The Netherlands as well as in Canada is treated by gamma irradiation before it becomes available to patients.

Gamma irradiation of herbal cannabis remains the recommended method of decontamination, although remains controversial among some consumers of medicinal cannabis. However, weighing the risks vs. the benefits currently keeps pointing toward the use of this decontamination procedure. After all, cannabis plants cannot (yet) be grown and processed under conditions aseptic enough to meet pharmaceutical standards, while infection risks are well documented in the medical literature and can be harmful or even fatal to seriously ill patients. Meanwhile, the main harm of gamma-irradiation seems to be limited to a reduction of some terpenes present in the cannabis, leading to a small quantitative effect, but keeping the terpene profile qualitatively essentially intact.

.The development of improved hygienic standards for cultivation and processing of medicinal cannabis may ensure that irradiation doses can be reduced to an absolute minimum. This is especially important when cannabis is prescribed to seriously ill and possibly immune-deprived patients, with an increased risk of suffering from microbial infection. In time, gamma-irradiation may eventually be replaced with other, more generally accepted, forms of reliable decontamination; see Hazekamp, Arno. "Evaluating the effects of gamma-irradiation for decontamination of medicinal cannabis." Frontiers in pharmacology 7 (2016): 108 which is incorporated herein as a reference.

Considering flowering time, traditionally, cannabis plants are harvested exclusively for their flowers, and cannabis flowers have an ideal maturity date which varies by Cannabis variety, including but not limited to Cannabis sativa and Cannabis indica. Depending on the variety, the harvesting time may be between 8 to 11 weeks. During this harvesting time, and sometimes a week or two prior, the Cannabis leaves start showing signs of nutrient deficiency as the Cannabis plant needs all of its available nutrients to support the Cannabis flowers see“Method of juicing cannabis plant matter”, patent application number US 20170274028 which is incorporated herein as a reference. Cannabis has a reputation for being pest-free. Actually, it is pest-tolerant Many pests have been found around Cannabis, but they rarely cause economic damage. The most common pests are arthropods; see Table 3 below.

Table 3 Common Cannabis pests, see McPartland, J. M. "Cannabis pests." Journal of the

International Hemp Association 3.2 (1996): 49-52 which is incorporated herein as a reference.

Diseases of Cannabis are caused by organisms or abiotic sources. Organisms include fungi (first and foremost), nematodes, parasitic plants, bacteria, and viruses. Abiotic (non-living) causes include nutrient deficiencies, pollutants and genetic diseases. Different diseases prevail in different crops (e.g., drug cultivars versus fiber cultivars). Disease prevalence is also modulated by geography and climate. The claim that Cannabis has no diseases is not correct, Cannabis suffers over 100 diseases, but less than a dozen are serious. Serious diseases include gray mold, hemp canker, damping off, assorted leaf spots, blights, stem cankers, root rots, nematode diseases, broomrape, macro and micronutrient deficiencies, and genetic diseases. Environmentally stressed plants become predisposed to diseases. Stress incudes drought, insufficient light, untoward temperatures, or growing plants in monoculture.

Few Cannabis diseases can be transmitted to humans, but there are exceptions. Some diseases prevail in Cannabis fiber and oil seed crops, other diseases predominate in drug crops. Disease prevalence shifts between greenhouse crops and outdoor crops. Disease prevalence alters as plants grow from seedlings to flowering adults, see Table 4 below. Table 4 Common Cannabis diseases, see McPartland, John M. "A review of Cannabis diseases."

Journal of the International Hemp Association 3.1 (1996): 19-23 which is incorporated herein as a reference.

In a recent study of a, cannabis sativa L. medicinal plant, a total of 30 different fungal endophytes were isolated from all the plant tissues. The most dominant species was Penicillium copticola that could be isolated from the twigs, leaves, and apical and lateral buds. The fungal endophytes were challenged by two host phyto-pathogens, botrytis cinerea and trichothecium roseum, See Kusari, Parijat, et al. "Endophytic fungi harbored in Cannabis sativa L.: diversity and potential as biocontrol agents against host plant-specific phytopathogens. " Fungal diversity 60.1 (2013): 137- 151, which is incorporated herein as a reference.

The recent discoveries of the medicinal properties of cannabis and the cannabinoids in addition to their potential applications in the treatment of a number of serious illnesses, such as glaucoma, depression, neuralgia, multiple sclerosis, Alzheimer's, and alleviation of symptoms of HIV/AIDS and cancer, have given momentum to the quest for further understanding the chemistry, biology, and medicinal properties of this plant.

Due to highly variability of chemical content of cannabis plants, concentration of cannabinoids, terpenes etc., long time to flowering and the high load of cannabis pants’ pests and disease, there is a long felt and unmet need to provide methods for in vivo modulating cannabis plant to overcome these hurdles, using efficacious modified non-thermal gas plasma field treatments and protocols which can be applied conveniently BRIEF DESCRIPTION OF THE FIGURES

Figure 1: Illustration of a method of regulating ratio of cannabis derived compounds

(cannabinoids, terpenes and phenolic compounds);

Figure 2: Illustration of a method for regulating cannabis plant's pests;

Figure 3: Illustration of the effect of a complete PMF treatment cycle on the leaf colour, whereas L* indicates lightness, a* is the red/green coordinate, b* is the yellow/blue coordinate, C* represents chroma, and h is the hue angle. Chroma and hue are calculated from a* and b* coordinates in L*a*b*; and

Figure 4A-C Photos depicting the results of Example 3 study.

SUMMARY OF THE INVENTION

One object of the invention is to disclose a method for increasing at least one of a parameter of at least one cannabis-derived compound to a range being at least 5% over normal in a non-treated plant; characterized by step of emitting a non-thermal plasma modified field (PMF) on a cannabis plant or a portion thereof; said parameter is selected from a group consisting chemical parameters.

Another object of the invention is to disclose the method as defined above, wherein said chemical parameter is selected from a group consisting of amount and concentration, weight percent and molar percent of said cannabis-derived compound in the plants or portions thereof.

Another object of the invention is to disclose the method as defined in any of the above, wherein said biological parameter is selected from a group consisting of accumulation rate of said cannabis- derived compound in the plants or portions thereof (i.e., either amount per time or concentration per time), pesticidic effect, including one or more effects selected from a group consisting activity as an insecticides, nematicide, molluscicide, piscicide, avicide, rodenticide, bactericide, insect repellent, animal repellent, antimicrobial, fungicide and disinfectant

Another object of the invention is to disclose the method as defined in any of the above, wherein said agro-technical parameter is selected from a group consisting of flowering rate and photosynthesis parameters.

Another object of the invention is to disclose the method as defined in any of the above, wherein said method is provided useful for increasing at least one chemical parameter of both at least one first cannabis-derived compound and at least one second cannabis-derived compound to a range being at least 5% over normal in a non-treated plant; characterized by step emitting a PMF on a cannabis plant or a portion thereof; said chemical parameter is selected from a group consisting of amount, concentration, accumulation rate {i.e., either amount per time or concentration per time), and said at least one first cannabis-derived compound to at least one second cannabis-derived compound (wt/wt) or (mole/mole) ratio.

Another object of the invention is to disclose the method as defined in any of the above, wherein said method is provided useful for increasing the ratio of at least one chemical parameter of at least one first cannabis-derived compound over at least one second cannabis-derived compound to a range being at least 5% over normal such a ratio in a non-treated plant; characterized by step emitting a PMF on a cannabis plant or a portion thereof; said chemical parameter is selected from a group consisting of amount, concentration, accumulation rate (i.e., either amount per time or concentration per time), and said ratio is selected from a group consisting of (wt/wt) and (mole/mole) ratio.

Another object of the invention is to disclose the method as defined in any of the above, wherein said cannabis-derived compound is selected from a group consisting of one or more cannabinoids, terpenes, phenolic compounds and any combination thereof.

Another object of the invention is to disclose the method as defined in any of the above, wherein said one or more cannabinoids are selected from a group consisting of Tetra-hydro-cannabinoids (d9- THC), Tetra-hydro-cannabinoids (d8- THC), Tetra-hydro-cannabinolic acid (THCA- d9), Tetra-hydro-cannabivarin (THCV/THC-C3), Cannabidiol (CBD), Cannabidiolic acid (CBDA), Cannabidivarin (CBDV), Cannabigerol (CBG), Cannabigerolic acid (CBGA), Cannabinol (CBN), Cannabidiolic acid (CBNA), Cannabichromene (CBC), Cannabichromenic acid (CBCA), and any combination thereof.

Another object of the invention is to disclose the method as defined in any of the above, wherein said one or more terpenes are either at least one mono-terpene or sesqui-terpene.

Another object of the invention is to disclose the method as defined in any of the above, wherein said one or more phenolic compounds are selected from a group consisting of O-glycoside Cannaflavin A, Cannaflavin B, Canabisin D, Friedelin and Epifreidelin. and any combination thereof. Another object of the invention is to disclose the method as defined in any of the above, wherein said method is provided useful for increasing the at least one first cannabis-derived compound (FCDC) to at least one second cannabis-derived compound (SCDC) ratio of a chemical parameter, said parameter is selected from a group consisting of concentration, amount, accumulation rate, % weight and % molar; said increase is to a range being at least 5% over normal such a ratio in a non-treated plant.

Another object of the invention is to disclose the method as defined in any of the above, wherein said method is provided useful for increasing the THC to CBD weight ratio to a range being at least 5% over normal such a ratio in a non-treated plant; characterized by step emitting a PMF on a cannabis plant or a portion thereof.

Another object of the invention is to disclose the method as defined in any of the above, wherein said method comprises steps of analyzing at least one first cannabis-derived compound (FCDC) to at least one second cannabis-derived compound (SCDC) ratio of said parameter; defining a desired FCDC to SCDC ratio; exposing said cannabis plant or a portion thereof to at least one first protocol of PMF emission, said protocol comprising a predefined measure of emission of PMF per time; analyzing resulted FCDC to SCDC ratio; if resulted FCDC to SCDC ratio equals said desired FCDC to SCDC ratio, harvesting the same; if FCDC to SCDC ratio does not equals said designated FCDC to SCDC ratio, exposing said cannabis plant or a portion thereof either to said at least one first protocol or to at least one second protocol of PMF emission; said first and said second protocols comprising a predefined measure of emission of PMF per time.

Another object of the invention is to disclose the method as defined in any of the above, wherein said method comprises a feedback and comprises steps of analyzing at least one first cannabis- derived compound (FCDC) to at least one second cannabis-derived compound (SCDC) ratio of a chemical parameter, said parameter is selected from a group consisting of concentration, amount, accumulation rate, % weight and % molar; defining a desired FCDC to SCDC ratio; exposing said cannabis plant or a portion thereof to at least one first protocol of PMF emission, said protocol comprising a predefined measure of emission of PMF per time; analyzing resulted FCDC to SCDC ratio; if resulted FCDC to SCDC ratio equals said desired FCDC to SCDC ratio, harvesting the same; if FCDC to SCDC ratio does not equals said designated FCDC to SCDC ratio, exposing said cannabis plant or a portion thereof either to said at least one first protocol or to at least one second protocol of PMF emission; said first and said second protocols comprising a predefined measure of emission of PMF per time; analyzing resulted FCDC to SCDC ratio (T_n=2 analysis); if resulted T FCDC to SCDC ratio equals said desired FCDC to SCDC ratio, harvesting the same; if FCDC to SCDC ratio does not equals said designated T FCDC to SCDC ratio, by means of a processor, calculating the function of said at least one first protocol or said at least one second protocol of PMF emission on said FCDC to SCDC ratio, thereby optimizing the required protocol of PMF emission to provide said desired FCDC to SCDC ratio o; and exposing said cannabis plant or a portion thereof to said hereto calculated optimized protocol of PMF emission.

Another object of the invention is to disclose the method as defined in any of the above, wherein analyzing THC to cannabinoid weight ratio; defining a desired THC to cannabinoid weight ratio; exposing said cannabis plant or a portion thereof to at least one protocol of PMF emission, said protocol comprising a predefined measure of emission of PMF per time; analyzing resulted THC to cannabinoid weight ratio; if resulted THC to cannabinoid weight ratio equals said desired THC to cannabinoid weight ratio, harvesting the same; if THC to cannabinoid weight ratio does not equals said designated THC to cannabinoid weight ratio, exposing said cannabis plant or a portion thereof either to said at least one first protocol or to at least one second protocol of PMF emission; said first and said second protocols comprising a predefined measure of emission of PMF per time.

Another object of the invention is to disclose the method as defined in any of the above, wherein said method comprises steps of analyzing THC to cannabinoid weight ratio (To analysis); defining a desired THC to cannabinoid weight ratio; exposing said cannabis plant or a portion thereof to at least one protocol of PMF emission, said protocol comprising a predefined measure of emission of PMF per time; analyzing resulted THC to cannabinoid weight ratio (T_n=1 analysis); if resulted THC to cannabinoid weight ratio equals said desired THC to cannabinoid weight ratio, harvesting the same; if THC to cannabinoid weight ratio does not equals said designated THC to cannabinoid weight ratio, exposing said cannabis plant or a portion thereof either to said at least one first protocol or to at least one second protocol of PMF emission; said first and said second protocols comprising a predefined measure of emission of PMF per time; analyzing resulted THC to cannabinoid weight ratio (T_n=2 analysis); if resulted THC to cannabinoid weight ratio equals said desired THC to cannabinoid weight ratio, harvesting the same; if THC to cannabinoid weight ratio does not equals said designated THC to cannabinoid weight ratio, by means of a processor, calculating the function of said at least one first protocol or said at least one second protocol of PMF emission on said THC to cannabinoid weight ratio, thereby optimizing the required protocol of PMF emission to provide said desired THC to cannabinoid weight ratio; and exposing said cannabis plant or a portion thereof to said hereto calculated optimized protocol of PMF emission.

Another object of the invention is to disclose the method as defined in any of the above, wherein the cannabinoid is selected from a group consisting of CBD, CBG, CBC and CBGA.

Another object of the invention is to disclose the method as defined in any of the above, wherein said method is provided useful for regulating terpenes to one or more members of a group consisting of FCDC, SCDC and FCDC to SCDC ratio (namely, "F,S,F/S"), comprising steps of selecting a cannabis plant or portion thereof, comprising an initial terpenes to F,S,F/S ratio; defining a desired terpenes to F,S,F/S ratio; exposing the same to a PMF emission in an emission protocol, thereby altering the content of either or both said terpene and said F,S,F/S; if terpenes to F,S,F/S equals said desired F,S,F/S ratio then harvesting the plant and plant portions; if terpenes to terpene F,S,F/S does not equal said designated terpenes to F,S,F/S, then further exposing the same to a PMF emission in PMF in said emission protocol or to another emission protocol.

Another object of the invention is to disclose the method as defined in any of the above, wherein said plant portion is selected from a group consisting root, leaf, flower, stem node, cola, stigma, pistil, bract, calyx, trichome and any combination thereof.

Another object of the invention is to disclose the method as defined in any of the above, wherein said method is provided useful for regulating cannabis plant's pest, comprising steps of selecting a cannabis plant or portion thereof, comprising an initial plant's pest content; defining a desired plant's pest content; exposing the same to a PMF emission, thereby altering plant's pest content in said cannabis plant or a portion thereof; If plant's pest content equals said desired plant's pest content, then harvesting the same; and If plant's pest content does not equal said designated plant's pest content, then further exposing the same to said NTP emission.

Another object of the invention is to disclose the method as defined in any of the above, wherein said regulating cannabis plant's pest is at least one of step of repelling, killing, destroying, treating, preventing, defending and any combination thereof.

Another object of the invention is to disclose the method as defined in any of the above, wherein said pest is botrytis cinerea.

Another object of the invention is to disclose the method as defined in any of the above, wherein said pest is selected from a group consisting of lepidopterous stem borers, hemp borers, beetle grubs, caterpillars, beetles, bugs, leafminers, piercing-sucking mouthparts, aphids, whiteflies, leafhoppers, mealybugs, mites, fungi, nematodes, parasitic plants, bacteria, molds, viruses and any combination thereof.

Another object of the invention is to disclose the method as defined in any of the above, wherein cannabis plant is selected from a group of cannabis Santhica, cannabis sativa L., cannabis indica Lam., cannabis ruderalis janisch, electric daisy, liverwort, or flax seeds, and any combination thereof.

One object of the invention is to disclose a method for producing at least one de novo cannabis- derived compounds by changing at least one parameter of a defined PMF treatment aimed at emitting PMF on a cannabis plant or a portion of it, said parameters are selected from a group consisting of the number of PMF emitting events, number of pauses in-between PMF emitting events, number of pulses used of the PMF emitting events, and any combination thereof.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

The following description is provided, alongside all chapters of the present invention, so as to enable any person skilled in the art to make use of said invention and sets forth the best modes contemplated by the inventor of carrying out this invention. Various modifications, however, is adapted to remain apparent to those skilled in the art, since the generic principles of the present invention have been defined specifically to provide method for in vivo affecting cannabis plant by application of non-thermal plasma to said plant. Moreover, present invention provides a method for modifying growth pattern or chemical content of cannabis plant using non-thermal plasma. Specifically, the present invention discloses a system configured to shorten life span of a cannabis plant to flowering time, or to increase concentration, or to repel microorganisms or pests of said cannabis plant.

As used herein after, the term "non-thermal plasma" or NTP generally refers hereinafter to any plasma which is not in thermodynamic equilibrium, either because the ion temperature is different from the electron temperature, or because the velocity distribution of one of the species does not follow a Maxwell-Boltzmann distribution. It is in the scope of the invention wherein the NTP is referred to by the specific technology used to generate it i.e. "gliding arc", "plasma pencil", "plasma needle", "plasma jet", "dielectric barrier discharge", "one atmosphere uniform glow discharge plasma", "atmospheric plasma", "ambient pressure non-thermal discharges", "non- equilibrium atmospheric pressure plasmas"; wherein those terms related to both: non-thermal plasma and plasma operated at or near atmospheric pressure. It is further in the scope of the invention, wherein the generated plasma is selected from a group consisting of positive ions, negative ions, electrons meta-stables, atoms, free radicals and photons.

The term "modified plasma" or "plasma modified field" or "PMF" as used herein refers to a plasma coupled to or modified or transformed by or generated by, at least one of ferroelectric means or elements, ferromagnetic means or elements, piezoelectric means or elements or by a combination of all elements or any partial combination thereof. According to a main embodiment, the plasma is further adjusted or influenced by a reflecting element as inter alia disclosed. It is within the scope of the present invention that a "plasma modified field" or "PMF" refers to plasma oscillations influenced by a coupling element selected from the group consisting of at least one ferroelectric element, at least one ferromagnetic element, at least one piezoelectric element or by any combination thereof, as well as by a reflecting element. The PMF is applied to a subject in a predetermined pulsed manner to induce a therapeutic or regenerative or beneficial effect. The PMF system used is disclosed in US 8,896,211 B2 ( by Ish -Yamini Tomer and Levin).

As used herein after, the term“cannabis-derived compounds” refers hereinafter to compounds found in cannabis plant, comprising cannabinoids, terpenes and phenolic compounds. As used herein after, the term“cannabinoids” refers hereinafter to a class of diverse chemical compounds which are ligands for cannabinoid receptors in cells that alter neurotransmitter release in the brain. Cannabinoids were primarily discovered in marijuana (cannabis flower) and hashish (compressed cannabis resin) from the plant of Cannabis sativa. This plant contains more than 80 phyto- cannabinoids. The main active constituent of marijuana is the psychoactive D9- tetrahydrocannabinol (D9-THC), which acts at cannabinoid 1 (CB1) and cannabinoid 2 (CB2) receptors as a partial agonist. Other important natural cannabinoids present in marijuana are the non-psychoactive cannabidiol (CBD), D9-tetrahydro-cannabivarin (D9-THCV) and cannabichromene (CBC) [1-3]. Among them CBD has attracted the greatest attention thus far. It was shown to antagonize the effects of CB 1/CB2 receptor agonists, to counteract the psychotropic and other negative effects of D9-THC and several data suggest that it behaves as an inverse agonist of CB1 and CB2 receptors. Some of these plant-derived cannabinoids are used in the medical practice, such as D9-THC (dronabinol) and its synthetic analogue, nabilone against chemotherapy- induced nausea and emesis, and as appetite stimulants (e.g. in AIDS patients). CBD combined with D9-THC (nabiximols) is used to relief neuropathic pain and spasticity in multiple sclerosis, and as an adjunctive analgesic treatment in advanced cancer pain.

As used herein after, the term "terpenes”, refers hereinafter to a large and diverse class of organic compounds, produced by a variety of plants. Terpenes are derived biosynthetically from units of isoprene, which has the molecular formula C₅H₈. The basic molecular formula of terpenes are multiples of that, (C₅H₈)n where n is the number of linked isoprene units. Terpenes are fragrant oils that give cannabis its aromatic diversity.

As used herein after, the term“pests” refers hereinafter to the organisms that cause injury and disease to cannabis plants, lepidopterous stem borers, hemp borers, beetle grubs, caterpillars, beetles, bugs, leafminers, piercing-sucking mouthparts, aphids, whiteflies, leafhoppers, and mealybugs, mites, fungi, nematodes, parasitic plants, bacteria, molds, viruses and any combination thereof.

As used herein after, the term“cannabis plant and plant portion thereof’ refers hereinafter to several structures, found in cannabis plants Cannabis grows on long skinny stems with its large, iconic fan leaves extending out from areas called nodes. The cannabis plant has, inter alia, several other portions: roots, Cola: A cola refers to a cluster of buds that grow tightly together. While smaller colas occur along the budding sites of lower branches, the main cola (sometimes called the apical bud) forms at the very top of the plant. Stigma and Pistil: The pistil contains the reproductive parts of a flower, and the vibrant, hair-like strands of the pistil are called stigmas. Stigmas serve to collect pollen from males. The stigmas of the pistil begin with a white coloration and progressively darken to yellow, orange, red, and brown over the course of the plant’s maturation. They play an important role in reproduction, but stigmas bring very little to the flower’s potency and taste. Bract and Calyx: A bract is what encapsulates the female’s reproductive parts. They appear as green tear-shaped“leaves,” and are heavily covered in resin glands which produce the highest concentration of cannabinoids of all plant parts. Enclosed by these bracts and imperceptible to the naked eye, the calyx refers to a translucent layer over the ovule at a flower’s base. Trichome: Despite their minute size, it’s hard to miss the blanket of crystal resin on a cannabis bud. This resin (or“kief’ when dry) is secreted through translucent, mushroom-shaped glands on the leaves, stems, and calyxes. Trichomes were originally developed to protect the plant against predators and the elements. These clear bulbous globes ooze aromatic oils called terpenes as well as therapeutic cannabinoids like THC and CBD. The basis of hash production depends on these trichomes and their potent sugar-like resin.

Cannabis plant is a genus of flowering plants in the family Cannabaceae. Three species are recognized: Cannabis sativa, Cannabis indica, and Cannabis ruder alis. Additionality, there are also legal cannabinoid-containing plants, comprising, electric daisy, liverwort, or flax seeds.

Electric Daisy: Acmella oleracea is a species of flowering herb in the family Asteraceae. Common names include toothache plant, paracress, sechuan buttonbuzz buttons, ting flowers and electric daisy. Electric daisies are a good example of legal plants that are similar to cannabis. These plants have many chemical properties that are surprisingly similar to cannabis, specifically chemical properties that are closely related to chemicals produced by the cannabis plant.

Electric daisies contain a variety of cannabinoids that affect the body’s endocannabinoid system. In particular, cannabinoids from this plant affect the CBD2 receptors in human body, exerting their effects in easing pain and reducing inflammation. In fact, electric daisies can be made into a powerful painkilling gel, making the electric daisy a natural ally of dentists everywhere.

The most important taste-active molecules present are fatty acid amides such as spilanthol, which is responsible for the trigeminal and saliva-inducing effects of products such as jambu oleoresin, a concentrated extract of the plant. It also contains stigmasteryl-3-O-b-D-glucopyranoside and a number of triterpenes; see Woelkart, Karin, and Rudolf Bauer. "The role of alkamides as an active principle of Echinacea." Planta medica 73.07 (2007): 615-623, which is incorporated herein as a reference.

Liverwort: The New Zealand plant called Liverwort or Radula contains a large amount of a cannabinoid called perrottetinenic acid. Scientists first discovered this cannabinoid in 2002.

The ether extract of the New Zealand liverwort Radula marginata afforded a new cannabinoid type bibenzyl compound named perrottetinenic acid, and two new bibenzyls, together with a known cannabinoid, perrottetinene. Their structures were established by two dimensional (2D) NMR spectral data. The structure of perrottetinenic acid was a similar to that of D1- tetrahydrocannabinol (THC). Cannabinoid type bibenzyls have been isolated from liverwort Radula perrottetii, and from the liverwort R. marginata. Much like THC, it’s a cannabinoid that acts on the CB1 receptors, which are part of the larger endocannabinoid system. The presence of cannabinoids means that liverwort can be used for medical purposes. So far, people have found ways to use the plant to treat a variety of illnesses, from bronchitis to bladder issues.See Toyota, Masao, et al. "New bibenzyl cannabinoid from the New Zealand liverwort Radula marginata." Chemical and pharmaceutical bulletin 50.10 (2002): 1390-1392., which is incorporated herein as a reference

Flax seeds: Flax plant is a valuable source of fibers, linseed and oil. In the course of analysis of fibers extract from previously generated transgenic plants overproducing phenylpropanoids a new terpenoid compound was discovered Cannabidiol (CBD)-like compound was found in flax fiber and tissues. Since CBD is one of the most important cannabinoids in the cannabis plant, this is a key similarity between the two plants.

Additionally, it was found that flax products can be a source of biologically active cannabinoid- like compounds that are able to influence the cell immunological response. These findings might open up many new applications for medical flax products, especially for the fabric as a material for wound dressing with anti-inflammatory properties. See Styrczewska, Monika, et al. "Cannabinoid-like anti-inflammatory compounds from flax fiber." Cellular & molecular biology letters 17.3 (2012): 479., which is incorporated herein as a reference.

As used herein after, the term "analytical methods” generally refers hereinafter to detection methods for analysis the content, amount and purity cannabinoids, terpenes and phenlic compounds. These detection methods comprises, detection methods as follows: HPLC- High- performance liquid chromatography, GC- gas chromatography, LC-MS- liquid chromatography- mass spectrometry, LC-MS/ MS- Liquid chromatography-tandem mass spectrometry GC-MS - gas chromatography-mass spectrometry, GC-TOF- gas chromatography -MS analyzer "Time of Flight", UPLC- MS- ultra performance liquid chromatography mass spectroscopy, LC-TOF- liquid chromatography -MS analyzer "Time of Flight", NMR- nuclear magnetic resonance, TLC - thin layer chromatography, DART- HPLC- direct analysis in real time coupled with high-performance liquid chromatography, SPME-HS- GC-MS- solid phase micro extraction headspace and gas chromatography-mass spectrometry and MALDI-TOF- matrix assisted laser desorption ionization-time of flight mass spectrometry As used herein the term "about" denotes ± 25% of the defined amount or measure or value. EXAMPLE 1

Goal The goal of the study is to examine the effects induced by exposing whole fresh cannabis plants to Modified Plasma Field (PMF ) in terms of physical and physiological parameters and the amount of the compounds (cannabinoids) produced.

The exposure of the plants to the PMF occurred by 3 stages treatment plus pause days referred to as a whole treatment cycle. The treatment stages were conducted on the I^st the 4^th and the 7^th day. The duration of each treatment was between 3 times: first exposure (for a few minutes) three days off, second exposure (for a few minutes) with three days off and final exposure (for a few minutes). The experiment included also a control group (non- treated/not exposed plants) (see Table 5).

Table 5: Design for studying the Treatment effects on Fresh

Plants Grown in Soil when using PMF (N=24)

The study used 12 plants for treatment with PMF and 12 control plants. Table 6: Design for studying the treatment effects on fresh plants grown in soil when using PMF on plants grown until having flowers (N=l2 selected from stage 1) .

Detailed Protocol Number and kind of Plants:

10 Santhica 27 (FR) plants grown in soil (12 for treatment of PMF treatment ; 12 for control)

Total Number of Plants required for experiment 1 is: 30

Treatment: A treatment is defined by its 3 stages to be implemented on: lst day; 4th day and

7th day (see design). The whole treatment including the pause day are 8 days (1, 4, 7) and 3+3 +1 pauses days.

Time length for each treatment stage = between 3.15 and 5.25 minutes (setting the pulse parameter on the device for between 4 and 8 pulses)

Environmental conditions for keeping plants between treatments (temperature, lightening water supply, humidity, pH) for treated and control samples are identical and are summarized in Table

7.

Table 7; Environmental conditions

The soil based plants were grown and tested in non- metal vessels

The PMF device was placed on a non-metal-based desk or metal surface

Following the treatment and the pause days in-door analysis of plants started in terms of physical and physiological parameters and also follow up on the plants growth /until flowers bloom or buds are immerged. Additionally, after dried freezing the leaves and roots parts, the samples were sent for analysis to assess the amount of cannabinoid compounds produced by the plants parts.

Physical Analysis Physiological and physical Analysis of fresh plants parts- is done in door analysis. Follow up on the physical growth treated and control plants was performed by photographic measurements once a week (stage 2).

Growth parameters Plant height of the treated and control plants was investigated using photographic documentation directly before and after exposure to OMPF.

The HunterLab-system was used to measure potential impact of the treatment on the leaf color directly before and after the treatment cycle with the OMPF device. A Minolta spectrophotometer (CM-2600D, Konica Minolta Inc., Osaka, Japan) was set at illuminant D65, 3 mm aperture, and 0° viewing angle. Color coordinates were recorded 5 times for 3 randomly selected leaves of the hemp plants.

Chlorophyll fluorescence imaging (CFI) Chlorophyll fluorescence is a non-invasive measurement of photosystem II (PSII) activity and is a commonly used technique in plant physiology. The sensitivity of PSII activity to abiotic and biotic factors has made this a key technique not only for understanding the photosynthetic mechanisms but also as a broader indicator of how plants respond to environmental change. Chlorophyll fluorescence imaging (CFI) was performed using a modular system (FluorCAM 700MF, PSI, Brno, Czech Republic), which measures sequences of fluorescence images with a user defined timing of set points, measurement intervals and irradiance. The initial basic fluorescence (FO) was induced by weak, non-actinic measuring-light pulses of two sets of 345 super-bright orange light emitting diodes (lmax = 620 nm). Maximum fluorescence (Fm) was excited by a short-term (1 s) saturation light pulse (max. 2500 mmol photons m-2 s-l), generated by a halogen lamp (250 W) equipped with an electronically controlled shutter. Fluorescence images were recorded by a CCD camera (12-bit, 512 x 512 pixels; maximum frame rate 50 images s-l) equipped with an Fl.2/2.8-6 mm objective and a short-pass filter system (high pass 695 nm, low pass 780 nm) synchronously with the weak, non-actinic measuring-light pulses. The system was controlled by Windows XP compatible software (FluorCAM 6, PSI, Brno, Czech Republic).

All measurements were performed after pre-darkening of the hemp plants for 15 min. Measurements were conducted directly before and after the treatment cycle with the OMPF device. The potential impact of the treatment on the physiological activity of the hemp plants was assessed by evaluating the maximum photon yield of electron transport through photosystem II (Fv/Fm; Fv = Fm - FO). This parameter is a valuable tool to determine both capacity and stability of photosynthesis and its response to biotic and abiotic constraints. Fv/Fm ranges between 0.84 in highly active plants and 0 in fully damaged (dead) tissues and Fv /Fm values below 0.1 were neglected.

V Chlorophyll Fluorescence Imaging (CFI) was performed using a modular system (FluroCAM 700MF, PSI, Brno, Czech Republic)

Samples for analysis: The samples of the fresh plants were dried frozen for analyzing the plants' parts compounds

Stage 1: Fresh Plants: 12 plants were assigned to a treatment group and 12 to control group. Of the 12 plants 6 were selected randomly to serve as the treated plants and 6 were selected in the stage 2 of the study (to follow up until flowers grow) and similarly: 6 samples were selected randomly from the 12 control plants and 6 were selected for stage 2 of the study ( for follow up until flowers grow) . Both leaves and roots were examined. The leaves and the roots were extracted from different places on the same plant. At this time leaves and roots were sent for cannabinoids analysis and physiological parameters were also observed and measured Stage 2: the 6 randomly selected samples and the 6 control samples were followed up and documented for growth until flowers form. At this time leaves and flowers were sent for analysis of cannabinoids analysis and physiological parameters were observed and measured.

Number of samples for analysis: From the 6 treated plants and 6 control plants 12 samples were formed -12 samples of leaves and 12 samples of roots parts: (12+12 treated samples for analysis) and similarly from the 6 control plants 12 samples were formed for the leaves and 12 for the roots parts forming 24 samples (12+ 12) for the analysis of leaves and for the roots.

Timing of Analysis: In-house analysis started immediately after the treatment ends (one day after the 3^rd treatment stage- see design). The follow up on the plants’ growth with photographic shots was done once a week.

For chemical analysis (analytics) of the cannabis-derived compounds, the samples were dried and frozen immediately after the treatment with its pause days, before the chemical analysis. Samples were studied using LC-MS/MS.

Outcome Variables The outcome variables examined were:

a. Plant Height ;

b. Leaf color;

c. Measurements derived from chlorophyll fluorescence image analysis (CFI - measuring capacity and stability of photosynthesis and its response to biotic and abiotic constraint. Fv/Fm ranges between 0.84 in highly active plants and 0 in fully damaged tissues); and d. Analytics of the type of plants’ products (cannabis-derived compounds such as cannabinoids, terpenes etc.) produced as well as the amount and concentration of the cannabis-derived compounds produced as a result of the PMF treatment.

Analytics included measuring the content (existence, amount, and purity) of the following cannabis derived compounds:

Cannabinoids d9- THC- Tetra-hydro-cannabinoids; THC total (THC+THCA); d8- THC neutral,- Tetra-hydro-cannabinoids; THCV etrahydrocannabivarin;

THCA- d9- Tetra-hydro-cannabinolic acid; THCA; THCV/THC-C3 - Tetra-hydro-cannabivarin

CBD - Cannabidiol and CBDA - Cannabidiolic acid; CBDV - Cannabidivarin CBG - Cannabigerol and CBGA - Cannabigerolic acid; CBN - Cannabinol ; CBNA - Cannabidiolic acid, CBC ; Cannabichromene ; and CBCA Cannabichromenic acid.

Terpens and Alkaloids Mono-terpens ; Sesqui- terpens; Friedelin and Epifreidelin; carvone and dihydricarvone; Cannabistivine and Anhydrocannabistivin; Sterole; and Choline

Phenolic Compounds: O-glycoside; Cannaflavin A; Cannaflavin B; and Canabisin D, Friedelin and Epifreidelin.

Results Results of Example 1 are summarized in Table 8.

Table 8: Growth parameters

of study as compared the heights at the end of the treatment (8 days after the first measurement) . ( p= 0.00015 for the treatment group ; while p= 2.96288E-07, for the control group) ; with no differences between the treated and the control group after the treatment ; indicating safety of the treatment.

The results also clearly show that although the heights of the treated plants were more variable at the begging of the study, these heights were significantly less variable at the end of the study for these treated plants (The difference between the variances are significant F=5.90072 s; p<0.03) ; suggesting the treatment led to decreased variability of the plants’ growth.

Leaf Color Figure 3 depicts the leaf color measurements of control compared to treated plants, at the beginning of the (before) and at the end of the treatment (after). No statistically significant effect of the treatment regarding the color of the leaves was detected.

Chlorophyll fluorescence Imaging (CFI)

The parameters measured are as follows:

F0= Minimal chlorophyll fluorescence intensity measured in the dark-adapted state, when all PSII reaction centres are open;

Fm=Maximal chlorophyll fluorescence intensity measured in the dark-adapted state during the application of a saturating pulse of light;

Fv= Variable chlorophyll fluorescence (Fm - FO) measured in the dark-adapted state, when non photochemical processes are minimum; and

Fv/Fm = Maximum quantum yield of PSII photochemistry measured in the dark-adapted state.

The results of the Chlorophyll fluorescence Imaging (CFI) show no significant effects of the treatment regarding measured plants’ characteristics, indicating safety of the PMF treatment.

Initial basic fluorescence (Fo) results are summarized in Table 9.

Table 9: Initial basic fluorescence (Fo) results

The results show no significant differences in Fo measurements between treated and control plants (t-Test: Two-Sample Assuming Unequal Variances). These results indicate safety and non toxicity of the treatment. This is specifically in view of the fact that increase in Fo represents any difficulty and degradation in photosystem II or any disruption in energy transfer into the reaction center ( see Calatayud A, Roca D, Martinez, PF. (2006) Spatial temporal variations in rose leaves under water conditions studied by chlorophyll fluorescence imaging. Plant Physiol Biochem

44:564-573).

Maximal chlorophyll fluorescence intensity (Fm) results are summarized in Table 10:

Table 10: Maximal chlorophyll fluorescence intensity (Fm) results

The results show no significant differences in Fm measurements between treated and control plants after treatment (t-Test for Two-Sample Assuming Unequal Variances t=-o.48l p<0.3)

The results of variable chlorophyll fluorescence (Fm - F0) measured in the dark-adapted state, when non-photochemical processes are minimum ( Fv) , are summarized in Table 11.

Table 11: Results of Variable chlorophyll fluorescence (Fm - F0) measured in the dark-adapted state, when non-photochemical processes are minimum), Fv.

The results show no significant differences in Fv measurements between treated and control plants after treatment (t-Test: Two-Sample Assuming Unequal Variances; t=-0.8267 p<0.21)).

Fv Physiological fitness

The results of Fv/Fm (Maximum quantum yield of PSII photochemistry measured in the dark- adapted state) are summarized in Table 12.

Table 12: The results of Fv/Fm (Maximum quantum yield of PSII photochemistry measured in the dark-adapted state).

The results show significant differences in Fv/FM ratio for treated compared to control plants after treatment (t-Test: Two-Sample Assuming Unequal Variances, t=-3.074 p<0.0066)); The results show significant difference within the treatment group between before and after treatment (t=2.027 p<0.049) and no significant difference was observed for the control group between before and after treatment (t=0.67 p<0.26).

Analytics results The aforementioned analytical results were obtained in the leaves of the examined plants, using LC-MS/MS technique, in control plants in plants treated according to the first treatment protocol. The results are summarized in Tables 13A- D. Table 13A: The cannabinoids content measured in control plants of example 1

Table 13B: The cannabinoids content measured in treated plants of Example 1

Table 13C: The CBS content measured in control and treated plants of Example 1.

The results of the CBS content measurements show less diversity, towards unified CBS content as a result of the PMF treatment.(F= 17.994 p<0.02).

Table 13D: The cannabinoids content’s ratios measured in control compared to treated plants of example 1.(* p< 0.05, t-Test: Two-Sample Assuming Unequal Variances).

The results show a significant difference between the CBGA content of the leaves of the treated group compared to the control group (p= 0.059, t-Test: Two-Sample Assuming Unequal Variances); indicating a significant effect of increasing the concentration of CBGA following PMF treatment. Additionally, the CBG content of the leaves of the treated group compared to the control group shows a trend line significant effect (p= 0.32, t-Test: Two-Sample Assuming Unequal Variances); indicating a marginal effect of increasing the concentration of CBA following PMF treatment.

Furthermore, when calculating the average amount of CBC in the plants’ leaves that show a measurable amount of CBC, the results indicate a significant increase in CBC concentration as results of the PMF treatment (p= 0.02, t-Test: Two-Sample Assuming Unequal Variances ).

Overall, these results show that the PMF treatment changes the concentration of at least one cannabinoid and changes the ratios among at least two cannabinoids, according to the set of parameters used in the study described in exmaple 1.

The analytical examination of the roots revealed no measurable amount of cannabinoids or terpenes. These results suggest that the treatment which was applied on the soil-based whole plant did not affect the roots in the soil. Since the roots are the lifeline of a plan they influence/effect the plant's growth and they are able to adjust their own growth to changing environments, The result therefore implies that the effects of the PMF applies directly on the leaves not through the roots. It also indicates safety of the treatment.

The study showed increased concentration of CBGA and CBC as well as a trend of increased concentration of CBG.

The study serves as a basis for further treating cannabis plants in order to regulate cannabinoids content or ratio see Fig. 1.

Reference is now made to Figure 1, illustrating a method (100) of regulating ratio of cannabis derived compounds (cannabinoids, terpenes and phenolic compounds) comprising steps of selecting a cannabis plant or portion thereof (110), defining a desired content or ratio of cannabis derived compounds (120), detection of content (existence, amount, and purity) of cannabis derived compounds (130), exposing the same to a PMF over time (140), thereby altering cannabis derived compounds in said cannabis plant or a portion thereof (150). If (170) content of cannabis derived compounds ratio equals said designated content (190), then harvesting the same (180) ; If content of cannabis derived compounds does not equal (160)said designated content, then further exposing the same to said PMF for additional time (140). EXAMPLE 2

Goal: Analytics of the type of plants’ products (the cannabis-derived compounds such as cannabinoids, terpenes etc.) produced as well as the amount and concentration of these cannabis- derived compounds produced as a result of a second treatment protocol of PMF treatment.

The design of the study of example 2 is depicted in Table 14.

Table 14: Design for studying the Treatment effects on Fresh

Plants Grown in Soil when using second protocol of PMF (N=6).

The treatment is defined: lst day treatment and 1 day pause; 2nd stage treatment and 1 day pause and 3rd day treatment and 1 day pause. Time length for each treatment stage (3 stages) is between 5 and 9 minutes (between 8 and 12 pulses). Treatment is applied vertically above the plant

Environmental conditions for treated and control samples are kept as in Example 1.

Results The results of the Example 2 study are summarized in the Tables no. 15A-B

Table 15A: The cannabinoids content measured in treated plants of example 2

Table 15B: The cannabinoids content measured in control plants of example 2.

The results of the study in example 2 show the existence and measurable amounts of CBC and THC (THCA and THCt); while these cannabinoids could not be detected in the control plants.

The results of this study show the effect of PMF in producing de novo new cannabinoids in cannabis plants in order to regulate cannabinoids content, and producing desired cannabinoids, which were not detected in the control (non-treated) cannabis plants. This might serve as an option to use less cannabis strains for the production of desired cannabis-based compounds as well as ratios between these cannabis- based compounds.

EXAMPLE 3

Goals

a. To examine the capability of PMF treatment to decontaminate whole cannabis plants grown in soil; and b. To demonstrate that after exposing fresh contaminated cannabis plants grown in soil to PMF, defined by a pre-determined treatment, the pathogen microorganisms will be exterminated or reduced.

The experiment examined the extermination and wiping out of the fungus botrytis, which contaminate cannabis plants. Plants grown in soil which were contaminated with the plant parasite botrytis were exposed to the PMF and following the exposure for a defined time length (a few minutes) 3 times according to the protocol described in Table 5), the parts of the plants were examined for existence (or no existence) of the fungi compared to non- exposed (control) plants.

The study design is as follows (see Table 16).

Table 16: Study design for Example 3

The physical appearance of treated plants was superior compared to infected non- treated plants

(see Figures 4A-B). Figure 4A depicts non- contaminated plants. Figure 4B depicts Non- contaminated treated (right) untreated (left). Figure 4C depicts contaminated treated (right) untreated (left).

EXAMPLE 4

Goals: a. To examine the defending nature of fresh cannabis plants (leaves on the plants) when exposed to modified plasma fields (PMF ): b. To demonstrate that after exposing fresh cannabis whole plants to PMF , the plants will reject contamination process by pathogenic microorganisms (fungus) on its leaves

The experiment examined the preventive/definitive nature of fresh cannabis plants by investigating the PMF induced rejection of pathogenic microorganisms on leaves of treated cannabis plants in comparison to non-treated plants.

For this study nine treated and nine control plants were used.

Prior and following treatment of the plants (definition of the treatment is identical to Example 1) the total viable count (TVC) of microorganisms on both treated and control plants was investigated (Pretest).

Subsequently following exposure to PMF treatment, nine treated and nine control plants were inoculated with botrytis cinereal using its micelles. The TVC of the microorganisms and the quantity of the botrytis cinereal on both treated and control plants were examined. Therefore each 6 leaves were removed from 3 treated and control plants. The plants were discarded afterwards

Following growing of the plants for at least 3 or more days, the plants, both treated and control were again analyzed (post-test). For their TVC of microorganisms and differences in fungal infestation ( botrytis cinereal). Conclusions regarding the specific effects of PMF treatment on the final infestation of the plants with fungal microorganism using botrytis Cinerea as an example is drawn in order to assess the ability of the PMF treatment to enable the treated plants to defend itself from infestation.

The existence and amount of pests including diseases caused by organisms was assessed and measured. Reference is now made to Figure 2, illustrating a method (200) for regulating cannabis plant's pest comprising steps of selecting a cannabis plant or portion thereof (210), comprising defining a desired plant's pest content (220) ; detection of the content of pests ( 230) ; exposing the same to an PMF over time (240), thereby altering plant's pest content (250) in said cannabis plant or a portion thereof; If (270) plant's pest content equals (280) said designated plant's pest content, then harvesting the same (290) ; If plant's pest content does not equal (260) said designated plant's pest content, then further exposing the same to said NTP for additional time (240).

Claims

1. A method for increasing at least one of a parameter of at least one cannabis-derived compound to a range being at least 5% over normal in a non-treated plant; characterized by step of emitting a PMF on a cannabis plant or a portion thereof; said parameter is selected from a group consisting chemical parameters,

2. The method of claim 1, wherein said chemical parameter is selected from a group consisting of amount and concentration, weight percent and molar percent of said cannabis-derived compound in the plants or portions thereof.

3. The method of claim 1 , wherein said biological parameter is selected from a group consisting of accumulation rate of said cannabis-derived compound in the plants or portions thereof (i.e., either amount per time or concentration per time), pesticidic effect, including one or more effects selected from a group consisting activity as an insecticides, nematicide, molluscicide, piscicide, avicide, rodenticide, bactericide, insect repellent, animal repellent, antimicrobial, fungicide and disinfectant

4. The method of claim 1, wherein said agrotechnical parameter is selected from a group consisting of flowering rate and photosynthesis parameters.

5. The method of claim 1, wherein said method is provided useful for increasing at least one chemical parameter of both at least one first cannabis-derived compound and at least one second cannabis-derived compound to a range being at least 5% over normal in a non-treated plant; characterized by step emitting a PMF on a cannabis plant or a portion thereof; said chemical parameter is selected from a group consisting of amount, concentration, accumulation rate (i.e., either amount per time or concentration per time), and said at least one first cannabis-derived compound to at least one second cannabis-derived compound (wt/wt) or (mole/mole) ratio.

6. The method of claim 1, wherein said method is provided useful for increasing the ratio of at least one chemical parameter of at least one first cannabis-derived compound over at least one second cannabis-derived compound to a range being at least 5% over normal such a ratio in a non-treated plant; characterized by step emitting a PMF on a cannabis plant or a portion thereof; said chemical parameter is selected from a group consisting of amount, concentration, accumulation rate (i.e., either amount per time or concentration per time), and said ratio is selected from a group consisting of (wt/wt) and (mole/mole) ratio.

7. The method of claim 1 wherein said cannabis-derived compound is selected from a group consisting of one or more cannabinoids, terpenes, phenolic compounds and any combination thereof.

8. The method of claim 7, wherein said one or more cannabinoids are selected from a group consisting of Tetra-hydro-cannabinoids (d9- THC), Tetra-hydro-cannabinoids (d8- THC), Tetra-hydro-cannabinolic acid (THCA- d9), Tetra-hydro-cannabivarin (THCV/THC-C3), Cannabidiol (CBD), Cannabidiolic acid (CBDA), Cannabidivarin (CBDV), Cannabigerol (CBG), Cannabigerolic acid (CBGA), Cannabinol (CBN), Cannabidiolic acid (CBNA), Cannabichromene (CBC), Cannabichromenic acid (CBCA), and any combination thereof.

9. The method of claim 7, wherein said one or more terpenes are either at least one mono- terpene or sesqui-terpene.

10. The method of claim 7, wherein said one or more phenolic compounds are selected from a group consisting of O-glycoside Cannaflavin A, Cannaflavin B, Canabisin D, and any combination thereof.

11. The method of claim 1, wherein said method is provided useful for increasing the at least one first cannabis-derived compound (FCDC) to at least one second cannabis-derived compound (SCDC) ratio of a chemical parameter, said parameter is selected from a group consisting of concentration, amount, accumulation rate, % weight and % molar; said increase is to a range being at least 5% over normal such a ratio in a non-treated plant.

12. The method of claim 11, wherein said method is provided useful for increasing the THC to CBD weight ratio to a range being at least 5% over normal such a ratio in a non-treated plant; characterized by step emitting a PMF on a cannabis plant or a portion thereof.

13. The method of claim 1, wherein said method comprises steps of

a. analyzing at least one first cannabis-derived compound (FCDC) to at least one second cannabis-derived compound (SCDC) ratio of said parameter; b. defining a desired FCDC to SCDC ratio;

c. exposing said cannabis plant or a portion thereof to at least one first protocol of PMF emission, said protocol comprising a predefined measure of emission of PMF per time; d. analyzing resulted FCDC to SCDC ratio;

e. if resulted FCDC to SCDC ratio equals said desired FCDC to SCDC ratio, harvesting the same;

f. if FCDC to SCDC ratio does not equals said designated FCDC to SCDC ratio, exposing said cannabis plant or a portion thereof either to said at least one first protocol or to at least one second protocol of PMF emission; said first and said second protocols comprising a predefined measure of emission of PMF per time.

14. The method of claim 1, wherein said method comprises steps of

a. analyzing at least one first cannabis-derived compound (FCDC) to at least one second cannabis-derived compound (SCDC) ratio of a chemical parameter, said parameter is selected from a group consisting of concentration, amount, accumulation rate, % weight and % molar;

b. defining a desired FCDC to SCDC ratio;

c. exposing said cannabis plant or a portion thereof to at least one first protocol of PMF emission, said protocol comprising a predefined measure of emission of PMF per time; d. analyzing resulted FCDC to SCDC ratio;

e. if resulted FCDC to SCDC ratio equals said desired FCDC to SCDC ratio, harvesting the same;

f. if FCDC to SCDC ratio does not equals said designated FCDC to SCDC ratio, exposing said cannabis plant or a portion thereof either to said at least one first protocol or to at least one second protocol of PMF emission; said first and said second protocols comprising a predefined measure of emission of PMF per time.

g. analyzing resulted FCDC to SCDC ratio (T_n=2 analysis);

h. if resulted T FCDC to SCDC ratio equals said desired FCDC to SCDC ratio, harvesting the same;

i. if FCDC to SCDC ratio does not equals said designated T FCDC to SCDC ratio, by means of a processor, calculating the function of said at least one first protocol or said at least one second protocol of PMF emission on said FCDC to SCDC ratio, thereby optimizing the required protocol of PMF emission to provide said desired FCDC to SCDC ratio o; and

j. exposing said cannabis plant or a portion thereof to said hereto calculated optimized protocol of PMF emission.

15. The method of claim 14, wherein said method comprises steps of

a. analyzing THC to a cannabinoid weight ratio;

b. defining a desired THC to said cannabinoid weight ratio;

c. exposing said cannabis plant or a portion thereof to at least one protocol of PMF emission, said protocol comprising a predefined measure of emission of PMF per time; d. analyzing resulted THC to said cannabinoid weight ratio;

e. if resulted THC to said cannabinoid weight ratio equals said desired THC to said cannabinoid weight ratio, harvesting the same;

f. if THC to said cannabinoid weight ratio does not equals said designated THC to a said cannabinoid weight ratio, exposing said cannabis plant or a portion thereof either to said at least one first protocol or to at least one second protocol of PMF emission; said first and said second protocols comprising a predefined measure of emission of PMF per time.

16. The method of claim 15, wherein said method comprises steps of

a. analyzing THC to a cannabinoid weight ratio (To analysis);

b. defining a desired THC to said cannabinoid weight ratio;

c. exposing said cannabis plant or a portion thereof to at least one protocol of PMF emission, said protocol comprising a predefined measure of emission of PMF per time; d. analyzing resulted THC to said cannabinoid weight ratio (T_n=1 analysis);

e. if resulted THC to CBD weight ratio equals said desired THC to said cannabinoid weight ratio, harvesting the same;

f. if THC to said cannabinoid weight ratio does not equals said designated THC to said cannabinoid weight ratio, exposing said cannabis plant or a portion thereof either to said at least one first protocol or to at least one second protocol of PMF emission; said first and said second protocols comprising a predefined measure of emission of PMF per time;

g. analyzing resulted THC to said cannabinoid weight ratio (T_n=2 analysis); h. if resulted THC to CBD weight ratio equals said desired THC to said cannabinoid weight ratio, harvesting the same;

i. if THC to said cannabinoid weight ratio does not equals said designated THC to said cannabinoid weight ratio, by means of a processor, calculating the function of said at least one first protocol or said at least one second protocol of PMF emission on said THC to said cannabinoid weight ratio, thereby optimizing the required protocol of PMF emission to provide said desired THC to said cannabinoid weight ratio; and j. exposing said cannabis plant or a portion thereof to said hereto calculated optimized protocol of PMF emission.

17. The method according to any one of claims 15 or 16, wherein said cannabinoid is selected from a group consisting of CBD, CBG, CBC and CBGA.

18. A method according to claim 1, wherein said method is provided useful for regulating terpenes to one or more members of a group consisting of FCDC, SCDC and FCDC to SCDC ratio (namely, "F,S,F/S"), comprising steps of

a. selecting a cannabis plant or portion thereof, comprising an initial terpenes to F,S,F/S ratio;

b. defining a desired terpenes to F,S,F/S ratio;

c. exposing the same to a PMF emission in an emission protocol, thereby altering the content of either or both said terpene and said F,S,F/S;

d. if terpenes to F,S,F/S equals said desired F,S,F/S ratio then harvesting the plant and plant portions;

e. if terpenes to terpene F,S,F/S does not equal said designated terpenes to F,S,F/S, then further exposing the same to a PMF emission in PMF in said emission protocol or to another emission protocol.

19. The method as defined in claim 1, wherein said plant portion is selected from a group consisting root, leaf, flower, stem node, cola, stigma, pistil, bract, calyx, trichome and any combination thereof.

20. The method as defined in claim 1, wherein said method is provided useful for regulating cannabis plant's pest, comprising steps of

a. selecting a cannabis plant or portion thereof, comprising an initial plant's pest content; b. defining a desired plant's pest content; c. exposing the same to a PMF emission, thereby altering plant's pest content in said cannabis plant or a portion thereof;

d. If plant's pest content equals said desired plant's pest content, then harvesting the same; e. If plant's pest content does not equal said designated plant's pest content, then further exposing the same to said NTP emission.

21. The method according to claim 19, wherein said regulating cannabis plant's pest is at least one of step of repelling, killing, destroying, treating, preventing, defending and any combination thereof.

22. The method according to claim 19, wherein said pest is botrytis cinerea.

23. The method according to claim 19, wherein said pest is selected from a group consisting of lepidopterous stem borers, hemp borers, beetle grubs, caterpillars, beetles, bugs, leafminers, piercing-sucking mouthparts, aphids, whiteflies, leafhoppers, mealybugs, mites, fungi, nematodes, parasitic plants, bacteria, molds, viruses and any combination thereof.

24. The method according to claims 1 , where cannabis plant is selected from a group of cannabis Santhica, cannabis sativa L., cannabis indica Lam., cannabis ruderalis janisch, electric daisy, liverwort, or flax seeds, and any combination thereof

25. A method for producing at least one de novo cannabis-derived compounds by changing at least one parameter of a defined PMF treatment aimed at emitting PMF on a cannabis plant or a portion of it, said parameters are selected from a group consisting of the number of PMF emitting events, number of pauses in-between PMF emitting events, number of pulses used of the PMF emitting events, and any combination thereof.