Cannabinoids and THC Part 1
Cannabinoids and THC
In this study we will examine two types of the several glands on Cannabis, the capitate sessile and capitate stalked forms. In addition, there are hairs on the surface of the plant. In our commentary we will refer to cannabinoids as the broad class of specialized compounds synthesized by Cannabis. Cannabinoids are dimers in that they are formed by condensation of terpene and phenol precursors (c).
Diagram 1 (Above). Cannabinoid pathway. Cannabinoids represent a dimer consisting of a terpene and a phenol component. Cannabigerol (CBG) is the first component of the pathway. It undergoes chemical change to form either cannabichromene (CBC), or cannabidiol (CBD). Delta 9-tetrahydrocannabinol (THC) is derived from CBD.
Cannabinoids include these more abundant forms:
THC, delta 9-tetrahydrocannabinol
CBD, cannabidiol
CBC, cannabichromene
CBG, cannabigerol
Another cannabinoid, cannabinol (CBN), is formed from THC and can be detected in some plant strains. Typically, THC, CBD, CBC and CBG occur together in different ratios in the various plant strains. In fiber strains CBD/CBC are in high concentrations and THC is at a low concentration; in drug strains, THC is high and CBD/CBC are low. An important area to be studied relates to the occurrence in the plant of potential precursor for cannabinoid formation and where synthesis occurs in the cell.
Distribution and types of glands
Glands cover entire surface of the above ground portion of both pistillate (female) and staminate (male) plants, but are most abundant on bracts of female plants (Fig. 1).
http://www.hempreport.com/issues/17/graphics/m1.gif Figure 1. (Above) Stalked and sessile glands on underside of a bract. Hairs also are present on bract. x35.
http://www.hempreport.com/issues/17/graphics/m2.gif Figure 2. (Above). Stalked gland showing large head (star) and abscission zone at base of gland head (arrow). Sessile glands are evident in background along with a portion of a hair. X300.
Legend for all figures: C, cuticle; D, disc cell; P, plastid; S, secretory material; T, secretory cavity; V, vesicle; W, disc cell wall.
There are two types of glands active in cannabinoid secretion on female plants (Figs. 1, 2):
a. Capitate sessile - most common in that it occurs on stems, leaves and bracts.
b. Capitate stalked - develops only after flower formation, and occurs especially on the bracts subtending a flower and seed. Both types are present on the bract subtending the seed, but some factor(s) that stimulates flowering also stimulates development of the stalk related to this gland. Thus this gland has evolved from the sessile type.
An abscission zone develops at the base of the head where the stipe cells attach to the disc cells resulting in abscission of glands upon attaining maturity (Fig. 2, arrow).
Cannabinoid, including THC, composition of glands
a. Effect of position and season on contents of gland.
We examined both gland types for their cannabinoid composition. Whole glands (20 glands) were removed from bracts and leaves of the same plant to compare their contents. They were extracted for cannabinoids and analyzed (Table 1).
We also separated them as to glands over a vein as contrasted to non-vein areas. Results showed that stalked glands over a vein on the bract contained more THC content, approximately 20 times more, than the sessile glands over a leaf vein. Similarly, stalked glands over a non-vein area contained much more THC than sessile glands over the leaf non-vein area.
Surprisingly, even stalked glands on a bract varied for THC content between the vein and non-vein area. We also found that sessile glands varied for contents between vein and non-vein areas on the leaf.
This table shows two intervals during the year, October and December, during which we compared levels of THC in glands. Comparison of October with December analyses showed again that stalked glands contained considerably more THC than sessile glands. For both types, however, the vein glands now showed less than the non-vein. Interestingly, the level of THC can decrease to a non-detectable level in the glands (leaf vein glands).
In similar analyses of these glands on other strains we found the same pattern, but the cannabinoid levels in the stalked glands were not as high as for this strain. In this strain the sampling was repeated during the following March and, again, the stalked glands on the bract contained higher concentrations of THC than sessile glands on the leaf. Thus, a repeatable pattern appears to occur in the plant in which stalked glands usually contained greater quantities of THC than sessile glands.
b. Effect of gland age on cannabinoid content
We also examined cannabinoid content of stalked gland by age to measure the major cannabinoid components in both a fiber and drug strain (Table 2). Glands, viewed under a microscope, can be classified according to their secretory phases from the color of their contents. Glands most active in secretion (mature) are translucent in appearance, aged glands are yellow in appearance and senescent glands are brown in color. Mature glands possessed the highest content of their major cannabinoid in both the fiber and drug strains. Senescent glands possessed low levels of cannabinoids. The concentration of some components, as CBD in the drug strains, may be so low that is was not detectable in our analysis; similarly, for THC and CBN in the fiber strain. It is unknown where the cannabinoids go during the aging process, but we suggest that it is possible they volatilize into the atmosphere along with the terpenes in glands, as noted later in this report. Nevertheless, this phenomenon of altered content in glands during aging is one that should be studied to gain a more complete understanding of the secretory process of cannabinoids in the cell.
http://www.hempreport.com/issues/17/graphics/mt5.gif Table 2. (Above) Cannabinoid content of capitate-stalked glands of different ages.
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Cannabinoids and THC Part 2
c. Content of secretory cavity
The gland consists of disc cells with their cytoplasm and a non-cellular intrawall secretory cavity. We examined the contents of this secretory cavity specifically to determine if cannabinoids occur in it. With microcapillary pipets we removed contents from only the secretory cavity without damaging the disc cells. Data show cannabinoids to be abundant in this cavity (Table 3). Each gland contained an average of 61 nanograms of cannabinoids. This strain was a hemp form with a CBD content. In these analyses each sample included the contents of approximately 100 or more glands. Some samples represent bracts of different lengths, ranging from 4 to 9 mm, the latter being the mature size. Those bract samples of small size, 4 mm., showed a range of variation in cannabinoid content per gland, but some showed the highest content (samples 12 and 16) while others showed a low content (sample 7). Similarly, samples from larger bracts showed relatively high content (sample 6) as well as low content (samples 1 and 2). Some samples also showed different ratios for CBD to THC even when total cannabinoid content was high (compare samples 12 and 16).
http://www.hempreport.com/issues/17/graphics/mt6.gif Table 3. Cannabinoid contents in secretory cavities of capitate-stalked glands of a hemp strain showing average cannabinoid content in a gland.
Although these difference perhaps should be expected in biological samples, it also emphasizes that the cellular component(s) which synthesize cannabinoids may not synthesize them in a constant ratio during development of an organ.
Summary:
a) Capitate stalked glands contained more THC (and total cannabinoids) than capitate sessile glands.
b) THC, and total cannabinoid, quantity in both gland types can vary during the year, and can decrease to very low levels.
c) Cannabinoids decrease with aging of glands.
d) Cannabinoids occur in the secretory cavity of the gland.
Location of cannabinoids in the gland
a. Development of the gland.
For both types of glands an epidermal cell enlarges and divides several times to form a tier of disc cells on a short stipe on the epidermis of the leaf or bract (Diagr. 2).
http://www.hempreport.com/issues/17/graphics/mt2.gif Diagram 2. (Above). Representation of mature secretory gland. Disc cells, attached to leaf or bract by stipe cells and basal cells (below stipe), release fibrillar wall matrix into secretory cavity where it contributes to thickening of subcuticular wall (wall) during enlargement of secretory cavity. Plastids (P) in disc cells produce secretions which accumulate outside plasma membrane, pass through cell wall as hyaline areas to form secretory vesicles (L) in secretory cavity. Vesicles in contact with subcuticular wall release contents to contribute to growth of cuticle during enlargement of secretory cavity. THC occurs in walls, fibrillar matrix and other contents surrounding the vesicles, but not in the vesicles; little THC is present in the disc cells. Nucleus = black; vacuole = V; vesicles = L; plastids = P; endoplasmic reticulum = ER.
The outer wall of the disc cells splits tangentially to initiate an intrawall cavity across the top of the entire gland surface. This cavity enlarges as secretions are accumulated in it (Fig. 3).
http://www.hempreport.com/issues/17/graphics/m3.gif Figure 3. Section where secretory cavity joins disc cell showing cuticle and subcuticular wall, vesicles in secretory cavity, secretions in disc cell just below the wall of the disc cell separating it from the secretory cavity. Bar = 0.5 µm.
The outer portion of the wall remains associated with the cuticle to form the subcuticular wall; the inner portion remains associated with the disc cells. Both the cuticle and subcuticular wall increase in thickness as the secretory cavity enlarges and, therefore, precursors for their growth must be present in the secretory cavity. Secretions as vesicles are evident in the secretory cavity. Note the vesicles are similar in density (grayness) to gray secretions in the disc cell. The mechanism that controls the thickening of cuticle and subcuticular wall are as yet unknown.
b. Secretion role of disc cells
It is pertinent to examine the organization of the disc cells because all contents in this cavity must be derived from the disc cells. Cannabinoids, or their precursors, are secretions from these cells. Another major group of secreted compounds are the terpenes (monoterpenes and sesquiterpenes). Monoterpenes are the more abundant of the two. Terpenes compose the "essential oils"; they contribute to the odors of the plant, and are sticky in character, as evident when one touches the plant. Different combinations of terpenes in different strains contribute to odor differences among the strains. Cannabinoids, and THC, are odorless to most humans.
http://www.hempreport.com/issues/17/graphics/m4.gif Figure 4. Portion of gland showing several disc cells each with numerous lipoplasts. Portion of secretory cavity is evident. Fibrillar matrix has separated from the wall (arrow). Bar = 0.5µm.
The tier of disc cells contains a typical cell complement including a large nucleus, plastids, mitochondria, endoplastic reticulum and abundant ribosomes, as well as vacuoles (Diagr. 2; Fig. 4). Plastids, however, represent the unique component of these cells. They are interpreted to be a source, perhaps the principal source, of secretions in the cavity. Plastids divide repeatedly and become very numerous in the disc cells. Plastids form a unusual central component, termed the reticulate body, derived from thylakoids (Figs. 4, 5). This body consists of thylakoids fused into a tubular array of light and dark areas in a hexagonal arrangement. This large body is somewhat like a prolamellar body that forms in chloroplasts when plants are grown in the dark because it has a lattice configuration similar to a prolamellar body. But it is unlike a prolamellar body in that it persists in both the light and the dark, and it does not contribute to formation of grana membranes as does the prolamellar body.
http://www.hempreport.com/issues/17/graphics/m5.gif Figure 5. Lipoplast showing presence of secretion on its surface and region where secretion joins the reticulate body (arrow). Bar = 0.2 µm.
The reticulate body was associated with secretory activity. Enlarged quantities of secretions accumulated on the surface of the plastids, and were continuous with the light zone in the reticulate body (Fig. 5). The association of the secreted mass with the light zone on the surface of the plastid supported an interpretation that the reticulate body contributes to the synthesis of these secretions (Fig. 5, arrow). Such secretions were evident on nearly all plastids and may become so voluminous as to surround a plastid, as it appears in an electron micrograph. These secretions are more or less round in appearance in these sections, but undoubtedly spherical in three-dimensions; since they are oily in composition they form spherical masses in the aqueous medium of the cell. These secretions are interpreted to be terpenes; plastids in other plants are reported to produce terpenes.
http://www.hempreport.com/issues/17/graphics/m6.gif Figure 6. Lipoplast near plasma membrane (arrow). Portion of secretion in contact with and passing through the plasma membrane (arrow). It will accumulate between the membrane and cell wall. Bar = 0.1 µm.
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Cannabinoids and THC Part 3
The secretions on the surface of plastids can be observed in contact with the plasma membrane (Figs. 5, 6). Similarly, other quantities of secretions of similar density, but not in contact with a plastid in a thin section, were observed in contact with the plasma membrane (Figs. 3, 7).
http://www.hempreport.com/issues/17/graphics/m7.gif Figure 7. Secretions (small light areas in wall) emerging as a vesicle (V) in the secretory cavity (T). Fibrillar matrix (arrow) appears to be released from the wall into the secretory cavity. In disc cell, secretions are evident in the cytoplasm adjacent to the plasma membrane and on the surface of lipoplasts. Bar = 0.2 µm.
Secretions in the secretory cavity
Secretions somehow passed through the plasma membrane and accumulated in the space between the plasma membrane and cell wall (Fig. 6, arrow and S). Secretions subsequently passed through the cell wall, often appearing as small light areas in the wall (Figs. 3, 5, 6).
Secretions emerged into the secretory cavity as small accumulations on the wall surface facing the cavity (Figs. 3). Small gray vesicles, partly embedded in the wall, at W and to the lower left of the large vesicle (V) near the "corner" of the secretory cavity. They have the same gray density as secretions in disc cell (Fig. 6). It appeared that the material passing through the wall accumulated in enlarged vesicles on the wall surface facing the secretory cavity. As vesicles emerge from the cavity they became surrounded with a surface feature about Ω the thickness of a typical membrane (Figs. 3, 7).
Vesicles are released from the wall surface to aggregate in the secretory cavity (Fig. 8). These vesicles are transported to the subcuticular wall surface where some of them released their contents into the wall (Fig. 8, curved arrow). The contents also moved through the subcuticular wall to the cuticle where the contents aided thickening of the cuticle (Figs. 3, 8). The irregular contour of the inner surface of the cuticle with dark fiber-like extensions in the cuticle, is derived from the fusion of quantities of vesicular material along with its surrounding surface feature.
http://www.hempreport.com/issues/17/graphics/m8.gif Figure 8. Outer sheath of secretory cavity consists of cuticle and subcuticular wall. Vesicles (light areas) are evident in the subcuticular wall (curved arrow). Vesicles and fibrillar matrix are evident in the cavity. Bar = 0.1 µm.
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Cannabinoids and THC Part 4
Fibrillar matrix is released from the surface of the wall into the secretory cavity. This matrix is evident at this wall as well as in the secretory cavity (Figs. 4 arrow, 7 arrow). The fibrillar matrix is transported to the subcuticular wall where it becomes incorporated thereby contributing to the thickening of this wall. The mechanisms controlling deposition of fibrillar matrix in the subcuticular wall, as well as the deposition of vesicular materials into the subcuticular wall and cuticle, remain to be studied. Since the sites of deposition are very distant from the origin of these materials, we speculate that the control mechanism somehow resides in the non-cellular secretory cavity.
Summary:
a) Plastids produce a major quantity of secretions that are released from their surface to pass through the plasma membrane and outer wall into the secretory cavity.
b) Secretions accumulate in the secretory cavity as secretory vesicles whose contents contribute to thickening of the cuticle.
c) Volatile components of secretory vesicles may volatilize into atmosphere and contribute to plant odor.
d) Fibrillar matrix and other material released from the wall surface surround the secretory vesicles with a surface feature of yet unknown composition.
Fibrillar matrix contributes to thickening of the subcuticular wall.
Localization of THC in the gland
Although cannabinoids were detected in the contents isolated from the secretory cavity, we do not yet know whether they occur in the disc cells. Further, we do not yet know where they are in the secretory cavity: in or around the vesicles, or elsewhere in the cavity.
For this phase of study fresh glands were frozen by high pressure cryofixation and then fixed (killed) by cryosubstitution to prevent movement of THC in the gland during the fixation process. We prepared a monoclonal antibody for THC as a probe by attaching gold particles to it so that it would be visible under the electron microscope. Then thin sections of glands were treated with the antibody probe; the antibody will attach to any THC in the tissue. Under the electron microscope we will see the electron dense gold particles, as dense black dots, where the antibody has attached to THC.
Upon examining the tissues we find that the THC, as indicated by the location of gold particles, is present in the cell wall facing the secretory cavity (W), and in the subcuticular wall (large arrowhead) under the cuticle (Fig. 9). It is also present in the fibrillar matrix being released into the cavity from the disc cell wall (open arrow). It was along the surface feature (small arrowhead) surrounding the large secretory vesicles in the cavity. It was also in the cuticle (arrow).
http://www.hempreport.com/issues/17/graphics/m910.gif Figure 9. (Above) Sites of THC accumulation are evident as black dots representing the gold attached to the THC antibody. THC is present in the disc cell wall (W), subcuticular wall (large arrowhead), along the surface features around vesicles in the cavity (small arrowhead), in fibrillar matrix being released from the disc cell wall (open arrow) and in the cuticle (arrow). No particles were evident outside the gland. Bar = 0.2 µm. Figure 10. THC is abundant along the surface features (arrow) of the numerous vesicles in the secretory cavity, but absent from the content in the vesicles. Where the surface feature of a vesicle is sectioned in surface view, THC appears over the entire surface. Bar = 0.1 µm.
Deep within the cavity we note, again, that THC was associated with the surface feature around the numerous secretory vesicles, but it was not inside the vesicles (Fig. 10 arrow). Some vesicles were cut with their surface feature in planar view (slightly gray appearance), and gold grains are associated with the area of the surface feature. The contents of these vesicles (clear area), presumably, are the monoterpenes (lipoidal materials).
Surprisingly, it is not in the cytoplasm of the disc cells (Fig. 9). We detected it only along the plasma membrane and in the wall proper of the disc cell. Only an occasional grain was detected in the cytoplasm, either over a plastid, or mitochondrion or among ribosomes. If cannabinoids are synthesized in the cytoplasm of disc cells, there should be abundant gold particles at sites of THC synthesis and accumulation.
In other cells, as epidermal cells, THC was present in the wall, but in smaller quantities than in walls of the disc cell. Few gold particles were in the cytoplasm or vacuoles of other cells.
Controls included sections treated with antibody alone, or treated with protein A-gold alone. Controls showed no antibody in the disc cell wall, around vesicles, in the subcuticular wall or in cuticle. No antibody was detected outside of the cells.
Possible site of cannabinoid synthesis
Our studies contribute several pieces to the puzzle on where cannabinoids are synthesized, yet we lack definitive information on the precise site of their formation. Data from the antibody probe showing that they are not evident in the cytoplasm of the disc cells suggest that they may be formed at or outside the plasma membrane surface. A working model for continued study on their synthesis is embodied in diagram 3.
http://www.hempreport.com/issues/17/graphics/mt3.gif Diagram 3. (Left) Representation of gland illustrating the possible process in cannabinoid localization in secretory cavity. A phenol glucoside is transported into a disc cell and stored as free phenol in the vacuole. Terpene is synthesized by the specialized plastid, lipoplast, in disc cells. These precursors, terpene and phenol, react to form cannabinoids at the plasma membrane surface or in the wall whereupon they appear in the secretory cavity.
As described in the cannabinoid pathway, these dimeric compounds consist of terpene and phenolic components. The abundant secretory activity of the disc cell plastids, and knowledge that this organelle does synthesize terpenes, suggests that they contribute the terpene component. Our detection, in previous studies, of abundant phenol in whole glands, and knowledge that phenols accumulate in vacuoles of cells, suggests that this cell feature may contribute the phenol component. Phenols are transported in the plant as glycosides and, when becoming localized in a cell vacuole, they accumulate there upon dissociation of the sugar moiety which returns to the cell cytoplasm.
We hypothesize that terpenes and phenols, when released from their respective sources, accumulate at the plasma membrane and cell wall interphase where enzymes dimerize these compounds into cannabinoids. It is necessary to determine enzymes involved in cannabinoid synthesis. Such an enzyme, when available, can be prepared as an antibody probe that can be used to identify more precisely the locus of cannabinoid, and THC, synthesis. Glands represent unique structures, and can be utilized to broaden our understanding of cannabinoid synthesis and aid in our effort to reduce the cannabinoid content of Cannabis strains for production of industrial hemp.
CONCLUSIONS
1. THC accumulated in abundance in the secretory cavity where it was associated with the: a) cell walls, b) surface feature of secretory vesicles, c) fibrillar material released from disc cell wall, and d) cuticle. It was not associated with the content of the secretory vesicles. The association of THC with structural components, particularly the wall, fibrillar matrix and surface feature of vesicles, suggests that it may be chemically bound to them rather than being free in the cavity. If THC and other cannabinoids are bound to components in the cavity, their presence and movement may require a source of energy in the cavity. Additional studies are necessary to determine their bound or free status.
2. Little or no THC was detected in the cytoplasm of the disc cells. This suggests that the terpene and phenol precursors, which must occur in the disc cells at some interval, may form the cannabinoids at the surface of the plasma membrane, or in the cell wall facing the secretory cavity.
3. Some THC was present in the cell walls of other cells. Genes for cannabinoid synthesis are present in all cells of the plant, but tissues other than glands produce low levels of these compounds.
4. Reduction or elimination of glands by mutation procedures will reduce significantly the quantity of THC in the plant. However, the pathway for cannabinoid synthesis is controlled genetically: glands are specialized to synthesize high levels of cannabinoids. Thus, a glandless plant can be expected to synthesize very low levels of cannabinoids. We do not know the roles of cannabinoids in the glands. They may be involved in some way with "protection", or other role. The absence of glands may or may not alter the functional role. Since other cells also synthesize these compounds, at very low levels, the quantity may be sufficient to perform the functional role. Therefore, a glandless mutant(s) would serve our purpose to reduce the THC concentration for utilization of such strains in the hemp industry.
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An elaboration on the phytochemical process that makes cannabis THC Part 1
An elaboration on the phytochemical process that makes cannabis THC
The resin exuded by the glandular trichome forms a sphere that encases the head cells.
When the resin spheres are separated from the dried plant material by electrostatic attraction and placed on a microscope slide illuminated with a 100W incandescent bulb, they appear very dark when observed through a 300X microscope. Since orange, red, and infrared are the component wavelengths of incandescent light, and since the absorption of light makes an object dark or opaque to the frequency of the incoming wave, one can conclude that these wavelengths are probably not directly involved in energizing the cannabinoid pathway.
However, the resin sphere is transparent to ultraviolet radiation.
The author found through trial and error that only one glandular
trichome exhibits the phytochemical process that will produce the amount of THC associated with pain relief, appetite stimulation and anti-nausea; euphoria and hallucinations are side-effects, however. This trichome is triggered into growth by either of the two ways that the floral bract is turned into fruit.
Of all the ways that optics are involved in the phytochemical production of THC, the most interesting has to be how the head cells and cannabinoid molecules are tremendously magnified by the resin sphere. These and other facts are curiously absent from the literature. The footnotes update the literature to include electrostatic separation of the resin sphere from the dried plant material and cannabis parthenocarpy.
(1) "For all spheres, a ray drawn perpendicular to the sphere's surface will intersect the center of the sphere, no matter what spot on the surface is picked, and the magnifying power(a) of a glass sphere is greater the smaller its size. A sphere of glass can also bring light that is heading to a focus behind it to a point within it, with freedom from two aberrations, spherial aberration and coma, but not from chromatic aberration. Chromatic aberration results when different wavelengths are focused on different planes and is the most difficult of the aberrations to correct. The human eye lens also exhibits chromatic aberration, but a yellow pigment(b) called the macula lutea in the fovea, an area at the rear of the eyeball, corrects this problem by the way it absorbs blue light."
(a)"The formula to calculate the magnifying power of a sphere is l=333/d, where l is the magnifying power and d is the diameter of the sphere expressed in mm."
(b)Interestingly, the resin exuded by drug-type flowering female cannabis plants has a yellow tint. Could this pigment work to correct chromatic aberration in the resin sphere like the macula lutea does in the fovea for the eyeball?
(2) Quoting from the Mahlberg and Kim study of hemp(a) "THC accumulated in abundance in the secretory cavity where it was associated with the following: cell walls, surface feature of secretory vesicles, fibrillar material released from disc cell wall, and cuticle. It was not associated with the content of the secretory vesicles."
The resin spheres contain the THC. It is not contained in the leaf or floral bract. After the resin spheres are dissolved in solvent or dislodged by electrostatic attraction, and a microscopic examination of the leaf or floral bract has revealed that only the glandular trichomes' stalks remain, no effect will be felt after smoking the dried plant material from which the resin spheres have been removed.
(3) The electrostatic collection of the resin spheres from dried cannabis plants with plenty of ripe seeds has been for hundreds of years the method indigenous people of North Africa and Lebanon have used to make hashish. Obtain a round metal can 8" or so in diameter x 3" or so in depth (the kind that cookies come in) with a smooth lid. Obtain 2 ounces of dried cannabis with plenty of ripe seeds in the tops. To remove the seeds and stems, sift the cannabis tops through a 10-hole-to-the-inch wire kitchen strainer into the can. Close the can with the lid and vigorously shake the closed can three or four times. This gives the resin spheres an excess negative charge. Let the can sit for a moment and then remove the lid. Opposites attract. The negative-charged resin spheres have been attracted to the metal surface of the can and lid which has a positive charge. Take a matchbook cover or credit card and draw the edge across the surface of the lid. Note the collected powder. Observed under 300X magnification, the collected powder from this "shake" is composed of resin spheres with an occasional non-glandular trichome. As the cannabis is shaken again and again, and more of the yellow resin spheres are removed from the plant material, the collected powder gradually becomes green-colored as the number of non-glandular trichomes increases in the collected powder. The greener the powder, the less the effect.
(4) "Cannabinoids represent a dimer consisting of a terpene and a phenol component. Cannabigerol (CBG) is the first component of the pathway. It undergoes chemical change to form either cannabichromene (CBC), or cannabidiol (CBD). Delta 9-tetrahydrocannabinol (THC) is derived from CBD."
(5) "Pate (1983) indicated that in areas of high ultraviolet radiation exposure, the UVB (280-320 nm) absorption properties of THC may have conferred an evolutionary advantage to Cannabis capable of greater production of this compound from biogenetic precursor CBD. The extent to which this production is also influenced by environmental UVB has also been experimentally determined by Lydon et al. (1987)."
The writer's own experience allow for a more specific conclusion: If the UVB photon is missing from the light stream(a), or the intensity as expressed in �W/cm2 falls below a certain level(b), the phytochemical process will not be completely energized with only UVA photons which are more penetrating but less energetic, and the harvested resin spheres will have mostly precursor compounds and not fully realized THC(c).
(a)Examples of an environment where the UVB photon would be missing from the light stream include all indoor cultivation illuminated by HID bulbs and in glass or corrugated fiberglass covered greenhouses.
(b)"The maximum UVB irradiance near the equator (solar elevation angle less than 25 deg.) under clear, sunny skies is about 250 �W/cm2. It was observed that the daily solar UVB in Riyadh, Saudi Arabia (N24.4Lat.) decreased from September to December by about 40% (Hannan et al. 1984). The further a person is from the tropics, the less UVB radiation there is: the average annual exposure of a person living in Hawaii is approximately four times that of someone living in northern Europe." Below are some UVB readings taken in Hoyleton, Illinois, on a clear sunny day in June by David Krughoff as reported in Reptile Lighting 2000.
7am: 12 microwatts/cm2
8am: 74 microwatts/cm2
9am: 142 microwatts/cm2
10am: 192 microwatts/cm2
11am: 233 microwatts/cm2
12pm: 256 microwatts/cm2
1pm: 269 microwatts/cm2
2pm: 262 microwatts/cm2
3pm: 239 microwatts/cm2
4pm: 187 microwatts/cm2
5pm: 131 microwatts/cm2
6pm: 61 microwatts/cm2
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An elaboration on the phytochemical process that makes cannabis THC Part 2
(c)Cannabinoid pathway: Anywhere in this pathway UVB does a better job than UVA in energizing a phytochemical reaction that will produce more fully realized THC because "all cannabinolic compounds show an absorption maximum between 270 and 280 nm in the ultraviolet region."
(6) Capitate-stalked glandular trichome.
(7) #1: The ovum has been fertilized and there is a seed developing: In the areas of the Northern Hemisphere where indigenous people have grown heterozygous drug-type cannabis for hundreds of years, pollination is used to trigger the growth of the capitate-stalked glandular trichome on the floral bract and concomitant leaves of the flowering females before the autumnal equinox(a) so the majority of seeds will be ripe(b) before November.
(7) #2: The floral bract has become parthenocarpic: Parthenocarpic fruits develop without fertilization and have no seeds. Except for transmutation and turning lead into gold, there has been more nonsense written about seedless cannabis than on any other subject. In cannabisparthenocarpy, the floral bract (the fruit) enlarges in size as though there were a seed growing inside, and the capitate-stalked glandular trichome is triggered into growth on the floral bract and concomitant leaves. "Most popular supermarket tomatoes are parthenocarpic which was induced artificially by the application of dilute hormone sprays (such as auxins) to the flowers." In a trial, cannabis parthenocarpy was not induced by the application of the spray used on tomatoes. Only the photoperiod(c) will trigger parthenocarpy in flowering female cannabis plants. Cannabis parthenocarpy occurring before the autumnal equinox is considered by the author to be "long-day" and cannabis parthenocarpy occurring after the autumnal equinox to be "short-day".
The longest photoperiod that will trigger parthenocarpy in unfertilized flowering homozygous(d) Indica female cannabis plants is 13:00 hours, give or take 15 minutes. This effect can be obtained in the month of August at N35Lat, and because the capitate-stalked glandular trichomes received plenty of UVB during this month at this latitude, the harvested resin spheres had fully realized THC. Rating: euphoria and hallucinations, major appetite boost and pain relief, deep dreamless sleep. These plants seldom grow taller than four feet but potency makes up for the reduced harvest.
The gene pool is heterozygous if a flowering female cannabis plant is not parthenocarpic by the end of the first week in September in the Northern Hemisphere. If this is the case, pollination is used instead of parthenocarpy to trigger the growth of the capitate-stalked glandular trichome before the autumnal equinox to obtain as much fully realized THC as possible in the harvested resin spheres by the time the majority of the seeds are ripe.
The longest photoperiod that will trigger parthenocarpy in unfertilized flowering heterozygous female cannabis plants is 11:00 hours, give or take 15 minutes: This effect can be obtained in the month of November at N35Lat. Because of the low intensity of UVB radiation at this latitude at sea level during November, the harvested resin spheres evidenced only slightly more THC than precursor compounds. Rating: mild to medium euphoria, appetite boost and pain relief, good snooze.
Thai cannabis falls into this 11:00 hour category, and its parthenocarpy is characterized by an inflorescence in which many floral bracts are attached to an elongated meristem. It is these elongated meristems that are harvested to become a THAI STICK. On the other side of the world, Mexican cannabis grown around the same latitudes (Michoacan, Guerrero, Oaxaca) also falls into this short-day parthenocarpic category and the unfertilized cannabis will become "sensimilla" in the 11:00 hour photoperiod which begins in mid-December in that region. The winter sunshine in those latitudes has enough UVB intensity to produce fully realized THC--unlike the winter sunshine at N35Lat.
All unfertilized flowering female cannabis plants will become parthenocarpic in a 9:00 hour photoperiod (15:00 hour dark period): This can be obtained in the month of December at N35Lat. At this latitude in this month there is not even enough UVB in sunlight for precursor vitamin D3 to develop in human skin. The phytochemical process will not produce fully realized THC when UVB falls below a certain level of intensity expressed in �W/cm2. Rating: no effect.
(a)In the Northern Hemisphere above the Tropic of Cancer, the key to all cannabis potency is this: The more days of sunlight the capitate-stalked glandular trichomes' resin spheres accumulate before the autumnal equinox the more fully realized THC.
(b)It is recognized in the indigenous world that drug-type cannabis with a majority of ripe seeds will produce more euphoria, hallucinations, appetite stimulation, pain relief, and sleep aid than with a majority of unripe seeds.
(c)The photoperiodic response is controlled by phytochrome. "Phytochrome is a blue pigment in the leaves and seeds of plants and is found in 2 forms. One form is a blue form(Pfr), which absorbs red light, and the other is a blue-green form(Pr) that absorbs far-red light. Solar energy has 10X more red (660nm) than far-red (730nm) light causing the accumulation of Pfr." The first and last hour of a day's sunlight is mostly red light because of the scattering effect on blue light. "So at the onset of the dark period much of the phytochrome is in the Pfr form. However, Pfr is unstable and returns to phytochrome Pr in the dark." The red light in sunrise returns the Pr to the Pfr form. "Phytochrome Pfr is the active form and controls flowering and germination. It inhibits flowering of short-day plants (the long night period is required for the conversion of Pfr to Pr) and promotes flowering of long day plants."
(d)In Nepal and nearby areas of India where the capitate-stalked glandular trichome is triggered into growth by parthenocarpy rather than by fertilized ovum, great care is taken to make sure that all male cannabis plants are destroyed as soon as they reveal their sex. This is because unfertilized Indica flowering females can have both stigma and anther protruding from the floral bract. In the Indica gene pool, female-produced pollen carries an allele for long-day parthenocarpy, and seeds resulting from this female-produced pollen will produce another generation of female plants that will also exhibit long-day parthenocarpy during flowering. But if pollen from male plants is introduced into this gene pool, the resulting seeds will produce a generation of females that will exhibit short-day parthenocarpy instead. The allele for long-day parthenocarpy in the female-produced pollen is carried into the gene pool by self-pollination and cross-pollination, and perhaps homozygous is used too loosely here to describe the genetic result.
(8) It appears that the resin sphere acts as an UVB receptor and magnifying lens. The latter apparently lets it gather in a lot more photons than would otherwise be possible; because a lens also acts as a prism, the resin sphere may prevent some wavelengths from being focused where the phytochemical processes are taking place because they could interfere with the phytochemical process that makes THC.
electrostatic collection of resin spheres and non-glandular trichome
