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Trichomes of Cannabis sativa L. (Cannabaceae)

Article  in  American Journal of Botany · May 1976

DOI: 10.2307/2441821

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Amer. J. Bot. 63(5): 578-591. 1976.

TRICHOMES OF CANNABIS SATIVA L. (CANNABACEAE)'

P. Da,vlNnNolN AND Pernn B. KLunvrlN

Department of Botany, University of Michigan, Ann Arbor 48104

ABSTR A C T
The diversity of non-glandular and glandular hairs of Cannabis sativa I:. (marihuana) are
described by scanning electron microscopy. The non-glandular hairs are of two major types,
as distinguished by size differences and locations, and all of them are highly silicified. The
presence of silica as well as cystoliths of calcium carbonate help in the identification of mari-
huana even in its ash residues. X-ray microanalyses of Cannabis hairs are compared with
those of Humulus lupulus and Lantana camera, whose hairs have been considered to resemble
those of marihuana. Glandular hairs are found to be of two major categories, One group con-
sists of glands whose heads are generally made up of eight cells and the other group whose
heads are generally made up of two cells but never more than four cells. All glands of both
categories are stalked. Some glands of the first category are massively stalked and these are
restricted solely to anthers and bracts of staminate and pistillate plants. The massive stalk is
considered to be made up of epidermal and hypodermal cells that have grown in response to
some stimulation during anthesis. Fine details of the shoot system of Cannabis, such as cuticu-
lar ridges on epidermal cells, warty protuberances on non-glandular hairs, and surface views
of glands in developing stages are also reported. Glandular hairs on the bracts of Humulus
Iupulus resemblethose of Cannabis.

Mosr or rHE AERIALparts of Cannabissativa L.
(the Indian hemp plant or marihuana) are covered
with different types of non-glandular and glan-
dular trichomes. In spite of its importance as an
economic plant of antiquity, the bbtany of Can-
nabis is still poorly understood (Schultes, 1970;
Schulteset al., 1974). This applies particularly
to the nature of glandular hairs which are con-
sideredto be the primary locations of the narcotic
cannabinoids.Briosi and Tognini (1894, 1897)
presenteda detailed description and illustrations
of the various trichomes of Cannabis and also
summarizedprevious works on Cannabis sative.
BesidesMohan Ram and Nath's U964) cursory
referenceto the ontogenyof the trichomes, most
'Received for publication 7 April 1975.
The authors wish to thank Dr. Erich Steiner, Direc-
tor, University of Michigan Matthaei Botanical Gardens
for the supply of specimens,and Dr. W. C. Bigelow and
his staff for use of the scanning electron microscope
facilities. We also thank Diane Hoefle, Frederick He-
bard, David Bay, and William Stein for their encourage-
ment and assistance. This work was supported in part by
NSF grant BMS 75-16359.
workers have describedsome aspectsof the ma-
ture trichomes only (Ballard and Phar, 1915;
Bouquet,1950; Shimomuraet al., 1967; Jackson
and Snowdon,1968). Recently,the needfor the
identification of marihuana in illicit trade and
possessionhas resulted in several publications,
forensicin nature, which mostlv describethe non-
glandular cystolith hairs. Theie studieshave em-
ployedeitherthe light microscope(Asahinaet al.,
1967; Shimomuraet al., 1967; Nakamura,1969;
Nordal, 1970; Thornton and Nakamura, 1972;
De Forest,Morton and Henderson.1974) or the
scanning electron microscope (Bradford and
Devaney, 1970; Devaney and Bradford, 1971;
Siegesmundand Hunter, 1971; Mitosinka, Thorn-
ton and Hayes, 1972). Such studieson the iden-
tification of marihuana have been carried out on
resin, dried leavesor ash residuesof Cannabisas
well as on the dried leavesof nearly 90 different
plants that are said to possesscystolith hairs thdt
could be confusedwith those of Cannabis.
Hammond and Mahlberg (1973) made use of
the scanningelectronmicroscopeto describethree

Fig. l-7. Surface features of regions of shoot system of Cannabis sativa. All figures obtained from fresh, live
specimens without any treatment. l. A lobe of a young leaflet with reduced stomata near the tip, and several non-
glandular trichomes. X 265, 2. Adaxial surface of a young leaf with cystolith trichomes of two sizes and a few
bulbous glands. Some trichome initials are seen in the groove. y 260. 3. Mature region of a leaflet tip showing
cuticular ridges and reduced stomata. X 265. 4. Surface of petiole showing non-glandular hairs, several capitate
glands and a few bulbous glands. X 85. 5. Abaxial leaf surface with trichomes and stomata. Hairs are devoid of
warty projections. Three capitate glands are seen to the left and two bulbous glands to the right. X 250. 6.
Adaxial leaf surface. Cuticular ridges on the epidermis are yet to develop. X 250, 7. Adaxial leaf surface show-
ing a capitate gland, long and short cystolith trichomes, and a ring of cells surrounding the long hair. X 250.
578
May*June, 19761 DAYANANDAN AND KAUFMAN-TRICHOMES OF CANNABIS 579

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 OF BOTANY
580 AMERICAN JOURNAL
[Vol. 63
May-June, 19761 DAYANANDAN AND KAUFMAN-TRICHOMES OF CANNABIS 581

types of glandularhairs that occur on the pistillate
plants, especiallyon the flowering bracts. In this
paper, we extend this study to the shoots of sta-
minate plants which bear as many different types
of glandular hairs as the pistillate plants. Using
an X-ray detecting systemthat is coupled to the
scanningelectronmicroscope,we also analysethe
pattern of silicification and calcificationin the non-
glandular hairs of Cannabissativa, Humulus lu-
pulus (hops, the only other genus in Cannaba-
ceae) and Lantana camera (Verbenaceae).X-ray
microanalysisdata on the above plants, as well as
on burnt ash residuesof marihuana, provide in-
formation of taxonomicvalue that make identifica-
tion of marihuanamore definitive. Becauseof the
variation that exists amongthe trichomes of Cqn-
nabis, their study should lead to a better under-
standingof the real taxonomic relationsthat exist
betweenwidely different forms of Cannabis that
have usually been grouped under one species,C.
sativa (Schulteset al., 1.974). Marihuana contains
about 20 different cannabinoids,including tetra-
hydrocannabinol,the major hallucinogeniccom-
ponent. The study of Cannnbisglandsis also es-
sential to understandtho biogenesis,distribution
and function of the different cannabinoids.
Mlrsnrars ANDMErHoos-A pistillate and a
staminate plant of Cunnabis which have been
growing in the University of Michigan Matthaei
Botanical Gardenswere used for most of the in-
vestigation. A few seedlingsfrom the Botanical
Gardenswere also employJd to stucly the differ-
entiation of trichomes during early stages of
growth of Cannabis. The plants employedfit the
usual description of typical Cannabis sativa
(Schulteset al., 1974) .
Most of the scanning electron microscopic
(SEM) studieswere done with fresh specimens.
Small segmentswere mounted on alumirium stubs
with carbon paint and examined imrrediately in
the SEM (JEOL model JSM-U3); At an ac-
celerating voltage of 15 KeV, fresh specimens
could be examinedfor about 20 min without muclt
damageto them. X-ray microanalyseswere also
done mostly on fresh specimensby coupling to
the SEM an X-ray detector and amplifier system
(KEVEX) and a multichannel analyser (North-
ern Scientific,Model 710). Setected'areasof the
specimenwere first photographedin the second-
ary emission mode followed by X-ray mapping
for calcium and/or silicon. This order was nec-
essarybecause. beam or vacuum_damage to the
specimenwas increasedwith the longer exposure
times that were used for X-rav analvsis.
Samples were also subject'edto- critical-point
drying in order to compare the structures with
fresh sample preparations. Materials were fixed
in FAA, dehydratedin ethanol and critical-point
dried with liquid COz as an intermediate. Prep-
arations thus obtained were as sood as those
that employed amyl acetate insteid of ethanol.
Critical-point dried samplesof Cannabis,as well
as air dried samples of Humulus lupulus and
Lantana camera (both obtained from the Mat-
thaei Botanical Gardens) were coatedwith a thin
layer of gold before examiningthem in the SEM.
Ash residueswere obtained frorn dried leaves
and flowering tops of pistillate and staminate
plants of Cannabis. A marihuana cigarette was
attachedto an aspirator through glassand rubber
tubings and the ash falling from the lit end was
collectedon clean piecesof aluminum foil. Small
quantities of the finest ash sampleswere spread
on scotch tapes and mounted on aluminum stubs
to be gold-coatedand examinedwiththe SEM.
Fresh samples of Cannnbis leaves that show
minimal damageduring examinationwith the SEM
are better suited for X-rav microanalvsis than
dried and gold-coatedsampl-es.Howevei, in Can-
rwbis, Humulus, Lantana, and ash residues of
Cannabis,silica and calcium deposition occur in
such large quantities that no interferenceis en-
countered even with gold-coated samplesduring
qualitative X-ray analysis for these elements.
While this study focuseson SEM aspectsof the
trichomes of Cqnnqbis, conventional microtech-
nical methods (microtome sections, acid-hydro-
lysed squashpreparations)were also employed
to corroboratefindings obtainedwith the SEM.
Os snnverroNs-Non-g IanduIqr hair s-The
stems,petioles,stipules,leaf blades,bracts, and
both surfacesof the tepals are all covered with
hairs that are strictly unicellular (Fig. 1-7, 30,
31). Figure 1 depictsa young leaf which shows
stagesin the developmentof hairs. The tip of a
leafletpossesses 6-15 stomata,probablymodified
to function as hydathodes. Immediately below
the tip are well-developed and elongating tri-
chomes. Warty protuberancescharacteristic of
thesetrichomesare alreadyvisible (Fig. 1-3). At
a still lower level, hair initials are seen among
fully differentiated hairs. Such an admixture of

Fig. 8-13. Secondary and X-ray images of silicon of non-glandular hairs of Cannabis.-Fig. 8-11. Fresh, live
specimens without any treatment. B, 9. Abaxial leaf surface. Note silicon localization only on hairs and also on a
contaminant, perhaps a soil particle. X 500. lO, ll. Two kinds of trichomes from adaxial leaf surface. Silicon
is localized in both types. X 520.-Fig. 12, l3,Trichome macerated in conc. nitric and chromic acid mixture, air
dried and gold-coated. 12, 13. Isolated hair from the adaxial leaf surface. The cystolith of CaCO, has been re-
moved but the region where it was attached still remains. Also seen is a broken piece of another hair. 1 950.
582 AMERICAN JOURNAL OF BOTANY
lVol. 63

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Fig. 30-33. Fresh, live and untreated specimensof male flowers of Cannabis. 30. Central region of a male
flower showing the bases of stamens. No pistillode is seen. Glandular ancl non-glandular hairs are visible. The
lattel have warty projections. The perfect spheresare pollen grains. X 180. 31. Basal region of outer tepal sur-
face. Typical hairs and capitate glands are found, but the epidermal cells lack cuticulal ridges. A11 hairs contain
Si. X 290. 32. Surface of anther towards the interior of flower with capitate glands arranged in a row. X 100.
33. Enlarged view of a capitate gland with a long stalk. Also note the stomata and cuticular ridges on epidermis.
x 400.
May-June, 19761 DAYANANDAN AND KAUFMAN_TRICHOMES
well-developed hairs and hair initials in early
stagesof developmentare also seenon young por-
tions of stems,petiole basesand regionsof the leaf
between the petiole and the bhdd 1Fig. 2, 34).
The leaflet tips, and the small epidermalcells be-
low them, are smooth and lack the characteristic
ridgesthat developlater (Fig. 3). Even though
glandular and non-glandular hairs are found at
this stage,these regionsstill lack stomata (Fig.
1). As apicalgrowih of the leafletceases,an'dis
stomatabegin to differentiate,the epidermal cells
ellarge, and the leaflet tips and the adaxial epi-
dermal cells develop the characteristiccuticular
ridges (Fig. 3,23). This pattern is particularly
striking on the leaflettips (Fig. 3).
Two major'of. types of non-glandularhairs cover
the shoots Calnnobis.Th; shorter (about 70-
125pm) and prominent cystolith-containinghairs
are generallyrestrictedto the adaxial leaf surfaces
(Fig. 7). The longer (250-370pm) and more
profuse hairs occur on the abaxial leaf surfaces,
stems, petioles and tepals (Fig. 5, 8). All the
hairs are pointed towards the tips of the stemsor
leaves and exhibit varying degreesof warty pro-
tuberances. The cystolith hairs on the adaxial
leaf surfacesexhibit differencesin size and wart-
iness (Fig. 6,7). Among the shorter (70r,m)
ones,somenever developthe warty protuberances
but they still containcystoliths(Fig. 10).
X-ray microanalysisrevealsthat hairs from any
part of the plant possesssilica (SiOz .nHrOi.
Silica is distributed more or less uniformly all
over the surfaceof the trichome(Fig. 8-13 ) . De-
position of calcium as CaCO3is confinedto the
enlargedbasalportionsof the cystolithhairs (Fig.
l4-I7). The cystoliths are very prominent in
hairs on the adaxialleaf surfaces.Some,but not
all the hairs from the stem, petiole, and abaxial
leaf surfacesalso show cystoliths of CaCOs. Be-
causeno particular concentrationof any element
is to be found on the warty prbtuberancesof the
trichomes, the protuberancesare perhaps.due to
local depositionsof cellulose,cutin or other wall
materials.
Remainsof marihuanaash revealthe existence
of both kinds of hairs as recosnizableand distinct
entities (Fig. 14-16). The cystolithsof CaCOs
often separateout as balls. X-ray mapping of the
ash further confirms the siliceous nature of the
hairs and the presenceof Ca in the cystoliths
(Fig. 16-18). Some Ca is probably distributed
throughout the inner cavity of the hairs. But the
enormous quantities of silica found on the cell
walls of the hairs mask the small quantitiesof Ca
(Fig.17).
A study of the hairs of Humulus lupulus re-
veals that some are roughly comparablewith the
two kinds of hairsfound in Cannabis@ig. 22,28,
29,42,43). However,the followingfeaturesread-
ily distinguishCannabis from hops. The abaxial
leaf surfaceof Humulus is totally devoid of hairs
OF CANNABIS 585
exceptover the midribs and major veins (Fig. 38).
The epidermis reveals intricate cuticular ridges
which are not found on the abaxial leaf surfacein
Cannqbis(Fig.5). The stem,inflorescence stalks,
petioles and major leaf vein surfaces possess
anvil-shapedunicellular hairs in hops (Fig. 39,
40). Hairs of this shapeare neverfound inCan-
nabis. X-ray microanalysis data also bring out
important differences. The hairs on the adaxial
leaf surfacesof Humulus are poorly silicified. In
contrast,a ring of epidermalcells around the base
of each hair is highly silicified (Fig. 2a-27).
Such basal rings of silicified cells are also found
associatedwith the anvil-shapedhairs (Fig. 40,
4r).
Lantqna cqmera is said to possesshairs that
are somewhatsimilar to that of Cannabis. The
hairs of this member of Verbenaceaehave been
described along with those of 90 other plants
as being of forensic interest in the identification
of illicit marihuana (Nakamura, 1969; Thornton
and Nakamura, 1972). Viewed through the
SEM, the upper leaf surface of Lantana only
superficially resemblesthat of Canrwbis, La:n-
tana leaf epidermal cells lack the cuticular ridges
and the hairs do not show warty protuberances
(Fig. 19) . Lantana has prominentrings of basal
cells. These cells are silicified while the hair
proper is calcified (Fig. I9-2L).
Glandulqr hairs-Glandular hairs also develop
along with the non-glandularhairs. The types of
glandular hairs that cover the aerial parts of the
pistillate plants, especially those on the bracts,
have been describedby Hammond and Mahlberg
(1973). The latter authorsmadethe first attempt
to classifythese glands as bulbous, capitate-ses-
sile, and capitate-stalked. Briosi and Tognini
(1897) describedthe bulbousand capitate-sessile
glands of Hammond and Mahlberg as almost ses-
sile, and the capitate-stalkedglands as stalked.
Our studiesindicate that the diversitv of the elan-
dular hairs has never been fully understoodand
properly classified. Pistillate plants possessthe
so-calledcapitate-stalkedglands only in the flow-
ering bracts. Glands that could be described as
capitate-stalkedalso occur on the staminateplants;
these are restricted solely to longitudinal t'ows
alongthe inner surfacesof anthers(Fig. 32,33).
Briosi and Tognini (1897) describedthe stalks
of the capitate-stalkedglands of the pistillate
plants as primarily derived from epidermal and
subepidermalcells at the bases of the glands.
These glandspossessglobular heads and multi-
cellular stalks that can attain a lensth of about
200pm. Some glands have much slorter stalks
while othersappearto be almostsessile(Fig. 32).
The importance of this gradation in stalk length,
and the true nature of the multicellular stalk, are
describedbelow.
Glands with large globular heads, and appar-
entlv without stalks. have been described as
AMERICAN JOURNAL OF BOTANY
lVol. 63
May-June, 19761 DAYANANDAN AND KAUFMAN_TRICHOMES OF CANNABIS 587

capitate-sessile by Hammond and Mahlberg eral-celledheads,the smaller glandsoften possess

(1973). These glands are the most abundant
type found, and they occur in especially high
numberson the abaxial leaf surfaces,petiolesand
young stems (Fig. 4-7, 34). The developmentof
these glands from epidermal initials to the fully
mature glands with a head of eight cells occurs
rapidly during early stagesof shoot growth (Fig.
34, 56-62). These glands are not sessile. Each
gland possesses a stalk that is only one cell in
length but 2-4 cells thick (Fig. 35). In fact, the
previously described capitate-stalkedglands are
similar in all respectsto the so-called capitate-
sessileglandsexceptfor the presenceof a massive
stalk. The massivestalk, however, is actually a
product of the elongationof hypodermalcells be-
low the glands. The cells that make up the heads
of the glands of both types form a disc-shaped
unit. fn early stagesthis cellular disc is convex
and at maturity it may become either a flat or
a concave disc (Fig. 36, 58-62). The disc is
about 30r,rmin diam and about 15um in heisht.
Because bt the accumulation of resinors se6re-
tions betweenthe outer surfaceof the disc and the
extended cuticular membrane, each gland gives
the appearanceof a sphericalstructure(Fi9.34,
45, 60). Some of the FAA-fixed and critical-
point dried specimensshowedsomeglandswhose
outer membraneshad ruptured, thus revealingthe
inner cellulardisc (Fis. 36).
Small glandular trTchomes(about 15-30pm)
describedas bulbous slandular hairs are consid-
ered by Hammond anl Mahlberg (1973) to be
least complex structurally. Our findings indicate
that these smaller glands are quite variable (Fig.
46-50). Somehaveheadsmadeup of singlecells
and they have stalks that are unicellular. Other
glands have four-celled headswith stalks that are
two cells thick. The majority of the small glands
possesstwo-celledheadsand.stalksthat are one
or two cells long and one or two cells thick (Fig.
51-55). This variability was observedamong
mature glands. Unlike the capitate glandsof sev-
chloroplastsin the stalk as well as in the head
cells (Fig. 51-55). Resinoussecretionsare found
to accumulatewithin the cells of these smaller
glands. With maturity the resin collects beneath
the outer membranesof the glands,giving the ap-
pearanceof nipple or flask-shapedunits (Fig, 47,
48, 53). That theseare not artifactsinducedby
the vacuum in the SEM was evidencedby the ob-
servation of such features in fresh specimensex-
amined with a light microscope. Figure 40 shows
a region of the outer membranethat is different
from the rest of the cuticular membrane. When
young leaves are hydrolysed in 1N HCI and
squashedbetween a slide and cover glass, this
resion is detached and comes off as a contact
leis-shapeddisc (Fig. 48,52).
DrscussloN-Although the descriptionthat fol-
lows and all but one fizure in this article are con-
cerned with the stamlnate plant, we found the
same types of glandular and non-glandularhairs
on the pistillate plants also. The exceptionsin dis-
tributions are to be found only among the male
and femaleflowersthemselves.
Both glandular and non-glandulartrichomes of
Cannabiscover the shootsystemof Cqwwbis fuom
the early seedling stagesto maturity. Massively
stalked glands (capitate-stalked), however, are
restrictedto the bracts of the pistillate plants and
anthers of the staminateplants. The assumption
that resinousglandsdo not developuntil the plant
is on the point of flowering is incorrect (Bouquet,
1950). Staminateplants do not possessa re-
stricted number of glandular types as stated by
Hammondand Mahlberg (1973). Both the sexes
bear similar types of glandular and non-glandular
hairs.
Non-glandular trichomes that cover the plants
are of two major types. The predominantly cys-
tolith-containins hairs are found on the adaxial
surfaceof the liaves. Differencesin size, as well
as the existenceof warty protuberances on them,

<-
Fig. 3443.-Fig. 34-37 . Cannabis sativa. 34. Details of a region that joins the petioles and leaf blade on the ab-
axial leaf surface. Well developed trichomes of grandular and non-glandular kinds are found in various stages of de-
velopment. Two and four-celled trichomes as well as mature capitate glands are seen. Arrows point to bulbous glands.
Non-glandular hair initials are also evident. X 230. 35. A light micrograph of a squash preparation made from a
young leaf. The eight cells that make up the head are seen in the periphery. The four cells in the center constitute
the stalk and foot cells. X 860. 36. Fixation arrd critical pcint drying have ruptured the outer cuticular membrane
revealing the inner disc of head cells that secrete resin. y 1125. 37. Cross-sectional view of a critical-point
dried leaf showing the two stalk cells and the head cells of a bulbous-type gland. Fixation has stabilized the
protoplasts as solid structures. 1 2405.-Fig. 38-43. Humulus lupulus. 38. Abaxial leaf surface of Humulus
lupulus. Only large glandular trichomes are found. Epidermal cells have intricate cuticular ridges-a feature
not found on the abaxial leaf surface of Cannabis. X 155. 39. Anvil-shaped trichome on a raised pedestal on
the surface of stem. X 175. 40, 41. Secondary electron image and an X-ray image for Si. This anvil-shaped
trichome from the stem surface possessesa ring of silicified cells similar to those on the adaxial leaf surface. Tilt-
ing of the specimen has cut off the X-ray image from some parts around the trichome. X345. 42, 4" . Large
lupulin gland and several bulbous type glands found on the outer surfaces of flowering bracts. These two types of
glands recall the capitate and bulbous glands of Cannabis. X 220.
588 AMERICAN JOURNAL
Fig. 44-50. Different types of glandular hairs of
Cannabis. All figures except Fig. 46, 49 and 50 were
prepared from fresh, live specimens without any treat-
ment. Fig. 46, 49 and 50 are from critical-point dried
preparations. 44. Capitate and bulbous glands over a
vein on the abaxial leaf surface. Both glands are stalked
even though the stalk of the capitate gland is not visible
in this view. The larger size of the capitate gland is
mainly due to the accumulated resin under the cuticle.
X 340. 45. Capitate gland with a prominent pseudo-
stalk on the surface of the anther wall that faces the cen-
ter of the flower. X 240. 46. Two bulbous glands on
the adaxial leaf surface. Both have stalks made up of
two cells each but the lower one has a head made of two
cells and the upper one has a four-celled head. X 1010.
47. A bulbous gland from a young adaxial leaf surface.
The tip over the head is resin-filled cuticle. \ 1125.
48. Bulbous gland from abaxial leaf surface. The stalk
and head are made up of two cells each. The tip of the
gland possessesa small disc shaped region below which
resin accumulates in the extended membrane. X 1125.
49. Bulbous gland from petiole surface. The head is
made of a single cell, and the stalk of two cells, one
above another. X 1690. 50. Bulbous gland from
abaxial leaf sur{ace with the head and stalk each made
of two cells. X 2250.
OF BOTANY
lVol. 63
61
-axJ-
.g
t6 I-
L
I
w'
56
(6lx
w, 57
Fig. 5l-62, Various gland types of Cannabis drawn
to scale.-Fig. 51-55. Bulbous glands. 51. Gland with
one cell each for head, stalk and foot. 52. One-celled
foot, stalk with two cells one above another and head
with one cell. The disc above represents the structure
that can be detached from the tip of some glands. 53.
Similar to gland in Fig. 52 but with two-celled head.
Resin accumulated within the cell. 54. Foot has one
cell but the stalk is two cells thick. Head has two cells.
55. Foot and stalk, each has two cells and the head
is of four cells.-Fig. 56-62. Many celled, capitate
glands. 56. Initiation of capitate gland. 57. Foot with
one cell, stalk with two cells, and head in the four
cell stage. 58, 59. Lateral and surface views of fully
developed gland before secretory activity. 60. Gland
with extended membrane within which resin accumu-
lates. 61. Gland after the membrane has ruptured and
the secretory activity has declined. 62. This gland from
a female bract shows the hypodermal and subepidermal
nature of the massive stalk. The gland proper still has
the same features that are found in such slands on leaves
and stems.
help to distinguish two kinds of cystolith hairs
(Fig. 2, 6, 7, 1.0). About 15-20 epidermal cells
form a circle at the base of the cystolith hairs
(Fig. 10, 23). In Humulus, such cells are filled
with silica bodies (Fig.22,24-27). A detailed
study of the non-glandular hairs among all the
different cultivars of Cannabis misht reveal fea.
tures that may be of help in solv-ing taxonomic
relationships of this genus.
The need for the identification of marihuana
from illicit samples has led to a study of dis-
tinguishing features that remain identifiable even
in marihuana ash (De Forest et al., 1974). This
has resulted in valuable comparative studies in-
volving some 600 species of plants that are known
to possess cystolith hairs (Nakamura, 1969;
Thornton and Nakamwa. 1972: Nakamura and
Thornton, L973). Because the scanninq electron
microscope is finding increasing use in-criminal-
istics, we have furthered its use in the identifica-
tion of marihuana by coupling the SEM to an X-
ray detector analyser system (see Methods).
May-June, 19761 DAYANANDAN AND KAUFMAN_TRICHOMES OF CANNABIS 589

Where chemical and light microscopic tests fail, first group (capitate) consists of glands whose
SEM and X-ray analysismay result in definitive headsare madeup mostly of 8 cells (Fig. 35, 36,
identification of marihuana frasments even in 45, 58-62). The headis supportedby a stalk that
ash remains (Fig. 14, 18). T[e characteristic is one cell in heisht and two to four cells in thick-
silica and calcium profiles readily distinguishCcr- ness(Fig. 58, 60J. The stalk is in turn supported
nabis from Humulus and Lantana, two plants by a foot which is one or two cells wide and is
whose hairs are known to resemblethe Cqnnabis level with the epidermal surface (Fig. 35, 56-
hairs (Fig. 14-18). Differencesthat exist in cal- 62). Glands of this category seem to be stim-
cium and silica X-ray profiles between Cannabis ulated, during anthesis,to produce massivestalks
and Humulus clearly distinguish these two gen- in flowering bracts of female flowers and anthers
era of Cannabaceae.A preliminary examination of male flowers (Fig. 33, 45, 62). Thesemassive
shows that members of Moraceae such as Ficus stalks,whether on bracts of pistillate plants or on
and Morus exhibit similar distinguishingpatterns anthers of staminate plants, are hypodermal and
in silicification and calcification. X-rav micro- epidermal in origin and are best described as
analysiswith the SEM thus promisesto b-ea valu- pusedo-stalks.
able tool in plant systematics. The secondgroup of glands(bulbous) is a col-
The silicified trichomes,especiallythe cystolith lection of different types of glands (Fig. 37, 46-
hairs, resemble curved, pointed glass needles. 50, 51-55). They are all generallystalked. Un-
Marihuana is often consumed orally in several like the capitate glands, the bulbous glands pos-
kinds of preparations. Such preparationsfor oral sesschloroplastsin their stalks as well as in their
consumptionreach their greatestdiversity in India heads. The head is madeup of one, two or four
(Chopra and Chopra, 1957). Is it possiblethat cells (Fig. 37, 5I-55). The stalksof theseglands
some of the adverseeffects of oral consumption are either one or two cells in heisht and when one
of marihuana, especially on the gastrointestinal cell high, the stalk can be one 5r two cells thick
system,may be due to irritations caused by the (Fig. 46-49, 5 1-55 ) . A foot madeof one or two
siliceoustrichomes? cellslie level with the epidermisand the cells have
Glandular hairs of different kinds have been staining properties different from the ordinary
describedunder different names by the previous epidermalcells (Fig. 51-55). A distinctionbe-
workers (Briosi and Tognini, 1894, 1897; Bou- tween the stalk cell and the foot cell is not always
quet, 1950; Jacksonand Snowdon,1968; Ham- possible(Fig. 37). Someof theseglandsalsopos-
mond and Mahlberg, 1,973). Some have de- sessunique disc-shapedregions on the cell wall,
scribed only the massivelystalked capitate glands near the apex of the glands (Fig. 48, 52). The
or apparentlystalklesscapitateglands. Hammond chemical nature, formation and function of these
and Mahlbere G973) classifythe glandularhairs discsare not known.
as capitate-stalked, capitate-sessileand bulbous. The bulbous glands appear to be smaller than
Our investigationsshow that better terminology the capitate glands (Fie. aa). This is due to the
is neededtd describetheseglands. For example, fact that the cuticular membranesof the capitate
the so-called capitate-sessileglands are in fact glands extend considerably to accommodatethe
not sessile;they possessstalks that are one cell resinoussecretionsand thus appear three to four
in height and generally twg cells in thickness. times larger than the bulbous glands (Fig. 36,
Again, the so-called capitatd-stalkedglands do 44) . The actualsizesof the headsof both typesof
in fact possessonly as many true stalk cells as the glands are quite comparable (Fie. 53-62). The
so-called capitate-sessileglands. The massive bulbous glands accumulateonly small quantities
stalk or pedicel of theseglands that occur on the of resin. Judging from the accumulationof resin
bracts of the pistillate plants appearto be derived and the resulting shapes,the bulbous glandsseem
from the epidermal and hypodermal cells below to be most active in their secretinsactivities dur-
the glands (Briosi and Tognini, 1897). These ing early stagesof shoot developmlnt.
could then be only pseudo-stalks.Also, some It is generally assumed that the inebriating
authors have implied that the various types of propertiesof maiihuana is due to the secretionsoT
glands are but different expressionsof one type the glandular hairs. Direct evidencefor this is
(Bouquet, 1950; Mohan Ram and Nath, 1964). meagre (Hammond and Mahlberg, 1973). Fu-
Briosi and Tognini (1897) classifiedthe glands jita et al. (1967) presentedgas-liquidchromato-
as almost sessile and stalked and included all graphic evidencefor the presenceof tetrahydro-
glands except the ones produced on the flower- cannabinolinside the cellular discsof the capitate
ing bractsin the first category. glands. Cannabispreparationsare known to con-
We have avoided proposing new terms to de- tain about 20 different related compounds, col-
scribe the different types of glands becauseof a lectivelycalledcannabinoids(Mechoulam,l97O).
lack of detailedontogeneticstudies. Instead,we Cannabidiolic acid is said to be the precursor
use the existing terminologies rather loosely to from which cannabidiol,tetrahydrocannabinoland
describe our observation. Our studies indicate cannabinol are derived, in that order. The bio-
that there are two major groups of glands. The logically active isomers of tetrahydrocannabinols
590 AMERICAN JOURNAL OF BOTANY lVol. 63

themselvesare said to be produced only during der that it is called a "deceptive weed," "a signal
drying and storageof plant material, or during ex- of misunderstanding" and "an example of taxo-
traction, or smoking (Schultes and Hofmann, nomic neglect."
1973). Recent studies have demonstratedthat
the staminateplants possessas much and as many LITERATURE CITED
cannabinoidsper unit fresh weight as the pistil-
Acuner-r. S. 1970. Constituents of male and female
late plants(Agurell,1970;Fettermanet alr,l97I). Cannabis, p. 57-59. In C. R. B. Joyce and S. H.
This correlateswell with our finding of as many Curry [ed.], The botany and chemistry of Canna-
different types of glandular hairs ln staminate bls. Churchill, London.
plantsasin pistillate plants. AsenrNe, H., M. ONo, K. T.exa.nesnr, aNo Y. ONo.
The precise location of the various types of 1967. Identification of Cannabis resin. Bull. Natl.
cannabinoidsin the different types of glandscalls Inst. Hyg. Sci. Tokyo 85: 123-125.
for the use of sensitivehistochemicalreasentsand Ber,renn. C. W.. AND D. PHAR. 1915. Notes on the
techniques.Reagentsused in Beam'stesi, Gham- histology of an American Cannabis. J. Am. Pharm.
rawy's test, Duquenois and Negm's reaction and Assoc. 4: 1299-1303.
R. J. 1950. Cannabis. Bull. Narcotics 2(4):
its modifications should be tested for their ef- Boueuer, 14-30.
ficiency and specificity as histochemicalreagents BRADFonD,L. W., eNo J. DBveNev. 1970. Scanning
(Duquenois, 1950; Thornton and Nakamura, electron microscopy applications in criminalistics.
1972). Variations in the activity of the different J. ForensicSci. 15: 110-119.
glands during different growth stages also need Bnrosr, G., eNo F. TocNrNr. 1894. Intorno alla anat-
to be studied to understandthe pharmacological omia della canapa (Cannabis sativa L.). Parte
activities of preparations from various stagesof prima: Organi sessuali. Atti Ist. Bot. Pavia, Ser.
plant growth. 2.3: 91-209.
AND -. 1897. Intorno alla anatomia
Humulus is the onlv other senus in the family
della canapa (Cannabis sativa L.). Parte seconda:
Cannabaceae.Humilus lupilus (hops) is ot Organi vegetativi. Atti Ist. Bot. Pavia, Ser. 2. 4:
economic importance because of the lupulin 155-329.
slandsfound on the flowerins bracts that are used Cuoene, I. C., lNn R. N. Cnopne. 1957. The use of
in the brewing of beer. TheG glandssuperficially of lhe Cannabrs drugs in India. 8u11. Narcotics 9
resemblethe capitate glands of Cannabisand are (1): 4-29.
distributed not only over the flowering bracts, De Fonrsr, P. R., C. V. MonroN, eNn R. A. HsNosn-
but alsoon the abaxialleaf surfaces(Fig. 38, 43). soN. 1974. Microscopic morphology of marijuana
A striking feature in hops is the profuse occur- ash. J. Forensic Sci. 19: 372-378.
J. R., eNo L. W. Bneorono. 1971. Applica-
rence of bulbous-type glands over the flowering DnveNnv, tions of scanning electron microscopy to forensic
bracts (Fig. 42, 43). These glands recall the science at Jet Propulsion Laboratory. 1969-1970,
bulbous glands of Cqnnabisthat have four-celled pt. 2, p. 561-568. In O. Johari and I. Corvin [ed.],
heads (Fig. 46, 55). A better understandingof Scanning electron microscope symposium, Scanning
the properties of all the different cannabinoids electron microscopy proceedings. IIT Research In-
from Cqnnabis and of the various secretionsfrom stitute, Chicago.
the different glands of Humulus may throw light Dueur,Nors, P. 1950. Chemical and physiological
on the inebriating properties and popularity of identification of Indian hemp. Bull. Narcotics 2
(3): 30-33.
beer. At present,no componentof'hops is shown
FnrrennalN, P. S., D. S. Knrrn, C. W. Werun, O. Gusn-
to possessproperties similar to tetrahydrocan- nrno, N. J. DooneNsos, lr.to M. W. Qurunv. 1971.
nabinol of marihuana. Mississippi-grownCannabts sativa L.i Preliminary
The genusCannabisitself is generallyassumedto observation on chemical definition of phenotype
be monotypic (Small, 1975). Schulteser al. (1974) and variations in tetrahydrocannabinol content ver-
contend that there are three species, namely, sus age, sex, and plant part. J. Pharm. Sci. 60:
C. sativa, C. indica and C. ruderalis. Someof the 1,246-1249.
discrepanciesin the accountsof the morphology Fu.rrre, M., S. Hnoro, E. Kunrverrl, M. Snlcnruno,
and anatomy,especiallyof the trichomes,and of AND M. Aru,su. 1967. Studies on Cannabis. IL
Examination of the narcotic and its related com-
the pharmacologicalactivity of different cultivars ponents in hemps, crude drugs, and plant organs by
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View publication stats
OF CANNABIS 591
chemistry of hallucinogens. Charles C. Thomas
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