C H A P T E R

1
The Cannabis Plant: Botanical Aspects
S. Farag, O. Kayser
Technical University Dortmund, Technical Biochemistry Dortmund, Dortmund, Germany

LIST OF ABBREVIATIONS
SUMMARY POINTS
• This chapter focuses on the botanical aspects of CBD Cannabidiol
Cannabis. CBDA Cannabidiolic acid
• Cannabis trichomes can come in glandular and CBN Cannabinol
nonglandular shapes, including oil resin. GPP Geranylpyrophosphate
• Resin glands are the main producer of GRIN Germplasm Resources Information Network
cannabinoids. ISSR Inter simple sequence repeat
NPGS National Plant Germplasm System
• Recently, hybrid cannabis strains have been RAPD Random amplified polymorphic
developed. RFLP Restriction fragment length polymorphism
• Modern hydroponic techniques, coupled with RH Relative humidity
selective artificial lighting, are used in order to RFLP Restriction fragment polymorphisms
solve the issue of low-potency cannabis. THCA Tetrahydrocannabinolic acid
• However, we argue that it is necessary to apply THC ∆9-Tetrahydrocannabinol
transgenic Cannabis plants to facilitate the USDA United States Department of Agriculture
metabolic pathway for cannabinoid production
or agronomic traits.
INTRODUCTION

Cannabis sativa L. (marijuana; Cannabaceae) is an

annual dioeciously flowering plant. The first appear-
KEY FA C T S ance of Cannabis was believed to be central Asia about
• Most popular varieties of Cannabis are a combination 5000 BC. For millennia, the plant has also been used for
of two or three of C. sativa, C. indica or C. ruderalis. fiber, oil production, and traditional uses. It contains a
• Cannabis cultivated for fiber or oil, or narcotics number of medicinally important compounds, such as,
production. cannabinoids (Appendino, Chianese, & Taglialatela-
• Cannabinoids are the main active ingredient. Scafati,  2011), terpenoids (Ross & ElSohly,  1996), flavo-
• Cultivation and breeding of narcotic strains is not noids (Vanhoenacker, Van Rompaey, De Keukeleire, &
permitted in most countries. Sandra,  2002), alkaloids (Turner & Elsohly,  1976), and
• Only female plants are economically important for others (Brenneisen, 2007). Cannabinoids are a unique
producing resin in narcotic strains. class of terpenophenolic compounds to Cannabis plants,
• Indoor horticultural lighting is a new system to mimic accumulated mainly in the cavity of trichomes (Kim
sunlight. & Mahlberg,  1997). More than 80 cannabinoids have
• Indoor hydroponic technology is used for maximizing been isolated from C. sativa (Elsohly & Slade, 2005). The
cannabinoids. main psychoactive compound is ∆9-tetrahydrocannabi-
nol (THC), with well-known medicinal effects (Elbatsh,

Handbook of Cannabis and Related Pathologies. http://dx.doi.org/10.1016/B978-0-12-800756-3.00001-6

Copyright © 2017 Elsevier Inc. All rights reserved.
3
4 1.  The Cannabis Plant: Botanical Aspects

Moklas, Marsden, & Kendall, 2012). At present, cultiva- erect stems. The stems are usually angular, furrowed,
tion and breeding of Cannabis is prohibited in most coun- branched, with woody interior, sometimes hollow in the
tries, except by permission for purposes of research and internodes, and vary from 1 to 6 m in height. The branch-
pharmaceutical uses (ElSohly, 2002). Cannabis plants are ing is either opposite or alternate. The roots are advanta-
usually propagated through the seed (sexual reproduc- geous, with branched taproot, generally 30–60 cm deep,
tion; outdoor cultivation) or by vegetative propagation, up to 2.5 m in loose soils, very near to the surface, and
using stem cuttings (asexual reproduction; indoor cul- more branched in wet soils. Leaves are green and pal-
tivation) (Potter, 2004). However, both techniques have mate (seven lobes). However, the size and shape of the
advantages and disadvantages. This chapter is dedicat- leaflets differs markedly, according to genetic origin. The
ed to botanical aspects, including morphology, taxono- leaf arrangement is either opposite, or alternate or spiral.
my, genetics, conservation, geographical distribution, The leaflets are 6–11 cm (length) and 2–15 mm (width).
and cultivation forms. Leaf margins are coarsely serrated. The adaxial and abax-
ial surfaces are green, with scattered, resinous trichomes.
Inflorescences consist of numerous flower heads that
BOTANY OF CANNABIS
can be found on long, leafy stems from each leaf axil.
The staminate (male flower) consists of five pale-green,
Macroscopical Features
hairy sepals about 2.5–4  mm long, and five pendulous
Information was published elsewhere, giving detailed stamens, with slender filaments and stamen. The pistil-
technical descriptions of Cannabis morphology (Clarke, late (female flowers) are almost sessile, and are in pairs.
1981; UNODC, 2009) Fig. 1.1. However, this information The fruit (seed), is an achene, contains a single seed with
has been simplified in the present text. C. sativa is an an- a hard shell tightly covered by the thin wall of the ovary,
nual, dioeciously (ie, male and female flowers are found and it is ellipsoid, slightly compressed, smooth, about
on separate plants), pollinated plant with strong taproot, 2–5 mm long, generally brownish and mottled.

FIGURE 1.1  (A) female C. sativa; (B) portion of the female flowers; (C) pistillate female flower (stigmas, style, perigonal bract, and stipule); (D)
portion of the female flowers show anther; (E) mature seed.

I.  Setting the scene, botanical, general and international aspects

 Botany of Cannabis 5

FIGURE 1.2  Microscopic photographs of C. sativa trichomes. (A) Trichomes on the flower; (B) capitate-stalked trichome; (C) capitate-sessile
trichome; (D) bulbous trichome; (E) trichomes on the bract; (F) trichomes on the stem; (G) trichomes on the adaxial surface of a floral leaf. A big
capitate-sessile trichome is indicated with an arrow; (H) trichomes on the abaxial surface of a leaf. Present abundant small capitate-sessile and
bulbous trichomes. Source: Adapted from Happyana et al. (2013).

Microscopical Features centuries, as the result of breeding and selection. How-

ever, the Cannabis processed by these methods creates
In general, Cannabis trichomes comprise a diverse set many debates about further botanical classification. So
of structures and different types of trichomes (eg, glan- far, there is no general agreement about the taxonomic
dular and nonglandular) on a single leaf, when viewed rank of various groups within the genus Cannabis, and
through a hand lens (Fig.  1.2). Cannabis trichome re- consequently its monospecific or polyspecific character,
searchers have commonly described two types of the since the time of Linnaeus (late 18th century) (Hazekamp,
nonglandular trichome that have not been associated Justin, Lubbe, & Ruhaak, 2010). UNODC (1956) divided
with terpenoid development (Table  1.1). Three types domesticated Cannabis into three different groups:
of glandular trichome have been described on female
plants, namely bulbous, sessile, and capitate stalked • fiber hemp, long, unbranched plants, with poor seed
(Happyana et al., 2013). Male plants have been found to production
exhibit a fourth type—the antherial glandular trichome, • oil seed hemp, short, early maturing plants, with rich
which has only been found on anthers (Fairbairn, 1972). seed production
Glandular trichomes are made from a series of differenti- • drug hemp, short, strongly branched plants, with
ated cells with different functional properties, namely the small dark green leaves.
secretory cells, and stalk cells (Kim & Mahlberg, 1991). Schultes, Klein, Plowman, & Lockwood (1974) distin-
guished three species within the genus: C. sativa L., C.
Classification of Cannabis indica Lam., and C. ruderalis. Other authors referred to
The first official publication which recorded the use the same taxa only at subspecific level within one single
of Latin binomials is Linnaeus’s Species Plantarum, and species, C. sativa (Hoffmann, 1961). Small and Cronquist
it can be dated back to the year 1753. Afterward, the in- (1976) divided the single species C. sativa into the sub-
ternational community acknowledged it as the starting species sativa and indica, each consisting of a domesti-
point for modern botanical nomenclature. The species cated (Table 1.2) and wild varieties. Within the subspe-
name Cannabis means “cane-like,” while the genus name cies sativa, the domesticated and the wild varieties are
“sativa” has the meaning “planted or sown,” and signifies C. sativa subsp. sativa var. sativa (domesticated), C. sativa
that the plant is propagated from seed, and not from pe- subsp. sativa var. spontama (wild), C. sativa subsp. indica
rennial roots (Raman,  1998). According to the modern var. indica (domesticated), and C. sativa subsp. indica
system of classification, Cannabis belongs to the family var. kafiristanica (wild). However, it is commonly ac-
of Cannabaceae, along with the Humulus genus (hops) cepted that Cannabis is monotypic, and consists only of
(Turner, Elsohly, & Boeren, 1980a,b). Different varieties of a single species: C. sativa (Brenneisen,  1983; Beutler &
Cannabis have been developed over the course of many Dermarderosian, 1978).

I.  Setting the scene, botanical, general and international aspects

6 1.  The Cannabis Plant: Botanical Aspects

TABLE 1.1 A Summary of Cannabis Trichomes Classification, Structure, Distribution, Timing of Development, and Lifespan
Trichomes
Timing of
development/
Classification Structure Distribution density Lifespan References

Nonglandular (1) Noncystolithic trichomes: Lower side of Decreases The viability and (Fairbairn, 1972;
trichomesa long, unicellular, smooth, vegetative leaves with age functioning Hammond &
curved, covering and pistillate secretion is Mahlberg, 1977; Turner
trichomes bracts correlated with et al., 1977, 1980b, 1981;
senescence of Croteau, 1988;
(2) Cystolithic trichomes: epidermal cells Werker, 2000; Guy &
more squat, unicellular, claw Stott, 2005; Happyana
shape, cystolith covering et al., 2013)
trichomes, containing
calcium carbonate

Glandular (1) Bulbous: Vegetative leaves

trichomesb with smallest gland and pistillate
bracts

(2) Capitate-sessile (unstalked):

the structure is commonly
simple, and the trichomes
head connected directly to
the mesophyll cells.

(3) Capitate-stalked: Bracts and floral Increases with

the structure more complex, leaves age
they developed resin
head (also known as
the glandular head)
that resembles a golf
ball sitting on a tee (the
trichome’s stalk).

Antherial sessile Large size, with a diameter Underside of the

trichomesc of approximately anther lobes
70–80 µm
a
Nonglandular trichomes lack cannabinoids.
b
Glandular trichomes are the principal or sole site of storage of most cannabinoids, the content of ∆9-THC in pistillate flowers ranged between 10 and 12%, and in leaves ranged
between 1 and 2%.
c
Male plants are of no consequence in medicine production because they develop few glandular trichomes and, consequently, produce few cannabinoids or terpenes.

TABLE 1.2 Synopsis of C. sativa Sectional Species, Subspecies, and Varieties Recognized Based on Chemical, Genetic, and
Morphological Variation
Section sativa Section spontanea
a
C. sativa (L.) C. ruderalis (L)a
C. chinensis (Delile) C. sativa subsp. spontanea (Serebr.)
C. gigantea (Delile) var. spontanea
C. americana (Houghton) var. ruderalis
C. sativa subsp. Intersita (So.)
Section indica
subsp. culta (Serebr) C. indica (Lam.)a
subsp. Sativa (L.) C. macrosperma (Stokes)
var. sativa C. sativa subsp. indica (Lam.)

var. praecox var. indica
var. monoica var. kif
var. gigantea var. afghanica
var. Chinensis var. kafiristanica
var. pedemontana
a
Includes the endemic and domesticated populations (Raman, 1998; Sytsma et al., 2002; Hillig, 2005).

I.  Setting the scene, botanical, general and international aspects

 Botany of Cannabis 7

The current scientific classification of Cannabis (Sytsma developing new varieties. Newly hybrid varieties have
et al., 2002) been developed as a result of the crossbreds, such as,
  Class   Hamamelidae “super-sativa” (Clarke & Watson, 2002; de Meijer, 2004).
   Subclass    Rosales Recently, varieties of Cannabis have been licensed to
    Order     Cannabaceae GW Pharmaceuticals Ltd, as part of indoor breeding
     Family      Cannabis programs (de Meijer & Hammond, 2005). In the United
      Genus       sativa States, the majority of Cannabis cultivars were selected
       Species
from single landrace sources, or from multihybrid prog-
enies made from different landraces (de Meijer,  2004).
The marijuana potency monitoring project at the Uni-
Other Recent Taxonomic Studies
versity of Mississippi (USA) is breeding Sinsemilla,
CHEMOTAXONOMIC CLASSIFICATION Skunk 1, Four Way, Four Way-F, Thai/Skunk, Terbag
Recently, chemotaxonomic classification splits the W1, K2, and MX Cannabis of hybrid varieties (ElSohly,
phenotypes based on the quantitative differences in the Holley, & Turner, 1985; Elsohly, Holley, Lewis, Russell, &
cannabinoid ratio of tetrahydrocannabinolic acid (THC), Turner, 1984). In the Netherlands, there are three differ-
cannabinol (CBN), and cannabidiol (CBD), in the ratio of ent Cannabis varieties from sativa: Bedrocan, Bedrobinol,
[THC] + [CBN]/[CBD]. If the ratio exceeded 1, plants are and Bediol, and one variety from C. indica is Bedica –
classified as “chemo-type,” otherwise as “fiber-type,” and all studied and registered by Bedrocan BV (Fischedick,
this was the first study to differentiate between the drug- Hazekamp, Erkelens, Choi, & Verpoorte,  2010). Nowa-
and fiber-type, by Fetterman et al. (1971). Therefore, this days, many Cannabis hybrid cultivars (Table  1.3) and
ratio was subsequently used to discriminate chemotype, some selected pure strains have been commercialized in
intermediate type, and fiber-type (Turner, Cheng, Lewis, many private companies, and there are up to 20 more
Russell, & Sharma,  1979). Hillig and Mahlberg (2004) or less well defined strains for either indoor or outdoor
split Cannabis into putative species and subspecies, us- cultivation, in The Netherlands, but a sufficient data set
ing multivariate data analysis. Moreover, it was reported is not available, due to illegal cultivation. Today, the cul-
that, depending on age, the Cannabis plant can be classi- tivation and production of hemp is restricted and con-
fied into different morphotypes, at different time points trolled because of its association with narcotic use. Most
of its development. Although this classification was not of the hemp breeders cultivate fiber hemp with the ul-
comprehensive enough to elucidate infrageneric taxo- timate goal to reduce THC content below 0.2%, or even
nomic structure, and does not define the contents of can- to get noncannabinoid plants by breeding and crossing
nabinoids for each chemotype, it provides a usable tool experiments (de ­Meijer, 1995).
for classification (Hazekamp et al., 2010).

MOLECULAR CLASSIFICATION
Genetics of Cannabis
Several molecular techniques have been evaluated to Genome of Cannabis sativa
establish the genetic relationship among different variet- The genome of Cannabis (2n = 18 + XX for female, and
ies of Cannabis plants. Some recent studies have classi- 2n = 18 + XY for male) has a karyotype composed of nine
fied and identified C. sativa samples that cannot be dif- autosomes and a pair of sex chromosomes (X and Y). Sex
ferentiated by HPLC analysis alone, by using genomic chromosomes changes during the developmental stages
DNA, random amplified polymorphic DNA (RAPD), are claimed to occur in many dioecious plants, as a strat-
and restriction fragment polymorphisms (RFLP) analy- egy for survival (Flemming et  al., 2007). The genome
sis, but little work appears to have been conducted with was measured in both female (XX) and male plants (XY)
marker types that would be usable for breeding (Gillan, (Vyskot & Hobza, 2015). The estimated haploid genome
Cole, Linacre, Thorpe, & Watson, 1995; Faeti, Mandolino, sizes are 818 Mb for female plants, and 843 Mb for males
& Ranalli, 1996). Recently, Kojoma, Iida, Makino, Sekita, (Sakamoto et al., 1998). The genomic resources available
and Satake (2002) reported that different Cannabis were for Cannabis are mainly confined to transcriptome infor-
identified by means of inter simple sequence repeat mation: the NCBI database contains 12,907 ESTs and 23
(ISSR). ISSR is a technique offering the reproducibility unassembled RNA-Seq datasets of Illumina reads (Marks
and simplicity of RAPDs with high reliability (Galvan, et al., 2009). The genetic basis of cannabinoid variation in
Bornet, Balatti, & Branchard, 2003). C. sativa showed that the amount of THC versus CBD is
likely governed by one locus with two codominant al-
leles, B(d) and B(t) (de Meijer et al., 2003). One possible
Current Cannabis Varieties
explanation for these results is that the two alleles en-
Recently, Cannabis growers have become more code either THCA or CBDA synthase so that homozy-
aware to create variations between different strains for gous plants would contain either tetrahydrocannabinolic

I.  Setting the scene, botanical, general and international aspects

8 1.  The Cannabis Plant: Botanical Aspects

TABLE 1.3 Origin of Hemp Varieties Were Reported in acid (THCA) or cannabidiolic acid (CBDA) as the major
Literaturea cannabinoid, and heterozygotes would have an approxi-
Variety Country mately equal mixture of the two (Fig. 1.3). Another ex-
planation is that THCA and CBDA synthases are closely
Finola Finland
linked genes, perhaps produced as a result of a gene
Glukhov 33, Kuban, Uso 11, Zenica, USO 13, USO Ukraine duplication event. A recent study analyzed the THCA
15, USO 31, YUSO 14, YUSO 16 synthase sequences from drug (high-THC) and fiber
Asso, Carmagnola, CS (Carmagnola Selezionata), Italy (low-THC) varieties, and found that the amino acid se-
Carmono, Carma, Codimono, Eletta Campana, quence of THCA synthase from high-THC varieties dif-
Ferrara, Ermes, Fibrimor fered by 37 major substitutions, compared to low-THC
Fibranova
varieties (Kojoma, Seki, Yoshida, & Muranaka, 2006).
Fasamo, Ferimon Germany

Santhica 27, Epsilon 68, Fedora 17, Fedora 19, France Geographical Distribution
Fedrina 74, Felina 32, Felina 34, Fibrimon 21,
Fibrimon 24, Fibrimon 56, Futura, Futura 77, Small and Cronquist (1976) state that genus Canna-
Futura 75, Santhica 23, Dioica 88 bis geographically grows to the north of latitude 30°N
Kompolti Sargaszaru, Kinai unisexualis, Kompolti, Hungary and south of latitude 60°N (Hillig,  2005). The genus is
Kompolti Hybrid TC, Kompolti Hyper, Elite, believed to have originated in the Northwest Himalayas,
Fibriko and occurs widely in Africa.
Fibramulta 151, Irene, Lovrin 110, Moldovan, Romania
Secuieni 1
Agricultural Status
Beniko, Bialobrzeskie, LKCSD, Dolnoslaskie Poland
Nowadays, fiber hemp is cultivated in a number
Chamaeleon, Dutch “Yellow” line Netherlands
of countries around the world, and China represents
Ermakovskaya Mestnaya Russia the largest producer of hemp with focus on fiber-type
Delta 405, Delta-llosa Spain (Mediavilla, Bassetti, & Leupin, 1999). Nevertheless, cul-
tivation of medicinal Cannabis is prohibited in most of
Kenvir Turkey
countries, except by permission for purposes of research
Swissmix Swiss and pharmaceutical uses.
Ratslaviska Czech

Silistrensi, Mecnaja copt Bulgaria Conservation Initiatives

Pesnica Slovenia Cannabis populations are facing the threat of genetic
Flajsmanova, Novosadksa, Novosadska plus, Former drift—which has a direct effect on the changes to the
Novosadska konoplja ­Yugoslavia phenotype and chemical profile, due to allogamous (de
Kinai Egylaki, Kinai Ketlaki China Meijer & Vansoest, 1992). The conservation of Cannabis
germplasm is divided into two main strategies: in situ
Kozuhura Zairai Japan
and ex situ.
a
Low THC cultivars, less than 0.2% dry weight.
Ex Situ Conservation in Gene Banks
The Cannabis gene bank at Vavilov Research Insti-
tute of Plant Industry (St. Petersburg, Russia) main-
tained about 200 accessions, for more than 50 years (de
Meijer,  1998). In addition, the Hungarian gene bank at
the Research Center for Agrobiodiversity (Tápiószele,
Hungary) maintained about 70 accessions. Collections of
up to 20 accessions are preserved in other depositories in
Germany, Turkey, and Japan. In comparison with other
crops, the available number of well-documented Cannabis
accessions is limited (de Meijer & Vansoest, 1992). Now-
adays, several accessions are maintained by the United
States Department of Agriculture (USDA)/National
FIGURE 1.3  Inheritance of chemical phenotype in C. sativa “co-
dominant monogenic control,” homozygous THC producing BtBt Plant Germplasm System (NPGS), and associated data
genotypes are typically selected for recreational use. Source: From de can be accessed from the Germplasm Resources Infor-
Meijer et al. (2003). mation Network (GRIN) database.

I.  Setting the scene, botanical, general and international aspects

 Botany of Cannabis 9
In Situ Conservation as In Vitro Gene Banks Indoor Cultivation
In vitro conservation of encapsulated microcuttings This method of breeding is used for increasing resin
of Cannabis shootlets was attempted under slow growth potency, and avoiding unwanted male plants (Chandra,
conditions between 5 and 15°C (Lata, Chandra, Khan, Lata, Khan, & ElSohly, 2010). The complete growth cy-
& ElSohly,  2008; Lata, Chandra, Mehmedic, Khan, & cle, quality, and quantity of biomass can be regulated
ElSohly,  2012), but adaptation to in vitro conditions under controlled environmental conditions (6–8 weeks).
could induce mutants of the offspring plants, causing The successful indoor system requires an effective hy-
genetic and chemical drift (Larkin & Scowcroft, 1981). droponic system to deliver nutrients and oxygen, and
support the plants’ growth (Fig. 1.4). However, there is a
Cultivation Techniques of Cannabis number of different techniques that have been proposed
for the indoor horticulture of Cannabis, for example, the
Outdoor Cultivation
standing aerated technique, nutrient film technique, and
Cannabis plant can be propagated from seeds, and aeroponics technique. In hydroponic growing, the nutri-
the life cycle is completed within 4–6 months, depend- ent solution is best at a pH within a certain range (5.5–
ing on the time of the plantation and the variety. It can 6.5) for maximum uptake and good plant growth (Argo
reach up to 5 m (16 ft.) in height, in a 4–6 months grow- & Fischer, 2002). Indoor Cannabis crop cultivation needs
ing season (Raman, 1998; Clarke & Watson, 2002). Her- artificial light and compressed CO2 gas for photosyn-
maphroditic varieties of this plant have been bred for thesis, and for controlling flowering and plant biomass
industrial hemp production, as this allows more uni- (Jones, 1997). Here, selective vegetative female plants are
form crops (Leggett, 2006). The process of germination is used for making clones. Later, all clones are kept under
usually completed in 3–7 days (Clarke & Watson, 2002). standard environmental conditions (light, temperature,
The seedling stage is completed within 2–3 months. Lat- RH, and CO2 concentration) in a growing room for vegeta-
er, the plant is characterized by increased biomass and tion (18 h/day photoperiod) and for flowering (12 h/day
total growth under long day time lengths (vegetative photoperiod).
growth). It is easy to recognize the male and female sex
at this stage. Later in summer, the reproductive phase In vitro Micropropagation
of Cannabis begins when the plant is exposed to short The micropropagation system offers a number of
day time lengths (less light per day than darkness) of 12– clear advantages, including (1) human-controlled
14 h or less, depending on its latitude and genetic origin method with fast propagation in a comparably short
(Brenneisen,  1983). Once the male flowers ripened and time, due to high potential multiplication rates, (2) it
pollinated, the female flowers died directly. The pro- is independent of seasonal factors like climate and ge-
duced seeds after flowering have combinations of traits ography, and (3) the produced plants are usually free
from two parents, as a result of cross fertilization (Clarke from ­microorganism-borne diseases (Zafar, Aeri, &
& Watson, 2002). This method is mostly used for the cul- ­Datta, 1992). On the other hand, in vitro propagation of
tivation of Cannabis for hemp fiber, or Cannabis seed with C. sativa through the seed is possible in most of cultivars,
less than 0.2% THC. although the greatest problem with such a method is the

FIGURE 1.4  Indoor cultivation of C. sativa. Source: Photo provided from Bedrocan BV, the Netherlands.

I.  Setting the scene, botanical, general and international aspects

10 1.  The Cannabis Plant: Botanical Aspects

FIGURE 1.5  In vitro micropropagation of leaf-derived calli from C. sativa L. (A) Callus culture, (B) meristemoid formation, (C) shootlets
multiplication on Gamborg’s B5 medium supplemented with various combinations of auxins and cytokinins. Source: Photos provided from Sayed
Farag PhD project, Technische Universität Dortmund.

high level of heterozygosity that could lead to a rapid Micropropagation  In vitro technique for multiplying plant tissues
and dramatic profile shift of secondary metabolites through in vitro culture, either indirectly (with intervening callus
stage) or directly (without an intervening proliferative stage). This
from one generation to the next (Chandra et  al.,  2010). is achieved by altering the concentration of growth regulators,
In fact, in vitro propagation using explants or somatic mainly auxins and cytokinins.
embryogenesis has been reported (Lata et al., 2002). Be- Random amplified polymorphic DNA (RAPD)  A molecular
sides the progress in the field of plant biotechnology, technique for the rapid assignation of DNA-based character states
very little progress has been made to date toward devel- for phylogenetic analysis. The technique uses the polymerase
chain reaction (PCR) to amplify any genomic region containing
oping an in vitro regeneration from C. sativa. Previous single primer of nucleotide arbitrary sequence.
reports on de novo organogensis of C. sativa emerged Restriction fragment length polymorphism (RFLP)  A molecular
in early 1980s (Fisse, Braut, Cosson, & Paris, 1981), and technique for genome mapping, and variation analysis (genotyping,
subsequently from callus of different genotypes and ex- forensics, paternity tests, hereditary disease diagnostics, etc.). The
plant sources, including cotyledons and stem (Wielgus, technique uses restriction of endonucleases to cut DNA at specific
(generally 4–6 bp) recognition sites.
Luwanska, Lassocinski, & Kaczmarek,  2008), young Trichome  Defined as hair-like structures that extend from the
leaves (Lata, Chandra, Khan, & ElSohly,  2010), inter- epidermis of aerial tissues; are present on the surface of most
nodes, and axillary buds and petioles (Slusarkiewicz- terrestrial plants.
Jarzina, Ponitka, & Kaczmarek, 2005), and roots (Ranalli
& Mandolino,  1999). Alternatively, the use of meriste- References
matic callus for micropropagation was studied recently Appendino, G., Chianese, G., & Taglialatela-Scafati, O. (2011). Can-
(Farag & Kayser, unpublished results, Fig. 1.5). nabinoids: occurrence and medicinal chemistry. Current Medicinal
Chemistry, 18(7), 1085–1099.
Argo, W. R., Fischer, P. R. (2002). Understanding pH management for
Recommendations for Future Action container-grown crops. Meister, Willoughby, Ohio.
Beutler, J. A., & Dermarderosian, A. H. (1978). Chemotaxonomy of
Given the high therapeutic and commercial value of Cannabis .1. Cross breeding between Cannabis sativa and Cannabis
Cannabis, legal indoor breeding started in some pharma- ruderalis, with analysis of Cannabinoid content. Economic Botany,
ceutical companies. The biotechnological research for ge- 32(4), 387–394.
netic improvement has been minimal to date. Researches Brenneisen, R. (1983). Psychotropic drugs. I. Cannabis sativa L. (Canna-
on transgenic Cannabis is still needed to facilitate the met- binaceae). Pharmaceutica Acta Helvetiae, 58(11), 314–320.
Brenneisen, R. (2007). Chemistry and analysis of phytocannabinoids
abolic engineering of cannabinoids and agronomic traits. and other Cannabis constituents. In M. A. Elsohly (Ed.), Marijuana
and the cannabinoids (pp. 17–49). Totowa: Humana Press.
Chandra, S. H., Lata, I., Khan, A., & ElSohly, M. A. (2010). Propagation
MINI-DICTIONARY of elite Cannabis sativa for the production of ∆9-Tetrahydrocannabinol
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