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

KEY FA C T S
• Most popular varieties of Cannabis are a combination
of two or three of C. sativa, C. indica or C. ruderalis.
• Cannabis cultivated for fiber or oil, or narcotics
production.
• Cannabinoids are the main active ingredient.
• Cultivation and breeding of narcotic strains is not
permitted in most countries.
• Only female plants are economically important for
producing resin in narcotic strains.
• Indoor horticultural lighting is a new system to mimic
sunlight.
• Indoor hydroponic technology is used for maximizing
cannabinoids.
annual dioeciously flowering plant. The first appear-
ance of Cannabis was believed to be central Asia about
5000 BC. For millennia, the plant has also been used for
fiber, oil production, and traditional uses. It contains a
number of medicinally important compounds, such as,
cannabinoids (Appendino, Chianese, & Taglialatela-
Scafati,  2011), terpenoids (Ross & ElSohly,  1996), flavo-
noids (Vanhoenacker, Van Rompaey, De Keukeleire, &
Sandra,  2002), alkaloids (Turner & Elsohly,  1976), and
others (Brenneisen, 2007). Cannabinoids are a unique
class of terpenophenolic compounds to Cannabis plants,
accumulated mainly in the cavity of trichomes (Kim
& Mahlberg,  1997). More than 80 cannabinoids have
been isolated from C. sativa (Elsohly & Slade, 2005). The
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-
tion and breeding of Cannabis is prohibited in most coun-
tries, except by permission for purposes of research and
pharmaceutical uses (ElSohly, 2002). Cannabis plants are
usually propagated through the seed (sexual reproduc-
tion; outdoor cultivation) or by vegetative propagation,
using stem cuttings (asexual reproduction; indoor cul-
tivation) (Potter, 2004). However, both techniques have
advantages and disadvantages. This chapter is dedicat-
ed to botanical aspects, including morphology, taxono-
my, genetics, conservation, geographical distribution,
and cultivation forms.
BOTANY OF CANNABIS
Macroscopical Features
Information was published elsewhere, giving detailed
technical descriptions of Cannabis morphology (Clarke,
1981; UNODC, 2009) Fig. 1.1. However, this information
has been simplified in the present text. C. sativa is an an-
nual, dioeciously (ie, male and female flowers are found
on separate plants), pollinated plant with strong taproot,
erect stems. The stems are usually angular, furrowed,
branched, with woody interior, sometimes hollow in the
internodes, and vary from 1 to 6 m in height. The branch-
ing is either opposite or alternate. The roots are advanta-
geous, with branched taproot, generally 30–60 cm deep,
up to 2.5 m in loose soils, very near to the surface, and
more branched in wet soils. Leaves are green and pal-
mate (seven lobes). However, the size and shape of the
leaflets differs markedly, according to genetic origin. The
leaf arrangement is either opposite, or alternate or spiral.
The leaflets are 6–11 cm (length) and 2–15 mm (width).
Leaf margins are coarsely serrated. The adaxial and abax-
ial surfaces are green, with scattered, resinous trichomes.
Inflorescences consist of numerous flower heads that
can be found on long, leafy stems from each leaf axil.
The staminate (male flower) consists of five pale-green,
hairy sepals about 2.5–4  mm long, and five pendulous
stamens, with slender filaments and stamen. The pistil-
late (female flowers) are almost sessile, and are in pairs.
The fruit (seed), is an achene, contains a single seed with
a hard shell tightly covered by the thin wall of the ovary,
and it is ellipsoid, slightly compressed, smooth, about
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-

In general, Cannabis trichomes comprise a diverse set
of structures and different types of trichomes (eg, glan-
dular and nonglandular) on a single leaf, when viewed
through a hand lens (Fig.  1.2). Cannabis trichome re-
searchers have commonly described two types of the
nonglandular trichome that have not been associated
with terpenoid development (Table  1.1). Three types
of glandular trichome have been described on female
plants, namely bulbous, sessile, and capitate stalked
(Happyana et al., 2013). Male plants have been found to
exhibit a fourth type—the antherial glandular trichome,
which has only been found on anthers (Fairbairn, 1972).
Glandular trichomes are made from a series of differenti-
ated cells with different functional properties, namely the
secretory cells, and stalk cells (Kim & Mahlberg, 1991).
Classification of Cannabis
The first official publication which recorded the use
of Latin binomials is Linnaeus’s Species Plantarum, and
it can be dated back to the year 1753. Afterward, the in-
ternational community acknowledged it as the starting
point for modern botanical nomenclature. The species
name Cannabis means “cane-like,” while the genus name
“sativa” has the meaning “planted or sown,” and signifies
that the plant is propagated from seed, and not from pe-
rennial roots (Raman,  1998). According to the modern
system of classification, Cannabis belongs to the family
of Cannabaceae, along with the Humulus genus (hops)
(Turner, Elsohly, & Boeren, 1980a,b). Different varieties of
Cannabis have been developed over the course of many
ever, the Cannabis processed by these methods creates
many debates about further botanical classification. So
far, there is no general agreement about the taxonomic
rank of various groups within the genus Cannabis, and
consequently its monospecific or polyspecific character,
since the time of Linnaeus (late 18th century) (Hazekamp,
Justin, Lubbe, & Ruhaak, 2010). UNODC (1956) divided
domesticated Cannabis into three different groups:
• fiber hemp, long, unbranched plants, with poor seed
production
• oil seed hemp, short, early maturing plants, with rich
seed production
• drug hemp, short, strongly branched plants, with
small dark green leaves.
Schultes, Klein, Plowman, & Lockwood (1974) distin-
guished three species within the genus: C. sativa L., C.
indica Lam., and C. ruderalis. Other authors referred to
the same taxa only at subspecific level within one single
species, C. sativa (Hoffmann, 1961). Small and Cronquist
(1976) divided the single species C. sativa into the sub-
species sativa and indica, each consisting of a domesti-
cated (Table 1.2) and wild varieties. Within the subspe-
cies sativa, the domesticated and the wild varieties are
C. sativa subsp. sativa var. sativa (domesticated), C. sativa
subsp. sativa var. spontama (wild), C. sativa subsp. indica
var. indica (domesticated), and C. sativa subsp. indica
var. kafiristanica (wild). However, it is commonly ac-
cepted that Cannabis is monotypic, and consists only of
a single species: C. sativa (Brenneisen,  1983; Beutler &
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
et al., 2002)
  Class   Hamamelidae
   Subclass    Rosales
    Order     Cannabaceae
     Family      Cannabis
      Genus       sativa
       Species
Other Recent Taxonomic Studies
CHEMOTAXONOMIC CLASSIFICATION
Recently, chemotaxonomic classification splits the
phenotypes based on the quantitative differences in the
cannabinoid ratio of tetrahydrocannabinolic acid (THC),
cannabinol (CBN), and cannabidiol (CBD), in the ratio of
[THC] + [CBN]/[CBD]. If the ratio exceeded 1, plants are
classified as “chemo-type,” otherwise as “fiber-type,” and
this was the first study to differentiate between the drug-
and fiber-type, by Fetterman et al. (1971). Therefore, this
ratio was subsequently used to discriminate chemotype,
intermediate type, and fiber-type (Turner, Cheng, Lewis,
Russell, & Sharma,  1979). Hillig and Mahlberg (2004)
split Cannabis into putative species and subspecies, us-
ing multivariate data analysis. Moreover, it was reported
that, depending on age, the Cannabis plant can be classi-
fied into different morphotypes, at different time points
of its development. Although this classification was not
comprehensive enough to elucidate infrageneric taxo-
nomic structure, and does not define the contents of can-
nabinoids for each chemotype, it provides a usable tool
for classification (Hazekamp et al., 2010).
MOLECULAR CLASSIFICATION
Several molecular techniques have been evaluated to
establish the genetic relationship among different variet-
ies of Cannabis plants. Some recent studies have classi-
fied and identified C. sativa samples that cannot be dif-
ferentiated by HPLC analysis alone, by using genomic
DNA, random amplified polymorphic DNA (RAPD),
and restriction fragment polymorphisms (RFLP) analy-
sis, but little work appears to have been conducted with
marker types that would be usable for breeding (Gillan,
Cole, Linacre, Thorpe, & Watson, 1995; Faeti, Mandolino,
& Ranalli, 1996). Recently, Kojoma, Iida, Makino, Sekita,
and Satake (2002) reported that different Cannabis were
identified by means of inter simple sequence repeat
(ISSR). ISSR is a technique offering the reproducibility
and simplicity of RAPDs with high reliability (Galvan,
Bornet, Balatti, & Branchard, 2003).
Current Cannabis Varieties
Recently, Cannabis growers have become more
aware to create variations between different strains for
I.  Setting the scene, botanical, general and international aspects
developing new varieties. Newly hybrid varieties have
been developed as a result of the crossbreds, such as,
“super-sativa” (Clarke & Watson, 2002; de Meijer, 2004).
Recently, varieties of Cannabis have been licensed to
GW Pharmaceuticals Ltd, as part of indoor breeding
programs (de Meijer & Hammond, 2005). In the United
States, the majority of Cannabis cultivars were selected
from single landrace sources, or from multihybrid prog-
enies made from different landraces (de Meijer,  2004).
The marijuana potency monitoring project at the Uni-
versity of Mississippi (USA) is breeding Sinsemilla,
Skunk 1, Four Way, Four Way-F, Thai/Skunk, Terbag
W1, K2, and MX Cannabis of hybrid varieties (ElSohly,
Holley, & Turner, 1985; Elsohly, Holley, Lewis, Russell, &
Turner, 1984). In the Netherlands, there are three differ-
ent Cannabis varieties from sativa: Bedrocan, Bedrobinol,
and Bediol, and one variety from C. indica is Bedica –
all studied and registered by Bedrocan BV (Fischedick,
Hazekamp, Erkelens, Choi, & Verpoorte,  2010). Nowa-
days, many Cannabis hybrid cultivars (Table  1.3) and
some selected pure strains have been commercialized in
many private companies, and there are up to 20 more
or less well defined strains for either indoor or outdoor
cultivation, in The Netherlands, but a sufficient data set
is not available, due to illegal cultivation. Today, the cul-
tivation and production of hemp is restricted and con-
trolled because of its association with narcotic use. Most
of the hemp breeders cultivate fiber hemp with the ul-
timate goal to reduce THC content below 0.2%, or even
to get noncannabinoid plants by breeding and crossing
experiments (de ­Meijer, 1995).
Genetics of Cannabis
Genome of Cannabis sativa
The genome of Cannabis (2n = 18 + XX for female, and
2n = 18 + XY for male) has a karyotype composed of nine
autosomes and a pair of sex chromosomes (X and Y). Sex
chromosomes changes during the developmental stages
are claimed to occur in many dioecious plants, as a strat-
egy for survival (Flemming et  al., 2007). The genome
was measured in both female (XX) and male plants (XY)
(Vyskot & Hobza, 2015). The estimated haploid genome
sizes are 818 Mb for female plants, and 843 Mb for males
(Sakamoto et al., 1998). The genomic resources available
for Cannabis are mainly confined to transcriptome infor-
mation: the NCBI database contains 12,907 ESTs and 23
unassembled RNA-Seq datasets of Illumina reads (Marks
et al., 2009). The genetic basis of cannabinoid variation in
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
explanation for these results is that the two alleles en-
code either THCA or CBDA synthase so that homozy-
gous plants would contain either tetrahydrocannabinolic
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
In vitro conservation of encapsulated microcuttings
of Cannabis shootlets was attempted under slow growth
conditions between 5 and 15°C (Lata, Chandra, Khan,
& ElSohly,  2008; Lata, Chandra, Mehmedic, Khan, &
ElSohly,  2012), but adaptation to in vitro conditions
could induce mutants of the offspring plants, causing
genetic and chemical drift (Larkin & Scowcroft, 1981).
Cultivation Techniques of Cannabis
Outdoor Cultivation
Cannabis plant can be propagated from seeds, and
the life cycle is completed within 4–6 months, depend-
ing on the time of the plantation and the variety. It can
reach up to 5 m (16 ft.) in height, in a 4–6 months grow-
ing season (Raman, 1998; Clarke & Watson, 2002). Her-
maphroditic varieties of this plant have been bred for
industrial hemp production, as this allows more uni-
form crops (Leggett, 2006). The process of germination is
usually completed in 3–7 days (Clarke & Watson, 2002).
The seedling stage is completed within 2–3 months. Lat-
er, the plant is characterized by increased biomass and
total growth under long day time lengths (vegetative
growth). It is easy to recognize the male and female sex
at this stage. Later in summer, the reproductive phase
of Cannabis begins when the plant is exposed to short
day time lengths (less light per day than darkness) of 12–
14 h or less, depending on its latitude and genetic origin
(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-
duced seeds after flowering have combinations of traits
from two parents, as a result of cross fertilization (Clarke
& Watson, 2002). This method is mostly used for the cul-
tivation of Cannabis for hemp fiber, or Cannabis seed with
less than 0.2% THC.
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
Indoor Cultivation
This method of breeding is used for increasing resin
potency, and avoiding unwanted male plants (Chandra,
Lata, Khan, & ElSohly, 2010). The complete growth cy-
cle, quality, and quantity of biomass can be regulated
under controlled environmental conditions (6–8 weeks).
The successful indoor system requires an effective hy-
droponic system to deliver nutrients and oxygen, and
support the plants’ growth (Fig. 1.4). However, there is a
number of different techniques that have been proposed
for the indoor horticulture of Cannabis, for example, the
standing aerated technique, nutrient film technique, and
aeroponics technique. In hydroponic growing, the nutri-
ent solution is best at a pH within a certain range (5.5–
6.5) for maximum uptake and good plant growth (Argo
& Fischer, 2002). Indoor Cannabis crop cultivation needs
artificial light and compressed CO2 gas for photosyn-
thesis, and for controlling flowering and plant biomass
(Jones, 1997). Here, selective vegetative female plants are
used for making clones. Later, all clones are kept under
standard environmental conditions (light, temperature,
RH, and CO2 concentration) in a growing room for vegeta-
tion (18 h/day photoperiod) and for flowering (12 h/day
photoperiod).
In vitro Micropropagation
The micropropagation system offers a number of
clear advantages, including (1) human-controlled
method with fast propagation in a comparably short
is independent of seasonal factors like climate and ge-
ography, and (3) the produced plants are usually free
from ­microorganism-borne diseases (Zafar, Aeri, &
­Datta, 1992). On the other hand, in vitro propagation of
C. sativa through the seed is possible in most of cultivars,
although the greatest problem with such a method is the
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
and dramatic profile shift of secondary metabolites
from one generation to the next (Chandra et  al.,  2010).
In fact, in vitro propagation using explants or somatic
embryogenesis has been reported (Lata et al., 2002). Be-
sides the progress in the field of plant biotechnology,
very little progress has been made to date toward devel-
oping an in vitro regeneration from C. sativa. Previous
reports on de novo organogensis of C. sativa emerged
in early 1980s (Fisse, Braut, Cosson, & Paris, 1981), and
subsequently from callus of different genotypes and ex-
plant sources, including cotyledons and stem (Wielgus,
Luwanska, Lassocinski, & Kaczmarek,  2008), young
leaves (Lata, Chandra, Khan, & ElSohly,  2010), inter-
nodes, and axillary buds and petioles (Slusarkiewicz-
Jarzina, Ponitka, & Kaczmarek, 2005), and roots (Ranalli
& Mandolino,  1999). Alternatively, the use of meriste-
matic callus for micropropagation was studied recently
(Farag & Kayser, unpublished results, Fig. 1.5).
Recommendations for Future Action
Given the high therapeutic and commercial value of
Cannabis, legal indoor breeding started in some pharma-
ceutical companies. The biotechnological research for ge-
netic improvement has been minimal to date. Researches
on transgenic Cannabis is still needed to facilitate the met-
abolic engineering of cannabinoids and agronomic traits.
MINI-DICTIONARY
Encapsulation  In vitro technique for the production of synthetic
seeds (Ca-alginate beads) for long-term storage of germplasm.
Genome  The complete set of chromosomal and extra-
chromosomal DNA/RNA of an organism, a cell, an organelle,
or a virus.
Inter simple sequence repeats (ISSR)  A molecular technique
for evolutionary biology. Its simple sequence repeats (SSR), also
known as microsatellites, are tandem repeats of a few base pairs
distributed throughout the genome.
I.  Setting the scene, botanical, general and international aspects
Micropropagation  In vitro technique for multiplying plant tissues
through in vitro culture, either indirectly (with intervening callus
stage) or directly (without an intervening proliferative stage). This
is achieved by altering the concentration of growth regulators,
mainly auxins and cytokinins.
Random amplified polymorphic DNA (RAPD)  A molecular
technique for the rapid assignation of DNA-based character states
for phylogenetic analysis. The technique uses the polymerase
chain reaction (PCR) to amplify any genomic region containing
single primer of nucleotide arbitrary sequence.
Restriction fragment length polymorphism (RFLP)  A molecular
technique for genome mapping, and variation analysis (genotyping,
forensics, paternity tests, hereditary disease diagnostics, etc.). The
technique uses restriction of endonucleases to cut DNA at specific
(generally 4–6 bp) recognition sites.
Trichome  Defined as hair-like structures that extend from the
epidermis of aerial tissues; are present on the surface of most
terrestrial plants.
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