Medicinal Ganja

Conservation and Cultivation of Elite

Varieties for Medicinal and Research
Applications

3rd Biennial Conference on Science and Technology

November 12, 2014

Ryan Francis (Presenter)

Jodi Spence
Gillian Rowe
 Outline
• Introduction
• In vitro Propagation of Cannabis sativa
• Hydroponic Propagation
• Greenhouse Propagation
• Conclusion
• References

2
 Introduction
• Cannabis sativa L. (Cannabaceace) originated in Asia,
spread through Europe and later introduced into the
America.
• Cannabis (Marijuana) is a dioecious annual herb.
• Produces equal number of male and female flower.
• This plant contains a unique class of terpenophenolic
compounds called Cannabinoids.
 Introduction
• More than 70 cannabinoids have been isolated from
this herb (Lata et al 2009; Russo 2013).

• Main biologically active compounds are:

Tetrahydro-cannabinol (THC) – Psychoactive.

• Cannabinol (CBD) – Nonpsycotophic (Mehmedic et

al 2010).
 Introduction

• Pharmacological and therapeutic potential

THC: Analgesic, anti-inflammatory, appetite stimulant,

depression, rheumatism, fatigue, migrane and anti-emetic
properties (Lata et al 2012).

CBD: Antimicrobial action, relieve anxiety, epilepsy,

multiple sclerosis, dystonia and other neurological
disorders (Clarke et al 1994).
 Introduction
• Ganja (all parts of the Cannabis sativa plant) is
one of the substances named in the Dangerous
Drugs Act of 1948.

• It is an offense to cultivate, sell, deal in,

transport, possess, import or export ganja.

• September 30, 2014 GOJ approved proposals for

amendments to the Act to facilitate medicinal
ganja and industrial hemp.

Sources: http://jis.gov.jm/law-amended-facilitate-medical-ganja-industry/
http://moj.gov.jm/sites/default/files/laws/The%20Dangerous%20Drugs%20Act.pdf
 Introduction
Breeding and Conservation challenges

• It is difficult to maintain pure strains of high THC or

CBD yielding elite plants under field or greenhouse
conditions.

• Plants are allogamous (cross-fertilization).

• Difficult to maintain elite cultivars by seed.

In Vitro Propagation of
Cannabis sativa
 Advantages of Tissue Culture
Propagation

• Facilitate the conservation of elite cultivars of

Cannabis sativa.

• Better storage method than seeds, which can be

highly variable and produce less efficacious
genotypes (Meijer et al 1992).
 Tissue Culture of Cannabis sp.
Materials and Methods

Plant material/explant
• Nodal segments with axillary buds (approx. 1cm)
• Obtained from healthy branches of high yielding
Cannabis sp.

Sterilization
• Surface sterilize with 0.5% NaOCl (15% v/v bleach) and 0.1 %
Tween 20 for 20 mins.
• 3 five mins rinses in sterile distilled water.

Lata et al 2009
 Tissue Culture
Initiation

Disinfected explants transferred to baby food jars containing

25 ml of sterile Murashige and Skoog’s (MS) medium
(Murashige and skoog 1992).

Supplemented with:

• 3% (w/v) sucrose
• 0.8% (w/v) agar
• 0.5 μM thidiazuron (TDZ)
• pH 5.7

Lata et al 2009b
 Tissue Culture
Multiplication
4-6 weeks after initiation emerging shoots should be
transferred to MS multiplication medium
supplemented with:

• 0.5 μM TDZ
• 7.0 μM Gibberellic acid (GA3)

Lata et al 2009b
 Tissue Culture
Rooting

This should be done on medium containing

• ½ strength MS medium, supplemented with

• 500 mg/L activated charcoal
• 5.0 μM Indole-3-butyric acid (IBA)

Lata et al 2009b
 Tissue Culture
Acclimatization

• Rooted plants should be transferred to Jiffy pots

containing potting mix.
• New growth after 2-3 weeks.

Lata et al 2009b
 Summary

Adopted from Lata et al 2009b

Hydroponic Cultivation
 What is hydroponic cultivation?
•Hydroponics is a method of growing
plants using mineral nutrient solutions
in water, without soil (Douglas 1985).

•Terrestrial plants may be grown with

their roots in the mineral nutrient
solution only or in an inert medium,
such as perlite, gravel, coco peat and
others.
Benefits of Hydroponics Over Soil
• No soil required.
• Lower water requirement as the water can be
reused.
• Allows full control over the plant nutrition levels.
• No fertilizer run off is released into the
environment.
• Stable and high yields.
• Reduces soil-borne pests and diseases.
• Ease of harvesting.
• The pH of nutrient solution can be controlled.
 Disadvantages of Hydroponics
• Requires more knowledge and expertise.
• System problems lead to rapid crop death.
• Requires constant monitoring.
• High humidity favours some pathogens.
• High maintenance costs for indoor systems.
 Types of Hydroponic Systems
• Static solution
• Continuous flow
• Ebb and flow
• Run to waste
• Aeroponics and fogponics
• Passive sub-irrigation
• Deep water culture
 Types of Hydroponic Systems

Source: http://www.nosoilsolutions.com/6-different-types-hydroponic-systems/
 Types of Substrates

• Sand • Perlite
• Expanded clay • Rockwool
aggregate • Growstones
• Coco peat • Rice hulls
• Gravel • Vermiculite
• Brick shards • Pumice
 Perlite
Growstones Rockwool

Expanded clay Pumice

Coco coir
Source: http://ezgrogarden.com/growing-organic/grorock-hydroponic-substrate/
Growing in a Greenhouse
 Greenhouse
A greenhouse is a structural building with different types of
covering materials, such as a glass or plastic roof and
frequently glass or plastic walls.

Purpose:
It heats up because incoming visible sunshine is absorbed
inside the structure.

Air warmed by the heat from warmed interior surfaces is

retained in the building by the roof and wall; the air that is
warmed near the ground is prevented from rising indefinitely
and flowing away.
 Growing Cannabis sp. in a Greenhouse

A Cannabis
sp. specific greenhouse typically has a blackout system to adjust
the photoperiod.

There are two phases of the greenhouse:

Vegetative Phase - The vegetative room offers plants 18 hours of
Flowering
light Phaseof–dark.
and 6 hours To induce flowering
The plants blackout
will grow curtains
forever. are
They grow
used
like to You
trees. maintain 12 hours
can keep strainslight
alive and 12 hoursOnce
for decades. dark.theInplants
4-8
weeks
reach flowers ripen.
a desirable (Grotenhermen
size, they are moved and Russo,2002).
to the flowering phase of
the greenhouse.
 Advantages of Greenhouse
• Ability to manipulate environment
• Protection

From:
a. Wind and pests
b. Extremely hot weather
c. Cross-pollination
d. Excess water/moisture

Source: http://www.maryjanesgarden.com/growing/greenhouse-growing
 Advantages of Greenhouse
Safety
• Plant disease suppression.
• Avoid workers exposing to severe weather
conditions.
• Structure with controlled access and
security features can be installed.
 Advantages of Greenhouse
Improve Growth

• Propagate new plants.

• Increase yields and quality.

Source: http://www.usatoday.com/story/news/nation/2014/08/12/colorado-marijuana-sales/13915859/
Disadvantages of Greenhouse
• Construction Costs
• High Energy/ Operation Costs
• Maintenance
• Tools and supplies – soil, pots etc.
• Human Resources
 Conclusion
•• Hydroponic
Tissue culture can be used
cultivation regenerate
of marijuana a large
offers number of
an alternative
Cannabis
to soil andsativa
has manyplants in a over
benefits shorttraditional
period of time
growing
(Slusarkiewicz-Jarzina
methods (Chandra et alet2010)
al 2005,
. Lata et al 2009a).
• These techniques can be used for germplasm exchange
between laboratories and in breeding to produce high
•• yielding
Tissue
Growingculture
marijuana
canforbethe
varieties inused
apharmaceutical
greenhouse
to conserveallows
elite cultivars
for control
industry. of
Cannabis
over growing
sativa conditions
in vitro (Latainetaal protected
2009b;2012). environment
(Chandra et al 2011).
 References
Angus, G. (2014, September 30). Law to be Amended to Facilitate Medical Ganja Industry. Jamaica Information Service.
Retrieved from http://jis.gov.jm/law-amended-facilitate-medical-ganja-industry/.
Chandra, S., Lata, H., Khan, I. A., & ElSohly, M. A. (2010). Propagation of elite Cannabis sativa for the production of D9-
Tetrahydrocannabinol (THC) using biotechnological tools. Medicinal plant biotechnology, 98-114.
Chandra, S., Lata, H., Khan, I. A., & ElSohly, M. A. (2011). Temperature response of photosynthesis in different drug and
fiber varieties of Cannabis sativa L. Physiology and Molecular Biology of Plants, 17(3), 297-303.
Clarke, R. C., & Pate, D. W. (1994). Medical marijuana. J. International Hemp Asso, 9-12.
Colorado firm partners with Jamaicans to set up medical ganja lab on island. (2014, October 5). The Jamaica Observer.
Retrieved from http://www.jamaicaobserver.com/business/Colorado-firm-partners-with-Jamaicans--to-set-up-medical-
ganja-lab-on-island_17680298.
Douglas, J. S. (1985). Advanced guide to hydroponics (No. new edition). Pelham Books.
Dorm, D. (September 26, 2013). Greenhouse Growing For Efficient Medical Marijuana Production. Retrieved from
http://www.medicaljane.com/2013/09/26/greenhouse-growing-efficient-medical-marijuana-production/.
Dreier, F. (March 31, 2014). Greenhouse Technology Shakes Up Economics of Cannabis Cultivation. In Marijuana Business
Daily. Retrieved from http://mmjbusinessdaily.com/the-basics-of-marijuana-greenhouse-technology/.
Espiritu, K. (2011). Hydroponic Media. Retrieved October 13, 2014, from http://www.epicgardening.com/hydroponic-
growing-media/.
Grotenhermen, F. and Russo, E. (2002). Cannabis and Cannabinoids: Pharmacology, Toxicology, and Therapeutic Potential.
Binghampton , N.Y. Haworth Integrative Healing Press.
 References
Growing Cannabis in a Greenhouse. (n.d.). Retrieved from http://www.dutch-
passion.nl/en/grow-info/growing-cannabis-in-a-greenhouse/.
Haze, N. (n.d.). Soil vs. Hydroponics. Retrieved October 2, 2014, from
http://www.growweedeasy.com/marijuana-soil-vs-hydroponics.
How to Grow Marijuana Hydroponically. (n.d.). Retrieved October 3, 2014, from
http://www.wikihow.com/Grow-Marijuana-Hydroponically
How to Grow Top Shelf Cannabis Using Greenhouses. (n.d.). Retrieved from
http://www.triqsystems.com/resources/grow-top-shelf-cannabis.html.
Hydroponics. (n.d.) Retrieved October 3, 2014, from
http://en.wikipedia.org/wiki/Hydroponics.
Lata, H., Chandra, S., Khan, I. A., & ElSohly, M. A. (2009a). Propagation through alginate
encapsulation of axillary buds of Cannabis sativa L.—an important medicinal plant.
Physiology and Molecular Biology of Plants, 15(1), 79-86.
Lata, H., Chandra, S., Khan, I., & ElSohly, M. A. (2009b). Thidiazuron-induced high-frequency
direct shoot organogenesis of Cannabis sativa L. In Vitro Cellular & Developmental Biology-
Plant, 45(1), 12-19.
Lata, H., Chandra, S., Mehmedic, Z., Khan, I. A., & ElSohly, M. A. (2012). In vitro germplasm
conservation of high Δ9-tetrahydrocannabinol yielding elite clones of Cannabis sativa L.
under slow growth conditions. Acta Physiologiae Plantarum, 34(2), 743-750.
 References
Mehmedic, Z., Chandra, S., Slade, D., Denham, H., Foster, S., Patel, A. S., & ElSohly, M. A.
(2010). Potency Trends of Δ9‐THC and Other Cannabinoids in Confiscated Cannabis
Preparations from 1993 to 2008*. Journal of Forensic Sciences, 55(5), 1209-1217.
Murashige, T and Skoog, F. (192). A revised medium for rapid growth and bioassays with
tobacco tissue culture. Physiologia Plantarum 15: 473-497.
Neita, L. (2014, October 5). The Greening of Jamaica. The Jamaica Observer. Retrieved from
http://www.jamaicaobserver.com/columns/The-greening-of-Jamaica_17676447.
Quimby M. 1974. Botany of Cannabis sativa. In: Mateos-Gomez JL (ed) Archivos be
investigacion medica/ El Instituto Mexicano del Seruto Social.
Russo, E. B. (2013). Cannabis and cannabinoids: pharmacology, toxicology, and therapeutic
potential. Routledge.
Slusarkiewicz-Jarzina, A. U. R. E. L. I. A., Ponitka, A., & Kaczmarek, Z. (2005). Influence of
cultivar, explant source and plant growth regulator on callus induction and plant
regeneration of Cannabis sativa L. Acta Biol Craco Series Bot, 47(2), 145-151.
THANK YOU!

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