A more scientific view of hemp cultivation

1. The Future of Cannabis Genetics
Jonathan Page

2. Cannabis has been used for millennia
Food and oil (hemp) Fibre (hemp)
Drug and Medicine
(“marijuana”)
O
OH
THC

3. O
OH
OH
HO
psychoac=v
ity
+ pain
relief,
appe=te,
an=spas=ci
ty…
O
OH
Cannabinoids
Terpenes
Cannabis produces a complex suite of bioac=ve
metabolites:
The mixture is the medicine

4. How are cannabinoids synthesized?
Cannabis sa(va
(marijuana, hemp)
?

5. Glandular trichomes on cannabis flowers

6. 100 µm

7. Cannabis trichomes Subcu5cular
cavi5es
Secretory
disc cells

8. Trichome-specific gene expression analysis
RNA
Protein

9. Cannabis is the first medicinal plant with a sequenced genome
Cannabis haploid genome is ~820 Mbp
Genome organiza=on
l 2n = 20
l Nine autosomes
l Two sex chromosomes (X and Y)
We sequenced three types:
1. Marijuana ‘Purple Kush’
• ~19% THCA
2. Hemp ‘Finola’
• Low THCA, 4% CBDA
3. Hemp ‘USO-31’
• Low to zero cannabinoids
Collabora5on with Tim Hughes (University
of Toronto)
Published in Genome Biology in November
2011
Ar5cle accessed 97,925 5mes!
www.genomevolu=on.org

10. The cannabinoid biosynthe=c pathway
ü
ü
ü
ü ü
ü
ü

11. Cannabinoid pathway bifurca=on: “Marijuana”
vs hemp
CBGA
CBDA
CBDA synthase
THCA synthase
Marijuana – 18% THCA
Hemp – 4% CBDA
THCA

12. 0
2
4
6
8
10
12
14
16
18
20
0 5 10 15 20 25 30 35
%CBD
% THC
Cannabis chemotypes result from simple gene=cs
• Chemotypes are chemical phenotypes
Chemotype I (high THC)
Chemotype II (THC:CBD)
Chemotype III (high CBD)
n
n

13. Gene=cs of cannabinoid biosynthesis
THCA synthase
THCA synthase
THCA synthase
CBDA synthase
CBDA synthase
CBDA synthase
Chemotype I
(high THC)
Chemotype II
(THC:CBD mix)
Chemotype III
(high CBD)
Fatty Acids
Olivetolic acid
CBGA
THCA CBDA
C10 terpene
THCA
synthase
Heat Heat
THC CBD
CBDA
synthase

14. Classifica=on of Cannabis is (s=ll!) unclear
GENUS
SPECIES?
SUBSPECIES?
FAMILY
Cannabis
sa2va
indica ruderalis sa2va
Cannabaceae
Hemp and marijuana
• many strains and cul5vars
• four chemotypes
Cannabis
Cannabaceae
indica ruderalis sa2va
Hemp and marijuana
• many strains and cul5vars
• four chemotypes

15. Pa5ents and growers differen5ate two general types:
“Indica”
• Short plant with broad leaves
• Seda5ve and “body stone”
“Sa=va”
• Tall plant with narrow leaves
• S5mula5ng and cerebral effects
Marijuana strains: Indica vs Sa=va
• Thousands of strains exist
• Most strains were developed for recrea5onal use
• Pa5ents report that strains differ in their effects

16. Cannabis genotyping
Three main ques=ons:
1. What is the gene5c structure of the genus Cannabis?
2. Are hemp and marijuana gene5cally dis5nct?
3. Are Indica and Sa5va gene5cally dis5nct?
• Collabora5on with Sean Myles (Dalhousie University)
• Accessed DNA samples represen5ng diverse cannabis types:
• 43 hemp
• 81 marijuana samples
• Analyzed using GBS (genotyping-by-sequencing)
• 14,031 single-nucleo5de polymorphisms (SNPs)

17. Cannabis genotyping: “marijuana” and hemp
Sawler et al, PLOS ONE 2015

18. Gene=c diversity is the basis of agricultural innova=on!
• Sources of new gene5c traits such as disease resistance
BUT
Cannabis is undergoing gene=c reduc=on
• Drug prohibi5on has destroyed plants e.g. feral hemp
• Legal exchange of seeds and plants is difficult
• “Dutchifica5on” is replacing landraces across the globe
• Reliance on vegeta5ve propaga5on (cloning)
• Long-term seed viability is limited
• MMPR restricts access to gene5cs
Preserving crop diversity requires seed banks (not just
seed companies…), sustained funding and research efforts
The problem of germplasm - is cannabis
endangered?

19. Cannabis breeding has been a huge success:
• THC levels and yields have increased; CBD too
But has led to a narrowing of gene pool with focus on indoor
cul=va=on and clonal propaga=on
• High THC product with lower levels of other cannabinoids
Gene=c traits for consumers:
• Novel cannabinoid profiles for medical use
• New terpene profiles
• Sensory proper5es
Gene=c traits for growers:
• Disease and pest resistance
• Higher yield, flowering 5me, plant architecture
The future of cannabis breeding

20. Analysis & predic5on
X
Select plants
from DNA & Chemotype 100s of seeds
back to breeder
100,000+
seeds
DNA sequencing
Genomics assisted breeding of cannabis

21. Cannabis patents
Q: Is paten=ng of cannabis strains possible in Canada?
A: No. Canadian law does not allow paten=ng of higher life
forms such as plants.
Registra=on of new varie=es through Plant Breeders Rights is
possible. PBR helps “protect the variety from exploita=on by
others”.
Varie=es must be:
• New
• Dis5nct
• Uniform
• Stable
Currently 6 registered hemp varie=es and 1 registered
marijuana variety

22. Cannabis needs for more science!

23. 23
• Science leadership
• Health Canada licensed lab
• Analy5cal tes5ng and QC
• Genotyping services
• Developing advanced cannabis strains
jpage@anandialabs.com

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