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The Future of Cannabis Genetics


Jonathan Page
Cannabis	has	been	used	for	millennia	
Food	and	oil	(hemp)	 Fibre	(hemp)	
Drug	and	Medicine	
(“marijuana”)	
O
OH
THC
O
OH
OH
HO
psychoac=v
ity	
+	pain	
relief,	
appe=te,	
an=spas=ci
ty…	
O
OH
Cannabinoids	
Terpenes	
Cannabis	produces	a	com...
How	are	cannabinoids	synthesized?	
Cannabis	sa(va		
(marijuana,	hemp)	
?
Glandular	trichomes	on	cannabis	flowers
100 µm
Cannabis	trichomes	Subcu5cular	
cavi5es	
Secretory	
disc	cells
Trichome-specific	gene	expression	analysis	
RNA	
Protein
Cannabis	is	the	first	medicinal	plant	with	a	sequenced	genome	
Cannabis	haploid	genome	is	~820	Mbp	
	
Genome	organiza=on	
l...
The	cannabinoid	biosynthe=c	pathway	
ü	
ü	
ü	
ü	 ü	
ü	
ü
Cannabinoid	pathway	bifurca=on:	“Marijuana”	
vs	hemp	
CBGA	
CBDA	
CBDA	synthase	
THCA	synthase	
Marijuana	–	18%	THCA	
Hemp...
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...
Gene=cs	of	cannabinoid	biosynthesis	
THCA	synthase	
THCA	synthase	
THCA	synthase	
CBDA	synthase	
CBDA	synthase	
CBDA	synth...
Classifica=on	of	Cannabis	is	(s=ll!)	unclear	
GENUS	
SPECIES?	
SUBSPECIES?	
FAMILY	
Cannabis	
sa2va	
indica	 ruderalis	sa2v...
Pa5ents	and	growers	differen5ate	two	general	types:	
	
	 	“Indica”		
•  Short	plant	with	broad	leaves	
•  Seda5ve	and	“body...
Cannabis	genotyping	
Three	main	ques=ons:	
1.  What	is	the	gene5c	structure	of	the	genus	Cannabis?	
2.  Are	hemp	and	marij...
Cannabis	genotyping:	“marijuana”	and	hemp	
Sawler	et	al,	PLOS	ONE	2015
Gene=c	diversity	is	the	basis	of	agricultural	innova=on!	
•  Sources	of	new	gene5c	traits	such	as	disease	resistance	
	
	B...
Cannabis	breeding	has	been	a	huge	success:	
•  THC	levels	and	yields	have	increased;	CBD	too	
	
But	has	led	to	a	narrowing...
Analysis	&	predic5on	
X	
Select	plants	
from	DNA	&	Chemotype	 100s	of	seeds	
back	to	breeder	
100,000+		
seeds	
DNA	sequen...
Cannabis	patents	
Q:	Is	paten=ng	of	cannabis	strains	possible	in	Canada?	
A:		No.	Canadian	law	does	not	allow	paten=ng	of	...
Cannabis	needs	for	more	science!
23
•  Science	leadership	
•  Health	Canada	licensed	lab	
•  Analy5cal	tes5ng	and	QC	
•  Genotyping	services	
•  Developing...
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The Future of Cannabis Genetics

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A more scientific view of hemp cultivation

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The Future of Cannabis Genetics

  1. 1. The Future of Cannabis Genetics Jonathan Page
  2. 2. Cannabis has been used for millennia Food and oil (hemp) Fibre (hemp) Drug and Medicine (“marijuana”) O OH THC
  3. 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. 4. How are cannabinoids synthesized? Cannabis sa(va (marijuana, hemp) ?
  5. 5. Glandular trichomes on cannabis flowers
  6. 6. 100 µm
  7. 7. Cannabis trichomes Subcu5cular cavi5es Secretory disc cells
  8. 8. Trichome-specific gene expression analysis RNA Protein
  9. 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. 10. The cannabinoid biosynthe=c pathway ü ü ü ü ü ü ü
  11. 11. Cannabinoid pathway bifurca=on: “Marijuana” vs hemp CBGA CBDA CBDA synthase THCA synthase Marijuana – 18% THCA Hemp – 4% CBDA THCA
  12. 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. 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. 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. 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. 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. 17. Cannabis genotyping: “marijuana” and hemp Sawler et al, PLOS ONE 2015
  18. 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. 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. 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. 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. 22. Cannabis needs for more science!
  23. 23. 23 •  Science leadership •  Health Canada licensed lab •  Analy5cal tes5ng and QC •  Genotyping services •  Developing advanced cannabis strains jpage@anandialabs.com
  24. 24. AFRIS. AsianFoodRegulationInformationService. We have the largest database of Asian food regulations in the world and it’s FREE to use. We publish a range of communication services, list a very large number of food events and online educational webinars and continue to grow our Digital Library. We look forward to hearing from you soon! www.asianfoodreg.com adrienna@asianfoodreg.com

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