The Future of Cannabis Genetics

The Future of Cannabis Genetics

Jonathan Page

Cannabishasbeenusedformillennia

Foodandoil(hemp) Fibre(hemp)

DrugandMedicine(“marijuana”)

O

OH

THC

O

OHOH

HO

psychoac=vity

+painrelief,

appe=te,an=spas=ci

ty…

O

OH

Cannabinoids

Terpenes

Cannabisproducesacomplexsuiteofbioac=vemetabolites:

Themixtureisthemedicine

Howarecannabinoidssynthesized?

Cannabissa(va(marijuana,hemp)

?

Glandulartrichomesoncannabisflowers

100 µm

CannabistrichomesSubcu5cularcavi5es

Secretorydisccells

Trichome-specificgeneexpressionanalysis

RNA

Protein

Cannabisisthefirstmedicinalplantwithasequencedgenome

Cannabishaploidgenomeis~820MbpGenomeorganiza=on

l  2n=20l  Nineautosomesl  Twosexchromosomes(XandY)

Wesequencedthreetypes:1.  Marijuana‘PurpleKush’

•  ~19%THCA2.  Hemp‘Finola’

•  LowTHCA,4%CBDA3.  Hemp‘USO-31’

•  Lowtozerocannabinoids

Collabora5onwithTimHughes(UniversityofToronto)PublishedinGenomeBiologyinNovember2011Ar5cleaccessed97,9255mes!

www.genomevolu=on.org

Thecannabinoidbiosynthe=cpathway

ü

ü

ü

ü ü

ü

ü

Cannabinoidpathwaybifurca=on:“Marijuana”vshemp

CBGA

CBDA

CBDAsynthase

THCAsynthase

Marijuana–18%THCA

Hemp–4%CBDA

THCA

0

2

4

6

8

10

12

14

16

18

20

0 5 10 15 20 25 30 35

% C

BD

% THC

Cannabischemotypesresultfromsimplegene=cs•  Chemotypesarechemicalphenotypes

ChemotypeI(highTHC)

ChemotypeII(THC:CBD)

ChemotypeIII(highCBD)

nn

Gene=csofcannabinoidbiosynthesis

THCAsynthase

THCAsynthase

THCAsynthase

CBDAsynthase

CBDAsynthase

CBDAsynthase

ChemotypeI(highTHC)

ChemotypeII(THC:CBDmix)

ChemotypeIII(highCBD)

Fatty Acids

Olivetolic acid

CBGA

THCA CBDA

C10 terpene

THCA synthase

Heat Heat

THC CBD

CBDA synthase

Classifica=onofCannabisis(s=ll!)unclear

GENUS

SPECIES?

SUBSPECIES?

FAMILY

Cannabis

sa2va

indica ruderalissa2va

Cannabaceae

Hempandmarijuana• manystrainsandcul5vars

• fourchemotypes

Cannabis

Cannabaceae

indica ruderalissa2va

Hempandmarijuana• manystrainsandcul5vars

• fourchemotypes

Pa5entsandgrowersdifferen5atetwogeneraltypes:

“Indica”•  Shortplantwithbroadleaves•  Seda5veand“bodystone”

“Sa=va”

•  Tallplantwithnarrowleaves•  S5mula5ngandcerebraleffects

Marijuanastrains:IndicavsSa=va•  Thousandsofstrainsexist•  Moststrainsweredevelopedforrecrea5onaluse•  Pa5entsreportthatstrainsdifferintheireffects

Cannabisgenotyping

Threemainques=ons:1.  Whatisthegene5cstructureofthegenusCannabis?2.  Arehempandmarijuanagene5callydis5nct?3.  AreIndicaandSa5vagene5callydis5nct?

•  Collabora5onwithSeanMyles(DalhousieUniversity)

•  AccessedDNAsamplesrepresen5ngdiversecannabistypes:•  43hemp•  81marijuanasamples

•  AnalyzedusingGBS(genotyping-by-sequencing)

•  14,031single-nucleo5depolymorphisms(SNPs)

Cannabisgenotyping:“marijuana”andhemp

Sawleretal,PLOSONE2015

Gene=cdiversityisthebasisofagriculturalinnova=on!•  Sourcesofnewgene5ctraitssuchasdiseaseresistance

BUT

Cannabisisundergoinggene=creduc=on

•  Drugprohibi5onhasdestroyedplantse.g.feralhemp•  Legalexchangeofseedsandplantsisdifficult•  “Dutchifica5on”isreplacinglandracesacrosstheglobe•  Relianceonvegeta5vepropaga5on(cloning)•  Long-termseedviabilityislimited•  MMPRrestrictsaccesstogene5cs

Preservingcropdiversityrequiresseedbanks(notjustseedcompanies…),sustainedfundingandresearchefforts

Theproblemofgermplasm-iscannabisendangered?

Cannabisbreedinghasbeenahugesuccess:•  THClevelsandyieldshaveincreased;CBDtoo

Buthasledtoanarrowingofgenepoolwithfocusonindoorcul=va=onandclonalpropaga=on

•  HighTHCproductwithlowerlevelsofothercannabinoidsGene=ctraitsforconsumers:

•  Novelcannabinoidprofilesformedicaluse•  Newterpeneprofiles•  Sensoryproper5es

Gene=ctraitsforgrowers:

•  Diseaseandpestresistance•  Higheryield,flowering5me,plantarchitecture

Thefutureofcannabisbreeding

Analysis&predic5on

XSelectplants

fromDNA&Chemotype 100sofseedsbacktobreeder

100,000+seeds

DNAsequencing

Genomicsassistedbreedingofcannabis

Cannabispatents

Q:Ispaten=ngofcannabisstrainspossibleinCanada?A:No.Canadianlawdoesnotallowpaten=ngofhigherlife

formssuchasplants.Registra=onofnewvarie=esthroughPlantBreedersRightsispossible.PBRhelps“protectthevarietyfromexploita=onbyothers”.Varie=esmustbe:

•  New•  Dis5nct•  Uniform•  Stable

Currently6registeredhempvarie=esand1registeredmarijuanavariety

Cannabisneedsformorescience!

23

•  Scienceleadership•  HealthCanadalicensedlab•  Analy5caltes5ngandQC•  Genotypingservices•  [email protected]

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