the future of cannabis genetics
TRANSCRIPT
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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