breeding and agronomical techniques to ...gianpaolo grassi crea-ci, rovigo’s branch station 15th...
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Gianpaolo Grassi
CREA-CI, Rovigo’s branch station
15th of November 2018 (Vienna)
BREEDING AND AGRONOMICAL TECHNIQUES TO PRODUCE MEDICAL CANNABIS
Universität Innsbruck
Institut für Pharmazie/Pharmakognosie
gianpaolo.grassi@crea.gov.it
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Italian pioneers of Cannabis sativa L. breeding
Dr. Domenico Allavena (Bologna 1910-1985)
Obtained the varieties: - Fibranova and - Carmagnola Selezionata (C.S.) On the left an original picture made by Allavena of the Pinnatofidofilla
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• Puglia,• Tuscany• Liguria,• Veneto,• Friuli Venezia Giulia• Abruzzo• Umbria• Marche• Sicily• Emilia Romagna• Trento and Bozen Aut. Reg.• Piedmont
Italian support the cost cannabinoids.
Regions (12/20) that have of the drugs with natural or synthetic
approved the law that
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Female flower
Cannabis sativa L.: Dioecious plant
Medicinal Cannabis origin
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Cannabis accessions collected in Italy (more than 300 strains)
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163
68
153
Cannabis germplasm collection in Italy
Ind. Hemp Landraces Drug type
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Some example of genotypes with interesting characters
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The only one area in open field admitted in Italy to grow Cannabis
with high THC concentration (2013-2018) about 2,000 sqm
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Cannabis genotypes with high THC grown in 2016
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Single female plant sex reverted
♂
Male side
♀
Female side
Breeding method is based on reversion of sex with Ag salt
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Natural photoperiod Controlled photoperiod
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Results achieved by the most advanced breeding by the leading pharmaceutical company in the
Cannabis sector
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In: Handbook of Cannabis: Edited by R. Pertwee, (2016),784 pp.
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Research programs running at the CREA- CI
1. Cannabis without cannabinoids
2. Cannabis with chemotype CBG prevalent
3. Cannabis with chemotype CBD prevalent
4. Cannabis with chemotype THC prevalent
5. Cannabis with chemotype THCV prevalent
6. Cannabis with chemotype CBDV prevalent
7. Cannabis with high content of GLA in seed
8. Cannabis with selected terpenes
9. Cannabis with high yield for indoor and
outdoor condition.
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Gas-chromatogram of Ermo’s extract
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The analysis of Ermo variety could be done using simple test
A colorimetric test allows to identify contamination
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Chemotype of Cannabis strain with THC prevalent
*
* = Internal Standard 0.01% conc.
THC
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Variability of THC content we have available in our accessions
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Mother plants of CINBOL variety registred in the CPVO and used as medicinal cannabis by the Italian state
Cuttings in rooting phase
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Chemotype of Cannabis strain with CBD prevalent
CBD
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Production of CBD in varieties of Italian cannabis grown in pots
*** Uso31 and Santhica, THC = 0
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Chemotype of Cannabis strain with CBG prevalent
CBG
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Chemiotipo di Cannabis con
CBDV/CBD prevalente
CBDV
CBD CBDV
THCV
CBD
THC
Int. Stand.
Ref.
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Chemotype of Cannabis strain with THC/THCV prevalent
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Indoor, hydroponic
SCROG
Vertical farming
How to grow Cannabis?
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THE EFFECT OF LIGHT SPECTRUM ON THE MORPHOLOGY AND CANNABINOID CONTENT
Light Treatments
• Valoya NS1 had the highest portion of blue light, including some UVA light
• Valoya AP673L spectrum had the most red light, spectrum designed for high vegetative growth
• HPS spectrum concentrated in the yellow-orange area of the visible spectrum. Very low blue light
content
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THE EFFECT OF LIGHT SPECTRUM ON
YIELD OF CANNABINOIDS
No significant differences found in the total yield of cannabinoids between the light
treatments or between the two experiments
• When comparing the yield of cannabinoids per Watts of electricity used, LED
treatments had the best results in both trial rounds and HPS the lowest values
Bar graphs (mean and standard deviation) of yield of cannabinoids per Watts (left) and yield of cannabinoids (right). Solid lines and lower case letters are for
trial 1 (blue), and dashed line and upper case letters are for trial 2 (green). Different letters show significant differences (Tukey's HSD, p
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Color change of
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Cannabinoid concentration in the flowers during the plant cycle
Total Cannabinoids. Top
Middle
Basal .
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Flower yield (dry weight) Variety Variety
G-170 G-301
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Accumulation of cannabinoids in Cannabis flowers
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Mature plant Principal fraction obtained
Leaves could be extracted with CO2 SCF
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Localization of the three fields where hemp with high CBD content has been grown
MEDICINAL CANNABIS IN OPEN FIELD
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Environment condition
Month
0,0
5,0
10,0
15,0
20,0
25,0
30,0
35,0
Feb Mar Apr May Jun Jul Aug Sep
Tem
p. °
C
Mean temperatures
Lecce
Bari
Foggia
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0,0
1,0
2,0
3,0
4,0
5,0
6,0
7,0
8,0
9,0
10,0
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20
CB
D (
% d
w)
CBD concentration in the 3 fields
Lecce
Foggia
Bari
Plant n°
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0,00
1,00
2,00
3,00
4,00
5,00
6,00
7,00
Lecce Foggia Bari
Co
nce
n. (
% d
w)
Cannabinoid variation CBD
0,0
0,10
0,20 % Limit
0,30
THC
5,5% CBD
Comparison between CBD and THC content
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Main tasks
Micro-cutting s
1. Fast multiplication 2. Virus eradication 3. Long term storage
Root phase
Adaptation to normal condition
Pots with cannabis plantlets
in vitro culture to improve breeding of Cannabis
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Cell with 2n=2X=20 2n=4X=40
Polyploidy strategy
2n=3X=30
Diploid Three-ploid Tetra-ploid
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Polyploid lines of Cannabis are available
Three chemotypes have been used CBD CBG THC
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First crop of medicinal Cannabis produced in Italy in the April 2015
Thank you for your attention !
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