Opium poppy, Papaver somniferum Alternative products – alternative uses. Phil Larkin CSIRO,...
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Transcript of Opium poppy, Papaver somniferum Alternative products – alternative uses. Phil Larkin CSIRO,...
![Page 1: Opium poppy, Papaver somniferum Alternative products – alternative uses. Phil Larkin CSIRO, Canberra.](https://reader035.fdocuments.us/reader035/viewer/2022081418/56649e4d5503460f94b431cb/html5/thumbnails/1.jpg)
Opium poppy, Papaver somniferumAlternative products – alternative uses.
Phil LarkinCSIRO, Canberra
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Australian poppy industry
• >50% of world’s legally traded opiates
• Demonstrated the power of genetics to transform the industry to new products– Thebaine poppies (top1 mutation)– Thebaine feedstock for high value painkillers, such as
buprenorphine, oxycodone.
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Top1 mutation
CO52 ThebaineControl mutant
Zero morphine and codeine.
Proprietary mutation of Tasmanian Alkaloids
Morphine-pathway block in top1 poppies.Millgate, Pogson, Wilson, Kutchan, Zenk, Gerlach, Fist, Larkin.Nature 431:413-414 (2004)
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Transgenic poppies
• Biotechnology is another strategy capable of transforming the industry:– Increasing morphinan content– Novel products
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Hypocotyl explants
Agrobacterium
Black primary callusWhite embryogenic
callusSomatic embryos
Transformation of Poppy
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Metabolic Engineering of Opium Poppy
Increasing alkaloid yield
thebaine
morphine
analgesics
Engineering novel products
reticuline
morphine
hpRNAi anticancerand antimalarialactivities
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0
1
2
3
4
5
6
NTg Tg
#1b
NTg Tg
#2
n=2
n=6
n=1
n=7
CO
R t
ran
sc
rip
t(n
orm
alis
ed a
rbit
rary
un
its
)
Transgenic over-expression of COR (codeinone reductase) increases yield
• COR gene transcript is increased
• Morphinan alkaloid content is increased …..
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Control - -
206-
4-1a
206-
4-1b
206-
4-2 -
206-
4-4
206-
4-7
206-
4-8
0.0
0.5
1.0
1.5
2.0
2.5
3.0
29%*** **
**
alka
loid
%D
W
Control 2
Control 3
Control m
ean
206-
4-1a
206-
4-1b
206-
4-2
206-
4-3
206-
4-4
206-
4-7
206-
4-8
0.0
0.5
1.0
1.5
2.0
2.5
3.0
27%
* *
alka
loid
%D
W
Control 2
Control 3
Control m
ean
206-
4-1a
206-
4-1b
206-
4-2
206-
4-3
206-
4-4
206-
4-7
206-
4-8
0.0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
28%* * **
* * *
alka
loid
%D
W
GH2002 GH2003
FIELD2003
Cor transgenics
GH2004
CO58-3
4
206-
4-1b
206-
4-1b
HOMa
206-
4-2
206-
4-2H
OMa
206-
4-2H
OMb
206-
4-2H
OMc
206-
4-3
206-
4-7
0.0
0.5
1.0
1.5
2.0
2.5
3.0
3.5 3 3 3 3 3 3 3
40%
Alk
alo
id %
DW
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COR over-expression – Field TrialField 2003% DW
Morphine Codeine Thebaine Total
CO58-34control
2.83 0.19 0.21 3.25
1a 3.43 * 0.24 0.22 3.91 *
1b 3.32 0.31 * 0.32 3.97 *
2 3.44 * 0.31 * 0.38 4.17 **
3 3.31 0.30 * 0.28 3.95 *
4 3.38 * 0.27 0.26 3.92 *
7 3.45 * 0.25 0.18 3.89 *
8 3.16 0.22 0.19 3.60* , P<0.05**, P<0.01
Control n=18, 20 plants per replicateTransgenics n=6, 20 plants per replicate
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Metabolic engineering for alternative products
• Zero opium poppy by genetic engineering
• CSIRO gene silencing technology to shut down all opiates.– Hairpin RNAi – E.g. Target gene: codeinone reductase– Accumulation of alternative non-opiate
product, reticuline.
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Opiate-free poppy
codeinone reductase
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Opiate-free poppy
Potential feedstock for other bioactives
Allen, RS et al. (2004) RNAi-mediated replacement of morphine with nonnarcotic alkaloid reticuline in opium poppy.Nature Biotechnology 22, 1559-66.
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Opiate-free poppy
T
O
C
M
LsLn
CmR
Transgenicpoppies
Control poppiesstandards
Morphine, codeine and thebaine replaced by reticuline, codamine, laudanine and laudanosine.
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Proposal for Afghan Poppy Industry
Phased transformation to a legal non-opiate industry:
• Phase 1. Biodiesel from poppy seed oil.• Phase 2. Higher value “soft” biodiesel.• Phase 3. Industrial and pharmaceutical products.
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Poppyseed oil for biodiesel.
• Current world diesel consumption 1087 billion litres p.a.• Biodiesel currently accounts for 0.4%.• Poppy yields 1.8 tonnes/ha seed (cf. 2.4 for canola)• 45-50% oil content (cf. 40% in canola)• 0.8 tonnes oil per ha.• Afghan poppy area in 2004 was 131,000 ha
– <2 % of arable land– Would produce 100,000 tonnes of oil – = 2.5% of current world biodiesel consumption.– Scope to expand production greatly.
• Protein “meal” byproduct as food and feed.
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Phase 1. Biodiesel from poppy seed oil.
• Replacement seed of opiate-free poppies mutation– Mutant cv. Sujata, opiate-free, opium-free poppy
(CSIR Lucknow, India)• Village-scale oil press and biodiesel processing• Local energy supply
– Transport fuel– Electricity generators
• Basis of a carbon-neutral export biodiesel industry.• Low tech and high volume• Requiring minimal adjustment from the poppy growing culture and
expertise.• Begins the process of legalising the industry:
– Agronomic support– Potential Carbon subsidies– Breeding support– Trade support
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Phase 2. Higher value “soft” biodiesel.
Value adding to poppy oil with biotechnology• Increased oil yield
– Overexpressing DAGAT2 (boost transfer of last acyl to glycerol)
• High oleic biodiesel.– Polyunsaturates can form oxidised polymers which reduce engine
performance.
– High oleic oils (monounsaturates) are preferred.
– Well attested CSIRO technology used to produce high oleic oils in soybean, canola, flax and cottonseed oils:
• RNAi silencing 12-desaturase (high oleic)• RNAi silencing FATB thioesterase (eliminate palmitic)
• High oleic poppy oil also a healthier food oil.
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