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Opium poppy, Papaver somniferum Alternative products – alternative uses. Phil Larkin CSIRO, Canberra. 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)

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opium poppy papaver somniferum alternative products alternative uses

Opium poppy, Papaver somniferumAlternative products – alternative uses.

Phil Larkin

CSIRO, Canberra

australian poppy industry
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.
top1 mutation
Top1 mutation

CO52 Thebaine

Control 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)

transgenic poppies
Transgenic poppies
  • Biotechnology is another strategy capable of transforming the industry:
    • Increasing morphinan content
    • Novel products
slide7

Transformation of Poppy

Hypocotyl explants

Agrobacterium

Black primary callus

White embryogenic callus

Somatic embryos

metabolic engineering of opium poppy
Metabolic Engineering of Opium Poppy

Increasing alkaloid yield

Engineering novel products

reticuline

thebaine

hpRNAi

morphine

morphine

anticancer

and

antimalarial

activities

analgesics

transgenic over expression of cor codeinone reductase increases yield
Transgenic over-expression of COR (codeinone reductase) increases yield
  • COR gene transcript is increased
  • Morphinan alkaloid content is increased …..
slide11

Cor transgenics

GH2002

GH2003

FIELD2003

GH2004

cor over expression field trial
COR over-expression – Field Trial

* , P<0.05

**, P<0.01

Control n=18, 20 plants per replicate

Transgenics n=6, 20 plants per replicate

metabolic engineering for alternative products
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.
opiate free poppy
Opiate-free poppy

codeinone reductase

opiate free poppy1
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 Biotechnology22, 1559-66.

opiate free poppy2
Opiate-free poppy

Transgenic

poppies

Control

poppies

Morphine, codeine and thebaine replaced by reticuline, codamine, laudanine and laudanosine.

standards

Ls

Ln

Cm

R

T

O

C

M

proposal for afghan poppy industry
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.
poppyseed oil for biodiesel
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.
phase 1 biodiesel from poppy seed oil
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
phase 2 higher value soft biodiesel
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.