Omega-3: new opportunities for the canola industry. James Petrie CSIRO
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1 Omega-3: new opportunities for the canola industry James Petrie CSIRO IRC, Saskatoon 2015
2 IRC, Saskatoon 2015 James Petrie, CSIRO
3 Long-chain vs short-chain omega-3? Marine: long-chain EPA, DPA, DHA (C20, 22) Strong health benefits Microalgal primary production Plants: short-chain ALA, SDA (C18) Limited health benefits Low conversion, especially to DHA IRC, Saskatoon 2015 James Petrie, CSIRO 3
4 Why long-chain omega-3? Increasing demand from: Ageing populations High-growth economies Dietary supplement markets (especially preventative health) Pharmaceutical pipeline IRC, Saskatoon 2015 James Petrie, CSIRO 4
5 Long-chain omega-3 supply vs demand Rx Prescribed Intake (3550mg/day) [VALUE] 000 tonnes Ocean capacity Japan model 1020 Mortality model 650 WHO model GOED and FAO data Supply Demand models IRC, Saskatoon 2015 James Petrie, CSIRO 5
6 How do you engineer oil profile? IRC, Saskatoon 2015 James Petrie, CSIRO 6
7 Genes Combinations Crop DNA Constructs Plant Transformation Field IRC, Saskatoon 2015 James Petrie, CSIRO 7
8 Genes Combinations Crop DNA Constructs Plant Transformation Field IRC, Saskatoon 2015 James Petrie, CSIRO 8
9 Long-chain omega-3 biosynthesis IRC, Saskatoon 2015 James Petrie, CSIRO 9
10 DHA biosynthesis organisms IRC, Saskatoon 2015 James Petrie, CSIRO 10
11 Genes Combinations Crop DNA Constructs Plant Transformation Field IRC, Saskatoon 2015 James Petrie, CSIRO 11
12 Rapid assessment of gene combinations 21st ISPL July 7-11, 2014
13 Finding the right gene combination IRC, Saskatoon 2015 James Petrie, CSIRO 13
14 Finding the right gene combination Australian National Algae Culture Collection IRC, Saskatoon 2015 James Petrie, CSIRO 14
15 Genes Combinations Crop DNA Constructs Plant Transformation Field IRC, Saskatoon 2015 James Petrie, CSIRO 15
16 A sustainable, land-based platform Canola: High quality and healthy oil Commercially available in multiple regions Canada/US/AUS >10M Ha Decades of breeding and commercial optimization Efficient plant, high oil yield IRC, Saskatoon 2015 James Petrie, CSIRO 16
17 Genes Combinations Crop DNA Constructs Plant Transformation Field IRC, Saskatoon 2015 James Petrie, CSIRO 17
18 Lots of possible construct designs Gene orientation and spacing Promoter strengths Promoter-gene combinations Timing of pathway expression Avoiding gene silencing How do you test the possibilities? 21st ISPL July 7-11, 2014
19 Rapid assessment of seed constructs 21st ISPL July 7-11, 2014
20 Testing seed constructs rapidly in leaf LEAFY COTYLEDON2 (LEC2) expression triggers embryogenesis Co-infiltrating a seed construct with LEC2 activates seed promoters in leaf Seed Construct Samples 35S mix Minus LEC2 Plus LEC2 SDA 2.0 ± ± ± 0.1 EPA 0.3 ± ± 0 DPA 2.3 ± ± 0.1 DHA 2.5 ± ± 0.2 IRC, Saskatoon 2015 James Petrie, CSIRO 20
21 Progressing from model species to crops Elements FP1: B. napus truncated napin FAE1: A. thaliana Fatty Acid Elongase 1 Cnl1, Cnl2: L. usitatissimum conlinin MAR: Matrix attachment regions TMV transcriptional leader as 5 UTRs IRC, Saskatoon 2015 James Petrie, CSIRO 21
22 Progression from research to crops 16:0 18:1 LA ALA SDA EPA DPA DHA Arabidopsis DHA (T3) % IRC, Saskatoon 2015 James Petrie, CSIRO 22
23 Progression from research to crops 16:0 18:1 LA ALA SDA EPA DPA DHA Arabidopsis DHA (T3) % Camelina DHA (T7) % IRC, Saskatoon 2015 James Petrie, CSIRO 23
24 Progression from research to crops 16:0 18:1 LA ALA SDA EPA DPA DHA Arabidopsis DHA (T3) % Camelina DHA (T7) % B. juncea DHA (T3) % IRC, Saskatoon 2015 James Petrie, CSIRO 24
25 Optimising the construct for Brassica napus IRC, Saskatoon 2015 James Petrie, CSIRO 25
26 Testing seed constructs rapidly in canola LEAFY COTYLEDON2 (LEC2) expression triggers embryogenesis Co-transforming a seed construct with LEC2 stimulates somatic embryogenesis Embryos are predictive of mature seed function and profile IRC, Saskatoon 2015 James Petrie, CSIRO 26
27 Stable, multi-gene construct design Elements FP1: B. napus truncated napin FAE1: A. thaliana Fatty Acid Elongase 1 Cnl1, Cnl2: L. usitatissimum conlinin MAR: Matrix attachment regions TMV transcriptional leader as 5 UTRs IRC, Saskatoon 2015 James Petrie, CSIRO 27
28 Genes Combinations Crop DNA Constructs Plant Transformation Field IRC, Saskatoon 2015 James Petrie, CSIRO 28
29 Germplasm selection is important 25 Copy number by Germplasm Copy number in primary transformants Germplasms IRC, Saskatoon 2015 James Petrie, CSIRO 29
30 Germplasm selection is important 25 Copy number by Germplasm 7 T 0 DHA by Germplasm (1-2 copy) Copy number in primary transformants DHA % Germplasms 0 Germplasms IRC, Saskatoon 2015 James Petrie, CSIRO 30
31 Gene dosage in complex constructs 6 T 0 DHA vs Copy number 18 DHA by Generation (1-2 copies) DHA % 3 DHA % T 0 T 1 T 2 Copy number by digital PCR Seed Generation IRC, Saskatoon 2015 James Petrie, CSIRO 31
32 The new canola oil profile 16:0 18:1 LA ALA SDA EPA DPA DHA Parent Parent IRC, Saskatoon 2015 James Petrie, CSIRO 32
33 The new canola oil profile: fish oil-like DHA 16:0 18:1 LA ALA SDA EPA DPA DHA Parent Parent Low copy % IRC, Saskatoon 2015 James Petrie, CSIRO 33
34 The new canola oil profile: plus a bit more 16:0 18:1 LA ALA SDA EPA DPA DHA Parent Parent Low copy % High copy % IRC, Saskatoon 2015 James Petrie, CSIRO 34
35 The new canola oil profile: is flexible 16:0 18:1 LA ALA SDA EPA DPA DHA Parent Parent Low copy % High copy % SDA/DHA % IRC, Saskatoon 2015 James Petrie, CSIRO 35
36 The new canola oil profile: is flexible 16:0 18:1 LA ALA SDA EPA DPA DHA Parent Parent Low copy % High copy % SDA/DHA % Total Omega-3: 48.1% Total LC Omega-3: 16.0% Total Omega-6: 7.4% Total LC Omega-6: 0.1% Omega-3:6 ratio: 6.5 : 1 LC Omega-3:6 ratio: 135:1 IRC, Saskatoon 2015 James Petrie, CSIRO 36
37 DHA is targeted to TAG sn-1,3 position(s) sn-1,3 DHA sn-2 DHA IRC, Saskatoon 2015 James Petrie, CSIRO 37
38 DHA synthesis during seed development mg / seed Total FA ALA DHA Days after flowering IRC, Saskatoon 2015 James Petrie, CSIRO 38
39 DHA canola seed lipid classes look normal Parent TAG DAG FFA PC PE PA PI MGDG DGDG Non-lipid IRC, Saskatoon 2015 James Petrie, CSIRO 39
40 DHA canola seed lipid classes look normal Parent DHA canola TAG DAG FFA PC PE PA PI MGDG DGDG Non-lipid IRC, Saskatoon 2015 James Petrie, CSIRO 40
41 Genes Combinations Crop DNA Constructs Plant Transformation Field IRC, Saskatoon 2015 James Petrie, CSIRO 41
42 Does it work in the field? Yes IRC, Saskatoon 2015 James Petrie, CSIRO 42
43 Does it work in the field? Yes R² = Field and glasshouse DHA levels are correlated. Field trials are ongoing including yield and oil content trials. Field Site 1 (% DHA) Glasshouse (% DHA) IRC, Saskatoon 2015 James Petrie, CSIRO 43
44 Does it work in the field? Yes 1 Ha of canola at 12% is the DHA yield from 10,000 fish IRC, Saskatoon 2015 James Petrie, CSIRO 44
45 CSIRO Surinder Singh* Allan Green Peter Nichols Pushkar Shrestha Xue-Rong Zhou Thank you James Petrie Nuseed Malcolm Devine Greg Buzza Wenxiang Gao Nelson Gororo Antonio Leonforte Jason McAllister
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