Re-wiring plant genetic circuits - a paradigm shift from metabolic - PowerPoint PPT Presentation

Re-wiring plant genetic circuits - a paradigm shift from metabolic engineering to synthetic biology in plants

Genes and standard components in plant engineering

In the dark 3 weeks Callus ready Immature Immature embryo (2,4 D) for bombardment embryo derived callus Corn seeds = 14 dap Callus tissue under Corn transformation using Donor plant phosphinothrici n selection direct DNA transfer 3-4 weeks darkness To the light Callus regenerating shoots Plant on rooting media Transgenic plant 3-4 weeks 3-4 cm tall

Combinatorial nuclear transformation generates a metabolic library for the investigation of complex and multi-step biosynthetic pathways and the accumulation of specific combinations of metabolites C Zhu T Capell Koreen Ramessar

psy1+crtI psy1+crtI+lycb+bch+crtW C Zhu T Capell Endosperm specific expression G Farre S Naqvi

High provitamin A corn; 100-200 g of grain provide full RDI of β- carotene (as a sole source of vitamin A)

Mo17 EZ6 EZ59 B73 A632 7300 kg/ha 5000 kg/ha 2100 kg/ha 6600 kg/ha 4100 kg/ha 11500 kg/ha 14900 kg/ha 9400 kg/ha 12400 kg/ha 10740 kg/ha 8100 kg/ha 8165.3 kg/ha 5300 kg/ha HC HCxMo17 HCxEZ6 HCxEZ59 HCxB73 HCxA632 Commercial hybrid %H: 0.27 %H: 2.33 %H: 3.06 %H: 1.57 %H: 1.70

R Alvez E Vilaprinyo

Workflow for data-driven modeling of carotenoid biosynthesis in modified maize lines. For each line, experimental data were collected and used to interpolate dense time series of gene expression and metabolite concentrations. These time series were used to estimate the parameter values for an optimized mathematical model that best fits the experimental results for that line. A sensitivity analysis of the optimized model identified the genes whose overexpression was most likely to increase the accumulation of each metabolite. The expression of these genes was doubled and simulations were run to determine the effects of that doubling on metabolite concentration.

Applications: Poultry feed

Chicken feeding trials performed by J Diaz-Gomez, C Nogareda, JA Moreno & E Angulo, UdL Animal Production Dept High carotenoid maize

Time course of egg development in laying hens BKT-keto diet HC-High Carotenoid diet

J Estany Higher and more mono-unsaturated intramuscular fat without increasing total fat content. Intramuscular to peripheral fat

The transition from metabolic engineering to synthetic biology in plants

Genome Editing in plants

What is the outcome of editing? NHEJ HR + donor DNA + donor DNA + ssODN + donor DNA + + + + small indels cassette insertion cassette insertion gene conversion genome reorganizations large deletion inversion translocation

Intervention strategies • Supplements • Fortification • Biofortification

Nutritional engineering • Most populations are simultaneously deficient for multiple nutrients • Solving one nutritional deficiency merely shifts the problem to another • The only way to avoid this is to simultaneously engineer staple crops to accumulate all essential nutrients – the holistic approach