Plant biotechnology applications Current status, socio -economic impact, legal aspects, and public perception Institute of Plant Biotechnology Outreach http://www.ugent.be/we/genetics/ipbo/ [email protected] 11 Course contents Plant biotechnology: history, state of the art Introduction to biosafety and regulatory aspects Food safety (case studies ) Environmental safety (case studies) Introduction to bioethics Public perception, risk and science communication vrijdag 30 november 2012 22 Biotechnology Medical Biotechnology Agricultural Biotechnology Industrial Biotechnology Marine Biotechnology vrijdag 30 november 2012 33 PLANTS ARE LIFE : Food Feed Oxygen Wood Paper Medicinal products Industrial products Ornamentals vrijdag 30 november 2012 44 Global Challenges of the 21st Century ENVIRONMENT OVERPOPULATION CLIMATE CHANGE POVERTY BIODIVERSITY POLLUTION FOOD SECURITY vrijdag 30 november 2012 55 Sustainable Development « A development which meets the needs of the present without compromising the ability of future generations to meet their own needs » World Commission on Environment and Development vrijdag 30 november 2012 66 The challenge of the 21 st Century Present Industrial Sustainable Agriculture Agriculture Yield oriented Multifunctional Monocultures Preservation of the High off -farm input resource base (water Groundwater pollution and soil) Soil degradation Preservation of the biodiversity Other environmental stresses Health farmers How to move towards sustainability maintaining production at secure levels? vrijdag 30 november 2012 77 Rethink the characteristics of fundamental and applied research and favour a strong and dynamic interaction between both “sciences” vrijdag 30 november 2012 88 Marc Van Montagu and Jozef Schell Biotech Crop History 1996 1904, Agrobacterium 1974, Ti 1990’s Commercial tumefaciens Plasmid, Gent genomics launch The discovery of plant transformation • Agrobacterium tumefaciens induces tumors in natural conditions. • These tumors are due to the transfer of a bacterial DNA-fragment = T-DNA into the plant chromosome. • No tumors are formed if the genes for plant hormone biosynthesis in the T-DNA are substituted by other genes, but the T-DNA is still transferred and integrated in plant DNA! vrijdag 30 november 2012 1111 Agrobacterium T-DNA Agrobacterium tumefaciens T-DNA borders Ti plasmid Tumor genes vrijdag 30 november 2012 1212 Agrobacterium T-DNA Agrobacterium tumefaciens T-DNA borders Ti plasmid marker Your favorite gene vrijdag 30 november 2012 1313 New DNA in plants • The introduced DNA is integrated by plant repair enzymes • Event-specific integration site >>> unique transitions • Primers can be designed as unique identifier (part of the dossier for GM-approval in Europe): traceability. T-DNA borders Plant DNA gene marker Construct DNA vrijdag 30 november 2012 1414 The essence of plant biotechnology • T-DNA integration has been studied very well. • Very precise technology: one gene can be isolated from one organism and introduced in another, this new gene is stably integrated and inherited as any other gene. • Other transformation methods have been developed using physical means. • Most of the current transgenic crops (corn, soybean,…) have been transformed using physical methods. vrijdag 30 november 2012 1515 CREATING VALUE WITH GENETIC ENGINEERING Input Traits •Biotic stresses: Insect resistance; Nematode resistance; Striga resistance; Tolerance towards bacterial, fungal and viral infections •Abiotic stresses: Better adaptation towards drought- salinity-cold, low nutrients, water logging, heavy metals vrijdag 30 november 2012 1616 CREATING VALUE WITH GENETIC ENGINEERING (2) Physical Traits Maturity; Plant architecture; Pod shattering; Shelf life Output Traits Content and quality of starch, protein, oil, nutritional elements vrijdag 30 november 2012 1717 Plant Genetics Systems 1982 Vaeck et al ., 1987 Insect resistant tobacco Plant Genetics Systems 1982 De Block et al ., 1987 BBasta resistant tobacco vrijdag 30 november 2012 2020 GM plants with insect resistance BtBt--proteinprotein toxic for insect larvae Bacillus thuringiensis biological pesticide environmentally safe Engineer the plant to produce BtBt--proteinprotein vrijdag 30 november 2012 2121 Slide courtesy of vrijdagRob 30 Fraley, november 2012 Monsanto2222 1995: ‘Roundup-Ready’ soya vrijdag 30 november 2012 2323 Enzyme A Round -up Enzyme A Enzyme A* vrijdag 30 november 2012 2424 vrijdag 30 november 2012 2525 Longer shelf life Flavr Savr Tomatoes vrijdag 30 november 2012 2626 vrijdag 30 november 2012 2727 How can biotechnology help for Striga? Farmers can’t afford herbicides and sprayers to spray over whole fields Approach: • Use herbicide tolerant maize • Treat crop seeds with herbicides instead of fields vrijdag 30 november 2012 2828 Papaya ringspot virus Worldwide, PRSV is the most deadly papaya virus in 1997 PRSV had decimated Hawaii’s fifth largest crop, papaya Production had fallen by nearly 40 percent Hawaii’s $17 million papaya industry was in danger vrijdag 30 november 2012 2929 Papaya ringspot virus Researchers inserted the virus coat protein gene in Papaya : gives virus resistance (similar to the way a vaccine makes people immune to disease) Research is also underway in other papaya growing regions to use biotechnology to combat papaya diseases. Four years after the introduction of a GM papaya with virus resistance, papaya production is back to old levels in Hawaii vrijdag 30 november 2012 3030 Second (and third) generation of GM plants multiple traits functional food (health food) eg omega-3 fatty acids, vitamins renewable and raw materials eg potatoes which modified starch content, corn with bioplastics, modification fatty acid composition, poplars with altered lignin content molecular pharming vrijdag 30 november 2012 3131 Molecular f(ph)arming: Growth of plants for the production of molecules: eg antibodies, edible vaccines, pharmaceuticals, industrial proteins vrijdag 30 november 2012 3232 vrijdag 30 november 2012 3333 Sustainable Industry Nature Biotechnol . 20:778 (2002) vrijdag 30 november 2012 3434 Tailoring the Plant to Fit the Soil • Growing nutrient-efficient plants on soils low in plant available nutrients • Improved match between cropping patterns and physical limitation of agricultural lands, to ensure long -term sustainability of production levels • Plant that are tolerant to toxic elements e.g Al vrijdag 30 november 2012 3535 Acacia albida, miracle tree of the Sahel vrijdag 30 november 2012 3636 Drought stress-inducible genes and their possible functions in stress tolerance and response FUNCTIONAL REGULATORY PROTEINS PROTEINS Membrane proteins Transcription factors (water channel (MYC, MYB, bZIP, protein, transporters) EREBP/AP2 ) Drought Proteinases (cytoplasm Protein kinases , chloroplast) stress (MAPK, MAPKKK, CDPK, S6K) Protection factors of macromolecules Protein phosphatases (Chaperones, LEA (PTP) proteins) PI turnover Osmoprotectant Detoxification (phospholipase C, synthases (proline, enzymes (GST, PIP5K, DGK, PAP) Glybetaine, sugar) sEH, SOD) ((Shinozaki and YamaguchiYamaguchi--Shinozaki,Shinozaki, 1999)vrijdag 30 november 2012 3737 Functional Food: Golden Rice Research Prof. Dr. Ingo Potrykus:: Golden Rice contains the genes required to activate the biochemical 2012 30 november vrijdag pathway leading to ß-carotene. The intensity of the color represents the concentration. 3838 Comparitive and Evolutionary GenomicsPhylogenetic Tree--Tree Tree Tree up to date as fromSeptember 4th 2012 CoGePedia http://genomevolution.org/wiki/index.php/Sequenced_plant_genomes BGI (formerly known as the Beijing Genomics Institute) has purchased 128 Illumina HiSeq 2000 sequencing system, which promises to generate up to 200 gigabases of sequence data per run. China's BGI plans to sequence the genomes of 1,000 important plant and animal reference species over the next two years as part of a new project that will solicit proposals from the scientific community. Acquisition puts Beijing Genomics Institute on path to become world’s largest sequencing facility The KBBE Need for sustainable supply of food, raw materials and energy Tremendous progress in life sciences and biotechnology , fundamental and applied knowledge Biotechnology (industrial) has been recognised as one of the key enabling technologies (KETs, 2009) Efficient use of plant genome data offers unprecedented potentials for crop improvement vrijdag 30 november 2012 4242 Plant Genomics 1. Structural genomics : Genome sequencing and EST’s 2. Functional genomics : • Expression analysis tools: micro-arrays and transcript profiling • Insertion mutagenesis • SNP mapping (Tilling) • Map-based cloning vrijdag 30 november 2012 4343 How Traitmill ® Achieves High Throughput assembly line approach automation & robotization continuous efficiency improvement Vector construction transformation plant evaluation vrijdag 30 november 2012 4444 Power of Novel Methods in Mass Spectrometry for Proteomics ••in micro sequencing ••in detecting protein modifications ••protein turnover ••biosynthetic pathway analyses Biocartis Innovative Systems Molecular Diagnostics Platform can detect and quantify multiple DNA-- or RNA--basedbased biomarkers in a wide variety of patient sample types with minimal user intervention vrijdag 30 november 2012 4747 Molecular Tools for Assessing Levels of Genetic Diversity DNA fingerprint • AFLP • Single -locus DNA fingerprinting • Nucleotide sequencing Proteomic profilling Metabolic profilling vrijdag