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Autar K. Mattoo USDA-ARS Sustainable Agriculture Systems Laboratory Beltsville, Maryland [email protected] Agricultural Biotechnology & Sustainability Princeton Symposium: April 29, 2009 Trends, Signs and Signatures from the Earth Present and Future World Population Trends 56% 10% 50% 120% -5% 42% 39% • Produce more:develop capacity to feed 10 billion people within the next 40 to 50 years • Declining trend in crop yields • Increasing water shortage worldwide Global Climate Change - Annual Cereal Yields – Declining trend Affects fruit production Source: VR Reddy et al Source: IFPRI Trends That Shape Our Future • Precision based genetic manipulation – Agricultural Biotechnology • Diets with pro-health nutrients – Preventive & Personalized ‘Medicine’ • Systems approach to crop production Sustainable Agriculture Production • Rational integration of GM & sustainable agriculture Agriculture for Health Engineering Tomato Fruit for Nutritional Attributes for Human Health ANTIOXIDANTS - NUTRIENTS Pro-Vitamin A - β-Carotene, Flavonoids, Phenolics, Lutein, Lycopene AMINO ACIDS Arginine, Glutamate, Glutamine, Isoleucine, Lysine, Methionine, Threonine MICRONUTRIENTS Calcium, Choline, Iron, Zinc WHAT IS NEW & EXCITING • Nutrition-enriched food potentially decreases incidence of diet-related diseases. • Multiple interactions among fruit nutrients positively influence antiproliferative activity compaed to an isolated antioxidant. • The nutritional molecules include: vitamins (B, C, E and β-carotene), folates, lycopene, flavonoids, isothyocyanates/glucosinolates, polyphenols, glutathione and minerals. HOWEVER, antioxidant levels present in fruits are low and influenced by genotype/ cultivar, growth condition and developmental stage Total antioxidant capacity of some fruits FRAP value1 B/C B RB SB 4.05.07 2.33.06 2.16 1.06-1. 6.88* 25 1Compare Peach Kiwi Plum Pineapple 0.911.33 Apricot Mango Tomato Apple Banana Pear Melon 0.52 <0.2 0.35 these values to ground cloves with FRAP = 125.55 nmol 100g-1 FW From Shukla & Mattoo 2009 Halvorsen et al. 2002 J Nutr Halvorsen et al. 2006 Am J Clin Nutr The HOPE • Genetic engineering - a refined tool to achieve antioxidant capacity to recommended levels. • GM makes new resource possible for studying relationships between diet, genetics and metabolism. • Interweaving biotech products and transcriptomeproteome-metabolome analysis should ease society’s concerns and open the GM horticulture market. • Precision-based engineering of biochem pathways have led to desired levels of antioxidants in fruits & other crops. Precision in Designed Gene Construct Ripening-Specific Expression of Yeast SAM Decarboxylase A. Gene Construct NOS promoter NPT II sequence B. Validation NOS terminator NOS E8 SAMdc promoter coding sequence terminator C. Developmental Regulation MG BR TU PK RD MG BR TU PK RD MG BR TU kb PK RD 1.5 SAMdc E8 exon1 Mehta et al. rDNA Nature Biotechnology (2002) 20: 613-618 1.5 Wild type 556HO 579HO Attributes of Transgenic Tomato Fruits Longer Vine Life – multiple harvesting Processing Quality – Higher by 50% Antioxidant Capacity & Lycopene Content Increased in Transgenic Tomato Fruit Genotype Azygous Control 579HO Transgenic FRAP value 0.57+0.11 0.95+0.08 Fold Lycopene Fold Increase mg-1 gFW Increase 25-50 1.67 125-155 2-3 Mehta et al. 2002. Nature Biotechnol; Fatima et al. 2009 Unpublished. Engineered value-added transgenic tomatoes • higher antioxidant levels • 50% improved juice quality, & • longer vine life Risk Assessment – Technologies Used Global gene expression Proton-NMR Spectroscopy DNA macroarrays to evaluate unusual Proteins – immunoblots resonances Metabolite profiles Srivastava et al. 2007 Plant Biotechnol Mattoo et al. 2007 J AOAC Intl Mattoo et al. 2006 Plant Physiol High antioxidant GM tomato fruit extends life span and protects against cardiovascular disease • Anthocyanin GM tomato fed to cancer-susceptible Trp53 knock-out mice extended their life span Butelli et al. 2008 Nat Biotechnol 26:1301 • Flavonoid-rich GM tomato fed to human Creactive protein transgenic (CRPtg) mice reduced basal human CRP concentration compared to the wild type tomato – Rein et al. 2006 J Nutr 136:2331 Note: The mice were fed a diet that had tomato quantity supplied at a level achievable in human diet. Conventional agriculture • New varieties and higher crop production HOWEVER, its heavy reliance on chemical inputs: synthetic fertilizer, pesticides & fossil fuel use Has negatively impacted World ecosystems; Human and animal health; Top soil; Soil fertility; Contamination of major natural resources, water and air ADDED CONCERNS: Rising energy costs; Global climate change & Increasingly scarce production resources AND, As if that is not enough….. We will be over 10 billion by 2050 in a much different climate than what we have today We need to produce enough goods and services in a sustainable way Building soil organic matter to build capacity for nutrients and water resources for crops • Build species diversity to manage pests & weeds • “Feed the soil, not the crop” Long-term systems approach to build the fertility & soil resilience to reduce synthetic chemical input for crop production Hairy vetch, an annual legume: • A beneficial cover crop • Fits well into different cropping rotations • Is capable of high nitrogen fixation • Produces substantial biomass Black Plastic 82 96 108 96 108 Hairy Vetch Mulch 82 Practice of using cover crop mulches: An alternative, eco-friendly production system 1. Enriches soil with high organic residues 2. Uses lesser chemical inputs, no tillage 3. Tomato plants are naturally tolerant to fungal pathogens & live longer % Defoliation Disease tolerance Growth, disease tolerance & fruit yield in HVtomato system is gene x environment related Signaling Foliage, Fruits Gene Class BP c. Chaperones Moisture Root health Microbes Soil biology Roots/soil ----- BP HV 0.6 0.9 0.6 1.1 0.3 1.0 0.5 1.2 1.0 1.5 chiB osmo 0.3 1.0 0.9 1.2 hsp70 BiP rRNA 0.2 0.9 0.5 1.1 rbcS a. Longevity & photosynthesis rbcL N-responsive G6PD GS1 Root to Shoot NiR Communications b. Defense related ----- HV (108/96) 82 96 108 82 96 108 Days after transplanting Kumar et al. PNAS 2004 Mattoo & Abdul-Baki 2006 Cellular Networks Linking Fruit Metabolites Mattoo et al. (2006) Plant Physiol. Mattoo & Handa (2008) Plant Sci. C:N Indicator Genes Phosphoenolpyruvate carboxylase & NADPdependent isocitrate dehydrogense are synergistically up-regulated in transgenic tomato fruit Neelam et al. 2008 JXB Mulch-based and Ripening-Specific Modulation in Metabolites in the Transgenic Fruit compared to the Azygous Control Metabolites whose levels increased (arrow head) or decreased (horizontal bar) at least by 50% in 556AZ-HV or 579HO-BP compared with 556AZ-BP • Genetic modification of crop plants to enhance nutritional quality and crop productivity is a promising technology for sustainable agriculture and boosting food production in the world. • Cultural practices that utilize alternative agriculture strategies including organic cultivation subscribe to sustainable agriculture by limiting chemical usage and reduced tillage. • Significant genotype x mulch-dependent interactions on fruit phenotype demonstrate synergistic modulation of nutrient pathways. • Synergism between HV mulch and transgenic tomato in upregulating N:C indicator genes PEPC and ICDHc demonstrates significant cross talk between plant organs. • Environment-dependent and transgenedependent changes coexist without any qualitative deviation from normal fruit metabolites. This observation bodes well for the incorporation of genetically engineered (transgenic) crops into alternative agriculture practices. USDA, ARS Tahira Fatima Vijaya Shukla John Teasdale Vinod Kumar Anil Neelam Aref Abdul-Baki Ravinder Goyal Purdue University Avtar Handa Alka Srivastava CNR, Rome Annalaura Segre† Anatoli Sobolev