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Biotecnologie Vegetali per Disegnare la Pianta del Futuro Antonio Michele Stanca U N A S A- UNIMORE ROVIGO 19 dicembre 2013 Nazareno Strampelli Mendel Norman Borlaug Neolitico 500 semi/m2 1‐1.5 q/ha Periodo Romano Rinasciment Pre‐Mendel 2.000 semi/m2 4‐5 q/ha 3.000 semi/m2 6‐8 q/ha Post‐Mendel 1970 Oggi 8.000 semi/m2 25 q/ha 14.000 semi/m2 20.000 semi/m2 90‐100 q/ha 50 q/ha Towards the molecular basis of heterosis. Phenotypic manifestation of heterosis Trends Plant Sci. 2007 Sep;12(9):427‐32 Model of the Breeding progress in the last 100 years TRAITS To cope with Heat shock Diseases Lodging Increasing CO2 Light Cold SALT Soil of low fertility WATER anoxia drought IONS acid alcaline Heavy metals GENETICA GENOMICA studia il Genoma (l’intero contenuto di DNA di una Cellula) • identify the POSITION of genes controlling useful traits and tag them with molecular markers; • ISOLATE the genes encoding for useful traits for further geneimprovement ACCGTTGGACCATGGATACTAATGG • The DNA based knowledge can lead to: Hv-WRKY38 Genomic Position Molecular Marker Technology MAS Molecular Assisted Selection MAS=Molecular Assisted Selection 12/06/0828/11/07 ORGANISMI GENETICAMENTE MODIFICATI – OGM INDIVIDUI CHE SI OTTENGONO MEDIANTE TRASFORMAZIONE GENETICA E PRECISAMENTE INSERIMENTO NEL GENOMA DELLA PIANTA TARGET MEDIANTE INGEGNERIA GENETICA DI UNA SEQUENZA DI DNA-GENE – PROVENIENTE DALLA STESSA PIANTA O DA ALTRI ORGANISMI FILOGENETICAMENTE VICINI O COMPLETAMENTE ESTRANEI. OGM DAL PUNTO DI VISTA LEGALE VIENE DEFINITO COME: “UN ORGANISMO IL CUI MATERIALE GENETICO E’ STATO MODIFICATO IN MODO DIVERSO DA QUANTO SI VERIFICA IN NATURA MEDIANTE INCROCIO O CON LA RICOMBINAZIONE GENETICA NATURALE” A) Malattie iperplastiche Agenti: A. tumefaciens e rhizogenes, P. syringae savastanoi e avellanea Transfer Il T-DNA è trasferito nel genoma della pianta Integration Expression 3'- LB- T-DNA -RB-5' PLANT TRANSFORMATION FUSARIUM spp LARVA PIRALIDE P66 Landriano (PV) , Az. Menozzi, settembre 2005 (foto Maggiore) P67 (Bt) Bt or non Bt? 5 Stati 14 anni Riduzione della popolazione di piralide sino al 70% nei campi convenzionali NON Bt Hutchison et al. 2010 Science Oltre i Transgenici Nature Biotec (marzo 2012) • L’ingegneria genetica di “prima generazione” ha permesso di inserire tratti di DNA a random nel genoma • Cellule vegetali ingegnerizzate per produrre molecole utili-non OGM• Zinc Finger Nucleasi-ZFN- Inserimento del gene in modo mirato mediante riconoscimento di siti specifici nel genoma • Cisgenesi ( non OGM?) e Intragenesi • GRAFTING • Modificazione Epigenetica ( metilazione-silenziamento) Seralini et al. 2012 Maize NK603 EFSA Insufficient Scientific Quality Trenta Anni di PGM -170 milioni di ettari -28 Paesi -11%superficie coltivata mondiale -90% Mais,Soia,Colza, Cotone Pochi caratteri Utili! GAS = Genomic Assisted Selection Centrodi Ricerca per la Genomica Selezione basata su migliaia/decine di migliaia di tratti di DNA , GENOTYPING PHENOTYPING Filmato RNA Genetic hypothesis to explain heterosis Hemizygous complementation in maize hybrids. Different maize inbred lines are characterized by a considerable loss of genetic colinearity, that is, the loss of particular genes. It is hypothesized that gene loss can be partly compensated in hybrids by hemizygous complementation, which also leads to an improved performance of hybrids versus inbred lines. The colored boxes A to D represent different genes. Different members of gene families are indicated by the same colour (e.g. A1 and A2 belong to a gene family). The loss of a member of a gene family can be partly compensated for by the action of other members of this gene family and leads only to minor phenotypic effects in inbred lines. When a hybrid is selfed over several generations inbreeding depression is observed because not all genes present in the hybrid will be maintained. To date, partial loss of colinearity has only been observed in maize. In other species as well as in maize, additional factors might also contribute to heterosis. Heterosis‐associated gene expression in maize Relative gene expression levels in hybrids and regulation of allele‐specific gene expression in hybrids. (a) Gene expression levels in hybrids are not strictly related to the genetic concepts of dominance and overdominance. (b) Effects of cis‐ and trans‐ regulation on the allelic contribution to gene expression in hybrids. Molecular mechanisms of polyploidy and hybrid vigor. A molecular clock model explains the basis of heterosis. In the hybrids, the allelic interactions between parent 1 (P1) and parent 2 (P2) induce epigenetic repression of CCA1 and LHY expression amplitudes (red dashed line) and upregulation of TOC1 expression amplitudes (green dashed line) relative to the expression values in the parents (solid red and green lines, respectively), whereas the periodicity of the clock remains the same because maintaining clock periodicity and rhythm is important for plant growth and fitness Trends Plant Sci. 2010 Feb;15(2):57‐71. FACE-Free-Air CO2 Enrichment- is considered the most advanced technology to investigate experimentally the impact of rising atmospheric CO2 on terrestrial ecosystems Effect of elevated CO2 on oat yield and quality traits? A rings of the FACE facility installed in Fiorenzuola d’Arda with its control unit Un genoma è come tante orchestre PLANT for the Future NUE e WUE MIRNA Potenzialità e Stabilità 25-30000 semi/m2 120-130q/ha METAGENOMIC PLANT-soil-microbeinteraction ROOTS SYSTEMS BIOLOGY and Beyond • Vibrant and Competitive Research • More SCIENCE in AGRICULTURE • SCIENCE FOR FARMING Breeding is not Art Breeding is SCIENCE Pontecorvo e la Scperta del Neutrino Giovani Studenti e Professori DOMANDA:Ma i neutrini cosa portano ai Kolkhoziani di RYAZAN? Risposta:Per ora il neutrino non dà nulla ai Kolkhoziani né a chiunque altro Modo per affermare che la ricerca spesso non dà un risultato immediato ai contadini e agli operai Modo elegante per affermare l’autonomia della ricerca SCIENZA e AGRICOLTURA Complementari……………………90.2% Incompatibili…………………..…….9.8% Chi SONO? 22 maggio 2009 BREEDING by DESIGN Integration of • • • • • • • GENETICS Molecular Genetics Molecular Breeding Physiology Food Technology Bioinformatics and Statistics Agronomy SYSTEMS BIOLOGY Model of the Breeding progress in the last 100 years Organismo geneticamente modificato (OGM) Un organismo il cui materiale genetico è stato modificato in modo diverso da quanto si verifica in natura mediante incrocio o con la ricombinazione genetica naturale 2005 MORE EFFICIENT AGRICULTURE Herbicide tolerance Insect resistance 2015 2025 HEALTHIER NUTRITION and QUALITY Aminoacids, oil, starch PLANTS AS FACTORIES Vitamins, long-chained fatty acids, omega 3 fatty acids, enzymes, PEST PROTECTION biopolimers, Virus, nematode, fungi, insects Pigments, pharmaceuticals, fiber. STRESS PROTECTION Cold, drought, salinity 1998 «tradizionale» COSTANZA «isogenico Bt» Lodi, azienda Bianchini-Elias (oggi Assessore all’Agricoltura in Lombardia) 5 ottobre (foto Maggiore) Ibridi Resa dt/ha U% Fori stocco n° 14% U Fori spiga Fori pedunc. n° n° Gallerie Fusarium Fumosi- cm/pianta spiga % na (ppb) ELGINA 141 23,5 0,8 48 0,06 CECILIA 110 21,3 6,31 P67 159 26,7 0,75 P66 111 21,3 9,54 130,25 23,2 4,35 0,44 0 6100 1,36 1,66 19,92 23 60 0,02 MEDIA 0,15 0,16 0,7 0 6300 3,26 4,7 2,58 1,14 68,53 22,4 29 26 3127 P66 P67 Visione del campo al momento della raccolta dopo aver eliminato il bordo centrale TRAITS To cope with Heat shock Diseases Lodging Increasing CO2 Light Cold SALT Soil of low fertility WATER anoxia drought IONS acid alcaline Heavy metals