NOVEDADES Biotecnológicas en cultivo y mejoramiento de SORGO

Presentación del tema: "NOVEDADES Biotecnológicas en cultivo y mejoramiento de SORGO"— Transcripción de la presentación:

1 NOVEDADES Biotecnológicas en cultivo y mejoramiento de SORGO
Todo Sorgo 2012 Rio Tercero, Septiembre 2012

2 Desarrollo tecnológico relativo
Promedios anuales, Argentina Fuente: SAGYP Maiz Sorgo Rto. grano (kg/ha) Año de cosecha

3 Progreso en el mejoramiento desde lo Convencional al ”Mundo molecular”
Mejoramiento “Convencional” Mutagenesis Mejoramiento asistido por marcadores: Fingerprinting Gene mapping In the previous slide we said that Yield gain for sorghum was doing OK What can we do to keep the momentum going??? This slide shows the progression of Plant Breeding from the conventional (visual world) to the Marker based Molecular genetics World) With Conventional Breeding – sometimes call Phenotyping – sometimes called Visual selection ---did we select for resitance or did the plant escape being attacked by the disease Genotyping – now we can select for the presence of a resistant gene at a molecular level – we know with a high degree of certainy when we make the selection that the plant is really resistant Molecular Assisted breeding Allows us to “Finger Print” our germplasm enabling us to to protect our Intelluctual Property Also allows us to map the location of a particular gene within the Sorghum Genome allowing us to develop and use Molecular Markers to easily identify the presense or absence of a particular trait

4 Germoplasma: Diversidad genética

5 Matriz de similaridades
Germoplasma: Diversidad genética Dendogram y ACPs Matriz de similaridades geneticas Analisis de polimorfismos SSRs (microsatelites) Lineas de sorgo In the previous slide we said that Yield gain for sorghum was doing OK What can we do to keep the momentum going??? This slide shows the progression of Plant Breeding from the conventional (visual world) to the Marker based Molecular genetics World) With Conventional Breeding – sometimes call Phenotyping – sometimes called Visual selection ---did we select for resitance or did the plant escape being attacked by the disease Genotyping – now we can select for the presence of a resistant gene at a molecular level – we know with a high degree of certainy when we make the selection that the plant is really resistant Molecular Assisted breeding Allows us to “Finger Print” our germplasm enabling us to to protect our Intelluctual Property Also allows us to map the location of a particular gene within the Sorghum Genome allowing us to develop and use Molecular Markers to easily identify the presense or absence of a particular trait

6 Una nueva Era para el Sorgo: Biotecnología
We have talked about technology available to sorghum Let’s get more specific and talk about BioTechnology advances for sorghum

7 Sequencing of sorghum genome completed
Fecha de publicación: 28-Ene Contacto: Joseph Blumberg Rutgers University Sequencing of sorghum genome completed Drought-resistant food, feed and biofuel source sequenced by international team                                                                          NEW BRUNSWICK, N.J. – En un trabajo publicado en la Revista Nature esta semana, Investigadores de Rutgers: Joachim Messing, Rémy Bruggmann, y un equipo internacional de colaboradores pudieron describir en el genoma del sorgo, un cultivo tolerante a sequia Africano. If you are involved in sorghum breeding anywhere in the world this announcement was really big news. We could now see where genes were located – height, maturity, disease and insect resistance and we could begin to get a handle on the “Holy Grail” of intrensic yield – not yield protection, but direct yield enhancement.

8 Arroz Sorgo Maiz Genoma del Sorgo: Comparacion de tamaño.
400 Mbp Mbp 3,000 Mbp Arroz Sorgo Maiz Mapping the genome also allowed us to compare the relative size of the genome among grain crops like Rice, Sorghum and Maize Rice and sorghum could be used to predict where genes were located within the much larger corn genome-this helped reduce the time required to find specific genes for corn scientists One of the reasons it is more difficult to find specific genes within the corn genome, in addition to sheer size, is due to the many multiple copies of genes or chromosome segments that are present within the genome

9 AH Paterson et al. Nature 457, 551-556 (2009) doi:10.1038/nature07723
Familias de genes ortologos entre: Sorgo, Arabidopsis, Arroz y Alamo. Mapping the genome also allowed us to compare sorghum to non-grain crops like Arabidopsis The take home message for this slide is that many crop species share genes with other unrelated crop species – this is the true definition of “odd couples” AH Paterson et al. Nature 457, (2009) doi: /nature07723

10 Genomica del Sorgo Disponibilidad de la secuenciación completa del genoma de sorgo mediante la integtracion de mapas recombinantes para desarrollar un mapa fisico This slide is displaying the 10 chromosomes of sorghum and the location of know genes. Ramu et al. Mol Breeding 2010, 26:

11 Post – Genomica del Sorgo
Mapeo de QTLs (usando SSR, DArT or SNP) Altura de planta y madurez. Control genetico de la fertilidad. Genes de resistencia a enfermedades fungicas y virosicas: (Downey mildew, Roya, Carbon, SCMV) Genes de resistencia a insectos. (Mosquita del sorgo, Pulgon verde) Genes y QTLs relacionados con estreses abioticos como: Sequía en post- floracion (stay-green), Brotado en pre-cosecha, Tolerancia a aluminio. Caracterizacion de factores de transcripcion Identificacion de factores de transcripcion de genes diferencial en respuesta inducciones bioticas y abioticas (ej. Ataque de pulgones, tratamientos de deshidratacion, alta salinidad, ABA, Ac Salicilico, etc.) This slide is displaying some of the activities now underway due to the knowledge gained by having access to the sorghum genome: QTL mapping (SSR, Darts or SNP) examples: Plant height & Flowering Time Genetic Control of fertility restoration Transcription Characterization Identifies which genes are behaving differently due to stress – an example would be stress caused by heat or drought, resulting in pre and /or post flowering stress which impacts the plants ability to maintain its green leaves – commonly called it ability to “stay green”. Association Mapping – can help us better understand “Traits” such as Height and maturity across environments Root angle Saccharification or Sugar yield Induced gene mutation - including screening and characterization of mutations for fitness and viability

12 Usos Actuales y resultados:
Selección por resistencia a Pulgon verde de los cereales biotipo I Se identificaron MM´s SSR asociados a QTLs que confieren resistencia. Lineas resistentes son seleccionadas tempranamente asistida por MM’s. This is an example of succuss as markers have become available within sorghum. Markers were used to identify lines resistance to Greenbug Biotype I, very early in the inbred development process Ensuring that the new greenbug biotype I greenbug resistant hybrids were selling today are in fact resistant to Greenbugs

13 Usos Actuales y resultados:
Conversiones limpias: Selección de BG mediante MMs para la introgresion de atributos de interes agronomico. This is an example of succuss as markers have become available within sorghum. Markers were used to identify lines resistance to Greenbug Biotype I, very early in the inbred development process Ensuring that the new greenbug biotype I greenbug resistant hybrids were selling today are in fact resistant to Greenbugs

14 Post – Genomica del Sorgo
Mapeo por asociacion Control de altura de planta y madurez en distintos ambientes. Angulo de raices. Rendimiento de azucar y contenido de azucar en jugo. Inducción de mutagenesis, screening, y caracterización. Identificacion de perdida o ganancia de genes funcionales. This slide is displaying some of the activities now underway due to the knowledge gained by having access to the sorghum genome: QTL mapping (SSR, Darts or SNP) examples: Plant height & Flowering Time Genetic Control of fertility restoration Transcription Characterization Identifies which genes are behaving differently due to stress – an example would be stress caused by heat or drought, resulting in pre and /or post flowering stress which impacts the plants ability to maintain its green leaves – commonly called it ability to “stay green”. Association Mapping – can help us better understand “Traits” such as Height and maturity across environments Root angle Saccharification or Sugar yield Induced gene mutation - including screening and characterization of mutations for fitness and viability

15 requieren un esfuerzo multidiciplinario
Nuevas tecnologías … requieren un esfuerzo multidiciplinario The advent of new technologies requires a multidisciplinary approach to solving problems The Lab and Field Teams must work together hand-in-hand Phenotypic data must be of extremely high quality for the labs to generate reliabe markers to help select for genes of interest This example reminds me of a phrase that a Professor in a Computer Coding class that i took in Graduate School Garbage in = Garbage out Gold in = Gold out 15

16 requieren un esfuerzo multidiciplinario
Nuevas tecnologias … requieren un esfuerzo multidiciplinario Nuevas tecnologias … 1. TRANSFORMACION Just as New technologies – requires multidisciplinary efforts New Technologies Require Transformation

17 Transformacion de sorgo ha sido lograda…
Primer reporte de trasnformacion de sorgo. (gene gun) Hasegawa’s PNAS 90, 11212 2000 Transformacion con Agrobacterium Zhao’s Hi-Bred Plant Mol Biol 44, 789 Transformation in Sorghum has been accomplished First reported in 1993 – Via the Gene Gun Reported again in 2000 – Via Agrobacterium

18 Pero, La eficiencia de transformacion en sorgo es (se considera) baja.
Año Publicacion E.T.% 1993 PNAS 90, 11212 0.3% 2000 Plant Mol Biol 44, 789 2% 2005 Plant Biotech J 3, 591 3% 2006 Plant Cell Rep 25, 784 2007 Plant Cell Tissue Organ Cult 91, 155 5% 2009 Plant Cell Rep 28, 429 7% Even though progress continues – success rates and efficiecy remain low relative to other crops like corn

19 La Historia de la Transformación esta ocurriendo en Argentina…
En el Laboratorio de Biotecnologia de Advanta en Balcarce, Buenos Aires ES REALIDAD proposal

20 Transformacion de embriones prematuros de sorgo con Agrobacterium…
2/3 dias ~2 h 5’ 2-3 dias Esterilizacion, aislamiento e infeccion Co-cultivo (24 C) 6-9 semanas 3/4 dias This slide is a summary of Agrobacterium Transformation of Sorghum immature zygotic embryos As you can see there are many steps involved in the process Starting from immature embryos as explants… cells may be cultured in the laboratory…. and regenerated into mature plants…. Carbenicillin 2,4-D Luz, kinetine, IAA 25 dias 2,4-D, selective agent carbenicilline Resto Regeneracion Seleccion (28 C) (28 C) (25 C) 20

21 Nuevas Tecnologías, uso practico… I NSP US / Texas A&M (Rooney et al
Nuevas Tecnologías, uso practico… I NSP US / Texas A&M (Rooney et al.) / Compañías de semilla. Practical uses of New Technologies

22 Rooney et al. Potenciación de heterosis genotipeado digital de germoplasma exótico
US based National Sorghum Producers Project Rooney et al - digital genotyping of exotic sorghum accesions for heterosis

23 Rooney et al. Potenciación de heterosis Composición alelica de 23 genotipos de sorgo.
This slide represents the allelic composition of 23 sorghum genotypes Haplotypes with Ma1 – critical locus that delimits photoperiod sensitivity Allows photoperiod sensitive genotypes to behave as non-photoperiod sensitive genotypes Enables photoperiod sensitive and photoperiod non-sensitive genotypes to flower at relatively the same time This will increase genetic diversity within the currently used germplasm base

24 Rooney et al. Potenciación de heterosis Cronograma de actividades y responsabilidades.
This chart is a list of activities, responsibilities and a time frame for the heterosis project

25 nuevas tecnologias, uso practico Sorgo Resistente a Herbicidas?
NSP US National Sorghum Producers: consorcio herbicida cletodim ( Select , Select Max ) Universidad de Kansas / Dupont Crop Protection. ALS herbicida sulphylurea ACC herbicida quizalofop Otros semilleros/ Otros herbicidas??? GMO No-GMO Based on Market Surveys from many parts of the world Farmers want herbicide resistant sorghum Good able to kill weeds in sorghum fields by spraying herbicides (just like in current corn and soya fields) Bad Sorghum will outcross to johnsongrass Do farmers want herbicide resistant johnsongrass Does it make a difference if it is a specific use herbicide (triazines) or a broad spectrum herbicide like glyphosate? Will regulatory/registration agencies approve the use of herbicide resistance sorghum It will be interesting to see how regulatory/registration agencies react to the idea of herbicide resistant sorghum

26 Nuevas Tecnologías, uso practico… II GMO : EUN
Practical uses of New Technologies

27 N 30% 70% Sorgo responde a la fertilización nitrogenada, pero aumentando parcialmente la absorción de N aplicado. El exceso de N se volatiliza y se transforma en un contaminante importante.

28 Nitrogeno, puede ser un contaminante atmosférico importante
Nitrogeno, puede ser un contaminante atmosférico importante. Representa un 5% de los gases de efecto invernadero. Fertilización Nitrogenada se incrementaría en el corto plazo. Source: J Bellarby et al. Cool Farming: Climate impacts of agriculture and mitigation potential. greenpeace.org MT of CO2-eq Fertilización N Año Millon Tons 2009/10 100 2014/15 (proyectado) 110 Source: P Heffer & M Prud’homme Fertilizer Outlook 2010 – th IFA Annual Conference 5% global GHG

29 Cultivos mas eficientes en el uso del N brindan beneficios económicos y ambientales que contribuyen a la sustentabilidad. 70% 30% N N Cultivos eficientes en el uso del N, rinden mas (biomasa y grano) y liberan menos N al ambiente.

30 NUE Control Rendimiento (grano) (kg/ha) 5000 4000 3000 2000 1000
Rendimiento (grano) (kg/ha) N aplicado (kg/ha) EUN en Canola en condiciones sub-optimas de crecimiento NUE – Example of Nitrogen use effeciency using Canola as the model crop More yield produced per unit of water available

31 Nuevas Tecnologías, uso practico… II GMO : Resistencia a Sequia
Practical uses of New Technologies

32 Limitaciones hídricas afectan a los cultivos e imponen fronteras de producción agrícola.
significant impact due to drought in Argentina 2012 season, significant negative drought impact in the US – corn yield will be significantly less in 2012 vs 2011 Terra Satellite - NASA

33 … El sorgo requiere menos agua?
… El sorgo requiere menos agua? S. Staggenborg, W. Barney Gordon, R. Vanderlip, K. Dhuyvetter Sorghum and Corn: Crop Management in Stress-Prone Environments. DONDE ESTAN LOS PERSONAJES AHORA (3) NECESIDADES DE AGUA

34 … El sorgo requiere menos agua? Resistencia a sequia Tolerancia Escape (ciclo, plasticidad fenologica) Eficiencia uso del agua DONDE ESTAN LOS PERSONAJES AHORA (3) NECESIDADES DE AGUA

35 Hahb4 es un factor de transcripción que induce la expresión de genes involucrados en una respuesta adaptativa a sequia Hahb4 Left plants not recovering from drought using Arabadopsis as the model plant Right plants recovering from drought Sequia Post-sequia

36 Eficiencia en el uso del N: Bajos costos. Altos rendimientos.
En Advanta intentamos buscar Sorgos que aseguren resultados cercanos a su potencial: Eficiencia en el uso del N: Bajos costos. Altos rendimientos. Bajo impacto ambiental. Tolerancia a sequia: Igual rendimiento con menos agua. Mas rendimiento por unidad de agua provista. DONDE QUEREMOS LLEVAR AL PERSONAJE

37 Para implementar este esfuerzo en el mejoramiento, un programa de ingeniería genética fue establecido en el centro de Biotecnología de Balcarce, con el objetivo de ampliar el espectro de genes disponibles en el pool de genes del sorgo.

38 Fueron licenciados genes de Arcadia y Bioceres para mejorar los sorgos…
Trait Gene Licensed from NUE Barley AlaAT Drought tolerance Sunflower Hahb-4 Water Use Efficiency IPT Salt Tolerance AtNHX1 This Advanta Genetic Engineering Project has in-licensed traits, genes from Arcadia and Bioceres to improve and protect yield potential Confidential 38

39 Resumen Mejoramiento convencional + Biotecnologia
Utilizacion de Nuevas Herramientas en Mejoramiento Conversiones limpias, Seleccion asistida por marcadores. Screening temprano de genes funcionales. Utilizacion de nuevas tecnologias Transformacion Objetivos Proteccion del rendimiento potencial del Sorgo Permitir el incremento de rendimiento de Sorgo mediante tecnicas de mejoramiento tradicional.

40 Gracias por su atencion.!!!
Pedro A. Pardo Investigación Sorgo – Venado Tuerto