A comparative light and electron microscope technique has been used to study the cytological changes accompanying teliospore (i.e. probasidium) germination in Ustilago hordei (Pers.) Lagerh. Special emphasis has been placed on determining the ultrastructural events involved in karyokinesls, especially meiosis, and cytokinesis. The thesis is divided into five parts, of which the first is concerned with pre-germinal differentiation. The great increase in microanatomical complexity which occurs during the pre-germinal stages is due largely to an increase in the amount of endoplasmic reticulum (ER) and to the formation of "primary hydration vacuoles." Evidently the nuclear envelope gives rise to the new ER which in turn dilates to form the vacuoles. This is accompanied by an increase in mitochondrial size and the development of patches of patches of "flocculent cytoplasm." Part II concerns the initiation and subsequent extension of the metabasidium (i.e. promycelium). Initiation involves the localized degradation of the inner spore wall, and deposition of new wall material. The ER and spherosome-like bodies seem to be associated with these activities. Once spore wall rupture has occurred the structural basis of promycelial extension is unknown but changes in the number, size, and distribution of the spherosome-like organelles appear to have profound effects on the differentiation of the organism. Septation, knee-joint formation, and budding are discussed in part III. Elaborate membrane complexes are associated with cross wall initiation. A membranous plate is completed across the cell before septal wall thickening begins. The initiation of sporidia (i.e. basidiospores) involves a localized plasti-cization of the promyoelial wall followed by degradation of the old wall and subsequent synthesis of new wall material. Bridge-formation results when two adjacent cells give rise to bud-like processes which grow together and subsequently fuse to produce a protoplasmic bridge. The structure and activities of the metabasidial nuclei and their associated structures are discussed in part IV, Both meiosis and mitosis are unusual in that the two chromatin bodies apparently remain attached to the centriolar-kinetochore-equivalent and at least one of the chromatin bodies in attached to the nucleolus throughout the division cycle. The results are compatible with Brown and Stack's (1971) model for somatic nuclear division in some fungi. Membrane complexes, resembling those which Initiate septa, form in association with prophase nuclei and maintain a specific relation with the nucleus throughout division. In part V the suggestion is made that these complexes form part of a mechanism controlling the positional relationships of nuclear and cell divisions in the promycelium. / Science, Faculty of / Botany, Department of / Graduate

Made available in DSpace on 2016-12-08T16:44:42Z (GMT). No. of bitstreams: 1 PGPV11MA101.pdf: 1302742 bytes, checksum: b7efc06d336ca4a49c0225d1d0b03cfb (MD5) Previous issue date: 2011-02-08 / The objectives of the work were: a) to obtain the damage function equations for multiple pathosystem to calculate the EDT to time fungicides application; b) to quantify the control and damage of the yield components, and c) To relate the brown spot incidence and severity in different growth stages. The experiments were carry out at the NBN Seeds Company during the 2009 and 2010 crop seasons, located in Muitos Capões county, Rio Grande do Sul state, Brazil and in the Agro science Center at Santa Catarina State University only in 2010. The cultivar BRS Cauê was used as susceptible cultivar to brown spot and powdery mildew. The experiments design was in randomized block with four replications. The nine treatments consisting of different rates (half and recommended rates) and fungicide applications number (one, two, three and four) of mixture strobilurin and triazole fungicides, generating the disease gradients intensity. The total area of 5.0 x 2.5 m was the experimental unit in both experiments. The fungicide applications and incidence and severity assessment were done at EC 22, EC 31, EC 39, EC 45 and EC 56 plant development stages. Plants from central rows of each plot were manually harvested and grain yield (GY), one-thousand grain weight (TGW) and granulometry (G) were evaluated. In the first chapter, the damage function equations between disease intensity and GY for each plant development stages, obtained by regression analysis, in both 2009-10 crop growing seasons were significant and negative indicative that increasing disease severity lead into decreasing grain yield. The damage coefficients of these equations can be used to calculate the economic damage threshold. In the second chapter, the values of disease intensity were used to calculate the area under disease progress curve (AUDPC). The GY, TGW, G and AUDPC values were tested using the mean comparison between treatments and the percentage of damage and disease control were determinated. The largest 2009 crop damage was 45.87%, 15.47% and 25.84% for GY, G and TGW, respectively. The ultimate control was 68.11% when considered the severity and four foliar applications, independent of the used dose. In 2010, the greatest damage were 31.16%, 14.02% and 10.76% in Muitos Capões and 39.44%, 23.59% and 45.88%, in Lages, for GY, 10 TGW and G, respectively. The highest percentage of control, based on the leaf severity were 71.63% and 73.96% for Muitos Capões and Lages, respectively. The greater control, independent of used dose were obtained with three and four applications in Muitos Capões and four applications in Lages. In the third chapter, brown spot incidence and severity data were subjected to regression analysis and correlation and the obtained values were significant and positive. The brown spot and powdery mildew diseases incidence and severity recommended by the Technique Indication of crop (TIC) to initiate fungicide applications is 20% and 5% respectively. The severity average values for initiating chemical control are 0.77% and 0.34% respectively for 2009 and 2010 crop seasons when the incidence diseases were substituted in the equations. These values are lower than TIC recommended values / Os objetivos do trabalho foram: a) obter equações de função de dano para patossistema múltiplo para calcular o LDE servindo como critério indicador de aplicação de fungicidas; b) quantificar controle e dano nos componentes de rendimento; e c) relacionar incidência e severidade foliar da mancha marrom em diferentes estádios fenológicos. Os experimentos foram conduzidos nas safras agrícolas 2009 e 2010 na NBN Sementes no município de Muitos Capões, RS; e, na safra de 2010 no Centro de Ciências Agroveterinárias no município de Lages, SC. Em todos os experimentos foi utilizada a cultivar BRS Cauê suscetível à mancha marrom e oídio. O delineamento foi blocos casualisados, com quatro repetições e nove tratamentos constituídos de diferentes doses (meia dose e dose indicada) e número (uma, duas, três e quatro) de aplicações de fungicidas triazóis e estrobilurinas para gerar os gradientes de intensidade das doenças. A área de cada unidade experimental correspondeu a 5,0 x 2,5 m. As aplicações e as avaliações da incidência e severidade foliar ocorreram nos estádios de crescimento (EC) EC 22, EC 31, EC 39, EC 45 e EC 56. A colheita foi manual cortando as plantas das linhas centrais de cada parcela. Foram avaliados rendimento de grãos (RG), massa de mil grãos (MMG) e granulometria (G). No primeiro capítulo, as equações de função de dano entre intensidade de doença e RG para cada EC, obtidas por análise de regressão, em ambas as safras agrícolas, foram significativas e negativas, ou seja, à medida que aumentou a intensidade de doença, diminuiu o RG. Os coeficientes de dano obtidos podem ser utilizados no cálculo do limiar de dano econômico. No segundo capítulo, os valores de intensidade das doenças foram usados para o cálculo da área abaixo da curva de progresso da doença (AACPD). Os valores de RG, MMG, G e AACPD foram submetidos ao teste de comparação de médias. Foram determinados o percentual de dano e de controle das doenças. Na safra 2009, os maiores danos foram 45,87%, 15,47% e 25,84% para RG, MMG e G, respectivamente. O controle máximo foi 68,11% considerando a severidade foliar e quatro aplicações, independente da dose usada. Em 2010, os maiores danos foram 31,16%, 14,02% e 10,76% em Muitos Capões e 39,44%, 23,59% e 8 45,88% em Lages, respectivamente para R, MMG e G. Com base na severidade obteve-se controle de 71,63% e 73,96% em Muitos Capões e Lages, respectivamente. Em Muitos Capões três e quatro aplicações e em Lages quatro aplicações, independente da dose, apresentaram maior percentual de controle. No terceiro capítulo, os dados de incidência e severidade foliar de mancha marrom foram submetidos à análise de regressão e correlação. As equações obtidas foram significativas e positivas. A incidência e a severidade foliar recomendado pela Indicação Técnica da cultura (ITC) para iniciar as aplicações de fungicidas é de 20% e 5%, respectivamente. Substituindo esse valor de incidência nas equações obtidas têm-se valores médios de severidade para iniciar o controle químico de 0,77% e 0,34% respectivamente para 2009 e 2010, inferior ao recomendado pela ITC

La cendrosa de l'ordi, causada pel fong Blumeria graminis (D.C.) E. O. Speer f. sp. hordei Em. Marchal, sin. Erysiphe graminis D.C.: Fr. f. sp. hordei Em. Marchal, anamorf: Oidium monilioides (Nees) Link, és la malaltia aèria més important en aquest conreu a Catalunya. El 1987 es van iniciar les investigacions sobre l'etiologia de les principals malalties dels cereals. Es realitzà una prospecció per tota la l’àrea cerealícola de la geografia catalana, que va permetre estimar la distribució i importància de les malalties que afecten l'ordi, el blat, el panís i l'arròs. Els resultats del període estudiat, 1987-1990, conclogueren que per a la cendrosa de l’ordi, la probabilitat d’una epidèmia severa (severitat final=60%5% en varietats susceptibles) varià entre p=0,2 i p=0,6 (entre dos i sis anys amb epidèmia severa de cada deu anys de cultiu), segons zones de conreu. Amb aquestes premisses es va anar elaborant un sistema de suport a la presa de decisions (SSPD) propi per a la cendrosa de l’ordi a l’àrea de Catalunya anomenat CENCONT, el qual prediu la malaltia tenint en compte que la cendrosa forma part d’un ‘complex de malalties’ i considerant la severitat (i l’àrea sota la corba epidèmica) com a variable independent i el possible efecte de la resta de malalties i plagues com a covariants. Per a l’aplicació pràctica de les anàlisis epidèmiques a CENCONT, es van estimar els valors dels paràmetres del model Logist com mitjanes d’un nombre de epidèmies representatives dels distints processos: de reacció varietal i de control químic. Aquests valors van resultar útils per a la predicció d’epidèmies i l’avaluació de CENCONT va demostrar la seva utilitat, tant pel que fa a la seva precisió com per a l’anàlisi econòmica fonamentada en el càlcul de la Funció de Guany. Però les assumpcions teòriques fetes en ajustar models tipus Logist, anomenats sintètics, a les dades epidèmiques de camp, han de permetre fer prediccions amb un marge d'error acceptable, en les condicions ambientals definides per cada predicció, la qual cosa s’ha de demostrar que és certa en una eina SSPD. Per aquesta raó els objectius que s’han formulat en aquest treball s’han basat resumidament en: 1) Fer una anàlisi matemàtica dels models sintètics, principalment els Logist i Richards, veient el seu comportament i estudiant les relacions y0, r i k mitjançant simulació, 2) Contrastar els resultats teòrics amb els de les epidèmies desenvolupades en condicions naturals, aplicant els models, en ajusts a epidèmies de camp de la cendrosa de l'ordi, amb la condició d'anar fixant els paràmetres o bé deixar-los lliures en el model, principalment l'asímptota k, i 3) Discutir la validesa de les assumpcions fetes en els models per ajustar les equacions a les dades experimentals, veient les interaccions entre paràmetres i les conseqüències que derivades de l'anàlisi comparativa d'epidèmies i la seva aplicació a sistemes predictius de la malaltia. Per assolir els objectius plantejats s’han utilitzat 113 processos epidèmics observats en camp en el període 1991-2002, avaluant principalment la intensitat de malaltia en el temps i basats en dos objectius experimentals: control químic i resistència varietal Paral•lelament, s’han analitzat des del punt de vista matemàtic mitjançant el programa MAPLE els models exponencial, Logist, Gompertz, Monomolecular, Von Bertalanffy-Richards i Weibull, i s’ha valorat la seva idoneïtat en la seva aplicació a malalties vegetals, especialment a la cendrosa de l’ordi. Després de l’anàlisi matemàtica, s’han aplicat els models mitjançant el programa Table Curve 2D©. En una primera fase s’han ajustat tots els models a les epidèmies de camp, excepte el model exponencial, i en una segona, de tots els models aplicats s’han seleccionat els que donaven millor rendiment estadístic pel que fa als diversos paràmetres d’avaluació. En aquest procés d’ajust, també s’han descartat les epidèmies que no aconseguien un ajust de suficient qualitat segons els criteris establerts. A més, s’han establert comparacions de les taxes relatives epidèmiques obtingudes en els ajusts mitjançant la utilització de la taxa mitjana ponderada absoluta. S’ha fet una anàlisi especial del comportament del paràmetre de forma del model Richards i s’han elaborat models de superfície de resposta per als paràmetres epidèmics y0, r i k dels models Logist i Richards. Entre les conclusions més importants s’ha trobat que el model Richards s’ajusta millor que Logist i aquest que Gompertz al conjunt d’epidèmies de cendrosa de l’ordi a Catalunya. La resta de models analitzats es van descartar per pitjor comportament. Quant als paràmetres dels models, en primer lloc no és el mateix fixar l’asímptota que no fixar-la, com a mínim des de la perspectiva estadística. En segon lloc, la taxa relativa epidèmica disminueix de valor segons si s’ajusten les dades a Richards, Logist o Gompertz respectivament, i això passa sempre respecte qualsevol variant dels models, asímptota lliure o bé asímptota fixa, i per a qualsevol objectiu epidèmic, control químic o bé resistència varietal. Respecte a la severitat inicial, no s’ha trobat una pauta de comportament definida. Sobre la predicció, es pot fer una millora d’aquesta en el sistema d’avisos CENCONT passant del model Logist al model Richards. El model Richards es pot utilitzar amb els tres paràmetres clàssics o bé amb quatre paràmetres, incorporant el paràmetre de forma però amb unes certes restriccions. Si s’agafa el model Richards triparamètric, serà necessari fixar el paràmetre de forma a un valor que raonablement pot ser inferior a 5. Aquest tipus de modelització és la recomanada per a incloure en Sistemes de suport a la presa de decisions com és el CENCONT. La millora en la predicció no implica necessàriament una millora en la presa de decisions del sistema d’avisos de la cendrosa de l’ordi a Catalunya, perquè les implicacions econòmiques no són en la major part dels casos prou importants com per provocar un canvi de decisió. La diferència màxima calculada de pèrdues associades estimades segons el model Richards, amb asímptota lliure i les associades segons el model Logist, se situa en el 2.9%, calculant-les a partir de l’ASCPM i de la severitat final. A efectes pràctics es pot considerar el mateix haver predit segons el model Richards, millor en el treball, o el Logist, el qual fou el que realment s’havia aplicat en el programa CENCONT. L’estimació Logist podria ser suficient per a la major part de les situacions (epidèmies) i que només en un petit percentatge hauria valgut la pena canviar de model. De tota manera, per al tipus de prediccions del present treball, no es veuria malament el fet d’adoptar el model Richards en un futur, amb la condició d’haver fixat el valor característic del paràmetre m per a la zona d’estudi. / El oídio de la cebada causado por el hongo Blumeria graminis (D.C.) E. O. Speer f. sp. hordei Em. Marchal, sin. Erysiphe graminis D.C.: Fr. f. sp. hordei Em. Marchal, anamorf: Oidium monilioides (Nees) Link, es la enfermedad aérea más importante en este cultivo en Cataluña. El año 1987 se iniciaron las investigaciones sobre la etiología de las principales enfermedades de los cereales. Se realizó una prospección por toda el área cerealista de la geografía catalana que permitió estimar la distribución e importancia de las enfermedades que afectan la cebada, el trigo, el maíz y el arroz. Los resultados del período estudiado, 1987-1990, concluyeron que para el oídio de la cebada, la probabilidad de una epidemia severa (severidad final=60%5% en variedades susceptibles) osciló entre p=0,2 y p=0,6 (entre dos y seis años con epidemia severa de cada diez años de cultivo), según zonas de cultivo. Con estas premisas se elaboró un sistema de soporte a la toma de decisiones (SSTD) propio para el oídio de la cebada en el área de Cataluña, llamado CENCONT, el cual predice la enfermedad teniendo en cuenta que el oídio forma parte de un ‘complejo de enfermedades’ y considerando la severidad (y el área bajo la curva epidémica) como variable independiente y el posible efecto del resto de enfermedades y plagas como covariables. Para la aplicación práctica de los análisis epidémicos en CENCONT se estimaron los valores de los parámetros del modelo Logist como medias de un número de epidemias representativas de los distintos procesos: reacción varietal y control químico. Estos valores resultaron útiles en la predicción de epidemias y la evaluación de CENCONT demostró su utilidad, tanto en lo que se refiere a su precisión como para el análisis económico fundamentado en el cálculo de la Función de Ganancias. Pero las asunciones teóricas realizadas en el ajuste de modelos tipo Logist, llamados sintéticos, a los datos epidémicos de campo, han de permitir predicciones con un margen de error aceptable, en las condiciones ambientales definidas para cada predicción, lo cual se debe demostrar como cierto en una herramienta SSPD. Por esta razón los objetivos que se han formulado en este trabajo se han basado resumidamente en: 1) Efectuar un análisis matemático de los modelos sintéticos, principalmente Logist y Richards, viendo su comportamiento y estudiando las relaciones y0, r i k mediante simulación, 2) Contrastar los resultados teóricos con los obtenidos de las epidemias desarrolladas en condiciones naturales, aplicando los modelos en ajustes a epidemias de campo del oídio de la cebada, con la condición de ir fijando los parámetros o bien dejarlos actuar libremente en el modelo, principalmente la asíntota k, y 3) Discutir la validez de las asunciones realizadas en los modelos para ajustar las ecuaciones a los datos experimentales, viendo las interacciones entre parámetros y las consecuencias derivadas del análisis comparativo de epidemias y su aplicación a sistemas predictivos de la enfermedad. Para conseguir los objetivos propuestos se han utilizado 113 procesos epidémicos observados en campo durante el período 1991-2002, evaluando principalmente la intensidad de enfermedad en el tiempo y basados en dos objetivos experimentales: control químico y resistencia varietal. Paralelamente se han analizado los modelos exponencial, Logist, Gompertz, Monomolecular, Von Bertalanffy-Richards y Weibul desde un punto de vista matemático mediante el programa MAPLE, y se ha valorado su idoneidad en la aplicación a enfermedades vegetales, especialmente al oídio de la cebada. Después del análisis matemático se han aplicado los modelos mediante el programa Table Curve 2D©. En una primera fase se han ajustado las epidemias de campo a todos los modelos excepto el modelo exponencial, y en una segunda fase de todos los modelos aplicados se han seleccionado aquellos que resultaban en un mejor rendimiento estadístico respecto a diversos parámetros evaluados. En este proceso de ajuste también se han descartado las epidemias que no conseguían un ajuste de suficiente calidad según los criterios preestablecidos. Además se han hecho comparaciones de las tasas relativas epidémicas obtenidas en los ajustes mediante la utilización de la tasa media ponderada absoluta. Se ha realizado un análisis especial del comportamiento del parámetro de forma del modelo Richards y se han elaborado modelos de superficie de respuesta para los parámetros epidémicos y0, r i k de los modelos Logist y Richards. Entre las conclusiones más importantes se destaca que el modelo Richards ajusta mejor que Logist y este que Gompertz respecto al conjunto de epidemias de oídio de la cebada en Cataluña. El resto de modelos analizados se descartaron debido a su peor comportamiento. Respecto a los parámetros de los modelos, en primer lugar no es lo mismo fijar la asíntota que no fijarla, como mínimo desde el punto de vista estadístico. En segundo lugar, la tasa relativa epidémica disminuye de valor según si se ajustan los datos a Richards, Logist o Gompertz respectivamente, sucediendo esto siempre respecto a cualquier variante de los modelos, asíntota libre o bien asíntota fija, y para cualquier objetivo epidémico, ya sea control químico o bien resistencia varietal. Respecto a la severidad inicial, no se ha encontrado una pauta de comportamiento definida. Sobre la predicción, se puede afirmar que se realizaría una mejora en el sistema de avisos CENCONT pasando del modelo Logist a utilizar el modelo Richards. El modelo Richards se puede utilizar con los tres parámetros clásicos o bien con cuatro parámetros, incorporando el parámetro de forma pero con ciertas restricciones. Si se escoge el modelo Richards triparamétrico, será necesario fijar el parámetro de forma hasta un valor que razonablemente puede ser inferior a 5. Este tipo de modelización es la recomendada para Sistemas de Soporte a la Toma de Decisiones como CENCONT. La mejora en la predicción no implica, sin embargo, una mejora en la toma de decisiones del sistema de avisos del oídio de la cebada en Catalunya, porque las implicaciones económicas no son en la mayor parte de los casos suficientemente importantes como para provocar un cambio de decisión. La diferencia máxima calculada de pérdidas asociadas, estimadas según el modelo Richards con asíntota libre y las asociadas según el modelo Logist se sitúa en el 2.9%, calculándolas a partir de l’ABCPE (Área Bajo la Curva de Progreso de la Enfermedad) y de la severidad final. A efectos prácticos se puede considerar como lo mismo el haber predicho según el modelo Richards, mejor en el trabajo, o el Logist, el cual fue el que realmente se había aplicado en el programa CENCONT. La estimación Logist podría ser suficiente para la mayor parte de las situaciones (epidemias) ya que solamente en un pequeño porcentaje habría valido la pena cambiar de modelo. De todas formas, para el tipo de predicciones del presente trabajo, no se vería como inconveniente el adoptar el modelo Richards en un futuro, con la condición de fijar un valor característico del parámetro m para la zona de estudio. / Powdery mildew caused by the fungus Blumeria graminis (D.C.) E. O. Speer f. sp. hordei Em. Marchal, sin. Erysiphe graminis D.C.: Fr. f. sp. hordei Em. Marchal, anamorf: Oidium monilioides (Nees) Link, is the most important disease on barley crop in Catalonia (Northeast of Spain). Surveys of cereal –barley, wheat, maize and rice– diseases in Catalonia were carried out on 1987 and provided the necessary data of the relative importance and crop losses of diseases on these crops. The results sowed that, in the period of 1987 to 1990, barley Powdery mildew disease had a probability of severe epidemic of p=0,2 and p=0,6 (final severity=60%5% in susceptible cultivars) depending on the sub-area. This means that frequency of critic disease is between two and six years in a ten years period of time. Those studies gave as a result the elaboration of a Decision Support System (DSS) on Powdery mildew in Catalonia called CENCONT. This DSS gives predictions of the disease as a “disease complex”, considering severity (and AUDPC) an independent variable and the rest of the diseases and pests as covariates. To build CENCONT parameters of the Logist model were estimated as means of a diversity of numerous epidemics obtained from chemical control and resistance cultivar trials. These values had showed to be useful on predicting epidemics and consequently evaluating CENCONT in the point of view of economical analysis. But theoretical assumptions of those synthetic models, as Logist model, must be sufficient to predict in an acceptable range of error in the environmental conditions of a particular prediction. This conclusion should be demonstrated. Thus, the objectives of this work had been: 1) working out of mathematical modelling using synthetic models, especially Logist and Richards, to study their behaviour and the relationships between y0, r and k by means of program simulation, 2) to check theoretical conditions applied to epidemics developed in natural conditions, adjusting models to data and testing the parameter behaviour using different starting fitting values, and 3) to validate the adjusting conditions to experimental data, knowing the interactions between parameters and their consequences in Comparative Epidemiology and Decision Support Systems. To achieve with the objectives there have been used 113 field epidemics obtained between 1991 and 2002, assessing the disease intensity vs. time and based on two experimental objectives: Chemical Control and Cultivar Resistance. Eventually, the Exponential, Logist, Gompertz, Monomolecular, Von Bertalanffy-Richards and Weibull models have been analysed mathematically using MAPLE computer program, and assessed their fitness on applied disease epidemics, particularly to powdery mildew of barley. Furthermore, the models have been applied to field epidemic data using Table Curve 2D computer program. First, all models except Exponential were tested to know their ability to fitness and later some of the models were discarded. To eliminate models statistical yield criteria on fitting were used. As well as the models some epidemics were also discarded in this process using the criteria of poor quality results of fitting. Once more, comparisons between relative epidemic rate data were done by testing the suitability of weighted mean absolute rate, and an additional analysis to study the particular behaviour of shape parameter of Richards’ model was found necessary. Finally, a 3D model of the surface response curve of epidemic parameters y0, r and k of Logist and Richards previous 2D models was developed. Main results of this work are that behaviour of Richards’ model is better than Logist model to the fitting to barley powdery mildew epidemics in Catalonia. Yields of the rest of the models had been worst and are discarded of the normal use in our conditions. With regard to parameters, it is stated that the use of fixed asymptote is statistically different in front of free asymptote. Besides, after fitting the relative epidemic rate values decrease in the order of Richards’, Logist and Gompertz models. This happens independently of considering the use of free or fixed asymptote and both chemical control or cultivar resistance objectives. Moreover, the initial severity of disease does not show any pattern of behaviour. As a consequence of the data obtained, it is obvious that should be convenient to use Richards’ model instead of Logist in the CENCONT computer program. Richards’ model could be used with his three or four parameter form, but if the second, the use of shape parameter should be controlled and restricted in a short range of values. Within the form parameter, a value les than 5 should be recommended. This modelling pattern is suitable to include in those DSS as CENCONT is. The improvement on prediction does not means that making decisions using this DSS should be better because the economic consequences are not enough frequent and relevant to decide changing the pattern. The maximum difference of crop losses calculated comparing the use of Richards’ model instead of Logist model is about 2.9%. These estimates are of final severity or AUDPC of the epidemic. In fact, the use of Richards’ model or the Logist model does not normally bear differences in the economic recommendations of CENCONT which uses Logist model. So actually the Logist model could be sufficient to the most of environmental circumstances (epidemics) and only few of them should carry a model change. Instead of this, the use of Richards’ model in next future should be considered as a clear improvement of the prediction, but fixing the shape parameter to the particular conditions of the area should be recommended as necessary in this hypothetical case.

Cereal loss due to fungal pathogens is an ongoing setback in agriculture. Elucidating plant&rsquo / s resistance and susceptibility mechanisms against these cereal killers, promises progress in agriculture. In the way of understanding barley resistance against fungus Blumeria Graminis we silenced FAS-Associated Factor 1 (FAF1) gene in its mRNA level with Virus Induced Gene Silencing (VIGS) technique. Previous research in our lab had shown an augmentation in mRNA levels of FAF1 gene in fungus infected wheat, suggesting a role of this gene in the resistance mechanism. We hypothesized that the apoptotic role of FAF1 protein in metazoan is conserved in plants by including FAF1 as a factor in hypersensitive response. Barley lines Pallas01 and Pallas03 which are respectively resistant and susceptible against fungus Blumeria graminis hordei 103 (Bgh103) were used for fungal inoculations after FAF1 silencing, to test if the hypersensitive response against fungus Bgh103 was prevented. In this aspect the formation of death lesions on the Pallas01 leaf due to fungal resistance was not prevented demonstrating that FAF1 silencing with VIGS in the resistant Pallas01 line of barley is not sufficient to stop apoptosis. On the other hand the FAF1-silenced barley susceptible line Pallas03 became more sensitive to fungal stress based on conidia (body part of the fungus) counting after trypan blue staining of the infected leaves. In the C-terminus of FAF1 an ubiquitin like domain-X (UBX) is found, which is the cause of stress sensitivity based on the reported data obtained about this domain&rsquo / s loss of function in other proteins. These results suggest that FAF1 is a catalyst in the hypersensitive response and its loss of function makes barley more susceptible to fungal stress. On the other hand a short mRNA homology was found among FAF1 and many pathogen disease related proteins making this homology a possible target site for VIGS of FAF1 generated siRNAs, which might cause some other protein to be responsible for the barley susceptibility against the fungus.

Barley is one of the most important crop species in Turkey and powdery mildew is one of the most common pathogen decreasing yield in barley. For this problem, agricultural biologists apply breeding technologies in order to select and propagate resistant barley cultivars. However, this is not a permanent solution since pathogens evolve rapidly to overcome plant resistance mechanisms. On the other hand, molecular plant pathologists are trying to understand basic mechanisms underlying plant-pathogen interactions by using molecular tools in order to develop long term solutions for preventing yield loss. In this thesis, miR159 mediated regulation of barley GAMyb transcription factor is studied. According to microRNA microarray results regarding to infection with powdery mildew pathogen Blumeria graminis f.spp hordei (Bgh) at different time points, miR159 expression level showed significant differences. Bioinformatics analysis revealed that miRNA159 targets GAMyb gene in barley. In order to investigate this relationsh&amp / #8223 / p, both miRNA and miRNA target were cloned into GFP containing expression vectors through Gateway cloning and resulting vectors were transformed into Nicotiana benthamiana through Agrobacterium mediated gene transfer. Observations based on GFP expression showed that miRNA159 targets and decreases the expression of GAMyb in vivo. v To conclude, this study can be evaluated as a distinctive study for two aspects / (i) it is the first study assessing a &ldquo / putative&rdquo / barley miRNA function biologically and (ii) developed a practical and effective functional assay for miRNA studies in plants.

Blumeria graminis f. sp. hordei is a biotroph pathogen that causes powdery mildew disease in barley. In this study, Pallas01 and Pallas03 barley lines having Mla1, Ml (Al2) and Mla6, Mla14 R-genes were inoculated with Bgh103(64/01) race of the Blumeria graminis f. sp. hordei having avirulence and virulence to Pallas01 and Pallas03, respectively. The proteins were isolated from the three biological replicates of 12, 24, and 48 hpi samples following the method in Rampitsch et al., 2006. These there biological replicates of three time points together with the mock inoculated plant proteins were separated on 2D-PAGE using IPG strips of 4-7 pH values as three technical replicates, resulting 108 gels. The gels were analyzed using PdQuest (Bio Rad) in order to assess up- or down-regulated protein spots by comparing against controls and the samples having resistance or susceptible responses with each other. According to the analysis, 36 proteins were found to be differentiated and among them 18 proteins were found up-regulated and 8 proteins were found down-regulated. The spots were manually v excised and subjected to the nano-LC-ESI-MS/MS analysis (Proteome Factory, Germany). The MASCOT algorithm was used for identification of the possible proteins. The experimental pI and MW values were used for selecting the differentiated proteins from the mass results. The relative abundance of each of the 38 identified polypeptides was calculated in terms of spot intensity. The majority of the most abundant proteins were found to be carbohydrate metabolism related. The relative distribution of the proteins into four main functional categories was taken into consideration. Statistical tests (Students&amp / #8223 / T-test) were carried among the identified proteins in order to reveal statistically significant proteins throughout the study. By making a WoLF PSORT search, subcellular localization of the proteins was predicted. Accordingly, most of the proteins were found to be located in cytoplasm or chloroplast.

Blumeria graiminis f. sp. hordei (powdery mildew) is an obligate biotroph infecting hordeum vulgare (barley). It is one of the most devastating pathogens of barley, decreasing barley yield in great extent. In order to decrease barley loss, numerous studies are being conducted for overcoming the disease from the sides of both pathogen and host. However the pathogen is evolving very rapidly preventing the effective use of pesticides such as fungisides or development of resistant barley varieties by crossing race-specific resistance varieties, varieties having R genes, with susceptible but high yield producing varieties. In order to understand the mechanism of pathogen-host interactions, and producing enduring solutions for the problem of yield loss in barley molecular tools need to be used. In this thesis study, Amplified Fragment Length Polymorphism (AFLP) molecular marker method is used in order to reveal the molecular characterization of Turkish Blumeria graminis f. sp. hordei varieties collected from &Ccedil / ukurova region in Turkey. Thirty-nine samples were analyzed with eigth universal races, of which virulence genes are studied. AFLP studies were conducted on LI-COR 4300 DNA Analyzer system. Bioinformatics analysis was performed with NTSYS program. By the help of this Numerical Taxonomic System, similarity, dissimilarity, clustering, dendograms, two-dimensional scatter plots, and three-dimensional perspective plots were obtained. By the light of these analyses Turkish Blumeria graminis f. sp. hordei varieties together with universal races are grouped into three clusteres. In conclusion, studying Turkish Blumeria graminis f. sp. hordei isolates and comparing them with universal races is a unique study in terms of characterizing the Turkish Bgh isolates for the first time, and can be used as a frontier study for studying Resistance genes, by reverse genetic tools.

Afin d'améliorer nos pratiques agricoles dans le contexte d'une agriculture durable, plusieurs agents de lutte biologique (ALB) ont été développés, testés et sont maintenant utilisés dans le monde pour combattre les pertes de rendements causées par les maladies. Blumeria graminis f. sp. hordei ( Bgh) est l'agent pathogène responsable du blanc de l'orge et peut réduire les rendements de cette culture jusqu'à 40%. Un champignon épiphyte, Pseudozyma flocculosa, a été découvert et identifié en 1987 en association étroite avec le blanc du trèfle. Les chercheurs ont alors remarqué que ce champignon exhibait une forte activité antagoniste contre le blanc en détruisant les structures de l'agent pathogène. Suite à d'autres travaux, il est apparu que ce comportement antagoniste était dirigé contre tous les membres des Erysiphales et semblait lié à la synthèse d'un glycolipide antifongique soit la flocculosine. Toutefois, on n'est toujours pas parvenus à associer l'efficacité de l'ALB avec la production de ce glycolipide. Ces observations suggèrent que d'autres facteurs seraient impliqués lorsque les deux protagonistes, l'ALB et le blanc, sont en contact. L'objectif principal de ce projet était donc de chercher d'autres mécanismes moléculaires pouvant expliquer l'interaction P. flocculosa-blanc et orge, en faisant une analyse transcriptomique complète des trois protagonistes en même temps. L'interaction tripartite a été échantillonnée à différents temps suivant l'inoculation de P. flocculosa sur des feuilles d'orge présentant déjà une intensité de blanc d'environ 50%. Les échantillons de feuilles prélevés ont ensuite été utilisés pour l'extraction de l'ARN qui ont été ensuite transformés en ADNc pour la préparation des librairies. Cinq répliquats ont été effectués pour chaque temps et le tout a été séquencé à l'aide de séquençage par synthèse Illumina HiSeq. Les séquences obtenues (reads) ont ensuite été analysées à l'aide du logiciel CLC Genomics Workbench. Brièvement, les séquences obtenues ont été cartographiées sur les trois génomes de référence. Suite à la cartographie, les analyses d'expression ont été conduites et les gènes exprimés de façon différentielle ont été recherchés. Cette étape a été conduite en portant une attention particulière aux gènes codant pour un groupe de protéines appelées CSEP pour “candidate secreted effector proteins” qui seraient possiblement impliquées dans l'interaction tripartite. Parmi les protéines exprimées de façon différentielle en présence du blanc ou en absence de ce dernier, nous avons pu constater que certaines CSEP étaient fortement exprimées en présence du blanc. Ces résultats sont prometteurs et nous offrent une piste certaine pour l'élucidation des mécanismes impliqués dans cette interaction tripartite.

Infection of barley (Hordeum vulgare) leaves with the rust fungus, Puccinia hordei, causes changes in the host protein synthesis. This thesis analyses these changes in the barley cultivar Triumph following inoculation of 7-day-old leaves with either a virulent or an avirulent race of P. hordei. The initial approach was to isolate membrane-bound polysomes from infected leaves, translate them in vitro and analyse the translation products. These products include the integral membrane proteins which were expected to be involved in the response of the host to the pathogen. A method based on differential centrifugation in the presence of a ribonuclease-inhibiting buffer was developed for separating membrane-bound polysomes from the rest of the cytoplasmic polysomes. Membrane-bound polysomes were found to comprise one fifth of the total polysomes in the leaves. Analysis of the translation products of membrane-bound polysomes by SDS-PAGE showed them to be of higher average molecular weight than those from free polysomes. Comparison of polypeptides produced by membrane-bound polysomes from healthy and inoculated plants showed some differences however the low yield of membrane-bound polysomes made it difficult to obtain conclusive results. Thus it was decided to isolate total polysomes by including 1% Triton X-100 in the extraction buffer. Polysomes were extracted from 12 to 72 h after inoculation. Infection caused a decline in yield of polysomes during this period when compared with healthy leaves of the same age. Polysomes isolated 16 h after inoculation with the virulent race were 20% less efficient at translation than polysomes from control leaves. In contrast polysome isolated from leaves inoculated with the avirulent race were 20% more efficient. Analysis of the labelled translation products by SDS-PAGE and fluorography showed relative increases in the synthesis of some proteins by 16 h after inoculation with either race when compared to products from healthy leaves. Protein synthesis in the infected plants was further analysed by in vivo labelling and one- and two-dimensional PAGE. The fluorographs revealed increased synthesis of a group of proteins from 58 to 116 kDa starting 12 h after inoculation with either race of P. hordei; confirming the results from the polysome translations. Two polypeptides with molecular weights of about 66 kDa were found to increase following infection only with the virulent race. By three days after inoculation with either fungal race the most obvious change in protein synthesis was a marked decrease in the synthesis of the two most prominent polypeptides with molecular weights of 15 and 51 kDa which were considered to be the subunits of ribulose bisphosphate carboxylase. The elicitor hypothesis, in attempting to explain cultivar-specific resistance in plants, postulates that resistance is controlled by the interaction of specific fungal elicitors and plant receptors and that this interaction which only occurs between resistant hosts and avirulent pathogens triggers specific gene expression leading to resistance. This hypothesis does not fit the situation in the barley-P. hordei interaction as protein synthesis showed similar changes following infection with either a virulent or an avirulent race.

Plant immune system is entirely based on the immunities of the individual cells in which systemic signals originate from the infection sites. Powdery mildew disease is one of the agents causing these infection sites, resulting in significant yield losses, if disease develops. Understanding the molecular basis of plant-pathogen interactions is the new trend for fighting against plant pathogens, since classical methods used in selection of resistant plants are becoming less and less efficient nowadays. Thus, finding out the genes which are responsible in plant&rsquo / s resistance is becoming very important. In this thesis, effect of &lsquo / General Control Nondepressible-2&rsquo / (GCN2) homolog protein in barley defense mechanism was aimed to be studied. The GCN2 of yeast was v previously identified in our laboratory as an interacting protein when the yeast cDNA library was screened with a putative yellow rust R gene (Yr10) fragment. There are reports available in the literature for the function of GCN2 protein, which makes it a good candidate for a role in disease resistance. Thus, the barley homologue of GCN2 might have a role in the R protein mediated early disease response of which may be proceeding via Programmed Cell Death (PCD). In order to observe such function of HvGCN2 in barley, silencing of its expression via Virus Induced Gene Silencing (VIGS) was investigated. Therefore, the GCN2 homologue was found to function as dampening the severity of the disease. The silencing with triple technical replicates was observed in 5 of the 6 samples, at an average of 43.2% by qRT-PCR analysis. The pathogen growth levels at different time points were analyzed under light microscope on the silenced and the control samples by measuring the primary and secondary hyphae lengths. The total of 24 seedlings and 292 individual spores were analyzed, and then the level of disease formation was quantitated with 603 primary hyphae and 106 secondary hyphae measurements. Up to 25% hyphae growth rate differences between the control and silenced groups were observed with a probability value less than 0.05 on t-test.