Thesis (MScAgric)--Stellenbosch University, 1998. / ENGLISH ABSTRACT: Eyespot is an important disease of spring wheat (Triticum aestivum L.). Four species of Ramulispora are associated with this disease, of which Tapesia yallundae and T. acuformis. are common. This thesis investigates the broader subjects of genetic variability, reproductive dynamics and fungicide resistance in Tapesia yallundae. Each of the chapters treats specific but related topics. T. yallundae, which is the only species thus far reported from South Africa, has been associated with yield losses of up to 50%. To enable the implementation of more accurate and effective control measures, understanding the dynamics of reproduction and the genetics of the pathogen is of utmost importance. Of the many plant disease control measures such as cultural practices, sanitation, biological control, etc., fungicide application is the most commonly resorted to measure in eyespot control. This thesis investigates the broader subjects of genetic variability, reproductive dynamics and fungicide resistance of Tapesia yallzll7dae. Fungicide application, however, is not without problems. The pathogen can build up resistance to fungicides. The most commonly used fungicides in eyespot control include the benzimidazole carbendazim, triazoles such as flusilazole, tebuconazole, propiconazole, bromuconazole, flutriafol, fenbuconazole, triademinol, and the imidazole, prochloraz. Cases of resistance to the groups listed above have been reported. Frequent monitoring for resistance is thus crucial to prevent wastage of fungicide and unnecessary impregnantation of the environment with potentially ineffective chemicals. In chapter 2 of this thesis 300 isolates of T. yallundae from 15 fields were evaluated for resistance against carbendazim, flusilazole, tebuconazole, propiconazole, bromuconazole, flutriafol and fenbuconazole. These results indicated that to some triazoles, such as fenbuconazole, a high level of resistance was already present in field populations. In a sexually reproducing fungus such as T. yallundae, knowledge pertaining to its ability to pass resistance factors to offspring is equally important. Mating studies were, therefore, also conducted with parental strains that showed signs of triazole resistance. Three generations were subsequently tested for resistance to five triazoles, namely flusilazole, tebuconazole, propiconazole, bromuconazole and flutriafol. Results of this study showed variable sensitivity in progeny, which indicated quantitative inheritance of resistance to triazoles. Although the sexual stage has not yet been observed in the field in South Africa, this knowledge lays the foundation for the long-term understanding of the population dynamics of the fungus. The ability of a heterothallic ascomycete population to reproduce sexually is dependent on the availability of its two mating types, MATI-I and MATI-2, their distribution, and female fertility amongst other factors. In the UK. the teleomorph is commonly observed in the field, which is in contrast to the situation in South Africa, where it has only been induced in the laboratory. A comparative study between the South African and the UK. populations was therefore undertaken. Isolates representative of the two populations were mated with tester strains as both sperm recipients and as sperm donors. This allowed the percentage of hermaphrodites to be determined. No difference in terms of female fertility was observed between the South African and the UK. populations, with both populations showing low effective population numbers. These data suggested, therefore, that the teleomorph would also occur more frequently in South Africa if the climate was more indusive to its development. The overall results of this study indicated that eyes pot could still be controlled by means of fungicide application in South Africa. Although a shift in sensitivity was observed towards fenbuconazole and flusilazole, no resistance was detected towards carbendazim. The latter might be due to the absen<.:eof the sexual stage in the field, coupled by the monocyclic nature of the pathogen and sensible fungicide regimes. The absence of T. acujormis makes the disease situation less complicated in terms of fungicide application and management. Continuous surveys will have to be conducted, however, to monitor this situation in future. / AFRIKAANSE OPSOMMING: Hierdie studie ondersoek die genetiese variasie, reproduksie dinamika en fungisied weerstand in Tapesia yallundae. Elke hoofstuk handel oor spesifieke maar verwante onderwerpe. Oogvlek is 'n belangrike siekte van lentekoring (Triticum aestivum L.). Vier spesies van Ramulispora word geassosieer met die siekte, waarvan Tapesia yallundae en T. acuformis mees algemeen voorkom. T. yallundae, wat tans die enigste spesie is wat in Suid-Afrika aangeteken is, het al verliese van tot 50% veroorsaak. Om meer akkurate en effektiewe beheermaatreels te implementeer, is dit noodsaaklik om die oorlewingsdinamika van die patogeen te verstaan. Van al die siektebeheermaatreels soos kulturele praktyke, sanitasie, biologiese beheer ens., bly fungisiedbehandeling die mees algemene maatreel vir die beheer van oogvlek. Fungisiedtoediening het egter ook verskeie probleme. Die patogeen kan weerstand opbou teen die fungisied. Die mees algemene fungisiedes wat vir oogvlekbeheer aangewend word sluit onder meer die benzimidasool karbendazim in, triasole soos flusilasool, tebukonasool, propikonasool, bromukonasool, flutriafol, fenbukonasool, triadimenol, en die imidasool, prochloraz. Weerstand is egter reeds teen hierdie middels bekend. Gedurige monitering vir weerstand is dus krities om die vermorsing van fungisied en besoedeling van die omgewing met oneffektiewe middels te beperk. In hoofstuk 2 van hierdie manuskrip word 300 isolate van T. yallundae van 15 lande geevalueer vir weerstand teenoor karbendazim, flusilasool, tebukonasool, propikonasool, bromukonasool, flutriafol en fenbukonasool. Resultate dui daarop dat teen sommige van hierdie triasole, soos bv. fenbukonasool, daar reeds 'n hoe vlak van weerstand teenwoordig was in veldpopulasies. In 'n seksueel reproduserende fungus soos T. yalluJ1dae, is dit noodsaaklik om te bepaal wat sy vermoe is om weerstandbiedenheid aan die nageslag oor te dra. Om die rede is paringstudies ook op ouers wat tekens van weerstand teenoor triasole getoon het uitgevoer. Drie generasies was gevolglik getoets vir weerstand teenoor vyf triasole, naamlik flusilasool, tebuconasool, propikonasool, brumukonasool en flutriafol. Resultate van die studie het 'n variasie in sensitiwiteit van die nageslag getoon, wat op 'n kwantitatiewe oorerwing van weerstand teen £riasole dui. Alhoewel die teleomorf nog nie in lande in Suid-Afrika opgemerk is nie, Ie hierdie kennis die fondament vir die langtermyn vertolking van die populasie dinamika van hierdie fungus. Die vermoe van 'n heterotalliese askomiseet populasie om seksueel voort te plant is afhanklik van die beskikbaarheid van sy twee paringstipes, MATI-I en MATl-2, hul verpreiding, vroulike vrugbaarheid en ander faktore. Alhoewel die teleomorf algemeen in lande in die Verenigde Koninkryk opgemerk word, is dit in kontras met die situasie in Suid-Afrika, waar hierdie stadium nog slegs in die laboratorium gelnduseer kon word. 'n Studie is dus onderneem om die Suid-Afrikaanse en V.K. populasies met mekaar te vergelyk. Isolate van die twee populasies is dus gepaar met paringsisolate as beide sperm ontvangers en sperm donors. Hierdie prosedure het dit moontlik gemaak om die persentasie hermafrodiete te bepaal. Geen verskille in vroulike fertiliteit is tussen die Suid-Afrikaanse en V.K. populasies bespeur nie, en beide populasies het ook 'n lae effektiewe populasie getal getoon. Hierdie data het dus voorgestel dat die teleomorf ook meer algemeen in Suid-Afrika sou voorkom as die klimaat meer geskik was vir teleomorf vormmg. Die resultate van hierdie studie het tot die slotsom gelei dat oogvlek steeds deur fungisiedbehandeling in Suid-Afrika beheer kan word. Alhoewel daar 'n merkbare verskuiwing in sensitiwiteit teenoor fenbukonasool en flusilasool was, was geen weerstand teenoor karbendazim waargeneem nie. Laasgenoemde kan dalk toegeskryf word aan die afwesigheid van die teleomorf in die veld, gekombineer met die monosikliese natuur van die patogeen en gebruik van alternerende fungisiedes. Die afwesigheid van T. acuformis maak die plaaslike siektetoestand minder gekompliseerd in terme van fungisied aanwending en bestuur. Voortdurende opnames sal egter uitgevoer moet word om hierdie situasie ook in die toekoms te monitor.

Doctor of Philosophy / Department of Plant Pathology / Megan Kennelly / Large patch, caused by the fungus Rhizoctonia solani anastomosis group (AG) 2-2 LP, is the most common and severe disease of zoysiagrass (Zoysia spp). Despite the importance of this disease, few studies have examined pathogen biology, cultivar susceptibility, cultural controls, and chemical controls. The objectives of this dissertation were: (1) Characterize large patch isolates based on anastomosis pairing, in-vitro mycelial growth rates, nuclear counts, virulence, PCR, and amplified fragment length polymorphism (AFLP); (2) Determine the effects of cultivation (aerification, verticutting, and sand topdressing) on disease severity; (3) Evaluate different fall and spring applications of the fungicides flutolanil, azoxystrobin, and triticonazole; (4) Evaluate the susceptibility of fifteen new zoysiagrass germplasm lines from parental crosses including Z. japonica, Z. matrella, and Z. pacifica. All the R. solani isolates from large patch-infected zoysiagrass from Kansas belonged to AG 2-2 LP. Variations were observed among the isolates in their average number of nuclei per cell, mycelial growth rates and virulence. There was also variation in the amplified fragment length polymorphism (AFLP) DNA fingerprints, suggesting possible underlying genetic differences of biological significance among members of AG 2-2 LP. Cultivation did not affect soil moisture or temperature. Cultivation also did not reduce patch sizes, nor influence turf recovery rate from large patch. From 2009 to 2011, spring and fall N fertility was consistently associated with lower percentages of diseased turf in both cultivated and non-cultivated plots at Manhattan and Haysville. In general, two fall applications of fungicide did not reduce disease compared to one fall application. Fungicides applied in the fall when thatch temperatures ranged from 17.8oC to 23.2oC reduced disease compared to untreated controls. Early spring applications reduced disease compared to later spring applications. In germplasm screening studies, all progeny had similar disease levels compared to Meyer in the growth chamber, but only 6 consistently had disease levels as low as Meyer in the field. Growth chamber results did not correlate to field results.

O fungo basidiomiceto Crinipellis perniciosa é o agente causal da vassoura-de-bruxa do cacaueiro (Theobroma cacao), podendo causar sérios perdas na produção. Este fungo é capaz de colonizar, além do cacau, várias outras plantas hospedeiras, onde pode se adaptar às novas condições, contribuindo, dessa forma, para o aumento da variabilidade genética desse microrganismo. Assim sendo, o objetivo desse trabalho foi isolar e identificar C. perniciosa de tecidos sadios de cacaueiro e comparar a sua variabilidade genética e fisiológica com isolados patogênicos por meio de análises moleculares e fisiológicas. Após a desinfecção e retirada da casca dos ramos, colônias morfologicamente similares a C. perniciosa foram obtidas de tecidos sadios de cacaueiro, os quais foram considerados como endófitos. A identificação preliminar desses isolados foi baseada nas observações morfológicas, tais como coloração da colônia e presença de grampos de conexão. Posteriormente, os isolados de C. perniciosa foram caracterizados geneticamente por RAPD, e sequenciamento de regiões do rDNA (18S+5.8S+28S), resistência a fungicidas, produção de exoenzimas e patogenicidade ao cacaueiro. A análise por marcadores RAPD separou os 37 isolados em 8 grupos (G1 a G8), sendo o grupo G1 dividido em 2 subgrupos (G1-1 e G1-2). Por esta análise foi possível observar que linhagens com 100 % de similaridade foram isoladas de locais diferentes, enquanto que isolados obtidos de uma mesma planta podem ser geneticamente diferentes. A análise da seqüência do rDNA dos isolados de C. perniciosa mostrou a formação de 6 grupos distintos (R1 a R6). Isolados obtidos de uma mesma planta não se agruparam, reforçando a hipótese de que isolados geneticamente diferentes podem ocupar a mesma planta hospedeira. Quanto resistência a fungicidas (tebuconazole, mancozeb, benomil e óxido cuproso), foi observado que tebuconazole foi o mais eficiente na inibição do crescimento miceliar, enquanto benomil apresentar menor taxa de inibição. Isolados endofíticos e patogênicos apresentaram comportamento similar em relação aos fungicidas. Resultado semelhante foi observado quanto a produção de exoenzimas (amilase, lipase, pectinase, exo e endoglicanase). Foi observada a produção de todas as enzimas avaliadas, sendo possível detectar variações entre os isolados. Entretanto, esta variação não foi correlacionada ao hábito endofítico ou patogênico. A patogenicidade de 8 isolados de C. perniciosa foi avaliada em mudas de cacau Catongo. A porcentagem de plantas infectadas com sintomas variou de 55% a 100%. Isolados endofíticos apresentaram baixa virulência sobre o cacaueiro, tendo o isolado endofítico 31 induziu sintomas em 60% das plantas inoculadas, resultado este provavelmente devido à alta pressão de inóculo. Este trabalho mostra pela primeira vez C. perniciosa como endófito em tecidos não meristemático do cacaueiro. / The basidiomycete fungus Crinipellis perniciosa (Stahel) Singer is the causal agent of Witches' Broom Disease of Cacao (Theobroma cacao L.) which is the main factor limiting cocoa production in the Americas. Pod losses of up to 90% are experienced in affected areas as evidenced by the 50% drop in production in Bahia province, Brazil following the arrival of the C. perniciosa in the area in 1989. The disease has proven particularly difficult to control and many farmers in affected areas have given up cacao cultivation. Any useful control strategy for Witches' Broom disease must be effective against in range strains of the pathogen. It is already known that pathogenic variation exists among isolates of Crinipellis perniciosa obtained from different areas and host plants. However, any study was developed about the variability between endophytic and pathogenic population. In order to evaluate the genetic variability of 35 isolates of endophytic and pathogenic populations of Crinipellis perniciosa, the RAPD technique, ITS sequencing and fungicide susceptibility were performed. Genetic variability between 35 isolates of C. perniciosa was analysed by the random amplified polymorphic DNA (RAPD) technique, which indicated that isolates from other host plants were more diverse than isolates obtained from cacao plants. Among cacao isolates was observed at least two groups, one formed mainly by endophytic isolates and other by pathogenic ones. Analysis by ITS sequence grouped isolates independently of endophytic or pathogenic status. Fungicide susceptibility showed that cupric oxide inhibits statistically more endophytic isolates than pathogenic ones, showing that could have physiological differences between these populations. The present study highlighted the possible genetic and physiological differences between endophytic and pathogenic population of C. perniciosa.

Les travaux de recherche présentés ici ont pour objectif la synthèse de fongicides innovants de la manière la plus éco-compatible qu’il soit. Pour cela, plusieurs axes de réflexion ont été étudiés. Tout d’abord, la synthèse d’un intermédiaire de fongicides bien connu, l’o-crésol1, a été réalisée à partir d’un substrat naturel, le salicylaldéhyde2. Les conditions expérimentales ont été éco-conçues afin de correspondre au mieux aux exigences de la chimie verte. Des solvants et des réactifs classés verts ont été utilisés, comme l’éthanol, l’eau et l’acide acétique3 par exemple, ainsi que des techniques alternatives comme l’activation par micro-ondes (Schéma 1). Dans un second temps, des synthèses éco-compatibles de nouveaux fongicides ont été réalisées. Le salicylaldéhyde, substrat naturel de référence, a été mis à réagir avec différents alcools afin de réaliser des réactions d’éthérifications réductrices. Les molécules obtenues sont composées d’une partie phénolique portant un groupement alcoxyle en position 2. Ce groupement alcoxyle se compose d’une longueur de chaîne variable ayant une influence sur les propriétés fongicides des produits synthétisés (Schéma 1). Troisièmement, des réactions d’homo- et d’hétéro-couplages pinacoliques ont été réalisées entre divers aldéhydes, aromatiques et aliphatiques, afin d’obtenir des diols vicinaux, appelés pinacols, symétriques ou non (Schéma 1). Dernièrement, les propriétés fongicides et anti-oxydantes de toutes les molécules obtenues de manière stable, référencées ou non, ainsi que les substrats de départ, ont été testés. Les tests microbiologiques ont été réalisés sur une gamme de diverses souches fongiques. / The purpose of this research is to synthesize innovative fungicides by the most ecocompatible way. Several hypotheses have been investigated. First, an extensively studied molecule, intermediate of fungicides, o-cresol1, has been achieved from a natural substrate, salicylaldehyde2. The experimental conditions have been eco-designed in the aim to best match green chemistry requirements. Green solvents and reactants have been used, as ethanol, water and acetic acid3 for example, as well as alternative technologies like microwaves activation (Schema 1). Secondly, novel fungicides green syntheses have been carried out. Salicylaldehyde, a natural substrate model, reacted with various alcohols in the aim to perform reductive etherification reactions. In this way, products were composed of phenolic part with an alkoxyl group at the 2nd position. These alcoxyl groups have various chain lengths, playing a key role in fungicidal ability (Schema 1). Then, homo and hetero pinacol coupling reactions have been achieved with various aromatic or aliphatic aldehydes to obtain symmetrical and asymmetrical vicinal diols, called pinacols (Schema 1). At last, fungidal and antioxidant properties of all stable obtained molecules, referenced or not, as well as starting materials, have been evaluated. Microbiological studies have been carried out with various fungal strains.

A alface é a hortaliça folhosa mais consumida no Brasil. Seu cultivo vem sendo severamente afetado pelo fungo Thielaviopsis basicola, que ocasiona podridão radicular e subdesenvolvimento de plantas. Não existem maiores estudos sobre o controle desse patógeno nas condições brasileiras. Os objetivos deste trabalho foram: a determinação da ocorrência de T. basicola em plantios comerciais de alface e outras folhosas do Estado de São Paulo, do efeito da temperatura sobre a patogenicidade in vitro dos isolados obtidos e a avaliação da viabilidade de seu controle através do emprego de cultivares de alface resistentes, de fungicidas e da solarização, com e sem incorporação de matéria orgânica. Quatorze isolados de T. basicola foram obtidos de raízes necróticas de alface, chicória e rúcula. O efeito da temperatura sobre a patogenicidade in vitro dos isolados foi determinado, inoculando-se sementes de alface semeadas em agar-água. Foram realizados dois ensaios. No primeiro, quatorze cultivares de alface foram inoculadas com o isolado L1, a 21 e 30ºC. No segundo, a cv. Elisa foi inoculada com os quatorze isolados de T. basicola, a 21 e 27ºC. O isolado L1 foi mais agressivo a 21ºC do que a 30ºC, ocasionando maiores severidade da doença e restrição do comprimento de plântulas em todas as cultivares. Todos os isolados causaram podridão radicular e reduziram o comprimento de plântulas a 21 e 27ºC, destacando-se o isolado A2 como o mais agressivo. De modo geral, os isolados foram mais agressivos a 27 que a 21ºC, exceto L1 e L2. A reação de cultivares de alface a três isolados de T. basicola foi avaliada in vivo. Primeiramente, treze cultivares foram confrontadas com o isolado L1 e, posteriormente, seis dessas cultivares foram inoculadas com os isolados A2 e CH. As cultivares do tipo lisa foram suscetíveis e as do tipo crespa, resistentes, exceto a cv. Verônica. Quanto às cultivares do tipo americana, houve variação na reação ao patógeno. O efeito de fungicidas sobre o crescimento micelial e a esporulação in vitro foi determinado para três isolados de T. basicola. Nove fungicidas foram testados para o isolado L1 e sete para os isolados A2 e CH, nas concentrações de 0; 0,1; 1, 10 e 100 ppm de ingrediente ativo. O fungicida carbendazim foi o mais efetivo. A eficiência de sete fungicidas foi avaliada no controle de T. basicola in vivo. Todos os produtos testados foram eficientes, destacando-se o tebuconazole e o triadimenol. O controle de T. basicola através da solarização, associada ou não à incorporação de 10 ton / ha de torta de mamona ou de cama de frango, também foi avaliado. A solarização do solo com e sem incorporação de matéria orgânica controlou efetivamente o patógeno. Os melhores resultados foram obtidos com a incorporação da torta de mamona seguida de solarização, que permitiu a produção de plantas com maiores comprimento e massa fresca e evitou o aparecimento de qualquer sintoma de necrose radicular. / Lettuce is the most used leafy vegetable in Brazil. Its crop is being severely affected by Thielaviopsis basicola, fungus which causes root rot and stunting in plants. There are no control studies about this pathogen for the brazillian conditions. This research aimed: to determine the T. basicola occurence in São Paulo State commercial lettuce and other leafy vegetables crops, the temperature effect on isolates pathogenicity in vitro and control viability evaluation by lettuce resistant varieties, fungicides and solarization with and without organic matter amendments. Fourteen T. basicola isolates were got from lettuce, chicory and arugula necrotic roots. Temperature effect on the isolates pathogenicity in vitro was determined in inoculated seeds sown on the surface of water agar medium. Two trials were carried out. In the first one, fourteen lettuce varieties were inoculated with L1 isolate at 21 and 30ºC. In the second, only cv. Elisa was inoculated with fourteen T. basicola isolates, at 21 and 27ºC. The isolate L1 was the most aggressive at 21ºC than 30ºC, causing higher disease severity and seedling length restriction of all varieties. All isolates induced root rot and reduced seedlings length at 21 and 27ºC, but A2 isolate was the most aggressive one. All isolates were more aggressive at 27 rather than 21ºC, except L1 and L2 isolates. Lettuce varieties reactions to three T. basicola isolates were evaluated in vivo. First about thirteen varieties were challenged with L1 isolate and later, six of them were inoculated with A2 and CH isolates. Butterhead types were susceptible and Grand Rapids types were resistant, except cv. Veronica. Iceberg type varieties varied in their reaction to pathogen. Fungicide effect on mycelial growth and sporulation in vitro was determined for three T. basicola isolates. Nine fungicides were tested for L1 isolate and seven for A2 and CH isolates, at concentrations of 0; 0,1; 1; 10 and 100 ppm of active ingredient. Carbendazim was the most effective. Seven fungicides efficacy was evaluated to control of T. basicola in vivo. All tested products were efficient, but tebuconazole and triadimenol were the best ones. The control of T. basicola through soil solarization with and without chicken litter and coarse castor bean meal amendments, at 10 ton / ha, also was evaluated. Soil solarization alone and combined with organic matter amendments were both effective ways to control the pathogen. The best results were got with incorporation of coarse castor bean meal followed by solarization, which allowed increase in fresh weight and length of plants, and avoided root rot symptoms.

Submitted by Sandra Pereira (srpereira@ufrrj.br) on 2017-05-12T14:19:07Z No. of bitstreams: 1 2010 - Henriqueta Talita Guimar?es Barboza.pdf: 10302674 bytes, checksum: 6daf4f88308c3c733615f0772d6db594 (MD5) / Made available in DSpace on 2017-05-12T14:19:07Z (GMT). No. of bitstreams: 1 2010 - Henriqueta Talita Guimar?es Barboza.pdf: 10302674 bytes, checksum: 6daf4f88308c3c733615f0772d6db594 (MD5) Previous issue date: 2010-03-04 / Twelve heterocyclic dialkylphosphorylhydrazones compounds were synthesized in this work. Dialkylphosphorylhydrazones were obtained through 3 steps of reaction. The first step consists on the synthesis of different dialkylphosphonates obtained by reaction of triethyl phosphate (PCl3) with 3 mols of the corresponding alcohol. The second step occurs by the reaction with dialkylphosphonates synthesized and hydrazine, producing dialkylphosphorylhydrazines. The third and last step is the condensation reaction of these dialkylphosphorylhydrazines with different heterocycle substituted aromatic aldehydes. The analysis of the 1H NMR spectra allowed to observe the stereoisomers E and Z, with the formation of E diastereisomer preferencially. All the compounds were characterized by 1H, 13C and 31P NMR, IR and mass spectroscopy. Tests in vitro with, Alternaria sp., Colletotrichum sp., Fusarium sp. e Fusarium solani, known as patogenic in papaya (Carica papaya L.) were performed in order to confirm if the compounds showed fungicide activity. The substance 6 B was the one with the best performance in the inhibition of all fungal growths. Finally, tests with acetylcholinesterase enzyme indicated that all substances did not inhibit this enzyme. Figure 2 - General structures of dialkylphosphoryhidrazones synthesized. R ? Ethyl; propyl; isopropyl e isobuthyl. X = O, Y, Z e W = CH; X = S, Y, Z e W = CH; X = NH, Y, Z e W = CH; X = NH, Y= CH, Z=CH e W = N; X = CH, Y= N, Z=CH e W = N; / Neste trabalho foram sintetizadas doze (12) novas dialquilfosforilidrazonas heteroc?clicas. Esses compostos foram obtidos utilizando-se tr?s etapas de rea??o. A primeira etapa consiste na s?ntese de diferentes fosfonatos de dialquila, obtidos por meio da rea??o do tricloreto de f?sforo (PCl3) com tr?s mols do ?lcool correspondente. A segunda etapa ocorre por meio da rea??o dos fosfonatos de dialquila sintetizados com a hidrazina, formando as dialquilfosforilidrazinas. A terceira e ?ltima etapa ? a condensa??o destas dialquilfosforilidrazinas com diferentes alde?dos heteroc?clicos. A an?lise dos espectros de RMN-1H indica a forma??o de diastereois?meros, E e Z, com predomin?ncia do diastereis?meros E. Todos os compostos obtidos foram caracterizados por RMN de H1 e C13 e P31, IV, an?lises de massas. Em seguida foram realizados testes in vitro para verificar se os mesmos possuem atividade fungicida sobre os fungos Alternaria sp., Colletotrichum sp., Fusarium sp. e Fusarium solani, conhecidos por serem pat?genos da cultura de mam?o (Carica papaya L.). O composto 6 B foi o que apresentou o melhor efeito fungicida sobre o crescimento de todos os fungos. Finalmente foram realizados testes com a enzima acetilcolinesterase a fim de observar a toxicidade dos compostos sintetizados frente a esta enzima. Figura 1- Estrututa gen?rica das dialquilfosforilidrazonas heteroc?clicas sintetizadas. R ? Etil; propil; isopropil e isobutil. X = O, Y, Z e W = CH; X = S, Y, Z e W = CH; X = NH, Y, Z e W = CH; X = NH, Y= CH, Z=CH e W = N; X = CH, Y= N, Z=CH e W = N.

Ce mémoire est subdivise en deux parties. La première partie concerne la synthèse de pyrazoles présentant un intérêt sur le plan agrochimique. En effet, le noyau pyrazole est présent dans de nombreux composes ayant des activités biologiques diverses et en particulier antifongique. Au cours de ce travail, nous avons développé diverses approches convergentes de pyrazoles diversement substitués au moyen de réactions de couplages croisés pallado-catalyses sélectifs et séquentiels à partir de pyrazoles possédant différents points d’encrages. Dans la deuxième partie, nous nous sommes intéressés à la synthèse de diverses furopyridones en tant qu’analogues de produits naturels possédant une activité antifongique, et notamment le Cladobotryal. Dans ce but, diverses alcynylpyridones ont été synthétisées et mises en jeu dans divers processus de cyclisation pour atteindre de manière divergente une série de furo[3,2-c]pyridin-4-ones, furo[3,2-c]pyridin-6-ones et furo[2,3-b]pyridin-4-ones / This thesis is subdivided into two principal parts. The first part is focussed on the synthesis of pyrazole derivatives of agrochemical relevance. Indeed, the pyrazole nucleus is found in numerous compounds possessing interesting biological properties, and notably antifungal activities. Various convergent approaches to diversely substituted pyrazoles have therefore been developed by means of site-selective palladium-catalyzed cross-coupling reactions conducted sequentially on pyrazole scaffolds. In the second part, we have been involved in the synthesis of furopyridones as simplified analogues of natural compounds possessing antifungal activities such as Cladobotryal. Toward this end, various alkynylpyridones have been synthesizes and involved in diverse cyclization processes to access a series of furo[3,2-c]pyridin-4-ones, furo[3,2-c]pyridin-6-ones, and furo[2,3-b]pyridin-4-ones

Integrated Pest Management (IPM) has long been promoted as a means of reducing reliance on pesticide inputs as compared to conventional farming systems. Reduced pesticide application could be beneficial due to the links between intensive pesticide use and negative impacts upon biodiversity and human health as well as the development of pesticide resistance. Work assessing the potential of IPM in cereal production is currently limited, however, and previous findings have generally covered the subject from the perspective of either field trial data or social science studies of farmer behaviour. This thesis attempts to help to address this knowledge gap by providing a more holistic assessment of IPM in Scottish spring barley production (selected because of its dominance in Scotland’s arable production systems), in relation to three of its most damaging fungal pathogens: Rhynchosporium commune, Blumeria graminis f.sp. hordei, and Ramularia collo-cygni. Several IPM techniques of potential relevance to the sector were identified, and the prospects of three in particular – crop rotation, varietal disease resistance, and forecasting disease pressure – were assessed in several ways. Preliminary analysis of experimental field trial data collected from 2011 – 2014 across Scotland found that the majority of spring barley trials in this period (65%) did not show a statistically significant impact of fungicide treatment on yield, with the average yield increase due to fungicide application being 0.62 t/ha. This initial analysis was expanded upon using stepwise regressions of long-term (1996 – 2014) field trial data from the same dataset. Here, the difference between treated and untreated yields could be explained by disease resistance, average seasonal rainfall (whereby wetter seasons saw an increased impact of fungicide use on yield), and high combined disease severity. Stakeholder surveying provided information about current practice and attitudes towards the selected IPM techniques amongst a group of 43 Scottish spring barley farmers and 36 agronomists. Stakeholders were broadly open to taking up IPM measures on farm; sowing of disease resistant varieties was most frequently selected as the best technique in terms of both practicality and cost, though individual preference varied. However, a disparity was seen between farmer perception of their uptake of IPM and actual, self-reported uptake for both varietal disease resistance and rotation. Farmers and agronomists also overestimated the impact of fungicide use as compared with the field trials results – the majority of stakeholders believed fungicide treatment to increase yields by 1 - 2 t/ha, while the majority of 2011 – 2014 field trials had a yield difference of under 1 t/ha. The reasons behind these differences between perception and practice are not currently known. Finally, an annual survey of commercial crops, gathered from 552 farms across Scotland (from 2009 – 2015), highlighted two gaps where IPM practice could be improved upon. Firstly, relatively few of the varieties listed in the commercial crops database were highly resistant to the three diseases – 26.1% were highly resistant to Ramularia, 14.2% to Rhynchosporium, and 58.1% to mildew. Secondly, 71% of the farms included in the database had planted barley in at least two consecutive seasons, indicating that crop rotation practices could be improved. The overarching finding of this project is that there is scope for IPM uptake to be improved upon and fungicide use to be reduced while maintaining high levels of yield in Scottish spring barley production. Incorporating experimental field data, stakeholder surveying, and commercial practice data offered a unique view into the potential for IPM in this sector, and provided insights which could not have been gained through the lens of a single discipline.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / This work describes the seasonal analysis of the yield and chemical composition of the essential oil (EO) for the native species Piper gaudichaudianum Kunth in a population of Santa Maria, in order to determine the best time for collection. The effect of leaves drying on the yield and chemical composition of EO was also verified, in order to evaluate the possible influence of this post-harvest process on the extractive features. The EO oil and its major constituent had their fungitoxicity assessed against phytopathogenic and wood decay fungi. EO from fresh and dried leaves, and reproductive organs (inflorescences and fruits) were extracted separately by hydrodistillation, and then had their yields determined (w/ w %). The chemical composition was analyzed by gas chromatography-mass spectrometry (GC-MS) and gas chromatography with flame ionization detector (GC-FID). Additionally, fungitoxic assays were performed by the agar dilution method using PSA (potato-sucrose-agar). The presence of reproductive organs (RO) did not affect the leaves (L) EO of P. gaudichaudianum on the observed year, with a minimum value of 1.32% and a maximum of 1.61% without significant difference (p > 0.05). Chemical analysis of L and RO EO revealed that these are predominantly formed by phenylpropanoid dilapiolle (59.2 to 87.8%), while myristicin was identified only in the EO of the RO. The chemical composition of EO oils had no influence of seasonality and phenophases, being possible to collect the plant material at any time of year. Drying of the leaves did not affect the yield and composition of EO. The L and RO EO of P. gaudichaudianum presented fungitoxic activity for phytopathogenic fungi (Fusarium moniliforme and Botryosphaeria rhodina) and also for wood decay fungi (Pycnoporus sanguineus and Gloeophyllum trabeum) at concentrations of 0.25-1.0 uL / mL. The results indicated that the major constituent dilapiolle is responsible for the activity. / Este trabalho descreve a análise sazonal do rendimento e composição química do óleo essencial (OE) da espécie nativa Piper gaudichaudianum Kunth em uma população de Santa Maria, RS, com o objetivo de determinar a época mais adequada para coleta. Também foi verificado o efeito da secagem de folhas sobre o rendimento e a composição química dos OE, a fim de avaliar a possível influência desse processo pós-colheita sobre estas características do extrativo. O EO bem como seu constituinte majoritário tiveram sua atividade fungitóxica avaliada contra fungos fitopatogênicos e apodrecedores da madeira. Os OE de folhas frescas, secas e órgãos reprodutivos (inflorescências e frutos) foram extraídos separadamente através do método de hidrodestilação e em seguida foram determinados seus rendimentos (m/m %). A composição química foi analisada por cromatografia gasosa acoplada a espectrometria de massas (CG-EM) e cromatografia gasosa com detector de ionização em chama (CG-DIC). Adicionalmente, os testes fungitóxicos foram realizados através do método de diluição em Ágar usando BSA (batata-sacarose-ágar). A presença de órgãos reprodutivos (OR) não alterou o rendimento do OE de folhas (F) de P. gaudichaudianum no ano observado, apresentando valor mínimo de 1,32% e máximo de 1,61% sem diferença significativa (p > 0,05). A análise química dos OE de F e OR evidenciou que estes são formados preponderantemente pelo fenilpropanóide dilapiol (59,2-87,8%) enquanto que a miristicina foi identificada apenas para os OE de OR. A composição química dos OE não teve influência da sazonalidade e das fenofases, sendo possível a coleta do material vegetal em qualquer época do ano. A secagem das folhas não alterou o rendimento e composição química dos OE. Os OE de F e OR de P. gaudichaudianum apresentaram atividade fungitóxica para os fungos fitopatogênicos (Fusarium moniliforme e Botryosphaeria rhodina) e também para os apodrecedores da madeira (Pycnoporus sanguineus e Gloeophyllum trabeum) em concentrações de 0,25-1,0 μL/mL. Os resultados indicaram que o constituinte majoritário dilapiol é o responsável pela atividade.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The sensitivity of ten isolated of Colletotrichum lindemuthianum the systemic fungicides was evaluated in vitro and in vivo, in experimental design was completely random with three replications. In the study in vitro, it was evaluated the percentage of germination of spores of the fungus when submitted to the benzimidazol fungicides: carbendazin and methyl tiophanate and the estrobilurins: azoxystrobin and pyraclostrobina. The methodology consisted in incorporate the fungicide to the agar at the final concentrations of 0; 0,1; 1,0; 10 and 100 ppm. With the germination data the DL50 (rate capable to inhibit 50% of the germination of spores) was calculated. For each fungicide was calculated the DL50, being observed differences in the sensitivity of isolated of C. lindemuthianum. Considering the fungicide carbendazin, race 65 presented low sensitivity while races 08, 81, 321 had been moderately sensible to this fungicide. In the case of the estrobilurins, all the isolated ones had revealed highly sensible to the fungicides azoxystrobin and pyraclostrobin, proving the effect of this chemical group on the germination of spores. In the in vivo evaluation, it was evaluated the DL50 having considered the injuries of C. lindemuthianum formed from the inoculation of a known concentration of spores in detached bean leaves submitted to the final concentrations of 0; 0,5; 1,0; 5,0; 10,0 and 50,0 ppm of tebuconazol and epoxiconazol. It was not observed difference among isolates, considering the classification of sensitivity proposed by Edgington al. (1971). However, it was verified great amplitude of values of DE50 between the isolated ones. Race 321 presented moderate sensitivity to the epoxiconazol, races 08 and 65 with relatively bigger values of DE50. The same occurred with race 81 with regard to epoxiconazol. The alteration in the sensitivity on C. lindemuthianum isolates, when submitted to the group of the benzimidazoles, was related to the frequency and pressure of selection imposed by repeated fungicides application. Regarding the triazoles, it was not observed differences in the classification of sensitivity between the isolates, showing that the use of these fungicides in the control of anthracnose did not produce a similar selective effect as observed with the fungicides tebuconazol and epoxyconazol. / A sensibilidade in vitro e in vivo de dez isolados de Colletotrichum lindemuthianum a fungicidas sistêmicos foi avaliada in vitro e in vivo, em delineamento experimental inteiramente casualizado com três repetições. No estudo in vitro foi avaliada a porcentagem de germinação de conídios do fungo quando submetidos aos fungicidas benzimidazóis: carbendazin e tiofanato metílico e estrobilurinas: azoxistrobina e piraclostrobina. Foi utilizada a metodologia do fungicida incorporado ao agar nas concentrações finais de 0; 0,1; 1,0; 10 e 100 ppm. Com os dados de germinação foi calculada a DL50 (dose efetiva capaz de inibir 50% da germinação de esporos) para cada fungicida, sendo observadas diferenças na sensibilidade dos isolados de Colletotrichum lindemuthianum com relação aos fungicidas testados. Considerando o fungicida carbendazin, a raça 65 apresentou baixa sensibilidade enquanto as raças 08, 81, 321 foram moderadamente sensíveis a este fungicida. No caso das estrobilurinas, todos os isolados mostraram-se altamente sensíveis aos fungicidas azoxistrobina e piraclostrobina, comprovando o efeito esporocida desse grupo químico. Na avaliação in vivo, foi avaliada a a DL50 considerando as lesões de C. lindemuthianum formadas a partir da inoculação de uma concentração conhecida de esporos em folhas destacadas de feijão submetidas a concentrações finais de 0; 0,5; 1,0; 5,0; 10,0 e 50,0 ppm de tebuconazole e epoxiconazole. Não houve diferença quanto à classificação de sensibilidade proposta na escala adaptada de Edgington et al. (1971), embora tenha sido verificada grande amplitude de valores de DE50 entre os isolados. A raça 321 apresentou moderada sensibilidade a epoxiconazole, ficando as raças 08 e 65 com valores de DE50 relativamente maiores. O mesmo ocorreu com a raça 81 com relação ao epoxiconazole. A alteração na sensibilidade de isolados de C.lindemuthianum quando submetidos ao grupo dos benzimidazóis está relacionada a origem, freqüência e pressão de seleção imposta pela aplicação repetida desses fungicidas em determinados locais onde predominam determinados grupos de raças fisiológicas. Com relação aos triazóis não houve diferenças na classificação de sensibilidade entre os isolados, mostrando que a utilização desses fungicidas no controle da antracnose não produziu um efeito seletivo similar ao observado com os fungicidas tebuconazole e epoxiconazole, considerando a mesma população do patogeno.