USAID / Se identifica la expresión diferencial de genes efectores tipo RXLR en dos cepas aisladas del centro de los andes peruanos de P. infestans EC-1 mediante secuenciamiento del transcriptoma de la interacción papa-P.infestans de los primeros días después de la infección, siendo confirmada por qRT-PCR. Los genes efectores fueron silenciados en una cepa para Avr-vnt1 en POX109 y para el homólogo Avh9.1 en POX067, pero expresados en la otra. Además, los resultados de transcriptoma fueron comparados con tres cepas adicionales del linaje EC-1. En el análisis de SNPs de Avr-blb1, Avr-blb2 y Avr-vnt1, la variabilidad alélica no tuvo predominancia frente a la variabilidad de expresión de genes. Asimismo, debido al silenciamiento génico de Avr-vnt1 se evaluó la expresión de estos en plantas transgénicas [Rpi-vnt1.1] a fin de encontrar si la resistencia transgénica era funcional. Encontrando que en ambas cepas en todos los eventos y en el control susceptible Yungay se expresan, a diferencia del resultado anterior. El descubrimiento de efectores silenciados en las poblaciones del patógeno pueden guiar al uso de genes R específicos en los programas de mejoramiento genético. Pudiendo el gen Rpi-vnt1 no ser recomendado. / Tesis

Oomycetes have long been considered as a separate class within the kingdom Fungi, but they are in fact closer to brown algae. They are currently classified in the Stramenopile eukaryotic kingdom, which includes heterokont algae and water molds. The major cell wall polysaccharides in Oomycetes are b-(1à3) and b-(1à6)-glucans, as well as cellulose, which has never been reported in any fungal species. Chitin - the major cell wall polysaccharide in fungi - occurs in minor amounts in the walls of some Oomycetes. Some Oomycete species are pathogens of great economical importance. For example, species of the genus Phytophthora are well studied plant pathogens that cause considerable economical losses in agriculture. Saprolegniosis, a fish disease caused by species from the genus Saprolegnia, is a major problem in the aquaculture industry and represents a threat to populations of salmonids in natural habitats. Currently, there are no chemicals available that are at the same time efficient Oomycete inhibitors, environmentally friendly and safe for human consumption of treated fishes. The biosynthesis of cellulose in Oomycetes is poorly understood, even though this biochemical pathway represents a potential target for new Oomycete inhibitors. In this work, cellulose biosynthesis was investigated in two selected Oomycetes, the plant pathogen Phytophthora infestans and the fish pathogen Saprolegnia monoica. A new Oomycete CesA gene family was identified. It contains four homologues designated as CesA1, CesA2, CesA3 and CesA4. The gene products of CesA1, 2 and 4 contain Pleckstrin Homology domains located at the N-terminus. This represents a novel feature, unique to the Oomycete CesA genes. CesA3 is the dominantly expressed CesA homologue in the mycelium of both S. monoica and P. infestans, while CesA1 and CesA2 are up-regulated in virulent life stages of P. infestans. CesA4 was expressed only in minute amounts in all investigated types of cells. Gene silencing by RNA interference of the whole CesA gene family in P. infestans lead to decreased amounts of cellulose in the cell wall. The inhibitors of cellulose synthesis DCB and Congo Red had an up-regulating effect on SmCesA gene expression, which was accompanied by an increased b-glucan synthase activity in vitro. In addition, these inhibitors slowed down the growth of the mycelium from S. monoica. Zoospores from P. infestans treated with DCB were unable to infect potato leaves and showed aberrant cell wall morphologies similar to those obtained by silencing the CesA gene family. Altogether these results show that at least some of the CesA1-4 genes are involved in cellulose biosynthesis and that the synthesis of cellulose is crucial for infection of potato by P. infestans. / QC 20101110

Made available in DSpace on 2015-03-26T13:39:59Z (GMT). No. of bitstreams: 1 texto completo.pdf: 425667 bytes, checksum: 1f8c842faf40449adebf9de5ae50ad48 (MD5) Previous issue date: 2013-07-17 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / The tomato production is considered a high risk economic activity, and of great agronomic complexity mainly because of the high number of pathogens that causes diseases in the culture. Among the pathogens of greatest importance, there is the oomycete Phytophthora infestans (Mont.) de Bary, causing late blight, which is considered the most destructive disease of tomato. There are no commercial tomato cultivars resistant and the control of this disease is highly dependent on the use of fungicides. Considering that the tomato is one of the most studied crop in genetic terms, the lack of resistant cultivars in the market reflects the difficulty in working this trait in breeding programs. The objectives with this present study was to select sources of resistance to late blight among accessions of Solanum lycopersicum, and estimate the overall and specific ability of combining six tomato accessions of Solanum lycopersicum selected for resistance to late blight, aiming at the selection of potential genitors for pre-breeding for resistance to this disease. Two experiments were carried out. In the first, twelve accessions of S. lycopersicum, from the Vegetable Germplasm Bank of the Federal University of Viçosa - BGH - UFV (BGH-973, BGH-1025, BGH- 2017, BGH-2093, BGH-2095, BGH-2102, BGH-2117, BGH-2127, BGH-2130, BGH- 2332, BGH-2333 and BGH-2343) were evaluated in a randomized block design with three replications. For the evaluation of the resistance, the plants were inoculated with a mixture of sporangia of P. infestans, in the concentration of 5x103 sporangia mL-1 , collected in different regions of the Zona da Mata Mineira. It was evaluated the percentage of plant tissue affected by the disease, or the percentage of severity, evaluated in the form of area under the disease progress curve (AUDPC). The accessions BGH-2102, BGH-2117, BGH-2127, BGH-2130, BGH-2332 and BGH-2343 obtained average values of AUDPC lower than the resistant controls, and were selected as a source of resistance to late blight. The selected accessions as resistant to late blight were used in a second experiment, where a balanced diallel of half table were realized, and fifteen F1 s hybrids were obtained. The six accessions of S. lycopersicum, BGH- 2102, BGH-2117, BGH-2127, BGH-2130, BGH-2332 and BGH-2343 together with the fifteen F1 s hybrid were evaluated in a randomized block design with three replications. For the evaluation of the resistance, the plants were inoculated with a mixture of sporangia of P. infestans, in the concentration of 5x103 sporangia mL-1 collected in different regions of the Zona da Mata Mineira. The percentage of plant tissue affected by the disease, or the percentage of severity, presented in the form of area under the disease progress curve (AUDPC) were evaluated. The diallelic analysis was carried out according to the model of GRIFFING (1956). Observed existence of additive genetic variability between the genitors and the predominance of additive gene effects involved in determining the AUDPC, besides the existence of bidirectional dominance deviations in the control of the character. The diallel analysis was efficient in selecting genitors aimed at a pre-breeding of resistance to late blight, highlighting the accessions BGH- 2117, BGH-2127 and BGH-2343 with the greatest frequency of favorable alleles and divergent, thus they can be included in crossings aiming the pre-breeding for resistance to late blight. / A tomaticultura é considerada uma atividade de elevado risco econômico e grande complexidade agronômica, principalmente pelo elevado número de patógenos que causam doenças na cultura. Entre os patógenos de maior importância, destaca-se o oomiceto Phytophthora infestans (Mont.) de Bary, causador da requeima ou mela, considerada a doença mais destrutiva do tomateiro. Não existem cultivares comerciais de tomate resistentes, e o controle da doença é altamente dependente do uso de fungicidas de caráter preventivo e/ou curativo. Considerando que o tomateiro é uma das plantas cultivadas mais bem estudadas em termos genéticos, a inexistência de cultivares resistentes no mercado reflete a dificuldade em se trabalhar essa característica nos programas de melhoramento. Os objetivos com o presente estudo foram selecionar fontes de resistência à requeima entre acessos de Solanum lycopersicum, e estimar as capacidades geral e específica de combinação dos acessos de S. lycopersicum selecionados quanto a resistência à requeima, visando selecionar potenciais genitores para o pré-melhoramento da resistência a esta doença. Foram realizados dois experimentos. No primeiro avaliou-se doze acessos de S. lycopersicum, do Banco de Germoplasma de Hortaliças da Universidade Federal de Viçosa BGH UFV (BGH- 973, BGH-1025, BGH-2017, BGH-2093, BGH-2095, BGH-2102, BGH-2117, BGH- 2127, BGH-2130, BGH-2332, BGH-2333 e BGH-2343) no delineamento em blocos casualizados, com três repetições. Para a avaliação da resistência, as plantas foram inoculadas com uma mistura de esporângios de P. infestans, na concentração de 5x103 esporângios mL-1, coletados em diferentes regiões da Zona da Mata Mineira. Avaliou-se a porcentagem do tecido vegetal afetado pela doença, ou seja, a porcentagem de severidade, avaliada sob a forma de área abaixo da curva de progresso da doença (AACPD). Os acessos BGH-2102, BGH-2117, BGH-2127, BGH-2130, BGH-2332 e BGH-2343 obtiveram valores médios de AACPD inferiores aos das testemunhas resistentes, sendo selecionados como fonte de resistência à requeima. Os acessos selecionados como resistentes à requeima foram utilizados em um segundo experimento, onde foi realizado um dialelo balanceado de meia tabela, e obtidos quinze híbridos F1 s. Os seis acessos de S. lycopersicum, BGH-2102, BGH-2117, BGH-2127, BGH-2130, BGH-2332 e BGH-2343 juntamente com os quinze híbridos F1 s foram avaliados em delineamento em blocos casualizados, com três repetições. Para a avaliação da resistência, as plantas foram inoculadas com uma mistura de esporângios de P. infestans, na concentração de 5x103 esporângios mL-1, coletados em diferentes regiões da Zona da Mata Mineira. Avaliou-se a porcentagem do tecido vegetal afetado pela doença, ou seja, a porcentagem de severidade, avaliada sob a forma de área abaixo da curva de progresso da doença (AACPD). Procedeu-se a análise dialélica de acordo com o modelo de GRIFFING (1956). Observou-se existência de variabilidade genética aditiva entre os genitores e predominância de efeitos gênicos aditivos envolvidos na determinação da AACPD, além da existência de desvios de dominância bidirecional no controle do caráter em questão. A análise dialélica foi eficiente na seleção de genitores visando o pré-melhoramento da resistência à requeima, destacando os acessos BGH- 2117, BGH-2127 e BGH-2343 como os de maior frequência de alelos favoráveis e divergentes, por isso devem ser incluídos em cruzamentos visando o pré-melhoramento da resistência à requeima.

Thesis (MSc)--Stellenbosch University, 2010. / ENGLISH ABSTRACT: Late blight caused by Phytophthora infestans on potato and tomato causes major economic losses worldwide. Until the 1980s, P. infestans populations outside its centre of origin (either central Mexico or the Andean region) only consisted of one mating type (A1), which prevented the pathogen from reproducing sexually. Pathogen populations outside the centre of origin most likely only consisted of a few genotypes prior to the 1980’s. Pan globally, these genotypes probably first consisted of genotype/s that had mitochondrial DNA (mtDNA) haplotype Ia, which was subsequently replaced by a mtDNA haplotype Ib genotype known as the US-1 lineage. This relative simple population structure of the pathogen changed almost worldwide in the late 1970s and early 1980s, when a second set of migrations took place from the centre of origin. These populations contained both A1 and A2 mating type isolates that consisted of several different genotypes, which were more virulent than the pre-1970s genotypes and resulted in the displacement of these genotypes almost worldwide. Some of the new genotypes were also resistant to metalaxyl, the fungicide that was most effective in controlling late blight. In Sub-Saharan Africa (SSA), the characteristics of P. infestans populations are not well documented in most countries except South Africa, Kenya and Uganda. Previous studies in SSA showed that populations were dominated by the US-1 lineage and its variants. The exceptions were reports of the presence of a few mtDNA haplotype Ia isolates in Rwanda and Ethiopia. The current study aimed to determine the population structure of P. infestans in eight selected SSA countries (Burundi, Kenya, Rwanda, Tanzania, Uganda, Malawi, Mozambique and South Africa), mainly on potato and on a limited scale on tomato and petunia, using ‘old’ markers (mating type determination, glucose-6-phosphate isomerase [Gpi] genotyping, mtDNA haplotyping, DNA fingerprinting with probe RG-57 and metalaxyl sensitivity). Populations were further also genotyped using seven recently published Simple Sequence Repeats (SSRs) markers. This information would help to define the population structure of P. infestans in SSA for the first time on a regional basis, and will also determine whether new migrations have taken place since the last characterization studies took place in 2001. A survey in the eight SSA countries yielded a total of 281 P. infestans isolates, mainly obtained from potato fields (Tanzania, Kenya, Uganda, Rwanda, Burundi, Malawi and South Africa), but also from tomato (Malawi, Mozambique and South Africa) and Petunia ´ hybrida (South Africa) that were characterized. Characterization of subsets of the isolates with the ‘old’ markers (176 isolates for mating type, 281 isolates for mtDNA, 70 isolates for [Gpi] and 49 isolates with restriction fragment length polymorphism analysis with probe RG-57), showed that most of the isolates belonged to the US-1 genotype or its variants (US-1.10 and US-1.11). The exception were isolates that belonged to genotype KE-1 (A1 mating type, mtDNA haplotype Ia, Gpi 90/100 and unique RG-57 genotype) that was identified in two fields in Kenya. Genotype KE-1, based on the ‘old’ marker data, is related to genotypes (RW-1 and RW-2) previously identified in Rwanda, and several Ecuadorean and European genotypes. Metalaxyl sensitivity testing of 64 isolates showed that metalaxyl resistant potato isolates were present in all the countries except Malawi, whereas all the tomato isolates were sensitive. Genotyping of 176 isolates with seven recently published simple sequence repeat (SSR) markers revealed a high number (79) of multi-locus genotypes (MLGs) in SSA. However, when locus D13, which was difficult to score, was excluded only 35 MLGs were identified. When locus D13 was excluded from analyses of molecular variance (AMOVA), (i) there was no significant genetic differentiation between populations from central-east Africa (Burundi, Kenya, Rwanda, Tanzania and Uganda), south-east Africa (Malawi and Mozambique) and South Africa, (ii) the KE-1 population was genetically differentiated (Fst = 0.33; P = 0.001) from the US-1 and US-1.10 populations and (iii) genetic differentiation between populations from potato and tomato was low (Fst = 0.07; P = 0.004). The study has expanded the worldwide genotypic database of P. infestans for SSA. Previously, no populations were characterized from Burundi, Malawi and Mozambique. The characterization work showed that migrations seem unlikely to have taken place in SSA, or if these did occur, it was on a very limited scale. The more severe epidemics in some SSA countries could be due to the presence of metalaxyl resistance. Furthermore, the occurrence of mutations or mitotic recombination might have resulted in more aggressive and/or better adapted genotypes, for example the US-1.10 lineage that was only detected in the Western Cape Province of South Africa. The significance of the discovery of the KE-1 genotype in Kenya needs further investigation since it might (i) be an asexual descendent of genotypes (RW-1 and RW-2) that were previously reported in Rwanda in the 1980s, (ii) previously have gone undetected due to the small surveys that were conducted in SSA, (iii) be a new migrant from countries other than SSA or (iv) have been introduced in the very first introductions into Kenya prior to the 1970s. The SSR results from the survey will allow comparison of the SSA late blight populations with other populations worldwide through the EucaBlight database in future studies. / AFRIKAANSE OPSOMMING: Laatroes, veroorsaak deur Phytophthora infestans op aartappel en tamatie, veroorsaak groot ekonomiese verliese wêreldwyd. Phytophthora infestans populasies buite hul kern van oorsprong (óf sentraal Meksiko óf die Andes area), het tot die 1980’s slegs uit een paringstipe (A1) bestaan, wat verhoed het dat die patogeen geslagtelik vermeerder. Patogeenpopulasies buite die kern van oorsprong, het heel moontlik vóór die 1980’s slegs uit ‘n paar genotipes bestaan. Wêreldwyd, het hierdie genotipes moontlik aanvanklik uit genotipe(s) bestaan wat mitokondriale DNS (mtDNS) haplotipe Ia bevat het, wat later met ‘n mtDNS haplotipe Ib genotipe, bekend as die US-1 genotipe, vervang is. Hierdie relatiewe eenvoudige populasiestruktuur van die patogeen, het omtrent wêreldwyd in die láát 1970’s en vroeë 1980’s verander, toe ‘n tweede stel migrasies vanaf die patogeen se kern van oorsprong plaasgevind het. Hierdie populasies het beide A1 en A2 paringstipe isolate ingesluit, wat uit verskeie verskillende genotipes bestaan het, wat meer virulent as die vóór-1970’s genotipes was, en wat die verskuiwing van hierdie genotipes omtrent wêrelwyd tot gevolg gehad het. Sommige van die nuwe genotipes was ook weerstandbiedend teen metalaksiel, die fungisied wat mees effektief in die beheer van laatroes was. Die kenmerke van P. infestans populasies is nie goed in die meeste lande in Sub- Sahara Afrika (SSA) gedokumenteer nie, behalwe vir Suid-Afrika, Kenia en Uganda. Vorige studies in SSA het aangedui dat populasies deur die US-1 genotipe en sy variante gedomineer word. Die uitsonderings was aantekeninge oor die teenwoordigheid van ‘n paar mtDNS haplotipe Ia isolate in Rwanda en Etiopië. Die huidige studie was daarop gemik om die populasiestruktuur van P. infestans in agt geselekteerde SSA lande (Burundi, Kenia, Rwanda, Tanzanië, Uganda, Malawi, Mosambiek en Suid-Afrika), hoofsaaklik op aartappel en op ‘n beperkte skaal op tamatie en petunia, vas te stel, deur die gebruik van ‘ou’ merkers (paringstipe-bepaling, glukose-6-fosfaat isomerase [Gpi] genotipering, mtDNS haplotipering, DNS fingerafdrukke met RG-57 en metalaksielsensitiwiteit). Die genotipe van populasies is verder ook bepaal deur gebruik te maak van sewe onlangs-gepubliseerde “Simple Sequence Repeats (SSRs)” merkers. Hierdie inligting sal help om die populasiestruktuur van P. infestans in SSA vir die eerste keer op ‘n streeksbasis vas te stel, en sal ook bepaal of nuwe migrasies sedert die laaste karakteriseringstudies wat in 2001 uitgevoer is, plaasgevind het. ‘n Opname in die agt SSA lande, het ‘n totaal van 281 P. infestans isolate opgelewer, hoofsaaklik vanaf aartappellande (Tanzanië, Kenia, Uganda, Rwanda, Burundi, Malawi en Suid-Afrika), maar ook vanaf tamatie (Malawi, Mosambiek en Suid- Afrika) en Petunia ´ hybrida (Suid-Afrika) wat gekarakteriseer is. Karakterisering van geselekteerde isolate met die ‘ou’ merkers (176 isolate vir paringstipe, 281 isolate vir mtDNS, 70 isolate vir Gpi en 49 isolate met restriksiefragment-lengte-polimorfismeanalise met RG-57), het aangetoon dat die meeste van die isolate aan die US-1 genotipe of sy variante (US-1.10 en US-1.11) behoort het. Die uitsondering was isolate wat tot die genotipe KE-1 behoort het (A1 paringstipe, mtDNS haplotipe Ia, Gpi 90/100 en unieke RG-57 genotipe) wat in twee velde in Kenia geïdentifiseer is. Genotipe KE-1, gebaseer op die ‘ou’ merkerdata, is aan genotipes (RW-1 en RW-2) verwant, wat voorheen in Rwanda, en verskeie Ekwadoreaanse en Europese lande geïdentifiseer is. Metalaksielsensitiwiteitstoetsing van 64 isolate het aangetoon dat metalaksiel-weerstandbiedende aartappel-isolate in al die lande teenwoordig was, behalwe vir Malawi, terwyl al die tamatie-isolate sensitief was. Genotipering van 176 isolate met sewe onlangs gepubliseerde “Simple Sequence Repeat” (SSR) merkers, het ‘n hoë aantal (79) multilokus genotipes (MLGs) in SSA aangedui. Met die uitsluiting van lokus D13, wat moeilik was om te evalueer, is slegs 35 MLGs egter geïdentifiseer. Met die uitsluiting van lokus D13 uit die analise van molekulêre variansie (AMOVA), was (i) daar geen betekenisvolle genetiese differensiasie tussen populasies van sentraal-oos Afrika (Burundi, Kenia, Rwanda, Tanzanië en Uganda), suid-oos Afrika (Malawi en Mosambiek) en Suid-Afrika nie, (ii) die KE-1 populasie geneties (Fst = 0.33; P = 0.001) van die US-1 en US-1.10 populasies gedifferensieerd en (iii) genetiese differensiasie tussen populasies vanaf aartappel en tamatie laag (Fst = 0.07; P = 0.004). Die studie het die wêreldwye genotipe-databasis van P. infestans vir SSA uitgebrei. Voorheen is geen populasies vanuit Burundi, Malawi en Mosambiek gekarakteriseer nie. Die karakteriseringswerk het aangetoon dat die waarskynlikheid klein is dat migrasies in SSA plaasgevind het, of indien dit wel plaasgevind het, dit op ‘n baie beperkte skaal plaasgevind. Die meer ernstige epidemies in sommige SSA lande kan die gevolg wees van die teenwoordigheid van metalaksiel-weerstand. Die voorkoms van mutasies of mitotiese rekombinasie kon verder meer aggressiewe en/of beter aangepaste genotipes tot gevolg gehad het, byvoorbeeld die US-1.10 genotipe wat slegs in die Westelike Kaapprovinsie van Suid-Afrika waargeneem is. Die betekenis van die ontdekking van die KE-1 genotipe in Kenia benodig verdere ondersoek aangesien dit (i) ‘n ongeslagtelike afstammeling van genotipes (RW-1 en RW-2) mag wees wat voorheen in die 1980’s in Rwanda aangeteken is, (ii) voorheen nie waargeneem is nie weens die klein opnames wat in SSA uitgevoer is, (iii) ‘n nuwe genotipe van lande buite die SSA kan wees of (iv) ingebring is tydens die heel eerste inkoms in Kenia vóór die 1970’s. Die SSR resultate van die opname sal vergelykings tussen die SSA laatroespopulasies en ander populasies wêreldwyd toelaat, deur gebruik te maak van die EucaBlight databasis in toekomstige studies.

Submitted by Gabriela Lopes (gmachadolopesufpel@gmail.com) on 2017-06-20T16:50:30Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertação Dediel.pdf: 766607 bytes, checksum: 1d7fec62b5b0587e0accd22056511b85 (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2017-06-21T18:58:18Z (GMT) No. of bitstreams: 2 Dissertação Dediel.pdf: 766607 bytes, checksum: 1d7fec62b5b0587e0accd22056511b85 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2017-06-21T18:58:18Z (GMT). No. of bitstreams: 2 Dissertação Dediel.pdf: 766607 bytes, checksum: 1d7fec62b5b0587e0accd22056511b85 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2012-03-30 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / A busca por alternativas no controle de doenças em substituição ao uso intensivo de agrotóxicos tem sido objeto da pesquisa agrícola. Rizobactérias são conhecidas pelo biocontrole de doenças e promoção de crescimento em diversos cultivos. Os objetivos deste trabalho foram avaliar a eficácia de seis rizobactérias, pré- selecionadas, no controle de Ralstonia solanacearum e Fusarium oxysporum f. sp. lycopersici (FOL), em casa de vegetação e relacionar este comportamento a produção de compostos “in vitro”, bem como avaliar a proteção de plantas de tomate contra doenças da parte aérea que ocorreram naturalmente em condições de campo. Foi avaliada a capacidade destas rizobactérias em produzir quitinases, amilases, lipases, compostos antibióticos e de solubilizar fosfato. A microbiolização das sementes com a rizobactéria DFs1420 (Bacillus sp.) reduziu a severidade da murcha bacteriana em 66% aos 14 dias após a inoculação, no primeiro ensaio e valores de AACPD em 70%, no segundo ensaio. Este controle pode ser associado à produção de compostos responsáveis pela antibiose observada “in vitro”. Streptomyces (DFs1296 e DFs1315) e Bacillus sp. (DFs1414) reduziram significativamente a murcha de fusário. O controle observado pode ser atribuído à atividade quitinolítica e/ou antibiótica por compostos voláteis. Foram instalados três ensaios de campo. A severidade das doenças foliares foi monitorada ao longo do tempo. As rizobactérias foram capazes de proteger as plantas contra Septoria lycopersici. DFs1414 e DFs1421 (Pseudomonas sp.) foram as mais estáveis, proporcionando proteção em dois ensaios consecutivos. As rizobactérias DFs1296 e DFs1420 foram capaz de controlar a requeima (Phytophthora infestans) em dois ensaios. A bactéria DFs1296 também apresentou capacidade de proteção contra estas e outras doenças quando pulverizada semanalmente na parte aérea. De modo geral, o controle alcançado por estas bactérias, microbiolizadas às sementes ou em aplicação foliar, não diferiu dos tratamentos utilizando produtos químicos recomendados para a cultura em aplicações semanais. / The search on alternative control of diseases to replace the use of pesticides has been the subject of agricultural research. Rhizobacteria are known by controlling diseases and promoting growth in several crops. The objectives of this work were to evaluate the efficacy of six rhizobacteria pre-selected for the control of Ralstonia solanacearum and Fusarium oxysporum f. sp. lycopersici (FOL) in greenhouse, and to relate this behavior with the production of compounds "in vitro", as well as evaluate the protection of tomato plants against foliar diseases under field conditions. It was evaluated the ability of these rhizobacteria to produce chitinases, amylases, lipases, antibiotic compounds and to solubilize phosphate. The seed microbiolization with rhizobacterium DFs1420 (Bacillus sp.) reduced the severity of the bacterial wilt 66% at 14 days after inoculation and AUDPC 70%, respectively, the first and second assays. This control may be associated with the production of compounds responsible for the antibiosis observed "in vitro". Streptomyces (DFs1296 and DFs1315) and Bacillus sp. (DFs1414) significantly reduced fusarium wilt. The control observed can be attributed to the chitinolytic activity and/or antibiosis in the presence of volatile compounds. Three field trials were carried out in field. The foliar diseases severity was monitored over time. The rhizobacteria were capable of protecting plants against Septoria lycopersici. DFs1414 and DFs1421 (Pseudomonas sp.) were the most stable, providing protection in two consecutive trials. The rhizobacteria DFs1296 and DFs1420 were able to control late blight (Phytophthora infestans) in two trials. DFs1296 also had the ability to protect against these and other diseases when sprayed weekly in the plant canopy.In general, the control achieved by these rhizobacteria, by seed microbiolization or by foliar application, did not differ from treatments using recommended chemical in weekly applications.

Submitted by Jorge Silva (jorgelmsilva@ufrrj.br) on 2017-02-09T19:14:10Z No. of bitstreams: 1 2009 - Evandro Silva Pereira Costa.pdf: 2009437 bytes, checksum: cf64a3e8f7e3e004841a52c65f4c6626 (MD5) / Made available in DSpace on 2017-02-09T19:14:10Z (GMT). No. of bitstreams: 1 2009 - Evandro Silva Pereira Costa.pdf: 2009437 bytes, checksum: cf64a3e8f7e3e004841a52c65f4c6626 (MD5) Previous issue date: 2009-09-21 / CNPq / Defining methods of quantification of tomato late blight, caused by Phytophthora infestans, is important to many researches about strategies of management of the disease. The aim of this present study was to assess and to validate the diagrammatic grading keys - Simplified, James-modified and Broad - and to determinate the possible correlations between the measures of severity and plants? physiological and nutritional state and biomass accumulation. Two simultaneous experiments were carried out: one in greenhouse and the other in field conditions. Two cultivars were studied in greenhouses, Super Sweet and ?Perinha ?gua Branca?, a resistant and a susceptible to late blight genotypes, respectively. The plants were inoculated or not with a sporangia suspension, and submitted to ten evaluations: 1, 2, 3, 4, 6, 7, 9, 10, 17 and 24 days after inoculation. The avaliations were performed in the fifth oldest leaf, and eighth and eleventh leaves, by visual estimation of severity, using three diagrammatic grading keys, Simplified, Broad and James-modified, but also measuring photosynthetic potential (Fv/Fm), by chlorophyll ?a? fluorescence emission. In field conditions, four cultivars were evaluated ? ?Perinha ?gua Branca?, Super Sweet, Santa Clara and Carmen F1. Periodically, late blight severity was quantified using the same three grading keys in leaves of different plant parts, at 51, 62, 74, 94 and 102 days after transplant (DAT). At the day following each severity disease evaluation, one plant was harvested per plot to measure the accumulation of biomass and macronutrients of different plant organs. Fruit yield was determined by summing fruits harvested between 68 and 114 DAT. In greenhouse conditions, the estimative of late blight severity with the three diagrammatic grading keys proved to be adequate to the disease?s quantification, enabling the discrimination between inoculation effects and differences between the cultivars in four and in nine days after inoculation, respectively, which were confirmed by measures of the photosynthetic potential of the leaves. In field conditions, the diagrammatic grading keys Simplified and Broad were more adequate to tomato late blight quantification. Quantifications in the youngest leaves, in the medium and superior third part of the plant, were the most representative of the disease progress and also discriminated the cultivars regarding disease resistance. Negative correlations were observed between severity, estimated with the three grading keys considering the whole plant and its medium third part, and leaf dry mass and leaf concentrations of K and P. The cultivar ?Perinha ?gua Branca? showed the lowest stem and leaf dry mass at the beginning of plant evaluations, and also late fruit yield, but had the higher stem and leaf mass and at the end of the experiment. The cultivar Perinha ?gua Branca and the hybrid Carmen F1 had higher commercial fruit yield. Based in the results, it is recommended the use of Simplified diagrammatic grading key and evaluations in the medium or superior tomato third to quantify the late blight disease. / A defini??o de m?todos de quantifica??o da requeima no tomateiro, causada por Phytophthora infestans, ? importante para v?rias pesquisas sobre estrat?gias de manejo da doen?a. Esse trabalho teve como objetivo aferir e validar as escalas diagram?ticas, Simplificada, James-modificada e Detalhada e determinar as poss?veis correla??es entre as medidas de severidade e o estado fisiol?gico e nutricional da planta e o ac?mulo de biomassa. Realizaram-se dois experimentos simult?neos, um sob condi??es de ambiente protegido e o outro sob condi??es de campo. Em casa de vegeta??o, utilizaram-se duas cultivares de tomate cereja, ?Perinha ?gua Branca? e Super Sweet, suscet?vel e resistente ? requeima, respectivamente. As plantas foram inoculadas ou n?o com suspens?o de espor?ngios de P. infestans e em seguida, avaliadas aos 1, 2, 3, 4, 6, 7, 9, 10, 17 e 24 dias ap?s a inocula??o. As avalia??es foram feitas na quinta, oitava e d?cima primeira folha, contadas do ?pice para a base, usando as tr?s escalas acima citadas e medi??es do potencial fotossint?tico (Fv/Fm) pela emiss?o da clorofila ?a?. No campo, utilizaram-se quatro cultivares, ?Perinha ?gua Branca?, Super Sweet, Santa Clara e Carmen F1. Periodicamente, quantificou-se a severidade da requeima utilizando-se as mesmas escalas acima citadas em folhas de diferentes partes da planta, aos 51, 62, 74, 94 e 102 dias ap?s o transplante (DAT). No dia seguinte a cada avalia??o de severidade, foi coletada uma planta por parcela para mensurar o ac?mulo de biomassa e de macronutrientes nos diferentes ?rg?os da planta. A produ??o de frutos foi determinada pelo somat?rio das colheitas realizadas entre 68 e 114 DAT. Em condi??es de casa de vegeta??o, a estimativa da severidade da requeima com as tr?s escalas se mostrou adequada para quantifica??o da doen?a tendo permitido discriminar efeito de inocula??o e diferen?as entre as cultivares aos quatro e nove dias ap?s a inocula??o, respectivamente, que foram confirmadas pelas medidas do potencial fotossint?tico das folhas. Em condi??es de campo, as escalas Simplificada e Detalhada, foram mais adequadas para a quantifica??o da requeima do tomateiro. Quantifica??es feitas nas folhas mais jovens, no ter?o m?dio e superior da planta, foram as que melhor representaram o progresso da doen?a e discriminaram as cultivares quanto a resist?ncia ? doen?a. Observaram-se correla??es negativas entre a severidade, estimada com as tr?s escalas, considerando a planta inteira e o ter?o m?dio, e a massa seca de folhas e os teores foliares de K e P nas quatro cultivares. A cultivar ?Perinha ?gua Branca? apresentou menor massa seca de caule e de folha no in?cio das avalia??es e produ??o de frutos mais tardia, por?m ao final do ciclo destacou-se pela maior massa seca de caule e de folha. A cultivar ?Perinha ?gua Branca? e o h?brido Carmen F1 apresentaram maior produ??o de frutos comerciais. Com base nos resultados pode-se recomendar o uso da escala simplificada e avalia??es no te?o m?dio ou superior do tomateiro para quantifica??o da requeima.

Phytophthora infestans (Mont) de Bary is a devastating pathogen in potato producing regions around the world. Populations of the organism in Canada shifted during the mid-1990’s as the US-1 strain (A1, metalaxyl-sensitive) was displaced by the highly aggressive, US-8 strain (A2, metalaxyl-insensitive). An increase in the incidence and severity of late blight has followed. Late blight is controlled by cultural practices aimed at eliminating disease sources and by the application of foliar fungicides. Tubers can become infected at harvest from contact with blighted vines leading to severe losses in storage. In many production areas, growers desiccate vines two to three weeks prior to harvest to reduce late blight tuber rot. However, in Manitoba, because of the loss of potential yield that results from vine killing prior to harvest in a late maturing cultivar such as Russet Burbank, growers are reluctant to adopt this practice. The objective of this study was to develop recommendations for preharvest management practices that reduce storage losses due to late blight. Field trials were conducted in 1997 and 1998 to investigate the effect of vine desiccation with diquat and/or a late season application of chlorothalonil and copper hydroxide on yield, processing quality, and disease development in storage. Desiccating vines with diquat two weeks prior to harvest reduced yield and tuber size. Compared to the untreated control, the largest reductions in marketable yield were observed for the early September harvest. By the late September harvest, however, the effect of vine killing in reducing marketable yield was less apparent. Specific gravity was lower in the vine killed treatment for all harvest dates in 1997 and in the early and mid September harvests in 1998. Vine killing did not contribute to elevated levels of reducing sugars or consistently darker fry colour at harvest or during storage. Skin-set was improved when vines were desiccated for all harvest dates in 1997 and at the early September harvest date in 1998. Vine killing reduced tuber rot in storage caused by Fusarium dry rot and Pythium leak for the early and mid September harvest dates in 1997. The incidence of late blight tuber rot was reduced in storage for the early September harvest in 1998 when vines were desiccated. The late-season application of chlorothalonil and copper hydroxide did not reduce tuber rot in storage in storage either year. Results from this study indicate that vine killing two weeks before an early to mid- September harvest is not recommended in Manitoba because of reductions in yield and specific gravity. Alternative management practices to reduce late blight tuber rot in storage should be investigated. / February 2005

Phytophthora infestans (Mont) de Bary is a devastating pathogen in potato producing regions around the world. Populations of the organism in Canada shifted during the mid-1990’s as the US-1 strain (A1, metalaxyl-sensitive) was displaced by the highly aggressive, US-8 strain (A2, metalaxyl-insensitive). An increase in the incidence and severity of late blight has followed. Late blight is controlled by cultural practices aimed at eliminating disease sources and by the application of foliar fungicides. Tubers can become infected at harvest from contact with blighted vines leading to severe losses in storage. In many production areas, growers desiccate vines two to three weeks prior to harvest to reduce late blight tuber rot. However, in Manitoba, because of the loss of potential yield that results from vine killing prior to harvest in a late maturing cultivar such as Russet Burbank, growers are reluctant to adopt this practice. The objective of this study was to develop recommendations for preharvest management practices that reduce storage losses due to late blight. Field trials were conducted in 1997 and 1998 to investigate the effect of vine desiccation with diquat and/or a late season application of chlorothalonil and copper hydroxide on yield, processing quality, and disease development in storage. Desiccating vines with diquat two weeks prior to harvest reduced yield and tuber size. Compared to the untreated control, the largest reductions in marketable yield were observed for the early September harvest. By the late September harvest, however, the effect of vine killing in reducing marketable yield was less apparent. Specific gravity was lower in the vine killed treatment for all harvest dates in 1997 and in the early and mid September harvests in 1998. Vine killing did not contribute to elevated levels of reducing sugars or consistently darker fry colour at harvest or during storage. Skin-set was improved when vines were desiccated for all harvest dates in 1997 and at the early September harvest date in 1998. Vine killing reduced tuber rot in storage caused by Fusarium dry rot and Pythium leak for the early and mid September harvest dates in 1997. The incidence of late blight tuber rot was reduced in storage for the early September harvest in 1998 when vines were desiccated. The late-season application of chlorothalonil and copper hydroxide did not reduce tuber rot in storage in storage either year. Results from this study indicate that vine killing two weeks before an early to mid- September harvest is not recommended in Manitoba because of reductions in yield and specific gravity. Alternative management practices to reduce late blight tuber rot in storage should be investigated.

Phytophthora infestans (Mont) de Bary is a devastating pathogen in potato producing regions around the world. Populations of the organism in Canada shifted during the mid-1990’s as the US-1 strain (A1, metalaxyl-sensitive) was displaced by the highly aggressive, US-8 strain (A2, metalaxyl-insensitive). An increase in the incidence and severity of late blight has followed. Late blight is controlled by cultural practices aimed at eliminating disease sources and by the application of foliar fungicides. Tubers can become infected at harvest from contact with blighted vines leading to severe losses in storage. In many production areas, growers desiccate vines two to three weeks prior to harvest to reduce late blight tuber rot. However, in Manitoba, because of the loss of potential yield that results from vine killing prior to harvest in a late maturing cultivar such as Russet Burbank, growers are reluctant to adopt this practice. The objective of this study was to develop recommendations for preharvest management practices that reduce storage losses due to late blight. Field trials were conducted in 1997 and 1998 to investigate the effect of vine desiccation with diquat and/or a late season application of chlorothalonil and copper hydroxide on yield, processing quality, and disease development in storage. Desiccating vines with diquat two weeks prior to harvest reduced yield and tuber size. Compared to the untreated control, the largest reductions in marketable yield were observed for the early September harvest. By the late September harvest, however, the effect of vine killing in reducing marketable yield was less apparent. Specific gravity was lower in the vine killed treatment for all harvest dates in 1997 and in the early and mid September harvests in 1998. Vine killing did not contribute to elevated levels of reducing sugars or consistently darker fry colour at harvest or during storage. Skin-set was improved when vines were desiccated for all harvest dates in 1997 and at the early September harvest date in 1998. Vine killing reduced tuber rot in storage caused by Fusarium dry rot and Pythium leak for the early and mid September harvest dates in 1997. The incidence of late blight tuber rot was reduced in storage for the early September harvest in 1998 when vines were desiccated. The late-season application of chlorothalonil and copper hydroxide did not reduce tuber rot in storage in storage either year. Results from this study indicate that vine killing two weeks before an early to mid- September harvest is not recommended in Manitoba because of reductions in yield and specific gravity. Alternative management practices to reduce late blight tuber rot in storage should be investigated.

Some species of Oomycetes are well studied pathogens that cause considerable economical losses in the agriculture and aquaculture industries. Currently, there are no chemicals available that are environmentally friendly and at the same time efficient Oomycete inhibitors. The cell wall of Oomycetes consists of b-(1à3) and b-(1à6)-glucans, cellulose and in some species minute amounts of chitin. The biosynthesis of cellulose and chitin in Oomycetes is poorly understood. However, cell wall synthesis represents a potential target for new Oomycete inhibitors. In this work, cellulose and chitin synthase genes and gene products were analyzed in the plant pathogen Phytophthora infestans and in the fish pathogen Saprolegnia monoica.   A new Oomycete CesA gene family was identified, containing four subclasses of genes designated as CesA1 to 4. The gene products of CesA1, 2 and 4 contain pleckstrin homology (PH) domains located at the N-terminus, which is unique to the Oomycete CesAs. Our results show that the SmCesA2 PH domain binds to phosphoinositides, F-actin and microtubules in vitro and can co-localize with F-actin in vivo. Functional characterization of the CesA genes by gene silencing in P. infestans led to decreased cellulose content in the cell wall. The cellulose synthase inhibitors DCB and Congo Red inhibited the growth of the mycelium of S. monoica and had an up-regulating effect on SmCesA gene expression. Zoospores from P. infestans treated with DCB were unable to infect potato leaves. In addition, two full-length chitin synthase genes (Chs) were analyzed from S. monoica.  Expression of SmChs2 in yeast yielded an active recombinant protein. The biochemical characterization of the in vitro product of SmChs2 confirmed that the protein is responsible for chitin formation. The chitin synthase inhibitor nikkomycin Z inhibited the SmChs2 both in vivo and in vitro.   Altogether these results show that at least some of the CesA1-4 genes are involved in cellulose biosynthesis and that synthesis of cellulose is crucial for infection of potato by P. infestans. The PH domain is involved in the interaction of CesA with the cytoskeleton. In addition, we firmly demonstrate that the SmChs2 gene encodes a catalytically active chitin synthase. / QC 20110531