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Magistrantūros studijų baigiamojo darbo tyrimų tikslas buvo ištirti cukrinių runkelių ligotumą, nustatyti grybinių lapų ligų paplitimą ir žalingumą cukriniuose runkeliuose bei nustatyti apsaugos priemonių, mažinančių ligų plitimą efektyvumą. Lauko eksperimentas buvo atliktas 2012 m. Lietuvos agrarinių ir miškų mokslų centro (LAMMC) Rumokų bandymų stotyje. Dirvožemis sekliai glėjiškas paprastasis išplautžemis (Haplic-Epihypogleyic Luvisol – Idg8-p). Darbo objektas - cukrinių runkelių (Beta vulgaris saccharifera) grybinės lapų ligos. Darbo metodai: Cukrinių runkelių lapų ligotumo, fungicidų biologinio efektyvumo, ligų žalingumo matavimai nustatyti tiesioginio matavimo, skaičiavimo ar svėrimo būdu pagal priimtas lauko eksperimento vykdymo metodikas (Šurkus, Gaurilčikienė, 2002). Tyrimų duomenys statistiškai įvertinti vieno veiksnio arba dviejų veiksnių kiekybinių požymių dispersinės analizės metodais, taikant kompiuterinę programą ANOVA (Tarakanovas, Raudonius, 2003). Darbo rezultatai: Tyrimų metais cukriniuose runkeliuose labiausiai plito cukrinių runkelių rudmargė ir baltuliai, kurių pažeidimo intensyvumas atitinkamai siekė 11,91 ir 1,81 proc. Didžiausias biologinis fungicido efektyvumas nuo cukrinių runkelių rudmargės nustatytas laukeliuose, kuriuose vieną ir du kartus buvo naudojamas fungicidas Maredo (vienkartinio purškimo norma 1,0 l ha-1, dukartinio - 0,75 l ha-1). Jo biologinis efektyvumas atitinkamai siekė 65,79 – 92,06 proc. ir 86,39 – 94,19 proc. Didžiausias... [toliau žr. visą tekstą] / The main objective of Master thesis research was to investigate the intensity of sugar beet diseases, determine the fungal disease prevalence and harmfulness of sugar beets, and determine the effect of chemical plant control against sugar beet diseases.The research was made in the Rumokai Experimental Station of the Lithuanian Research Centre for Agriculture and Forestry in 2012. The soil type – Haplic-Epihypogleyic Luvisol – Idg8-p. Object of the research – sugar beet (Beta vulgaris saccharifera) fungal leaf diseases. Working methods: biological effectiveness of fungicides and disease harmfulness measurements were determined while using direct measurements, calculations or weighing according field experimental techniques (Šurkus, Gaurilčikienė, 2002). Research data have been statistically evaluated with a single factor or two factor analysis of variance of quantitative methods of evidence, using a computer program ANOVA (Tarakanovas, Raudonius, 2003). Research results: The most effective fungicide against Cercospora leaf spot disease in the once and twice treatment field of sugar beet plants was Maredo (once treatment rate 1,0 l ha-1, twice treatment rate 0,75 l ha-1). Its efficiency was highest and reached 65,79 – 92,06 % and 86,39 – 94,19 %. The most effective fungicideagainst Ramularia leaf spot disease in the once and twice treatment field of sugar beet plants was also Maredo. Its efficiency reached 75,41 – 93,52 % and 82,76 – 93,89 %. The highest significant root... [to full text]

Submitted by BELISA CRISTINA SAITO null (belisasaito@gmail.com) on 2017-04-20T18:20:53Z No. of bitstreams: 1 Tese Belisa Saito.pdf: 1868613 bytes, checksum: 40d639682537f01c2cc01e7a92791445 (MD5) / Approved for entry into archive by Luiz Galeffi (luizgaleffi@gmail.com) on 2017-04-25T17:59:26Z (GMT) No. of bitstreams: 1 saito_bc_dr_ilha.pdf: 1868613 bytes, checksum: 40d639682537f01c2cc01e7a92791445 (MD5) / Made available in DSpace on 2017-04-25T17:59:26Z (GMT). No. of bitstreams: 1 saito_bc_dr_ilha.pdf: 1868613 bytes, checksum: 40d639682537f01c2cc01e7a92791445 (MD5) Previous issue date: 2017-03-03 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / O milho é uma das culturas mais extensamente cultivadas em todo mundo. A incidência e a severidade de doenças têm aumentado significativamente nos últimos anos acarretando perdas no rendimento e afetando a qualidade dos grãos. Muitos trabalhos têm sido desenvolvidos na tentativa de identificar híbridos resistentes às principais doenças que acometem a cultura do milho, mas poucos são os relatos de estudos com linhagens. Dessa forma, o objetivo deste estudo foi: 1) identificar linhagens resistentes e susceptíveis com base na área abaixo da curva de progresso de doenças (AACPD) para os sintomas de ferrugem tropical (TR), ferrugem polissora (SR), cercosporiose (GLS), helmintosporiose (NLB), mancha marrom (PBS) e mancha branca (PLS); 2) identificar linhagens resistentes e suscetíveis com base nos parâmetros de adaptabilidade e estabilidade fenotípica para os sintomas de cercosporiose, helmintosporiose, mancha marrom e mancha branca; 3) identificar as melhores datas de semeadura, com a maior ocorrência das doenças, para fins de avaliação de linhagens e outros genótipos para resistência. Cinquenta linhagens, derivadas de populações com grãos flint e dent, foram avaliadas em blocos casualizados com três repetições, aos 45, 60, 75 e 90 dias após a semeadura em duas épocas, para medição da AACPD. Para a análise de adaptabilidade e estabilidade, 41 linhagens foram avaliadas em blocos casualizados com três repetições, 30 dias após o florescimento feminino, em onze épocas de semeadura, usando o método de análise de regressão. Foram atribuídas notas de 1, 2, 3, 4, 5, 6, 7, 8 e 9 correspondendo a 0, 1, 10, 20, 30, 40, 60, 80 e > 80% de área foliar com sintomas de doença. Para a AACPD, a análise de variância conjunta foi significativa para TR, SR, GLS e PLS e a interação linhagens x épocas foi significativa para ferrugem tropical e polissora. Para GLS e NLB as 41 linhagens foram classificadas como resistentes, sendo que as maiores severidades de doenças ocorreram nas semeaduras entre Junho e Setembro. As linhagens IVF1-3, IVF1-7, IVF1 -9, IVF1-10, IVF1 -11, IVF1 -25, IVF1-230, IVD1-2, IVD1 -2-1, IVD1-3, IVD1-9, IVD1 -12, 2F, 3F, 6F, 9F, 10F, 4C, 2D e 7D foram classificadas como resistentes para as doenças estudadas, sendo indicadas para o desenvolvimento de sintéticos. Para a mancha marrom e mancha branca, as semeaduras de Abril, Junho, Julho e Agosto apresentaram maiores severidades de doenças. As linhagens IVD1-9, IVD1-10, 7D, 10D e 2F podem ser indicadas no desenvolvimento de sintéticos resistentes. / Corn is one of the most widely cultivated crops in the worldwide. The incidence and severity of diseases affecting crops have increased significantly in the past years, leading to yield losses and affecting grain quality. Many studies have been carried out with the attempt to identify hybrids that are resistant to the main diseases, but few reports have studied inbred lines. Therefore, the objectives of this study were: 1) identify resistant and susceptible inbred lines based on the area under disease progress curve (AUDPC) for tropical rust, southern rust, gray leaf spot, northern leaf blight, physoderma brown spot and phaeosphaeria leaf spot; 2) identify resistant and susceptible inbred lines based on adaptability and stability parameters for symptoms of gray leaf spot (GLS), northern leaf blight (NLB), physoderma brown spot (PBS) and phaeosphaeria leaf spot (PLS); 3) identify the best planting dates, with the highest occurrence of diseases, for the purpose of evaluating inbred lines and other genotypes for resistance. For AUDPC, fifty inbred lines, derived from populations with flint and dent grains, were evaluated in randomized block designs with three replications, at 45, 60, 75 and 90 days after planting in two seasons. For the analysis of adaptability and stability, forty-one inbred lines were evaluated in randomized blocks with three replications, 30 days after silking, in eleven planting dates, using regression analysis method. The scale of scores from 1, 2, 3, 4, 5, 6, 7, 8 and 9 corresponding to 0, 1, 10, 20, 30, 40, 60, 80 and > 80% of leaf area with disease symptoms was used. For AUDPC, the joint analysis of variance was significant for TR, SR, GLS and PLS, while the interaction inbred lines x environments, was significant for TR and SR. For GLS and NLB, forty-one inbred lines were classified as resistant and the highest severities of diseases occurred in planting dates between June and September. The inbred lines IVF1-3, IVF1-7, IVF1 -9, IVF1-10, IVF1 -11, IVF1 -25, IVF1-230, IVD1-2, IVD1 -2-1, IVD1-3, IVD1-9, IVD1 -12, 2F, 3F, 6F, 9F, 10F, 4C, 2D and 7D were classified as resistant to the diseases studied and are indicated to produce synthetics. For PBS and PLS, the plating dates of April, June, July and August showed higher disease severity. The inbred lines IVD1-9, IVD1-10, 7D,10D and 2F may be indicated to produce synthetics.

Orientador: João Antonio da Costa Andrade / Resumo: O milho é uma das culturas mais extensamente cultivadas em todo mundo. A incidência e a severidade de doenças têm aumentado significativamente nos últimos anos acarretando perdas no rendimento e afetando a qualidade dos grãos. Muitos trabalhos têm sido desenvolvidos na tentativa de identificar híbridos resistentes às principais doenças que acometem a cultura do milho, mas poucos são os relatos de estudos com linhagens. Dessa forma, o objetivo deste estudo foi: 1) identificar linhagens resistentes e susceptíveis com base na área abaixo da curva de progresso de doenças (AACPD) para os sintomas de ferrugem tropical (TR), ferrugem polissora (SR), cercosporiose (GLS), helmintosporiose (NLB), mancha marrom (PBS) e mancha branca (PLS); 2) identificar linhagens resistentes e suscetíveis com base nos parâmetros de adaptabilidade e estabilidade fenotípica para os sintomas de cercosporiose, helmintosporiose, mancha marrom e mancha branca; 3) identificar as melhores datas de semeadura, com a maior ocorrência das doenças, para fins de avaliação de linhagens e outros genótipos para resistência. Cinquenta linhagens, derivadas de populações com grãos flint e dent, foram avaliadas em blocos casualizados com três repetições, aos 45, 60, 75 e 90 dias após a semeadura em duas épocas, para medição da AACPD. Para a análise de adaptabilidade e estabilidade, 41 linhagens foram avaliadas em blocos casualizados com três repetições, 30 dias após o florescimento feminino, em onze épocas de semeadura, us... (Resumo completo, clicar acesso eletrônico abaixo) / Doutor

The filamentous ascomycete fungus Magnaporthe oryzae is a pathogen of over 50 genera of grasses. Two important diseases it can cause are gray leaf spot in Lolium perenne (perennial ryegrass) and blast in Oryza sativa (rice). The telomeres of M. oryzae isolates causing gray leaf spot are highly variable, and can spontaneously change during fungal culture. In this dissertation, it is shown that a rice-infecting isolate is much more stable at the telomeres than an isolate from gray leaf spot. To determine the molecular basis of telomere instability several gray leaf spot isolates telomeres were cloned, which revealed two non-LTR retrotransposons inserted into the telomere repeats. The elements have been termed Magnaporthe oryzae Telomeric Retrotransposons (MoTeRs). These elements do not have poly-A tails common to many other non-LTR retrotransposons, but instead have telomere like sequences at their 5’ end that allow them to insert into telomeres. Intact copies of MoTeRs were restricted to the telomeres of isolates causing gray leaf spot. Surveys for the presence of these elements in M. oryzae showed they were present in several host-specialized forms including gray leaf spot isolates, but were largely absent in the rice blast isolates. The absence of MoTeRs in rice blast isolates, which are relatively stable by comparison, suggested that the telomere instability in gray leaf spot isolates could be due to MoTeRs. Analyzing spontaneous alterations in telomere restriction fragment profiles of asexual progeny revealed that MoTeRs were involved. Expansion and contraction of MoTeR arrays were observed and account for some telomere restriction profile changes. New telomere formation in asexual progeny followed by MoTeR addition was also observed. Based on this evidence, MoTeRs are largely responsible for the high variability of telomere restriction profiles observed in GLS isolates.

O caquizeiro (Diospyros kaki L.) é uma planta caducifólia pertencente à família Ebenaceae, tendo como centro de origem a região central da China. Entre os estados brasileiros, São Paulo é o que apresenta maior produção de caquis com 49.727 t. A cercosporiose (Pseudocercospora kaki), também conhecida como mancha angular, é a principal doença foliar da cultura. Alguns autores atribuem à cercosporiose a queda precoce de folhas, maturação antecipada dos frutos e a redução da produção na safra seguinte. Entretanto, não existem trabalhos com o patossistema P. kaki - D. kaki que deem suporte a essas pressuposições. Diante disso, os objetivos desse trabalho foram comparar a susceptibilidade de quatro cultivares de caquizeiro e avaliar o efeito da cercosporiose na fisiologia da planta. Os experimentos foram conduzidos em condições naturais de campo, com as cultivares Fuyu, Rama Forte, Taubaté e Giombo, por serem as mais cultivadas no estado de São Paulo. Nos experimentos desenvolvidos buscou-se conhecer o grau de resistência das quatro cultivares à cercosporiose, avaliando a incidência, severidade e queda de folhas ao longo de dois ciclos de cultivo, assim como avaliar o controle químico na redução da doença em cvs. Taubaté e Rama Forte e, consequentemente, no acúmulo de reservas na raiz e na qualidade dos frutos colhidos. Foram avaliadas também os danos na fotossíntese em folhas das quatro cultivares com diferentes intensidades de cercosporiose. Como resultados pode-se constatar, que a cv. Fuyu é a mais susceptível à cercosporiose, seguida com resistência média \'Giombo\' e \'Rama Forte\' e com maior resistência a cv. Taubaté. A intensidade da cercosporiose, para severidades da doença de até 7% não influencia na queda de folhas. O controle químico reduziu a intensidade de doença nas cvs. Taubaté e Rama Forte, entretanto, entre as intensidades de cercosporiose avaliadas em plantas tratadas e não tratadas, não há redução no acúmulo de reservas no sistema radicular, assim como na qualidade físico-química dos frutos colhidos. Mesmo a cercosporiose afetando a fotossíntese, no processo fotoquímico, tanto na área lesionada como na área adjacente à lesão, a intensidade da doença observada no campo não foi suficiente para causar redução da produção à cultura. / The persimmon (Diospyros kaki L.) is a deciduous plant of the botanical family Ebenaceae, native to central China. Among the Brazilian states, São Paulo has the highest production of persimmon with 49,727 ton/year. The angular leaf spot of persimmon (Pseudocercospora kaki), also known as cercosporiosis, is the main leaf disease of the crop. Some authors attribute to the cercosporiosis the premature defoliation, early maturation of the fruits and the reduction of the yield in the next crop cycle. However, there are no previous studies with the Pseudocercospora kaki pathosystem that support these hypotheses. Therefore, the objectives of this study were to compare the susceptibility of four persimmon cultivars and evaluate the effect of angular leaf spot on plant physiology. A field experiment was conducted with Fuyu, Rama Forte, Taubaté and Giombo cultivars, the four most cultivated in the state of São Paulo. The degree of resistance to cercosporiosis was determined by the incidence and severity of the disease and by defoliation of the plants in two crop cycles. Chemical spraying for the disease reduction was also evaluated in cvs. Taubaté and Rama Forte. The reduction of the carbohydrate reserve accumulation in the roots and the quality of the fruits harvested were tested. The photosynthesis reduction was also evaluated in leaves of the four cultivars with different intensities of cercosporiosis. The Fuyu cultivar was the most susceptible to cercosporiosis, followed by moderate resistance of \'Giombo\' and \'Rama Forte\' and high resistance of cv. Taubaté. The intensity of cercosporiosis, for disease severity up to 7% does not influence leaf drop. Though the chemical control reduced the disease intensity in cvs. Taubaté and Rama Forte, treated and untreated plants showed no differences in the reserves accumulation in the roots, as well as in the physicochemical quality of the harvested fruits. Even though cercosporiosis affects photosynthesis in the photochemical process on both the necrotic and remaining green leaf area, the intensity of the disease observed in the field was not enough to cause yield reduction to the culture.

Foram conduzidos quatro experimentos com a cultivar 'Carioca' do feijoeiro (Phaseolus vulgaris), objetivando a possibilidade de implementação de um sistema de manejo integrado da ferrugem e mancha angular do feijoeiro após a intervenção com os fungicidas oxycarboxin (Hokko Plantvax 750 - 700 gramas/ha) e trifenil hidróxido de estanho (Mertin â - 660 ml/ha), respectivamente, aplicados em diferentes épocas. Foi realizada a quantificação de danos nesses patossistemas, verificando por meio de regressões lineares e não lineares as possíveis relações existentes entre as variáveis integrais AUDPC (área sob a curva de progresso de doença)¸ HAD (duração da área foliar sadia) e HAA (absorção da área foliar sadia) e suas correspondentes HLAI (índice de área foliar sadio) e HRI (absorção da área foliar por unidade de tempo) com a produção (gramas/planta). Os melhores resultados foram obtidos com as variáveis HAD e HAA, com coeficientes de determinação (R 2 ) variáveis entre 0,19 e 0,56 para HAD e 0,23 e 0,49 para HAA. Quando as variáveis HLAI e HRI foram relacionadas com a produção, os coeficientes de determinação variaram entre 0,24 e 0,57 para HLAI e 0,28 e 0,56 para HRI. A partir das avaliações semanais de HLAI e HRI foram realizadas regressões lineares entre estas variáveis e a produção (gramas/planta). Os coeficientes angulares foram relacionados com os estádios de desenvolvimento observando-se a estabilização das curvas entre os estádios R5-R8 para o patossistema ferrugem - feijão e entre os estádios V4-R8 para o patossistema mancha angular - feijão para HLAI e HRI. Nesses experimentos foram também realizadas avaliações do Índice de Área Foliar (LAI) utilizando-se medições diretas (avaliações manuais) e indiretas com ceptômetro. As medições obtidas com o este equipamento apresentaram sensibilidade suficiente para captar as variações no índice de área foliar sadio e portanto podem ser usadas em sistemas de manejo integrado da ferrugem e da mancha angular do feijoeiro. / Four trials were carried out on dry beans (Phaseolus vulgaris), variety 'Carioca' with goal to verify the possibility of implementation of an integrated management system for rust and angular spot after the application with of oxycarboxin (Hokko Plantvax 750 - 700 g/ha) and trifenil stannic hydroxide (Mertin â - 660 ml/ha) fungicides, applied in different timings. The damage quantification in these pathosystems were done analyzing the possible existing relation among the integral variables AUPDC (Area Under Disease Progress Curve), HAD (Healthy Area Duration) and HAA (Healthy Area Absorption) and its correspondents HLAI (Healthy Leaf Area Index) and HRI (Healthy Radiation Intercept) with yield (grams/plant) through linear and non linear regression. The best results were obtained using the variables HAD and HAA with the determination coefficients (R 2 ) from 0.19 to 0.56 for HAD and 0.23 to 0.49 for HAA. When HLAI and HRI were related with yield, the determination coefficient ranged from 0.24 and 0.57 for HLAI and 0.28 to 0.56 for HRI. From weekly assessments of HLAI and HRI, linear regression analysis were done between those variables and yield (grams/plant). The angular coefficient were related with the different crop stages and the stability of curves was observed between R5-R8 stages for rust - dry bean pathosystem and between V4-V8 stages for angular spot - dry bean pathosystem when HLAI and HRI were analyzed. In these experiments, Leaf Area Indice (LAI) was also assessed with direct (manual) and indirect measures with a ceptometer. The measures obtained using this set provided necessary sensibility to catch HLAI variation and, therefore, may be used in an integrated management system of rust and angular spot in dry beans.

Lily leaf spot disease, caused by the fungal phytopathogen, Pseudocercosporella inconspicua, infects Lilium canadense and L. grayi. The disease is currently ubiquitous in populations throughout the range of L. grayi. To determine the historical prevalence of the disease, lily specimens from eight herbaria were examined visually and microscopically, and a search for records of the pathogen was conducted using mycology databases and relevant literature. Of 516 herbarium specimens, two L. canadense and one L. grayi had the characteristic leaf lesions that contained diagnostic conidia of P. inconspicua. All three diseased specimens were collected prior to 1950. Mycological collections included two North American records of P. inconspicua on L. canadense, two on L. michiganense, one on L. philadelphicum var. andinum, and one on a cultivated Eurasian lily hybrid. Interestingly, the earliest diseased herbarium specimens were from the northeastern US with a later appearance in the southern Appalachians, a pattern also present in mycological collections of P. inconspicua. The rarity of historical specimens with disease, the temporal geographic pattern of occurrence, and the ubiquity of P. inconspicua in current populations of L. grayi suggest the spread of lily leaf spot disease in North America may threaten the viability of native Lilium host species.

In the past two decades a new barley threat has emerged as the disease Ramularia leaf spot (RLS) became more prevalent in temperate regions worldwide. This disease, first identified in the late 19th century, is caused by the filamentous fungus Ramularia collo-cygni (Rcc) and can cause substantial yield losses as well as reduce grain quality. RLS typically occurs late in the growing season and characteristic disease symptoms are usually seen after the crop has flowered. Expression of RLS lesions is thought to be associated with the action of fungal secondary metabolism products. The one group of secondary metabolites (SMs) characterised to date from Rcc, the anthraquinone toxins rubellins, are known to cause necrosis to plant tissues in a non-host specific manner. Therefore, it appears that fungal secondary metabolism might be a key component in understanding the interaction between Rcc and its host. In this study, more than 23 core genes involved in the biosynthesis of SMs belonging to the polyketide and non-ribosomal peptide pathways were identified in the genome of Rcc. Putative clusters containing genes with a predicted function relating to secondary metabolism were identified by in silico genome walking in the genetic loci adjacent to Rcc SM core genes. Two gene clusters containing no SM core gene were also identified. Five of the putative SM clusters exhibited similarity to the known fungal SM biochemical pathways involved in gliotoxin, monodictyphenone, ferricrocin, betaenone and chaetoglobosins biosynthesis. Several gene clusters exhibited similarity to SM clusters from fungal species where the SM pathway is uncharacterised. Changes in transcript abundance of selected SM core genes during RLS development in artificially inoculated barley seedlings were tested. Transcript levels were found to be the highest at an early stage of disease development, typically during the asymptomatic and early lesions formation stages and declined over time, suggesting that the associated SMs in Rcc, may not necessarily be involved in symptoms appearance. The in planta mode of action of the non-host specific photoactivated toxin rubellin D was studied in the model plant Arabidopsis thaliana. Rubellin-induced cell death appeared phenotypically reminiscent of programmed cell death (PCD). Full expression of rubellin D-induced cell death required the host salicylic acid (SA) pathway and the host proteasome supporting the PCD response to this fungal SM. However, a clear correlation between toxin sensitivity and disease susceptibility could not be found, suggesting a potential alternative role for rubellin in disease symptom development.

Made available in DSpace on 2015-03-26T13:39:29Z (GMT). No. of bitstreams: 1 texto completo.pdf: 355166 bytes, checksum: fca33886e7049d30889ecd53a351d550 (MD5) Previous issue date: 2009-02-17 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / The infectious process of Ramularia areola, the causal agent of ramularia leaf spot, on leaves of cotton, Gossypium hirsutum L. (Malvaceae), was studied by using the scanning electron microscope. Conidia started to germinate at 12 hours after inoculation (hai) and no appressoria were observed at this time. In most cases, conidia germinated and produced two germ tubes that grew towards the stomata where the penetration occurred. Fungus sporulation occurred at both adaxial and abaxial leaf epidermis through the stomata. Fungal hyphae suscefully colonized the mesophyll cells both inter and intra cellularly. Disease symptoms appeared at 12 dai with severity level of 1%, but reached 32% at 22 dai. / O processo infeccioso de Ramularia areola, o agente causal da mancha da ramulária, em folhas de algodoeiro, Gossypium hirsutum L. (Malvaceae), foi estudado utilizando a microscopia eletrônica de varredura. Os conídios iniciaram a germinação 12 horas após a inoculação (hai) e nenhum apressório foi observado nesse período. Na maioria dos casos, os conídios germinaram e produziram dois tubos germinativos que cresceram através dos estômatos onde a penetração ocorreu. A esporulação do fungo ocorreu em ambas as faces adaxial e abaxial da epiderme foliar através dos estômatos. As hifas do fungo colonizaram com sucesso as células do mesófilo inter e intra celularmente. Os sintomas da doença apareceram em 12 dai com um nível de severidade de 1%, porém alcançaram 32% em 22 dai.