This diploma thesis is focused on study of potential production of extracellular hydrolytic enzymes. The theoretical part deals with characterization of selected hydrolytic enzymes, their catalytic properties, the possibility of extracellular hydrolase production by fungi and their applications. In experimental part production strains Aureobasidium pullulans, Fusarium solani and Phanerochaete chrysosporium were used. Productions of cellulase, amylase, xylanase, lipase, protease and lignin-degraded enzymes (laccase, manganese- dependent peroxidase, lignin peroxidase) were observed. Cultivations were carried out in submersed mode in mineral medium supplemented by waste co-substrates such as wheat bran, corn bran, rice bran and oat bran, sawdust, rice, apple fiber, egg pasta and egg-free pasta. Production of enzymes depended on the substrate type and time of cultivation. The highest cellulase, xylanase and amylase activities were measured in the first period of cultivation (3 to 7 day). Lignin-degraded enzymes and proteases were produced at the end of cultivation (7 to 10 days). Lipolytic activity was detected only in A. pullulans, where the activity increased with time of cultivation. The highest value was determined during cultivation on wheat bran (3.6 nmol/ml.min). The highest xylanase and celulase activity (170.3 nmol/ml.min, 248.0 nmol/ml.min) were determined during cultivation of F. solani on corn bran. The highest amylase activity (111.8 nmol/ml.min) was reported in P. chrysosporium during the cultivation on rice. The highest protease activity (68.0 nmol/ml.min) was determined in F. solani grown on wheat bran. The best producer of laccase was A. pullulans, the highest production was recorded for egg-free pasta (27.0 nmol/ml.min). The maximum lignin peroxidase activity (12.5 nmol/ml.min) was measured during the cultivation of F. solani on egg pasta, while the highest yield of Mn-dependent peroxidase (7.7 nmol/ml.min) was achieved during the cultivation of A. pullulans on wheat bran. Lignin-degraded enzymes behaved as inductive, while the other enzymes were produced in mineral medium too. Activity of cellulase in the mineral medium was in A. pullulans strain higher than in media with waste substrates. Enzymes produced into A. pullulans medium were purified by ultrafiltration, ion exchange chromatography and gel filtration.

The diploma thesis concerns observing the health conditions of pea plants. The experiments were established in 2016–2017 in cooperation with AGRITEC, research, breeding and services, s.r.o. company in Šumperk. There were 9 varieties and 3 lines selected for the experimentations. The observation for each experiment was divided into two timing terms. The resistance to pea powdery mildew caused by pathogen Eryshipe pisi, was observed in greenhouse conditions. The highest level of resistance occurred line 1109/13 and variety FRANKLIN. The lowest level of resistance had variety ARVIKA. However this variety also occured to have the highest level of resistance in laboratory tests to resistance to Fusarium solani and Fusarium oxysporum. The pathogens causing the complex of root rot and black stem deseases were observed in conditions of infectious field. Variety ATLAS was infected the least.

Rhizoctonia blight (sensu lato) is a common and serious disease of many turfgrass species. The most widespread causal agent R. solani consists of several genetically different anastomosis groups (AGs) and subgroups. Though anastomosis or hyphal fusion reactions have been used to group Rhizoctonia species, they are time consuming and sometimes difficult to interpret. Anastomosis reactions are incapable of identifying isolates belonging to different AG subgroups within an AG. This study evaluated molecular techniques in comparison with traditional anastomosis grouping (AG) to identify and group isolates of Rhizoctonia. More than 400 Rhizoctonia isolates were collected from diseased turfgrass leaves from eight geographic areas in Virginia and Maryland. A random sample of 86 isolates was selected and initially characterized by colony morphology, nuclei staining and anastomosis grouping. Molecular identification was performed by analysis of rDNA-ITS region and DNA fingerprinting techniques universally primed PCR (UP-PCR) and amplified fragment length polymorphism (AFLP). The cladistic analysis of ITS sequences and UP-PCR fragments supported seven clusters. Isolates of R. solani AG 1-IB (n=18), AG 2-2IIIB (n=30) and AG 5 (n=1) clustered separately. Waitea circinata var. zeae (n=11), and var. circinata (n=4) grouped separately. A cluster of six isolates (UWC) did not fall into any known Waitea group. Most of the binucleate Rhizoctonia-like fungi (BNR) (n=16) grouped separately. AFLP grouping also largely agreed with the above results. However, UWC isolates clustered into two groups. Molecular analyses corresponded well with traditional anastomosis grouping by clustering isolates within an AG or AG subgroup together. UP-PCR cross-hybridization could distinguish closely related Rhizoctonia isolates to their infraspecies level. Genetically related isolates belonging to the same AG subgroups cross-hybridized strongly, while isolates of different AGs did not cross-hybridize or did so weakly. Sequence-characterized amplified region (SCAR) markers were generated from UP-PCR products to identify isolates of major pathogenic groups AG 1-IB and AG 2-2IIIB. Specific primer pairs successfully distinguished isolates of AG 1-IB and AG 2-2IIIB from isolates of other AGs. Sensitivity of Rhizoctonia species and AGs was tested in vitro to commercial formulations of iprodione, triticonazole and pyraclostrobin. W. circinata isolates were moderately sensitive to iprodione while isolates of R. solani and BNR were extremely sensitive. Isolates of AG 2-2IIIB showed less sensitivity to triticonazole than other Rhizoctonia isolates. W. circinata var. zeae isolates were moderately sensitive to pyraclostrobin while most of the other isolates were extremely sensitive. / Ph. D.

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.

Nas últimas décadas, a podridão vermelha das raízes da soja (PVR), causada pelo Fusarium solani f.sp. glycines (FSG), tornou-se uma doença séria nas regiões brasileiras onde já foi constatada, sendo a utilização de cultivares resistentes um componente fundamental de um sistema integrado de controle. Este trabalho teve por objetivo pesquisar estratégias de seleção de genótipos resistentes a PVR. Várias metodologias foram implementadas: avanço de progênies até a geração F7:2 e seleção em campo infestado, de progênies selecionadas em F2. Avaliou-se o cruzamento IAC 4 x Conquista em casa de vegetação (geração F3:2) e em campo infestado (gerações F3:2, F4:2 e F5:2), através de estudo de estabilidade e adaptabilidade das progênies. As metodologias de infecção em vasos de barro e bandejas de isopor em casa de vegetação foram comparadas para melhorar a eficiência de seleção; também foi feita análise de repetibilidade dos sintomas foliares da PVR, para otimização do número de avaliações necessárias para classificar o genótipo de soja quanto à reação ao FSG. Cultivares brasileiras de soja foram avaliadas em campo infestado. Em campo, as plantas foram avaliadas no estádio R5-6, com notas variando de 1 (ausência de sintomas) a 5 (100% da raiz principal com sintomas). Em casa de vegetação, as plantas foram avaliadas aos 35 dias pós-semeadura para sintomas radiculares e da parte aérea. Houve eficiência da seleção para resistência a PVR tanto em F2 (sintomas foliares em casa de vegetação) quanto em F7:2 (sintomas radiculares em campo naturalmente infestado). A seleção praticada nas gerações intermediárias também foi eficiente para aumentar a produtividade de grãos das progênies. Para o cruzamento IAC 4 x Conquista nas gerações F3:2, F4:2 e F5:2 em campo naturalmente infestado e na F3:2 em casa de vegetação, a metodologia de Annicchiarico possibilitou a seleção de genótipos com maior estabilidade e adaptabilidade para os diferentes ambientes; com o estudo genético destas gerações, demonstrou que há efeitos gênicos aditivos e dominantes no controle genético da PVR. Além disso, constatou-se que os genes responsáveis pela resistência estão dispersos nos genitores, provavelmente agrupados em blocos gênicos. A presença de dominância indica que a seleção deve ser postergada para gerações com maior homozigose (linhagem pura); esta idéia foi reforçada pela baixa herdabilidade ao nível de plantas. O uso de bandejas de isopor foi significativamente mais eficiente do que vasos de barro para inoculação de FSG em casa de vegetação. Pela análise de repetibilidade, detectou-se que quatro avaliações foram suficientes para discriminar se o genótipo era realmente suscetível ao FSG. A correlação da geração F3:2 (IAC 4 x Conquista) entre campo infestado e casa de vegetação foi praticamente nula; no entanto, 54% das progênies selecionadas em campo também foram selecionadas em casa de vegetação. A seleção de genótipos superiores para resistência ao FSG não é tarefa fácil, mas pode ser aprimorada pelo uso conjugado de metodologias suplementares que aumentem a eficiência de seleção. / During the last decades, soybean sudden death syndrome (SDS), caused by Fusarium solani f.sp. glycines (FSG), has become a serious disease in the regions where it was encountered. The use of resistant soybean cultivars has been an important component of an integrated management system. The aim of this work was to research selection strategies of SDS-resistant genotypes. A number of methodologies were used: self pollination up to generation F7:2 and progeny selection in infested field of selected F2. The crossing IAC 4 x Conquista was studied in greenhouse (generation F3:2) and in a naturally infested field (generations F3:2, F4:2 and F5:2), where the progeny stability and adaptability study was carried out. The methodologies of infection in clay pots and polystyrene trays in greenhouse were compared to improve selection efficiency. An analysis of the repeatability of foliar SDS symptoms was also performed in order to optimize the number of evaluations needed to classify a soybean genotypes reaction to FSG. Brazilian soybean cultivars were evaluated in a naturally infested field, where plants were evaluated at stage R5-6 with scores varying from 1 (absence of symptoms) to 5 (main root 100% symptoms). In the greenhouse, the plants were evaluated 35 days after sowing for root symptoms, while the leaf symptoms was only evaluated after the appearance of symptoms. The selection done in F2 (leaf symptoms in greenhouse) as well as F7:2 (root symptoms in infested field) for FSG were efficient. The selections performed in intermediary generations were efficient in increasing the grain yield of the progeny. Studies of the F3:2, F4:2 and F5:2 generations of the IAC 4 x Conquista cross in an infested field and F3:2 in greenhouse, enabled through Annicchiaricos methodology, the selection of genotypes with greater stability and adaptability to different environments. The genetic study of these generations demonstrated there are additive and dominant genetic effects on the genetic control of SDS. Moreover, it was noted that the genes responsible for resistance are dispersed on the genitors, probably grouped into genic blocks. The presence of dominance indicates that the selection should be passed onto generations with greater homozygosity. There is also a clear indication that breeders should work with progeny lines in order to assist selection due to the low inheritability at the individual plant level. For the experiments with different FSG inoculation methodologies (clay pots and polystyrene trays) in greenhouse the use of trays was significantly more efficient; by studying the repeatability analysis, it was found that four evaluations is enough to discriminate if the genotype is really susceptible to FSG. The correlation of the F3:2 generation (IAC 4 x Conquista) between infested field and greenhouse was practically null; however, 54% of the progeny selected in the field were selected in greenhouse. As observed, the selection of superior genotypes for FSG resistance is not an easy task, but may be improved by means of new methodologies and the conjugated use of methodologies which improve selection efficiency.

Submitted by (lucia.rodrigues@ufrpe.br) on 2017-03-16T12:53:24Z No. of bitstreams: 1 Eddy Enrique Barraza Andrion (1).pdf: 947495 bytes, checksum: aa5e0f6002d19bd71d54a71eae9dc237 (MD5) / Made available in DSpace on 2017-03-16T12:53:24Z (GMT). No. of bitstreams: 1 Eddy Enrique Barraza Andrion (1).pdf: 947495 bytes, checksum: aa5e0f6002d19bd71d54a71eae9dc237 (MD5) Previous issue date: 2009-07-31 / The cowpea (Vigna unguiculata L.) is one of the main crops in the Northeast of Brazil especially for the small farmers. The Fusarium wilt and Rhizoctonia canker caused by Fusarium oxysporum f.sp. tracheiphilum and Rhizoctonia solani, respectively are the cowpea diseases showing more frequency and intensity in the Northeast of Brazil. This work aimed to evaluate the natural suppressiveness of 66 soils of this region to the Fusarium wilt and Rhizoctonia canker, and to analyze the physical, chemical and biological characteristics of this soils associated with disease suppressiveness or conducivity. The evaluated soils were grouped from highly suppressive to highly conducive in relation to Fusarium wilt and Rhizoctonia canker severities. The main variables involved in Fusarium wilt suppressiveness were high levels of phosphorus and potassium, basal respiration (CO2 evolution) and indexes of microbial diversity and equitability. For Rhizoctonia canker important correlations were determined with levels of phosphorus, potassium and sodium, basal respiration and enzymatic activity of fluorescein diacetate. There was no correlation between physical factors and suppressiveness to Fusarium wilt, but it was possible to correlate the levels of sand, clay and silt with suppressiveness and/or conducivity of Rhizoctonia canker. Three soils previously classified as highly supressive to Rhizoctonia canker were evaluated in relation to eight strains and three inoculum densities of R. solani. There was significant difference among soils and strains in relation to levels of disease severity. In the three soils the severity levels induced by the strain CMM-1053 were similar to those observed in former studies. Most of the strains showed different behavior in relation to soils, except for CMM-1064 and CMM-1066. There was significant difference among disease severity levels and different inoculum densities. The three soils presented good stability in relation to the different R. solani strains, but the inoculum density may be a limiting factor in the implementation of the natural soil suppressiveness or the supressivity induction in conducive soils. / O caupi (Vigna unguiculata L.) é uma das culturas mais importantes da região Nordeste do Brasil, principalmente na economia de pequenos produtores rurais. A murcha-de-fusário e a rizoctoniose, causadas pelos fungos Fusarium oxysporum f.sp. tracheiphilum e Rhizoctonia solani, respectivamente, são as doenças mais freqüentes e de maior intensidade em caupi no Nordeste brasileiro. Esta tese teve como objetivos avaliar a supressividade natural de 66 solos do Nordeste brasileiro à murcha-de-fusário e rizotoniose do caupi, analisar as características físicas, químicas e biológicas dos solos associadas com a supressividade ou conducividade às doenças, bem como avaliar a estabilidade da supressividade de três solos à rizoctonose do caupi, considerando diferentes isolados e densidades de inóculo de R. solani. Em relação à severidade da murcha-de-fusário e da rizoctoniose do caupi, os solos avaliados foram agrupados desde fortemente supressivos a altamente conducivos. As principais variáveis envolvidas na supressividade da murcha-de-fusário foram elevados teores de fósforo e potássio, respiração basal (CO2 evoluído) e os índices de diversidade e eqüitabilidade microbiana. Para a rizoctoniose, foram determinadas correlações importantes com os níveis de fósforo, potássio e sódio, respiração basal e atividade enzimática de diacetato de fluoresceína. Não foram correlacionados fatores físicos com a supressividade à murcha-de-fusário, porém foi possível correlacionar os teores de areia, argila e silte com asupressividade e/ou conducividade da rizoctoniose. Três solos classificados como fortemente supressivos à rizoctoniose foram avaliados em relação a oito isolados e três densidades de inoculo de R. solani. Houve diferença significativa entre os solos e os isolados quanto aos níveis de severidade da doença. Nos três solos os níveis de severidade induzidos pelo isolado CMM-1053 foram similares aos verificados nos estudos prévios. A maioria dos isolados apresentou comportamentos diferente em função dos solos, com exceção dos isolados CMM-1064 e CMM-1066. Foi verificada diferença significativa entre os níveis de severidade da doença e as diferentes densidades de inoculo. Os três solos evidenciaram estabilidade em relação aos diferentes isolados de R. solani, porém a densidade de inóculo pode ser um fator limitante na implementação da supressividade natural dos solos ou da indução da supressividade em solos conducivos.

No description available.

The soil-borne fungi Rhizoctonia solani Kuhn and Sclerotinia sclerotiorum De Bary are ubiquitous plant pathogens with a wide host range. They are among the most widespread and destructive diseases of many crops, including sunflowers. Although in many cases, the use of chemicals appears to be the most economical and efficient means of controlling plant pathogens, their environmental concerns and the development of tolerance in pathogen populations have led to drastic reduction in their usage and increased the need to find alternative means of disease control. The potential benefits of applying Trichoderma harzianum Rifai and silicon (Si) nutrition to plants have been extensively reviewed. In this study, the ability of T. harzianum (Eco-T®), soluble silicon, and their combination was evaluated on sunflower (Helianthus annuus L.), for their potential to suppress pathogenic strains of R. solani and S. sclerotiorum. The ability of this crop to take up and accumulate Si in different plant parts was also investigated. In vitro assessment of fungal responses to Si in PDA showed that both R. solani and S. sclerotiorum were inhibited in the presence of Si. More inhibition was observed as the Si concentration increased with a relative increase in pH. Maximum growth inhibition was observed at 3000 mg ;-1 – 6000 mg ;-1 of PDA. No difference in inhibition between the two pathogens was observed, thus confirming the fungitoxic/suppressive ability of high Si concentrations to fungal growth. In addition, in vivo trials showed that the Si concentration of 200 mg ;-1 applied weekly significantly increased the dry weight of plants inoculated with R. solani and S. sclerotiorum and was therefore considered the optimum concentration. Assessments on in vitro antifungal activities of Eco-T® on R. solani and S. sclerotiorum, showed that Eco-T® significantly inhibited mycelial growth, in both dual culture methods and volatile and non-volatile compounds produced by Eco-T®. In addition, the combination of Eco-T® and Si was most effective in suppressing damping-off and increasing plant dry weight of sunflower seedlings in the greenhouse. The combination of Si and Eco-T® significantly increased percentage germination, number of leaves and head dry weight of the sunflower cultivars tested. Silicon alone increased growth but was unable to control R. solani and S. sclerotiorum effectively. Rhizotron studies showed that S. sclerotiorum infected the host through the roots and the stem, whereas R. solani only infected the host through the roots. A study on Si uptake and distribution showed that sunflower accumulates Si in various plant tissues. Analysis of plant tissues revealed that more Si was accumulated in leaves > stems > roots, with the Si levels in leaves being significantly higher than in stems and roots. In conclusion, Si alone could be used to increase growth but was unable to control R. solani and S. sclerotiorum. However, Si together with Eco-T® provides an environmentally friendly alternative for the control of R. solani and S. sclerotiorum, and enhanced plant growth and yield. / Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2009.

Inoculation with free-living diazotrophic bacteria is well documented to enhance vegetative growth and yield increases of various crops coupled with suppression of sublethal pathogens. The use of microbial inoculants has been identified as an alternative or supplement to use of nitrogenous fertilizers and agrochemicals for sustainable agriculture. The search for effective free-living diazotrophic bacterial strains for formulation as biofertilizers has been on going since the 1970’s and a number of inoculant biofertilizers have been developed and are commercially available. In the current study, 250 free-living diazotrophic bacteria were isolated from soils collected from the rhizosphere and leaves of different crops in different areas within KwaZulu-Natal, province, Republic of South Africa. These were evaluated for plant growth-promotion and biological control of damping-off fungi initially by in vitro screening. The growth promotion traits tested included, phosphate-solubilization, production of indole-3-acetic acid, production of ammonia and acetylene reduction. Biocontrol traits evaluated included siderophore-production, antibiosis, and production of hydrogen cyanide (HCN). Biochemical and molecular bioassay tests were conducted to identify the twenty most promising isolates selected in the in vitro study. The twenty isolates were further tested in combination with various levels of nitrogenous fertilizer for growth-promotion of maize (Zea mays L.) and wheat (Triticum aestivum L.) under greenhouse conditions. The five most promising isolates identified for growth promotion under greenhouse conditions for each crop were assessed for their effects on the germination of maize in vitro and growth and yields of two maize and two wheat cultivars, when combined with a low dose of nitrogenous fertilizer in field trials. The five Bacillus subtilis (Ehrenberg) Cohn isolates that suppressed the growth of a wide range of pathogenic fungi in vitro were tested for their efficacy against damping-off of wheat caused by Rhizoctonia solani Kühn in the greenhouse. These isolates were further studied for their modes of action against R. solani in vitro. The modes of action tested included antibiosis, production of siderophores, extracellular enzymes, production of hydrogen cyanide (HCN) and antibiotic resistance. The twenty most promising bacterial isolates identified from the in vitro screening reduced acetylene to ethylene, produced indole-3-acetic acid and siderophores, one isolate solubilized phosphate, and 9 inhibited the growth of R. solani. These twenty isolates enhanced growth of maize and wheat above the Uninoculated Control under greenhouse conditions. The growth enhancements varied with bacterial isolate x crop species interactions, which identified five different isolates for each of the two crops. Relative to the Uninoculated Control, the best five isolates significantly (P = 0.001) enhanced the growth of maize and wheat at all fertilizer levels for a number of growth parameters: increased chlorophyll levels and heights of maize, shoot dry biomass of maize and wheat, and enhanced root development of maize in the greenhouse. Inoculation of maize and wheat with the two most promising isolates identified from the field trial for each crop, in combination with 65% and 50% of the recommended amount of nitrogenous fertilizer for maize and wheat, respectively, caused the same increases in shoot biomass as the Fully Fertilized Control. Application of a combination of the best bacterial isolates and 35% nitrogenous fertilizer resulted in the same or greater shoot dry biomass and yields of both maize and wheat under field conditions. Shoot dry biomass of wheat increased by 75% above the Uninoculated Control and 30% above the Fully Fertilized Control. The wheat yield increased by 95% above the Uninoculated Control and 43% above the Fully Fertilized Control. Seed inoculation with the best isolates combined with 35% N increased yields of maize by 41% above the Uninoculated Control and 15% above the Fully Fertilized Control. The best isolates significantly (P < 0.001) increased plant height, chlorophyll levels and shoot biomass of maize relative to the Uninoculated Control. There was a positive correlation between chlorophyll level and yield, chlorophyll level and shoot dry biomass, height and shoot dry biomass and height and yield of maize at P = 0.01 with r values of 0.87, 0.77, 0.92 and 0.81, respectively. The isolates that exhibited multiple plant-growth promoting traits in vitro, increased shoot biomass of both maize and wheat in the greenhouse and field, and caused yield increases in the two crops under field conditions. Five B. subtilis isolates inhibited the growth of some of the pathogenic fungi tested in vitro up to 95%. Seed inoculation with the same isolates significantly (P = 0.001) suppressed R. solani damping-off of wheat under greenhouse conditions and exhibited multiple mechanisms of disease control in vitro. The use of microbial inoculants in combination with low doses of nitrogenous fertilizers can enhance crop production without compromising the yields. The B. subtilis isolates obtained in this study can effectively control R. solani damping-off of wheat, fix nitrogen and enhance plant growth. The use of microbial inoculants can contribute to the integrated production of cereal crops with reduced nitrogenous fertilizer inputs, as a key component of sustainable agriculture. Key words: Free-living bacteria; plant growth-promotion; diazotrophs; biological nitrogen fixation; phosphate-solubilization; siderophores; indole-3-acetic acid; biocontrol; damping-off / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2013.

Submitted by Marco Antônio de Ramos Chagas (mchagas@ufv.br) on 2017-04-19T13:51:31Z No. of bitstreams: 1 texto completo.pdf: 481806 bytes, checksum: 719ba3466811da8459b6597dd508ae83 (MD5) / Made available in DSpace on 2017-04-19T13:51:31Z (GMT). No. of bitstreams: 1 texto completo.pdf: 481806 bytes, checksum: 719ba3466811da8459b6597dd508ae83 (MD5) Previous issue date: 2001-11-24 / Avaliou-se a eficiência de diferentes indutores de resistência nas espécies de tomateiro Lycopersicon esculentum cv. Miller (suscetível) e L. hirsutum v. glabratum introdução CNPH 417 (resistente) ao agente etiológico da “pinta preta” do tomateiro, Alternaria solani. Os indutores de resistência testados foram quitosana, ácido salicílico, benzotiadiazol e ácido acetil salicílico tamponado. Verificou-se que somente o ácido salicílico e o benzotiadiazol foram capazes de induzir resistência. Em todos os ensaios, a espécie resistente apresentou um número médio de lesões por folíolos inferior ao da espécie suscetível, tratada ou não tratada. O número médio de lesões por folíolos apresentados pela espécie resistente não foi, em nenhum dos ensaios, afetado por quaisquer tipos de indutores de resistência. Realizou-se a histopatologia comparativa da interação hospedeiro suscetível x A. solani, tratado ou não com os indutores de resistência ácido salicílico, benzotiadiazol e ácido acetil salicílico tamponado e da interação hospedeiro resistente x A. solani, não induzido. Coletaram-se os materiais para exame 24, 48 e 72 horas após a inoculação. Este estudo foi realizado com o material diafanizado em cloral hidratado saturado. Verificou-se que as substâncias indutoras de resistência interferiram na adesão foliolar dos esporos e, ou, tubos germinativos, reduzindo o número médio de lesões de esporos nas primeiras 48 horas. Em CNPH 417, observou-se que o número médio de lesões era menor até o último intervalo estudado, de 72 horas. As barreiras morfológicas observadas por contraste diferencial de interferência Nomarski, como modificações de paredes, modificações citoplasmáticas e modificações morfológicas observadas em pontos distantes aos do sítio de penetração, de modo geral, foram as mesmas observadas na testemunha. Entretanto, o contraste diferencial não fornece nenhum indicativo sobre a composição química das barreiras. Portanto, é possível afirmar que, apenas como estruturas físicas, as modificações estruturais da parede e do citoplasma não foram eficientes para reduzir a penetração do fungo e não diferiram entre plantas induzidas e testemunha, embora pudessem apresentar efeito de retardamento da penetração e da colonização. / The synthetic chemicals chitosan, salicylic acid, benzothiadiazol and buffered acetyl salicylic acid were evaluated as inducers of resistance to Alternaria solani in Lycopersicon esculentum cv. Miller (susceptible) and L. hirsutum var. glabratum CNPH 417 (resistant). The treatments with salicylic acid and benzothiadiazol reduced lesion numbers in susceptible tomato. There was no addititional effect of the chemical treatments in reducing lesion numbers in the resistant species. Chitosan and buffered acetylsalicylic acid were ineffective as resistance inducers. For histological study, leaflets of susceptible tomato L. esculentum cv. Miller were treated with salicylic acid, buffered acetylsalicylic acid or benzothiadiazol, and sample tissues were cleared in chloral hydrate and examined by differential interference contrast microscopy. The chemical treatments reduced numbers of spore and of germtubes attached to the cell epidermis. Cytoplasmic aggregation, changes in the cell wall and other modifications of the host cell were also observed. These modifications might contribute as structural defense mechanisms impeding host cell penetration and retarding fungal growth. No differences in frequency were found in treated and nontreated tomato tissues. However, these structural modifications as inducing mechanisms of resistance cannot be discarded yet since further clarifications are needed, such as those provided by cytological and biochemical studies. / Tese importada do Alexandria