Submitted by (lucia.rodrigues@ufrpe.br) on 2017-03-14T15:38:58Z No. of bitstreams: 1 Katia Cilene da Silva Felix (1).pdf: 376089 bytes, checksum: 45b421012fb39c4ea534d1182006970e (MD5) / Made available in DSpace on 2017-03-14T15:38:58Z (GMT). No. of bitstreams: 1 Katia Cilene da Silva Felix (1).pdf: 376089 bytes, checksum: 45b421012fb39c4ea534d1182006970e (MD5) Previous issue date: 2009-02-18 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / Bacterial wilt caused by Ralstonia solanacearum, race 1, biovars 1 and 3 causes severe damage to pepper growth in Agreste region of state of Pernambuco in Brazil. The pathogen survival in infected tissues of stem and roots incorporated to the soil at 0, 5 and 15 cm; and at 10 different soil types of Pernambuco without host plant was studied by using a spontaneous mutant resistant to 100 mg l-1 of rifampicin (R. solanacearum A1-9Rif). Pathogen survival only showed difference (P<0.05) in relation to vegetal tissue. Pepper root tissues showed higher survival duration (DUR) (17.1 d), area under population curve (AACPOP) (430x104) and population at 7 (POP7) (5.0x104 UFC g-1 tissue) and 21 days (POP21) (3.1x104 UFC g-1 tissue) than stem tissues, which were respectively, 7.0 d; 3.8 x104; 0.35 and 3.1x104 UFC g-1 tissue. On the other hand stem tissues presented higher decomposition index (81.3%) and pH (7.7) than root tissues, respectively 68.9% and 6.8. The previous soil microbiological analysis did not finddifferences among populations of actinomycetes, copiotrophic, bacteria, total bacteria and total fungi respectively 5.02; 3.63; 5.23 and 4.19 log UFC g-1 dry soil. Populations of oligotrophyc bacteria, Bacillus spp., fluorescent Pseudomonas spp. and Trichoderma spp. were not detected. The pathogen was isolated from the soil adjacent to infected tissues of stem and roots at the three depths six weeks after experiment establishment. The analysis of AACPOP in relation to locals (counties) of soil sampling, type of soil coverage at the sampling time and soil texture was not significant by Kruskal-Wallis test (P≤0.05). Among the 10 studied soils, seven were classified as suppressive showing low intervals of DUR (42 to 49 days), AACPOP (0.77 to 4.05), POP14 (5.59 to 6.18 log UFC g-1 soil) and POP42 (5.44 to 6.31 log UFC g-1 soil). The soils S3, S4 and S6 were evaluated as conducive. Considering all soils together or only the suppressive, DUR,AACPOP and POP42 only showed significant correlation with soil physical and chemical characteristics, positive for clay, residual humidity and useful water and negative for pH. The population of copiotrophic bacteria, actinomycetes, total bacteria, total fungi (Fusarium spp., Aspergillus spp., Penicillium spp. and Rhizopus spp. were identified) and Bacillus spp. did not differ among the 10 soil types. / A murcha-bacteriana do pimentão é causada por Ralstonia solanacearum, raça 1, biovares 1 e 3 e causa grandes prejuízos à cultura desta solanácea. Este trabalho teve por objetivo estudar a sobrevivência do patógeno em tecidos infectados de caule e raiz incorporados ao solo às profundidades de 0, 5 e 15 cm; e em 10 diferentes tipos de solo na ausência da planta hospedeira. Foi utilizado um mutante resistente a 100 mg l-1 de rifampicina (R. solanacearum A1-9Rif). A sobrevivência do patógeno diferiu significativamente (P<0,05) apenas em relação aos tecidos vegetais analisados isoladamente. Desta forma, tecidos de raiz de pimentão apresentaram maior duração da sobrevivência (17,1 d), área abaixo da curva da população (AACPOP) (430x104) e população aos 7 (POP7) (5,0 x 104 UFC g-1 tecido) e 21 dias (POP21) (3,1 x 104 UFC g- 1 tecido) que os de caule, que foram respectivamente, 7,0 d; 3,8; 0,35 x104 e 0,48 x 104UFC g-1 tecido. Por outro lado, os tecidos do caule apresentaram maior índice de decomposição (81,3%) e pH (7,7) do que os de raízes, respectivamente 68,9% e 6,8. A análise microbiológica prévia deste solo não evidenciou diferenças significativas entre as populações de actinomicetos, bactérias copiotróficas, bactérias totais e fungos totais, que foram de 5,02; 3,63; 5,23 e 4,19 log UFC g-1 solo seco, respectivamente. Não foram encontradas populações de bactérias oligotróficas, Bacillus spp., Pseudomonas spp. fluorescentes, e Trichoderma spp. Entre os 10 tipos de solos estudados, sete foram classificados como supressivos por apresentarem baixas duração da sobrevivência (42 a 49 dias), AACPOP (0,77 a 4,05), POP14 (5,59 a 6,18 log UFC g-1 solo) e POP42 (5,44 a 6,31 log UFC g-1 solo). Os solos S3, S4 e S6 foram avaliados como conducivos. Neste estudo, apenas algumas características físicas e químicas apresentaram correlaçãosignificativa com duração da sobrevivência, AACPOP e POP42, considerando todos ossolos em conjunto ou apenas os supressivos, destacando-se argila, umidade residual e água disponível que apresentaram correlação positiva e o pH com correlação negativa.

Submitted by (lucia.rodrigues@ufrpe.br) on 2017-03-14T16:51:26Z No. of bitstreams: 1 Leonardo Tavares de Souza (1).pdf: 547012 bytes, checksum: 3ce24d8a338d375481b7eaa6393560c4 (MD5) / Made available in DSpace on 2017-03-14T16:51:26Z (GMT). No. of bitstreams: 1 Leonardo Tavares de Souza (1).pdf: 547012 bytes, checksum: 3ce24d8a338d375481b7eaa6393560c4 (MD5) Previous issue date: 2009-02-18 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / The Fusarium wilt caused by Fusarium oxysporum f. sp. lycopersici, is an important disease of tomato (Solanum lycopersicon L.) in Northeastern Brazil. Order to select genotypes with potential for use in the disease management, 60 strains were evaluated (F7 generation) from the crossing of access BHRS-2, 3 and Viradoro cultivar in relation to isolates from the physiologic races 2 and 3 of F. oxysporum f. sp. lycopersici. Seedlings 21 days old were inoculated using the method of cutting the roots and soak in the suspension of conidia of the pathogen. The evaluation was performed after 21 days, with the scale of grades ranging from 1 to 5. The genotypes were grouped into five classes of reaction. Most genotypes (73.3%) behaved as highly resistant to the race 2 isolate, while 45.0% were classified as susceptible and 28.3% as highly susceptible to the race 3 isolate. Only the L-1 strain showed high resistance reaction to both isolates. The stability of this line of resistance was evaluated on five isolates ofeach race (2 and 3). The line L-1 showed high levels of resistance to all race 2 isolates, therefore indicating high stability of resistance. However, for race 3 isolates, this strain showed three distinct classes of reaction, ranging from highly resistant to susceptible, indicating instability of resistance to this race. / A murcha-de-fusário, causada por Fusarium oxysporum f. sp. lycopersici, é uma importante doença do tomateiro (Solanum lycopersicon L.) no Nordeste brasileiro. Visando selecionar genótipos com potencial de utilização no manejo da doença, foram avaliadas 60 linhagens (geração F7) oriundas do cruzamento entre o acesso BHRS-2,3 e a cultivar Viradoro, em relação a isolados das raças fisiológicas 2 e 3 de F. oxysporum f. sp. lycopersici. Mudas com 21 dias de idade foram inoculadas pelo método do corte de raízes e imersão na suspensão de conídios do patógeno. A avaliação foi realizada após 21 dias, com o auxílio de escala de notas de 1 a 5, para agrupamento dos genótipos em cinco classes de reação. A maioria dos genótipos (73,3%) se comportou como altamente resistente ao isolado da raça 2, enquanto 45,0% foram classificados como suscetíveis e 28,3% como altamente suscetíveis ao isolado da raça 3. Somente a linhagem L-1 apresentou reação de alta resistência aos dois isolados de ambas as raças. A estabilidade da resistência dessa linhagem foi avaliada em relação a cinco isolados de cada raça (2 e 3) do patógeno. A linhagem L-1 apresentou reação de alta resistência a todos os isolados da raça 2, evidenciado estabilidade da resistência. No entanto, em relação aosisolados da raça 3, essa linhagem apresentou três classes de reação distintas, variando de altamente resistente a suscetível, indicando instabilidade da resistência à essa raça.

Standoff detection of targets using volatiles is essential when considering substances that are hazardous or dangerous, or for which the presence or location is unknown. For many invasive biological threats, their presence is often not realized until they have begun visibly affecting and spreading through crops or forests. The fungus Raffaelea lauricola is a biothreat vectored by the invasive beetle Xyleborus glabratus, or redbay ambrosia beetle (RAB), whose presence in avocado groves is currently detectable by visual inspection. Once visually identified, the affected trees must be removed and destroyed to protect those remaining trees. However, if the fungus is identified via standoff volatile detection, there is anecdotal evidence that it can be treated with propiconazole and saved from progression to the fatal laurel wilt disease. As a result of the rapid spread of R. lauricola and the quick death of trees, early detection through standoff methods is essential. The only current method of pre-symptomatic identification is canine detection. Canines are sensitive and selective biological detectors that can trace odors to their source, despite the presence of a variety of background odors. The present research evaluated the volatile organic compounds (VOCs) of the laurel wilt disease and R. lauricola using headspace solid phase microextraction-gas chromatography-mass spectrometry (HS-SPME-GC-MS). Additionally, a new method for odor collection and presentation to trained detection canines was developed. Knowledge of the disease and standoff volatile detection capabilities are improved using this information.

Oak wilt is a fatal disease of oaks caused by the fungus Ceratocystis fagacearum. Loss or degradation of habitat due to the disease may negatively affect the federally endangered golden-cheeked warbler (Setophaga chrysoparia). To assess the impact of oak wilt on golden-cheeked warblers, I investigated its influence on habitat selection and quality. I used remote sensing to estimate the amount of potential golden-cheeked warbler habitat currently affected by oak wilt, to predict the amount of potential habitat likely to be affected in the near future, and to assess the current probability of warbler occupancy in areas affected by oak wilt historically. I also quantified vegetative characteristics to assess overstory vegetation and regeneration in areas affected by the disease. I found proportional occupancy and territory density in unaffected areas to be, respectively, 3.5 and 1.8 times that of affected areas. Pairing success was 27% lower for territories containing oak wilt but fledging success was not affected. I estimated that 6.9% of potential golden-cheeked warbler habitat and 7.7% of the total area within my study region was affected by oak wilt in 2008. By 2018, I predicted that 13.3% of potential golden-cheeked warbler habitat and 16.0% of the study region would be affected by the disease. Using historical imagery, I found that areas affected by oak wilt in the past are less likely to be classified as current potential warbler habitat than areas never affected by the disease. I found no differences between the understory vegetation of affected and unaffected areas but that oaks were more common in the overstory than in the understory, suggesting that species composition in affected areas may shift in the years following an outbreak of the disease. My results suggest that the presence of oak wilt negatively influences habitat selection and quality for golden-cheeked warblers, likely due to reduced canopy cover in susceptible oak species. Additionally, oak wilt frequently occurs in golden-cheeked warbler habitat and will continue to spread into warbler habitat in the coming years. Future management efforts should address the threat oak wilt poses to golden-cheeked warblers by incorporating applicable preventative measures.

Infectious disease models can be used to understand mechanisms of the spread of diseases and thus, may effectively guide control policies for potential outbreaks. Deardon et al. (2010) introduced a class of individual-level models (ILMs) which are highly flexible. Parameter estimates for ILMs can be achieved by means of Markov chain Monte Carlo (MCMC) methods within a Bayesian framework. Here, we introduce an extended form of ILM, described by Deardon et al. (2010), and compare this model with the original ILM in the context of a simple spatial system. The two spatial ILMs are fitted to 70 simulated data sets and a real data set on tomato spotted wilt virus (TSWV) in pepper plants (Hughes et al., 1997). We find that the modified ILM is more flexible than the original ILM and may fit some data sets better.

Inhibition of Fusarium oxysporum f. sp. cubense race 4 by Burkholderia cepacia was evident when grown on various media (TSA, PDA, PSA, YM, KMB, PPM, NYGA, LA) with different carbon sources and under various pH and temperature conditions. In addition, B. cepacia was able to inhibit several fungal pathogens in vitro. Antagonism of B. cepacia against F. oxysporum f. sp. cubense occured at high levels of Fe³+, which may suggest that antagonism by B. cepacia did not involve siderophore production. Thin layer chromatogram (TLC) examination showed that B. cepacia produced several substances, one of which had similar R[f] value to that described for pyrrolnitrin. Cell-free supernatant of a 4-day culture of 6. cepacia was applied to an Amberlite XAD-2 column and inhibitory activity co-eluted with the 95% methanol (pH 9.5) fraction. The concentrated activated fractions showed inhibitory activity against F. oxysporum f. sp. cubense. A GC-MS chromatogram indicated numerous components in the antifungal extracts. The only compound identified in the Wiley 138 library, was 1,2- Benzenedicarboxylic acid, bis (2-Ethylhexyl) ester. Observations by light microscopy indicated that B. cepacia inhibited spore germination in F. oxysporum f. sp. cubense race 4 and retarded the mycelial growth. The interaction between the endophytic bacterium, B. cepacia and F. oxysporum f. sp. cubense race 4 was investigated with aid of scanning and transmission electron microscopy. This demonstrated that the bacterium was able to colonize the surface of hypha and macrospore of F. oxysporum f. sp. cubense. Mycelial deformation, terminal and/or intercalary swelling were evident. At later stages, hyphae of F. oxysporum f.sp. cubense, colonized by B. cepacia, were found to have collapsed. Further studies in vivo confirmed that B. cepacia colonized the hypha of F. oxysporum f. sp. cubense which had invaded banana roots. TEM observation showed that in the banana plant B. cepacia was closely associated with the healthy banana roots and a matrix was frequently found to be present between the bacterium and the plant surface. In addition, B. cepacia exists mainly in the intercellular space of the banana roots. UV irradiation treatment of B. cepacia resulted in a mutant that had lost inhibitory activity against F. oxysporum f. sp. cubense on TSA agar. Transposon mutagenesis of B. cepacia was performed by Tn5 insertion. Six mutants which had lost or had reduced inhibitory activity against F. oxysporum f. sp. cubense were generated. These mutants showed no inhibitory zones on TSA medium in the presence of the fungus. It was observed that one mutants. cepacia :: Tn5-188 appeared to lose the ability to colonize the fungal hypha, whilst a different mutant B. cepacia ::Tn5 - 217 was still able to colonize the fungal hyphae. TLC analyses showed that there was a decrease in antibiotic production in mutants B. cepacia :: Tn5 - 217 and B. cepacia - UV - 34, compared with the wild type. GC- MS analyses showed that there was no evidence of the peaks at 14.62 minutes, 20.0 minutes and 20.46 minutes in both chromatograms of mutants B. cepacia :: Tn5 -217 and 8. cepacia -UV - 34, compared with the wild type B. cepacia. No PCR products were detected using primers that were developed from sequences within the biosynthetic loci for Phi of P.fluorescens Q2-87(GenBank accession no. U41818) and PCA of P. fluorescens 2-79 (GeneBank no. L48616). Colony hybridization suggested that genomic DNA from B. cepacia could contain both Phi- and PCA probes. It was found that hybridization of genomic DNA digested with Cla-I of B. cepaca with Phl2a probe only occurred at low stringency. A hybridization signal was detected from a Cla-l fragment of approximately 2800bp. / Thesis (M.Sc.)-University of Natal, Pietermaritzburg, 1997.

Verticillium wilt, caused by Verticillium dahliae Kleb, is one of the most economically important diseases of woody hosts such as ash (Fraxinus spp.), sugar maple (Acer saccharum), and redbud (Cercis canadensis). The causal agent has a broad host range, including not only woody hosts but also important vegetable and field crops, and it is distributed worldwide. Diagnosis of V. dahliae in infected woody hosts is often based on the occurrence of vascular discoloration and time-consuming isolation. However, not all woody hosts exhibit vascular discoloration symptoms, and not all vascular discoloration symptoms are due to infection by V. dahliae. In this study, real-time PCR-based assays were evaluated and employed for rapid and accurate detection of V. dahliae in different woody hosts. DNA was extracted in large quantities from presumptively infected woody hosts by collecting drill-press shavings from sample tissue, bead-beating, and extracting using a CTAB method. Six published primer sets were evaluated against genomic DNA of V. dahliae as well as selected negative controls, and two sets (VertBt-F/VertBt-R and VDS1/VDS2) showed promise for further evaluation using DNA extracts from field samples. The VertBt primers amplified a species-specific 115-bp fragment of the expected size, while the VDS primers amplified the expected specific 540-bp fragment. However, the VertBt primer set exhibited higher sensitivity in detection of V. dahliae even in asymptomatic trees. The PCR-based methods developed here could be used as rapid tools for pathogen detecting and monitoring, thus informing plant pathogen management decisions.

Fusarium oxysporum f. sp. vasinfectum (Fov) was first identified in Australia in 1993, and has since become one of the most significant threats to the country’s thriving cotton industry. The interaction between a unique Australian biotype of Fov and cotton hosts with varying susceptibilities to Fusarium wilt was studied. This research described the infection process and associated host defence mechanisms of two commercial cotton varieties after inoculation with Fov, and quantified their subsequent accumulation of antimicrobial terpenoids. / A rapid, reliable glasshouse bioassay that correlated with field resistance was developed for the study of Fusarium wilt of cotton. Detailed observations of the infection process obtained through light microscopy were used to formulate the disease cycle of Australian Fusarium wilt cotton. Using pathogen growth assays, varietal differences in root exudates and vascular tissues in the cotton hosts were documented. Root diffusate from the most susceptible cotton variety to Fusarium wilt, Siokra 1-4, contained a lipophilic compound that promoted the germination of Fov microconidia. On the other hand, a lipophilic compound present in diffusate from the least susceptible variety, Sicot 189, inhibited the growth of Fov germ tubes. / A bioassay using inoculated whole plants showed that Fov colonisation of the vascular tissues of Sicot 189 was restricted after 3 days. The basis for this inhibition was investigated further using light and transmission electron microscopy. Infection induced the reorganisation of contact cells in host vascular tissue, including an increase in cytoplasmic content and the partitioning of vacuoles, which was concurrent with the accumulation of materials in adjacent vessel lumens, via pits. Histochemical analysis indicated these globular materials secreted into the vessels were terpenoids. These structural and terpenoid responses in Siokra 1-4 and Sicot 189 were similar, however, they were more intense and rapid in the latter, less susceptible variety. The responses in Sicot 189 also corresponded to the time period that pathogen inhibition was observed. Thus, a correlation was demonstrated between the rapid and intense induction of both structural and biochemical responses with decreased susceptibility to Fusarium wilt. Detailed HPLC analysis of vascular tissues confirmed that terpenoids accumulated more rapidly and at higher concentrations in the less susceptible cotton variety. These findings provided strong evidence for the involvement of antimicrobial terpenoids in the determination of Fusarium wilt susceptibility of Australian cotton varieties. / This work represents the most complete survey to date of the interaction of Australian biotypes of Fov with cotton. These insights can contribute to future cotton breeding efforts and cultural management of Fusarium wilt in the field. Thus, each part of this study has advanced complementary facets of our understanding of Fov, and has provided a framework from which future studies on phytoalexins and other putative cotton defences can be studied.

Thesis--Auburn University, 2009. / Abstract. Vita. Includes bibliographical references.

Thesis (Ph. D.)--University of Missouri-Columbia, 1998. / Typescript. Vita. Includes bibliographical references (leaves 130-145). Also available on the Internet.