Thesis (M.Sc.(Chemistry))--University of Pretoria, 2001. / Includes abstracts in English and Afrikaans. Includes bibliographical references.

Thesis (Ph. D.)--Illinois State University, 1974. / Title from title page screen, viewed Oct. 26, 2004. Dissertation Committee: Anthony E. Liberta (chair), T.I. Chuang, H. Huizinga, A. Richardson, E. Willis. Includes bibliographical references (leaves 87-90) and abstract. Also available in print.

Thesis (M.Sc.)(Biochemistry)--University of Pretoria, 2006. / Includes summary. Includes bibliographical references. Available on the Internet via the World Wide Web.

Fusarium verticillioides (=Fusarium moniliforme) a common fungal contaminant of maize throughout the world has been associated with diseases in both man and animals. The structure of the fumonisins, a family of structurally related mycotoxins isolated from cultures associated with the high incidence of human oesophageal cancer in the Transkei region in South Africa and with equine leucoencephalomalacia, a neurological disorder in horses and donkeys, has been established. The main mycotoxin, fumonisin B₁ consists of the diester formed by the C(14) and C(15) hydroxy groups of (2S,3S,5R,10R,12S,14S,15R,16R)-2-amino-12,16-dimethyleicosane-3,10,14,15-pentaol with the Si carboxy group of propane-1,2,3-tricarboxylic acid. A comparison of the structures of the 28 known fumonisins isolated since 1988 reveals that they share a common structural motif for the C(11)–C(20) unit, and probably also the same stereochemistry for the 4 stereogenic centres present in this part of the C20 backbone. Disconnection of the C(9)–C(10) bond in a retrosynthetic analysis of the fumonisins identifies (3S,5S,6R,7R)-3,7-dimethylundecane-1,5,6-triol as a common building block for the synthesis of any of the fumonisins. In the dissertation the retrosynhetic analysis of this 3,7-dimethylundecane-1,5,6-triol building block identifies a simple precursor, ethyl 2-heptenoate, as the starting material for the proposed synthetic route toward this target. The Sharpless asymmetric epoxidation reaction plays a pivotal role in this synthetic route as all 4 stereogenic centres present in the 3,7-dimethylundecane-1,5,6-triol target are generated by this methodology at three different stages of the proposed synthesis. The epoxy alcohol formed at each stage was subjected to regioselective ring opening followed by a protective group strategy which allowed for the protection of the secondary hydroxyl group as the benzyl ether and left the primary hydroxyl group, available after oxidation to the aldehyde, for a two-carbon chain extension to an α,β-unsaturated ester. This ester in turn was reduced to an allylic alcohol which formed the starting material for a second cycle of reactions. In this manner a synthetic route towards the target compound was developed and problems associated with the route investigated. The dissertation shows that a viable route was developed with complete stereochemical control in the formation of the stereogenic centres, even though the final product, the protected 3,7-dimethylundecane-1,5,6-triol was not obtained due to time constraints and material shortages. / Dissertation (MSc (Chemistry))--University of Pretoria, 2007. / Chemistry / MSc / unrestricted

The consequences of toxigenic fungi associated with stored seed have stimulated these investigations aimed at developing treatments to minimise this mycoflora, without significantly reducing seed quality or viability. The effects of immersion in water at 55, 57 and 60 QC for durations of 5 to 60 min were assessed for maize (Zea mays L.) seed in terms of fungal status, water uptake, electrolyte leakage, germination and seedling establishment. These assessments were conducted immediately after treatment, after re-dehydration for 2 days in an ambient air stream, and following a 1 month storage period under either cold (4 QC) or ambient (25 QC) conditions (33% and 91% RH, respectively). In all cases, the results are compared with those of control seeds and seeds pre-imbibed for 4 h at ambient temperature. The level of internal contamination, represented almost entirely by Fusarium moniliforme Sheldon, declined significantly when assessed immediately after treatment, the efficacy of which increased with increasing temperature and duration of treatment. Seeds immersed in water at 55 QC for a duration of 15 min exhibited an 85% reduction in infection levels, when compared with those of the control, while those treated at 57 and 60 QC (same duration) were uninfected. Immersing seeds in hot water, however, resulted in a lag in germination rate and drop in germination totality, the degree of which was enhanced by increasing duration and temperature of treatment, suggesting the status of the manipulation to be an accelerated ageing treatment. The electrolyte leakage studies indicated that the reduced germination performance of these seeds was not due to plasmalemma disorganisation. These deleterious effects, however, were counter-balanced as seeds treated at 55, 57 and 60 QC for durations up to 60, 30 and 10 min, respectively, produced plants of superior quality than those of the control, which is ascribed to the reduction of systemically transmitted pathogens. The efficacy of the hot water treatment in reducing the levels of seed infection and improving seedling quality was enhanced by subsequent re-dehydration. The reduction in seed-associated mycoflora was maintained following storage for 1 month at both 4 QC (33% RH) and 25 QC (91% RH). However, both seed and seedling quality were adversely affected following storage even under cold, dry conditions, which may be a consequence of the pre-treatment history of the seeds, which had been cold-stored for two years prior to the experiments. Applied as a pre-sowing treatment, therefore, hot water treatment shows promise for producing a crop of superior quality, less prone to fusarial pathogenesis. This treatment may be of particular importance to Third-World subsistence communities. / Thesis (M.Sc.)-University of Natal, 1995.

Fusarium verticillioides (= Fusarium moniliforme) a common fungal contaminant of maize throughout the world has been associated with diseases in both man and animals. The structure of the fumonisins, a family of structurally related mycotoxins isolated from cultures associated with the high incidence of human oesophageal cancer in the Transkei region in South Africa and with equine leucoencephalomalacia, a neurological disorder in horses and donkeys, has been established. The main mycotoxin, fumonisin B1 consists of the diester formed by the C(14) and C(15) hydroxyl groups of (2S,3S,5R,10R,12S,14S,15R,16R)-2- amino-12,16-dimethyleicosane-3,10,14,15-pentaol with the Si carboxy group of propane- 1,2,3-tricarboxylic acid. A comparison of the structures of the 28 known fumonisins reveals that they share a common structural motif for the C(11)-C(20) unit, and probably also the same stereochemistry for the 4 stereogenic centres present in this unit. Disconnection of the C(9)–C(10) bond in a retrosynthetic analysis of the fumonisins C20 backbone (C19 in the fumonisin C series)identifies (3S,5S,6R,7R)-3,7-dimethylundecane-1,5,6-triol as a common building block for the synthesis of any of the fumonisins. In the dissertation the retrosynthetic analysis of this 3,7-dimethylundecane-1,5,6-triol building block identifies (3S,4R,5R)-5-methylnonane-1,3,4-triol as a viable target which in turn could be derived from a simple starting material trans-4-hexen-3-one. Key reactions identified to realise the required transformations leading to the identified target included kinetic enzymatic resolution of the racemic alcohol obtained from trans-4-hexen-3-one, and a pivotal role for both the [2,3]-Wittig rearrangement and the use of Sharpless asymmetric epoxidation methodology as these reactions generated the requisite stereogenic centres present in (3S,4R,5R)-5-methylnonane-3,4-diol. In this manner a synthetic route from trans-4-hexen-3- one to (2R,3R,4R,5R,6E)-4-(benzyloxy)-2,3-epoxy-5-methylnon-6-en-1-ol using appropriate functional group transformations and protective group strategies, with complete stereochemical control, were developed in this work. Alternative strategies to overcome problems encountered during the synthesis are presented for future work. The conversion of the 4-(benzyloxy)-2,3-epoxy-5-methylnon-6-en-1-ol intermediate to the protected 5- methylnonane-1,3,4-triol target could not be carried out due to time constraints and material shortages. / Dissertation (MSc)--University of Pretoria, 2011. / Chemistry / unrestricted

Fusarium é um grande gênero de fungos filamentosos amplamente distribuídos que são importantes patógenos de plantas, animais e humanos. As doenças causadas por Fusarium spp. geram grandes perdas econômicas na produção de frutas, legumes, cereais e de celulose. Em humanos, o controle da fusariose com apenas um antifúngico é ineficaz, apresentando uma elevada taxa de mortalidade, especialmente em pacientes imunocomprometidos. A maior resistência aos fungicidas utilizados atualmente tem estimulado o desenvolvimento de novas tecnologias mais eficazes para o controle de fungos patogênicos. Assim, é necessária a busca de alternativas para o controle de microrganismos em ambas as áreas, clínica e agrícola. A inativação fotodinâmica antimicrobiano (IFA) é uma promissora plataforma antifúngica alternativa que pode ser utilizada para controlar o inóculo de fungos em mamíferos e no meio ambiente. A IFA baseia-se na utilização de um fotossensibilizador (FS) que se acumula na célula fúngica alvo. A exposição do FS à luz com um comprimento de onda apropriado, inicia um processo fotoquímico que produz espécies reativas de oxigênio (EROs), principalmente o oxigênio singleto, causando um dano oxidativo não especifico levando a morte da célula fúngica, sem dano significativo às células do hospedeiro. Em comparação com os fungicidas utilizados atualmente, a multiplicidade dos danos causados pelas EROs, reduzem a chance de selecionar microrganismos tolerantes. No presente trabalho, avaliamos o efeito da IFA com a combinação de luz vermelhas com fluências de 10 e 15 J cm-2 e cinco FS fenotiazínicos, azul de metileno (MB), azul de toluidina O (TBO), novo azul de metileno N fórmula sem zinco (NMBN), novo azul de metileno N formula com zinco (NMBN Zn) e um novo fenotiazínico pentacíclico S137, em microconídios não germinados e 4 h-germinados de Fusarium oxysporum, F. moniliforme e F. solani. A IFA com NMBN Zn resultou em uma redução de aproximadamente 5-logs na sobrevivência dos microconídios não germinados (quiescentes) e 4 h-germinados (metabolicamente ativos) das três espécies de Fusarium quando expostas a luz na fluência de 15 J cm-2. A lavagem dos microconídios não germinados e 4 h-germinados para retirar o excesso de FS antes da exposição à luz reduziu, porém não impediu a morte provocada pela IFA. A IFA com todos os FS e fluência aumentou a permeabilidade da membrana celular dos microconídios nas três espécies de Fusarium. O dano oxidativo causado pelas EROs produzidos durante o processo fotoquímico da IFA foi avaliado nos lipídios, proteínas e DNA dos microconídios das espécies de Fusarium. Foi observado um aumento da peroxidação lipídica em microconídios das três espécies de Fusarium após a IFA com NMBN Zn e S137. Observamos o aumento na carbonilação de proteínas em microconídios de F. oxysporum após IFA subletal com todos os FS. O aumento no dano do DNA em microconídios não germinados e 4 h-germinados foi observado apenas para o S137 na fluência de 0, 10 e 15 J cm-2. Nossos estudos expandem a compreensão da inativação fotodinâmica de fungos filamentosos / Fusarium is a large genus of filamentous fungi widely distributed wich are important pathogens of plants, animals and humans. Crop diseases caused by Fusarium generate great economic losses in the production of fruit, vegetables, cereals, and cellulose. In humans the control of progression of fusariosis by single-agent antifungal therapy is problematic, leading to a high mortality rate, especially with immunocompromised patients. The increased tolerance to currently used fungicides has stimulated the development of novel and effective technologies to control pathogenic fungi. Thus, the search for alternatives to control microorganisms is necessary in both, clinical and agricultural areas. Antimicrobial photodynamic treatment (APDT) is a promising alternative antifungal platform that can be used to control fungi both in mammalian hosts and in the environment. APDT is based on the use of a photosensitizer (PS) that accumulates in the target fungal cell. The exposure of the PS to light of an appropriate wavelength starts a photochemical process that produces reactive oxygen species (ROS), especially singlet oxygen, leading to non-specific oxidative damage causing the death of the fungal cell without significant harm to the host cells. In comparison with currently used fungicides, the multiple and variable targets of ROS reduce the chance of selecting tolerant microorganisms. In the present study, we evaluated the effect of APDT with the combination of red light with fluence of 10 and 15 J cm-2, and five phenotiazinium photosensitizer, methylene blue (MB), toluidine blue O (TBO), new methylene blue N zinc free form (NMBN), new methylene blue N zinc chloride double salt (NMBN Zn) and a novel pentacyclic phenothiazinium S137, on ungerminated and germinated microconidia of Fusarium oxysporum, F. moniliforme and F. solani. APDT with NMBN Zn resulted in a reduction of approximately 5 logs in the survival of the quiescent ungerminated microconidia and metabolically active germinated microconidia of the three Fusarium species when the light fluence of 15 J cm-2 was applied. Washing out the PS from both ungerminated and germinated microconidia before light exposure reduced but did not prevent the killing effect of APDT. APDT with all the PS and fluences increased cell membrane permeability for the three Fusarium species. The oxidative damage caused by ROS produced during the photochemical process of APDT, was evaluated in the lipids, proteins and DNA present in the microconidia. Increases in lipid peroxidation in microconidia of the three Fusarium species were observed only after APDT with NMBN Zn and S137. Proteins oxidative damage was observed by the increase in protein carbonylation in microconidia of the F. oxysporum after APDT with all PS. The increases in DNA damage from both ungerminated and germinated microconidia was observed only for S137 at fluence of 0, 10 and 15 J cm-2. Our study expands the understanding of photodynamic inactivation in filamentous fungi

Fusarium é um grande gênero de fungos filamentosos amplamente distribuídos que são importantes patógenos de plantas, animais e humanos. As doenças causadas por Fusarium spp. geram grandes perdas econômicas na produção de frutas, legumes, cereais e de celulose. Em humanos, o controle da fusariose com apenas um antifúngico é ineficaz, apresentando uma elevada taxa de mortalidade, especialmente em pacientes imunocomprometidos. A maior resistência aos fungicidas utilizados atualmente tem estimulado o desenvolvimento de novas tecnologias mais eficazes para o controle de fungos patogênicos. Assim, é necessária a busca de alternativas para o controle de microrganismos em ambas as áreas, clínica e agrícola. A inativação fotodinâmica antimicrobiano (IFA) é uma promissora plataforma antifúngica alternativa que pode ser utilizada para controlar o inóculo de fungos em mamíferos e no meio ambiente. A IFA baseia-se na utilização de um fotossensibilizador (FS) que se acumula na célula fúngica alvo. A exposição do FS à luz com um comprimento de onda apropriado, inicia um processo fotoquímico que produz espécies reativas de oxigênio (EROs), principalmente o oxigênio singleto, causando um dano oxidativo não especifico levando a morte da célula fúngica, sem dano significativo às células do hospedeiro. Em comparação com os fungicidas utilizados atualmente, a multiplicidade dos danos causados pelas EROs, reduzem a chance de selecionar microrganismos tolerantes. No presente trabalho, avaliamos o efeito da IFA com a combinação de luz vermelhas com fluências de 10 e 15 J cm-2 e cinco FS fenotiazínicos, azul de metileno (MB), azul de toluidina O (TBO), novo azul de metileno N fórmula sem zinco (NMBN), novo azul de metileno N formula com zinco (NMBN Zn) e um novo fenotiazínico pentacíclico S137, em microconídios não germinados e 4 h-germinados de Fusarium oxysporum, F. moniliforme e F. solani. A IFA com NMBN Zn resultou em uma redução de aproximadamente 5-logs na sobrevivência dos microconídios não germinados (quiescentes) e 4 h-germinados (metabolicamente ativos) das três espécies de Fusarium quando expostas a luz na fluência de 15 J cm-2. A lavagem dos microconídios não germinados e 4 h-germinados para retirar o excesso de FS antes da exposição à luz reduziu, porém não impediu a morte provocada pela IFA. A IFA com todos os FS e fluência aumentou a permeabilidade da membrana celular dos microconídios nas três espécies de Fusarium. O dano oxidativo causado pelas EROs produzidos durante o processo fotoquímico da IFA foi avaliado nos lipídios, proteínas e DNA dos microconídios das espécies de Fusarium. Foi observado um aumento da peroxidação lipídica em microconídios das três espécies de Fusarium após a IFA com NMBN Zn e S137. Observamos o aumento na carbonilação de proteínas em microconídios de F. oxysporum após IFA subletal com todos os FS. O aumento no dano do DNA em microconídios não germinados e 4 h-germinados foi observado apenas para o S137 na fluência de 0, 10 e 15 J cm-2. Nossos estudos expandem a compreensão da inativação fotodinâmica de fungos filamentosos / Fusarium is a large genus of filamentous fungi widely distributed wich are important pathogens of plants, animals and humans. Crop diseases caused by Fusarium generate great economic losses in the production of fruit, vegetables, cereals, and cellulose. In humans the control of progression of fusariosis by single-agent antifungal therapy is problematic, leading to a high mortality rate, especially with immunocompromised patients. The increased tolerance to currently used fungicides has stimulated the development of novel and effective technologies to control pathogenic fungi. Thus, the search for alternatives to control microorganisms is necessary in both, clinical and agricultural areas. Antimicrobial photodynamic treatment (APDT) is a promising alternative antifungal platform that can be used to control fungi both in mammalian hosts and in the environment. APDT is based on the use of a photosensitizer (PS) that accumulates in the target fungal cell. The exposure of the PS to light of an appropriate wavelength starts a photochemical process that produces reactive oxygen species (ROS), especially singlet oxygen, leading to non-specific oxidative damage causing the death of the fungal cell without significant harm to the host cells. In comparison with currently used fungicides, the multiple and variable targets of ROS reduce the chance of selecting tolerant microorganisms. In the present study, we evaluated the effect of APDT with the combination of red light with fluence of 10 and 15 J cm-2, and five phenotiazinium photosensitizer, methylene blue (MB), toluidine blue O (TBO), new methylene blue N zinc free form (NMBN), new methylene blue N zinc chloride double salt (NMBN Zn) and a novel pentacyclic phenothiazinium S137, on ungerminated and germinated microconidia of Fusarium oxysporum, F. moniliforme and F. solani. APDT with NMBN Zn resulted in a reduction of approximately 5 logs in the survival of the quiescent ungerminated microconidia and metabolically active germinated microconidia of the three Fusarium species when the light fluence of 15 J cm-2 was applied. Washing out the PS from both ungerminated and germinated microconidia before light exposure reduced but did not prevent the killing effect of APDT. APDT with all the PS and fluences increased cell membrane permeability for the three Fusarium species. The oxidative damage caused by ROS produced during the photochemical process of APDT, was evaluated in the lipids, proteins and DNA present in the microconidia. Increases in lipid peroxidation in microconidia of the three Fusarium species were observed only after APDT with NMBN Zn and S137. Proteins oxidative damage was observed by the increase in protein carbonylation in microconidia of the F. oxysporum after APDT with all PS. The increases in DNA damage from both ungerminated and germinated microconidia was observed only for S137 at fluence of 0, 10 and 15 J cm-2. Our study expands the understanding of photodynamic inactivation in filamentous fungi

A cana-de-açúcar é uma das principais culturas do Brasil e nos últimos anos está recebendo especial atenção devido ao crescente aumento da área cultivada e produção de etanol para uso como biocombustível. Considerando-se sua importância econômica e a possibilidade do uso de plantas geneticamente modificadas, essa cultura tem se tornado foco de pesquisas relacionadas à produtividade e sustentabilidade. Neste contexto, o estudo de comunidades microbianas associadas à cana-de-açúcar é de fundamental importância, pois além dessas comunidades desempenharem importante papel funcional na interação com a planta, os estudos realizados nas condições tropicais são limitados. Comunidades de fungos e bactérias associadas à cana-de-açúcar geneticamente modificada IMI-1 e sua isolinha convencional SP80-1842 foram sistematicamente isoladas de plantas cultivadas em área experimental em Piracicaba, SP, Brasil. Por meio de isolamento e PCR-denaturing gradient gel electrophoresis (PCR-DGGE) foi verificado que a transgenia não afeta a diversidade da comunidade fúngica associada à cana-deaçúcar. A diversidade dessas comunidades foi descrita e caracterizada molecularmente. O fungo Fusarium moniliforme apresentou alta freqüência de isolamento e também foi observado que o gene da endopoligalacturonase, pgIII, desempenha um importante papel no tipo de associação, endofítica ou patogênica, do F. moniliforme e a planta. O complexo Burkholderia cepacia constitui uma importante fração da comunidade de bactérias associada à cana-de-açúcar no Brasil e isolados deste complexo são capazes de inibir o crescimento do patógeno F. moniliforme. Análises filogenéticas indicaram que os isolados de bactérias endofíticas de Burkholderia são proximamente relacionados com linhagens-tipo do complexo B. cepacia isoladas de pacientes de fibrose cística. / In Brazil, the sugarcane is one of the most important cultivated crops. The sugarcane has received increased interest in the last years because of increase of the cultivated area and ethanol production to be used as biofuel. Considering its economical importance and the possibility of the use of the genetically modified plants; this crop has become the aim of current research for productivity and sustainability. In this context, the work on microbial communities associated with sugarcane is remarkable, because both, these communities play important functional role in the interaction with the plant, and studies performed in tropical conditions are limited as well. Fungal and bacterial communities associated with genetically modified sugarcane IMI-1 and its conventional isoline SP80-1842 were systematically isolated from plants cultivated in an experimental area in Piracicaba, SP, Brazil. The fungal communities associated with sugarcane were accessed by means of cultivation approach and PCR-denaturing gradient gel electrophoresis; the results revealed that these communities are not affected by transgeny. The microbial communities\' diversity was characterized and identified by using molecular tools. The fungus Fusarium moniliforme showed high frequency in association with the plant and it was observed that the endopolygalacturonase gene, pgIII, plays important role in order to determine the sort of association, either endophytic or pathogenic, between F. moniliforme and the host. The Burkholderia cepacia complex is an integral part of the endophytic bacterial community of sugarcane in Brazil and isolates of this complex are able to control F. moniliforme growth. Phylogenetic analyses indicated that the endophytic Burkholderia are closely related to clinical isolates of the B. cepacia complex isolated from cystic fibrosis patients.

A cana-de-açúcar é uma das principais culturas do Brasil e nos últimos anos está recebendo especial atenção devido ao crescente aumento da área cultivada e produção de etanol para uso como biocombustível. Considerando-se sua importância econômica e a possibilidade do uso de plantas geneticamente modificadas, essa cultura tem se tornado foco de pesquisas relacionadas à produtividade e sustentabilidade. Neste contexto, o estudo de comunidades microbianas associadas à cana-de-açúcar é de fundamental importância, pois além dessas comunidades desempenharem importante papel funcional na interação com a planta, os estudos realizados nas condições tropicais são limitados. Comunidades de fungos e bactérias associadas à cana-de-açúcar geneticamente modificada IMI-1 e sua isolinha convencional SP80-1842 foram sistematicamente isoladas de plantas cultivadas em área experimental em Piracicaba, SP, Brasil. Por meio de isolamento e PCR-denaturing gradient gel electrophoresis (PCR-DGGE) foi verificado que a transgenia não afeta a diversidade da comunidade fúngica associada à cana-deaçúcar. A diversidade dessas comunidades foi descrita e caracterizada molecularmente. O fungo Fusarium moniliforme apresentou alta freqüência de isolamento e também foi observado que o gene da endopoligalacturonase, pgIII, desempenha um importante papel no tipo de associação, endofítica ou patogênica, do F. moniliforme e a planta. O complexo Burkholderia cepacia constitui uma importante fração da comunidade de bactérias associada à cana-de-açúcar no Brasil e isolados deste complexo são capazes de inibir o crescimento do patógeno F. moniliforme. Análises filogenéticas indicaram que os isolados de bactérias endofíticas de Burkholderia são proximamente relacionados com linhagens-tipo do complexo B. cepacia isoladas de pacientes de fibrose cística. / In Brazil, the sugarcane is one of the most important cultivated crops. The sugarcane has received increased interest in the last years because of increase of the cultivated area and ethanol production to be used as biofuel. Considering its economical importance and the possibility of the use of the genetically modified plants; this crop has become the aim of current research for productivity and sustainability. In this context, the work on microbial communities associated with sugarcane is remarkable, because both, these communities play important functional role in the interaction with the plant, and studies performed in tropical conditions are limited as well. Fungal and bacterial communities associated with genetically modified sugarcane IMI-1 and its conventional isoline SP80-1842 were systematically isolated from plants cultivated in an experimental area in Piracicaba, SP, Brazil. The fungal communities associated with sugarcane were accessed by means of cultivation approach and PCR-denaturing gradient gel electrophoresis; the results revealed that these communities are not affected by transgeny. The microbial communities\' diversity was characterized and identified by using molecular tools. The fungus Fusarium moniliforme showed high frequency in association with the plant and it was observed that the endopolygalacturonase gene, pgIII, plays important role in order to determine the sort of association, either endophytic or pathogenic, between F. moniliforme and the host. The Burkholderia cepacia complex is an integral part of the endophytic bacterial community of sugarcane in Brazil and isolates of this complex are able to control F. moniliforme growth. Phylogenetic analyses indicated that the endophytic Burkholderia are closely related to clinical isolates of the B. cepacia complex isolated from cystic fibrosis patients.