Resíduos produzidos em atividades industriais podem trazer danos significativos ao ambiente, principalmente quando estes não são degradáveis (ou são dificilmente degradáveis) por processos naturais. Polímeros sintéticos estão entre os principais resíduos descartados na natureza, permanecendo inertes por séculos e interferindo em processos ecológicos. Diante deste contexto, explorar o potencial biodegradável de plásticos e uma alternativa na substituição de embalagens descartáveis. Para a construção e otimização de materiais biodegradáveis, faz-se necessária a compreensão da ação da microbiota natural em processos de biodegradação natural. Neste trabalho, buscou-se compreender a dinâmica de microrganismos do filo Bacteroidetes no processo de biodegradação de blenda polimérica Poli (ɛ-caprolactona) / Amido/ Proteína Isolada de Soja (PCL/A/PIS), considerando solos e materiais plásticos distintos, como também a presença de genes notadamente associados a degradação biológica. Para tanto, partindo de amostras de DNA obtidas do solo ao longo do processo de biodegradação, foram utilizadas técnicas de PCR e PCR-DGGE (Reação de Polimerização em Cadeia - Eletroforese em Gel com Gradiente Desnaturante). Observou-se que: Bacteroidetes estão presentes em todos os momentos avaliados do processo de biodegradação da Blenda e do PCL; o solo Arenoso, onde a biodegradação e mais efetiva, apresenta comunidades com perfis bem delimitados, que se diferenciam na degradação da Blenda e do PCL; a dinâmica no solo Argiloso não exibe um padrão claro de influencia dos plásticos sobre a comunidade, apontando, porem para uma convergência de similaridade das comunidades sobre blenda e PCL no momento de maior atividade degradativa. Chitinophagaceae e uma família presente na biodegradação, sendo Chitinophaga pinensis uma das espécies presentes; genes exclusivos a esta espécie e outras da mesma família que codificam lipases e proteases, associados a degradação de polímeros pela literatura cientifica, estão presentes nas amostras de solo avaliadas, sendo possíveis atores na biodegradação dos plásticos. Bacteroidetes aparenta ser um filo importante em processos naturais de biodegradação de polímeros em solos, e explorar o potencial deste grupo e da família Chitinophagaceae pode oferecer subsídios para a construção de plásticos mais eficientes em sua biodegradação. / The human productive activity has brought significant damage to the environment, particularly by the disposal of non-degradable waste (or poorly degradable) by natural processes. Synthetic polymers are among the main materials discarded, remaining inert for centuries, causing damage to the natural wildlife or interfering with ecological processes. Given this context, the potential use of biodegradable plastics is an alternative, both for agriculture and for the replacement of disposable packaging. For the construction and optimization of biodegradable materials it is necessary to understand the action of natural microbial communities in the process. Microorganisms have predominant and diversified roles in the biodegradation process, with complex ecological relationships in their responses, either by their phylogenetic relationships or the enzymes used during the process. In this work, we sought to understand the dynamics of Bacteroidetes phylum in the biodegradation process of polymer blend poly (ɛ-caprolactone) / starch / Soy Protein Isolate (PCL/ A/ SPI), considering different soils and plastics, and also the presence of genes associated with biological degradation. Therefore, starting from DNA samples taken from the soil during the biodegradation process, PCR techniques and PCR-DGGE (Polymerization Chain Reaction - Denaturing Gradient Gel Electrophoresis) were used. It was observed that: the Phylum Bacteroidetes is present in all periods evaluated of the biodegradation process of Blend, PCL and even soils without the addition of plastics; the sandy soil, where the biodegradation is more effective, presents communities with well defined profiles that differ in the degradation of blends and PCL; the dynamics in clay soil does not show a clear pattern of plastics influence over the community, pointing to a convergence of communities similarity in blends and PCL over greater degradation activity. Chitinophagaceae is a family present in biodegradation, Chitinophaga pinensis being one of the species present; genes unique to this species and others of the same family encoding lipases and proteases, associated with the degradation of polymers reported in the scientific literature, are present in the evaluated soil samples, potentially acting on plastics biodegradation. Bacteroidetes appears to be an important phylum in natural processes of polymer biodegradation in soils, and to explore the potential of this taxon and of Chitinophagaceae family can shed light on the construction of more efficient biodegradable plastics.

O estudo buscou avaliar a diversidade de genes funcionais microbianos em solos da Amazônia, submetidos a diferentes formas de uso, empregando o Geochip. Este é um microarranjo com sondas para genes relacionados a diversos processos funcionais, incluindo ciclos biogeoquímicos. Foram avaliados solos de floresta primária, pastagens e floresta secundária. A conversão de floresta para pastagem causou alteração da estrutura funcional da comunidade e redução da diversidade de genes, e o crescimento da floresta secundária restabeleceu a estrutura e diversidade original. Fatores físico-químicos do solo correlacionaram significativamente com a estrutura da comunidade, indicando importância dos mesmos para o perfil observado. A análise de associação de genes aos ambientes revelou diferentes respostas, e um maior número de genes associados às duas florestas. Os resultados mostraram que o uso do solo para pastagem causa impactos sobre a diversidade e a abundância de genes de importância ambiental e um restabelecimento do potencial funcional na floresta secundária. / The functional gene diversity of Amazon soils under different land use systems was accessed by Geochip, which is a microarray targeting genes related to a variety of processes, including biogeochemical cycles. Soils from a primary forest, three pastures and a secondary forest were analyzed. The forest to pasture conversion led to functional structure changes and functional gene diversity loss, whereas the secondary forest growth promoted the recovery of a profile found in primary forest. Correlation of soil physical-chemical properties with the community structure was found, suggesting that soil characteristics may be driving the biological profile observed. The association index showed a range of responses and a greater number of genes associated to both forests. The results revealed an impact of pasture establishment on the diversity and abundance of environmentally important genes, and a recovery of the functional potential with secondary forest growth.

A pesquisa sobre bactérias diazotróficas associadas à cultura do trigo tem demonstrado a necessidade de associar bactérias eficientes a genótipos responsivos ao nitrogênio, os quais mais se beneficiariam dessa associação. Em um experimento com parcelas subdivididas instalado em condições de campo, em Mococa (SP), empregando os tratamentos: 3 doses de N (0, 60 e 120 kg ha -1 ) x 3 genótipos de trigo (IAC-24, ITD-19 e IAC-355), foi avaliada a ocorrência de microrganismos diazotróficos endofíticos em raízes desinfestadas superficialmente, utilizando-se 3 meios de cultivo distintos, NFb, semi-específico para Azospirillum spp, JNFb, semi-específico para Herbaspirillum spp., e LGI-P, semi-específico Gluconacetobacter diazotrophicus. O genótipo IAC-355 apresentou a menor quantidade de bactérias diazotróficas endofíticas. Além disso, para este genótipo, houve um ajuste linear ascendente da quantidade de bactérias diazotróficas com o aumento na quantidade de N adicionada, o que comprova a influência do genótipo da planta na associação com essas bactérias. Nenhuma bactéria pertencente aos gêneros Azospirillum ou Herbaspirillum foi isolada do genótipo IAC-355. Nas condições estudadas, não foi identificado nenhum isolado de Gluconacetobacter diazotrophicus nas raízes do trigo. Foram obtidos 8 isolados do gênero Azospirillum e 12 do gênero Herbaspirillum. Esses isolados foram testados "in vitro", no genótipo do qual foram originalmente isolados. Todos os isolados testados no genótipo ITD-19 causaram maior crescimento radicular que a testemunha e apenas 1 isolado de Herbaspirillum sp. propiciou aumento significativo do teor de N na parte aérea. A colonização micorrízica no genótipo IAC-355 foi maior que nos demais genótipos independente da dose de N, comprovando a influência do genótipo na colonização. A colonização micorrízica se correlacionou com a massa da matéria seca, e com o teor e a quantidade acumulada de P e N na parte aérea, assim como com a produtividade. A atividade da biomassa microbiana foi alterada na ausência de N, obtendo-se correlação entre o qCO2 e a relação Cmic:Corg, indicando que na ausência de N, houve perdas de C no solo cultivado com o genótipo IAC-24, enquanto que na presença do IAC-355, houve maior eficiência na utilização do C do solo pelos microrganismos. Na contagem de bactérias nitrificadoras obteve-se um ajuste linear ascendente em relação à quantidade de N adicionada, provavelmente devido à maior disponibilidade de substrato. A quantidade de microrganismos nitrificadores se correlacionou com a massa da matéria seca, e com o teor e a quantidade acumulada de N na parte aérea, assim como com a produtividade. A interação FMA-bactéria diazotrófica não propiciou benefícios para cultura do trigo. A interação FMA-bactéria diazotrófica demonstrou ser bastante especifica. As plantas associadas a Glomus, quando em presença dos isolados bacterianos apresentaram maior crescimento, acúmulo e aproveitamento dos nutrientes. Confirmou-se que o fungo micorrízico realmente é um agente transmissor de bactérias diazotróficas endofíticas, sendo que Acaullospora causou maior colonização radicular. As plantas dos tratamentos em que somente a bactéria foi inoculada apresentaram o dobro da produção de matéria seca, da quantidade acumulada e do índice de eficiência de utilização de N e P na parte aérea em relação à testemunha. A especificidade da interação planta-bactéria diazotrófica associativa indica que é possível obter benefícios desta associação, explorando bactérias e cultivares locais. / The research on diazotrophic bacteria associated to wheat has demonstrated the need to associate efficient bacteria to N-responsive genotypes, which would be more benefited from this association. A field experiment was carried out in Mococa, state of São Paulo-Brazil, with 3 genotypes of wheat (IAC-24, ITD-19 and IAC-355) under 3 nitrogen doses (0, 60 e 120 kg ha -1 ). The occurrence of diazotrophic bacteria was evaluated in three media, namely, NFb semi-specific for Azospirillum spp., JNFb semi-specific for Herbaspirillum spp., and LGI-P semi-specific for Gluconacetobacter diazotrophicus, using surface-sterilized roots. Regardless the nitrogen dose, the population of diazotrophic bacteria established poorly in the genotype IAC-355, but the infection increased with the addition of nitrogen for the same genotype, proving the influence of the host genotype for its association with these bacteria. Azospirillum spp. or Herbaspirillum spp. could not be isolated from the surface-sterilized roots of IAC-355. In the field experiment G. diazotrophicus was not found in any of the wheat-genotype roots. It was obtained 12 Herbaspirillum spp. isolates and 8 Azospirillum spp isolates. These strains were tested under gnotobiotic conditions, using the genotype from which they had been originally isolated. Inoculated ITD-19 plants showed an increase in root length, even though, only one strain showed a significant increase on shoot N accumulation. In the genotype IAC-355 mycorrhizal colonization was higher, proving the influence of the plant genotype. Mycorrhizal colonization showed significant correlation to shoot dry matter, shoot N and shoot P concentration and accumulation, as well as to the grain yield. In the absence of added N, the activity of microbial biomass was affected. The correlation between the qCO2 and biomass C-to-N ratio, showed that in absence of N, soil-C loss under IAC-24 cultivation, but a greater efficiency in the use of the soil-C by the microorganisms under IAC-355 cultivation. The populations of nitrifying bacteria increased with N addition, probably due to the N-rich substrate availability. The nitrifying bacteria showed significant correlation to shoot dry matter, shoot N concentration and accumulation, as well as to grain yield. There was no benefit from the AMF-diazotrophic bacteria co-inoculation on wheat plants. The specificity of AMF-diazotrophic bacteria interaction was demonstrated, it was confirmed that AMF indeed is a transmitting agent of endophytic diazotrophic bacteria, Acaullospora caused higher endophytic-bacteria root colonization. Plants inoculated with single strain doubled shoot dry matter, shoot N and P concentration and accumulation as compared to the control. Plant-bacteria interaction specificity demonstrates the possibility of getting benefits from this association by exploring both bacterial strains and plant genotypes from the same location.

Uma alternativa de manejo das doenças de citros causadas por Phytophthora spp. é o uso de matéria orgânica. Com o objetivo de avaliar os efeitos da incorporação de lodo de esgoto ao solo na indução de supressividade a Phytophthora nicotianae Breda de Haan (1896) em plântulas de limão cravo (Citrus lemonia (L.) Osbeck), foram realizados diversos experimentos em laboratório, casa de vegetação e campo. O aumento nas doses de lodo de esgoto, nos experimentos em laboratório, em casa de vegetação e em campo, resultou na redução do pH e aumento da condutividade elétrica do solo; aumento da atividade microbiana do solo (avaliada pela hidrólise de diacetato de fluoresceina – FDA e pela respiração microbiana); além de uma redução na recuperação de P. nicotianae, tanto do substrato e do solo como das raízes de plântulas e mudas. Em alguns experimentos, a recuperação do patógeno correlacionou-se significativa e negativamente com a atividade microbiana do solo(FDA) e com a condutividade elétrica. Um melhor desenvolvimento de plântulas e mudas foi observado com a incorporação de lodo até 20%. Esses resultados indicam um efeito supressivo do lodo de esgoto a P. nicotianae, nas condições avaliadas, explicado por fatores químicos e biológicos. Dentre os fatores químicos destacam-se o aumento da condutividade elétrica e a inibição do crescimento das colônias do patógeno em meio de cultura com extratos ácidos de lodo. Os fatores biológicos envolveram o aumento da atividade microbiana do solo e a presença de fungos (Aspergillus sp. e Trichoderma sp.) e actinomicetos antagonistas a P. nicotianae. / Soil organic matter amendments may provide an alternative for the management of citrus soil diseases caused by Phytophthora spp. The effects of incorporating residential sewage sludge into the soil in order to induce suppressiviness to Phytophthora nicotianae Breda de Haan (1896) in seedlings and plantlets of rangpuor lime (Citrus limonia (L.) Osbeck) were evaluated. For this, several experiments under laboratory, greenhouse and field conditions were conducted. In almost all experiments, while sewage sludge doses enlarged, soil pH values decreased and soil electrical conductivity increased; soil microbial activity (evaluated by the fluorescein diacetate hydrolysis – FDA and microbial respiration) increased, and P. nicotianae recovery from soil and roots tended to decreased. In some experiments, significantly and negative correlations were observed between P. nicotianae recuperation and microbial activity (FDA), and between P. nicotianae recuperation and soil electrical conductivity. Seedlings and plantlets development improved at a maximum of 20% v/v sewage sludge soil incorporation. These results suggest a sewage sludge suppressive effect on P. nicotianae, explained by chemical and biological factors, under the experimental conditions of the tests performed. Among chemical factors, the increase of the soil electrical conductivity and colony growth inhibition of P. nicotianae on culture media amended with an acid sewage sludge extract, were indicated. The biological factors involved in suppressiviness were the increase of soil microbial activity and the presence of fungi (Aspergillus sp. and Trichoderma sp.) and actinomyces antagonistic to P. nicotianae.

<div>In fire-prone forests, tree taxa and burn temperature are the major controllers of the chemical and physical properties of pyrogenic organic matter (PyOM), the aromatic carbon-rich product of the incomplete combustion of plant biomass, that accumulates in soil in such settings. These controls also dictate how soil microbes can degrade plant C once it enters into soil as previous studies demonstrate that increased fire temperature results in low PyOM degradability but also impacts the decomposition of the original soil. However, we know little about how taxa and temperature of C-inputs impact the production and accumulation of cellular residues from soil microbes, which can be the dominant source of stabilized soil organic matter in many ecosystems. </div><div>This work presents the results of the analysis of soil microbial amino sugars, as proxies for soil microbial necromass, from long-term soil incubation experiments, 180 and 600 days, that were amended with jack pine (JP) and red maple (RM) wood or their PyOM produced at 300°C or 450°C. Both wood taxa amendments resulted in an increase in microbial sugar residues compared to non-amended soils but RM, the taxa with the highest proportion of soluble sugars and low tannin content, exhibited the highest percentage increase. Soils amended with PyOM exhibited lower amino sugar content as compared to their wood but no difference compared to controls (non-amended soils). There was no difference in soil amino sugars observed between the PyOM derived from the two taxa nor between the temperature of pyrolysis, possibly due to only small amounts of bioavailable C and N in the PyOM. Total amino sugar concentrations varied significantly between PyOM and fresh wood treatments, with PyOM treatments yielding 659 – 730 μg/ g soil while wood treatments yielded 757 – 930 μg/ g soil early in incubations. While fungal-derived amino sugars were dominated in all treatments, longer soil incubation time, 600 days vs 180 days, resulted in a proportionately greater decrease bacterial-derived amino sugars. Overall, at 180 days, PyOM treatments exhibited 19-27% of soil N and 4-5% of soil C quantifiable as amino sugars while wood treatments exhibited 27-28% of total soil N and 6-7% of total soil C as amino sugars. This work shows, for the first time, that on a per C or per N basis, PyOM versus fresh wood addition to soils will result in a net depletion of microbial residues. The variable response in amino sugars between treatments and incubation time highlights the importance and dynamic nature of the physicochemical characteristics of organic matter input to soil in controlling the contribution of soil microbial residues to that soil. </div><div><br></div>

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No description available.

Master of Science

Thesis (M. Tech. Agriculture) -- Central University of Technology, Free state, 2012 / A series of incubation studies and greenhouse experiments were conducted to evaluate the use of EM seed treatments, at different application levels, handling techniques and soil conditions on germination and seedling vigour of selected cultivars of maize, sorghum and sunflower. Two incubation studies were conducted to evaluate the germination and seedling vigour of maize, sorghum and sunflower seeds treated with M-EM from three different suppliers, multiplied at two different ratios (1% and 3%) and diluted at three different levels (0.01%, 0.1% and 1.0%) compared to a control treated with pure water. Results revealed no significant differences under optimum germination conditions, while seedlings under cold stress indicated that M-EM treatments positively affected germination and seedling vigour compared to the control treatments. Two incubation studies were also conducted to evaluate the germination and seedling vigour of maize, sorghum and sunflower seeds treated with M-EM from three different suppliers, multiplied at two different ratios (1% and 3%) and exposed to the influences of irradiation and temperature fluctuation. From the results became clear that the correct storage and handling is essential in optimizing the effect of M-EM on seeds. Even though M-EM was exposed to irradiation and temperature fluctuation, M-EM still had positive effects on germination and seedling vigour. Pot experiments were conducted to determine the effect of EM as seed treatment, at different dilutions, on germination, seedling vigour and dry mass of maize, sorghum and sunflower at different planted depths. Germination were not affected by the M-EM treatment, while shoot length results indicated that seed treated with M-EM could have significant effect on seedling survival. A greater effect was visible on the shoot length of shallow planted seeds, than on deeper planted seeds. From the results no single company, ratio or dilution could be prescribed as paramount. To further investigate the effect of M-EM subjected to the influences of irradiation and temperature fluctuation; maize, sorghum and sunflower seeds were treated with M-EM from three different suppliers, multiplied at two different ratios (1% and 3%) and exposed to the influences of irradiation and temperature fluctuation and planted in soil. M-EM treatments only benefited the germination of deeper planted sorghum seeds compared to the control treatments. The shoot lengths of deeper planted maize and sunflower seed were positively increased by the M-EM treatments while also resulting in significant results for the overall shoot length of sorghum. The third pot study was conducted to determine the influence of EM as a seed treatment on maize, sorghum and sunflower planted in three different soils, namely: sterilized soil, soil treated with M-EM and Fusarium containing soil. Germination and seedling vigour results of the sterilized and M-EM treated soil revealed to be superior to that of the Fusarium containing soil. From the results was concluded that M-EM treatments will probably improve early seedling growth of maize, sorghum and sunflower compared to untreated seed and that M-EM seed treatment and a pre-plant EM soil treatment might assist seeds in unfavourable germination and growth conditions.

Thesis (MSc)--University of Stellenbosch, 2004. / ENGLISH ABSTRACT: Numerous workers studied the impact of pollutants and agricultural chemicals, containing heavy metals such as copper (Cu), on soil microbes. It was found that elevated soil Cu levels do have a detrimental effect on soil bacterial populations however the filamentous fungi seemed to be less affected. Most of these studies were conducted in soils containing already relatively high Cu levels and the effect of this heavy metal on the nonfilamentous fungi (i.e. yeasts) was never investigated. The aim of this study was therefore to determine the impact of elevated Cu levels on filamentous fungi and yeasts occurring in soils containing relatively low natural Cu levels. A synthetic selective medium containing glucose as carbon source, thymine as nitrogen source, vitamins, minerals and chloramphenicol as anti-bacterial agent (TMV-agar), was used to enumerate ascomycetous and basidiomycetous Cu resistant yeasts in a sample of virgin soil containing ~ 2ppm Cu. Media that were used to enumerate Cu resistant filamentous fungi were malt extract agar, malt extract agar with streptomycin sulfate, maltyeast- extract-peptone agar with chloramphenicol and streptomycin sulfate, benomyl–dichloran-streptomycin medium for the enumeration of hymenomycetous fungi and two selective media for the isolation of mucoralean fungi. Cu resistant fungi able to grow on all of the above mentioned solid media supplemented with 32 ppm Cu occurred in the soil sample. To obtain an indication of the level of Cu tolerance of fungi present in this soil sample, a number of fungal isolates were screened for the ability to grow on a series of agar plates, prepared from glucose-glutamate-yeast extract agar, containing increasing concentrations of Cu. It was found that filamentous fungi and yeasts that were able to grow on this agar medium containing up to 100 ppm Cu were present in the soil. A series of soil microcosms was subsequently prepared from the soil sample by experimentally contaminating the soil with increasing amounts of copper oxychloride, were after fungal populations in the microcosms, including Cu resistant fungi, were monitored using plate counts. At the end of the incubation period, after 245 days, fungal biomass in the microcosms was compared by determining the concentrations of the fungal sterol, ergosterol, inthe soil. Generally, Cu had little impact on the numbers of filamentous fungal colony forming units on the plates, as well as on the ergosterol content of the soil. The numbers of filamentous fungi in the soil, including the Mucorales and hymenomycetes, seemed to be less affected by the addition of copper oxychloride than the numbers of soil yeasts able to grow on TMV-agar. The focus of the next chapter was on the response of yeasts in different soils to elevated levels of Cu in the soil. TMV-agar was used to enumerate yeasts in soil microcosms prepared from four different soil samples, which were experimentally treated with copper oxychloride resulting in Cu concentrations of up to 1000 ppm. The selective medium supplemented with 32 ppm Cu was used to enumerate Cu resistant yeasts in the microcosms. The results showed that the addition of Cu at concentrations ≥ ~1000 ppm did not have a significant effect on total yeast numbers in the soil. Furthermore, it was found that Cu resistant yeasts were present in all the soil samples regardless of the amount of Cu that the soil was challenged with. At the end of the incubation period, yeasts in the microcosms with zero and ~1000 ppm additional Cu were enumerated, isolated and identified using sequence analyses of the D1/D2 600-650bp region of the large subunit of ribosomal DNA. Hymenomycetous species dominated in the control soil, while higher numbers of the urediniomycetous species were found in the soil that received Cu. These observations suggest that urediniomycetous yeasts may play an important role in re-establishing overall microbial activity in soils following perturbations such as the addition of Cu-based fungicides. / AFRIKAANSE OPSOMMING: Vele navorsers het al die impak van besoedelingstowwe en landbouchemikalieë wat swaarmetale soos koper (Cu) bevat, op grond-mikrobes bestudeer. Dit is gevind dat verhoogde Cu vlakke ‘n nadelige effek het op grond-bakteriese populasies, maar dat die filamentagtige fungi geneig is om minder geaffekteer te word. Meeste van hierdie studies is gedoen met gronde wat alreeds relatief hoë Cu vlakke bevat het en die effek van hierdie swaarmetaal op die nie-filamentagtige fungi (d.i. giste) is nooit ondersoek nie. Die doel van hierdie studie was dus om die impak van verhoogde Cu vlakke op filamentagtige fungi en giste in gronde, wat natuurlike lae vlakke van Cu bevat, te bepaal. ‘n Sintetiese selektiewe medium wat glukose as koolstofbron, timien as stikstofbron, vitamiene, minerale asook chloramfenikol as anti-bakteriese agent bevat (TMV-agar), is gebruik om askomisete en basidiomisete Cu weerstandbiedende giste in ‘n monster ongeskonde grond, bevattende ~ 2dpm Cu, te tel. Media wat gebruik is om Cu weerstandbiedende filamentagtige fungi te tel, was mout-ekstrak agar, moutekstrak agar met streptomisiensulfaat, benomiel-dichloran-streptomisien medium vir die tel van hiemenomiseetagtige fungi en twee media vir die isolasie van mukoraliese fungi. Cu-weerstandbiedende fungi wat op al die bogenoemde media, aangevul met 32 dpm Cu, kon groei, het in die grondmonster voorgekom. Om die mate van Cu-weerstandbiedendheid van fungi wat in die grondmonster voorkom, te bepaal, is ‘n getal fungus-isolate op agarplate, voorberei met glukose-glutamaat-gis ekstrak agar, bevattende verhoogde konsentrasies Cu, nagegaan. Daar is gevind dat daar filamentagtige fungi en giste in die grond voorkom wat die vermoë het om op media bevattende 100 dpm Cu te groei. ‘n Reeks grond mikrokosmosse is dus voorberei vanaf die grondmonster deur om dit eksperimenteel te kontamineer met verhoogde hoeveelhede koper oksichloried, waarna die fungus-populasies asook die Cu-weerstandbiedende fungi in die mikrokosmos gemoniteer is deur middel van plaattellings. Aan die einde van die inkubasie periode, 245 dae, is die fungus biomassa in al die mikrokosmosse bereken deur die konsentrasie van die fungus sterool ergosterool te bepaal en dit met mekaar te vergelyk. Oor die algemeen het Cu min impak ten opsigte van diegetal filamentagtige fungi kolonie vormende eenhede die plate, asook op die ergosterool inhoud van die grond gehad. Dit wil voorkom of die getal filamentagtige fungi in die grond, insluitende die Mucorales en die hymenomisete, minder geaffekteer is deur die toediening van koperoksichloried as die aantal grondgiste wat op die TMV-agar kan groei. Die fokus van die volgende hoofstuk was dus op die reaksie wat giste in verskillende grondtipes gehad het op verhoogde Cu in die grond. TMV-agar is gebruik om die getal giste te bepaal in die grond mikrokosmosse van die vier verskillende grondmonsters, wat voorberei is deur om dit eksperimenteel met koper oksikloried te kontamineer tot en met Cu konsentrasies van 1000 dpm. Die selektiewe medium wat gesupplementeer is met 32 dpm Cu, is gebruik om Cu weerstandbiedende giste in die mikrokosmosse te bepaal. Die resultate toon dat die toevoeging van Cu by konsentrasies ≥ ~1000 dpm nie enige beduidende effek op die totale gis getalle gehad het nie. Daar is ook gevind dat daar Cu weerstandbiedende giste in die grond monsters voorkom gekom het ten spyte van die hoeveelheid Cu wat tot die grond toegevoeg is. Aan die einde van die inkubasie periode is die giste wat die die mikrokosmosse bevattende nul en ~1000 dpm Cu getel, geïsoleer en geïdentifiseer deur gebruik te maak van DNA volgorde bepaling van die D1/D2 600-650 bp areas geleë in die groter subeenheid van die ribosonale DNA. Hymenomisete spesies het in die grond kontrole gedomineer, terwyl hoër getalle uredinomisete spesies in die grond met addisionele Cu gevind is. Die resultate dui daarop dat uredinomisete giste dalk ‘n belangrike rol kan speel in die hervestiging van die oorwegende mikrobiese aktiwiteit in grond na skoktoestande soos die aanwending van Cu-gebaseerde fungisiede.