There is accumulating evidence that arbuscular mycorrhizal fungi (AMF) produce a glycoprotein called glomalin, which has the potential to increase soil carbon (C) and nitrogen (N) storage, thereby reducing soil emissions of carbon dioxide (CO2) and nitrous oxide (N2O) into the atmosphere. However, other soil microorganisms such as plant growth-promoting rhizobacteria (PGPR) that interact with AMF could indirectly influence glomalin production. The objectives of this study were to determine the effects of AMF and PGPR interactions on glomalin production and identify possible combinations of these organisms that could enhance C and N storage in the rhizosphere. The effects of AMF and PGPR interactions on pea (Pisum sativum L.) growth and correlations between glomalin production and plant growth also were assessed.<p> A series of growth chamber and laboratory experiments were conducted to examine the effect of fungal and host plant species on glomalin production by comparing the amounts of glomalin produced by Glomus clarum, G. intraradices, and G. mosseae in association with corn (Zea mays L.), in addition to examining differences in the ability of corn, pea, and wheat (Triticum aestivum L.) to support glomalin production by G. intraradices. There were no significant differences in glomalin production [measured in the rhizosphere as Bradford-reactive soil protein (BRSP)] by the three AMF species, whereas host plant significantly affected glomalin production. Specifically, higher BRSP concentrations were found in the rhizosphere of corn as compared to pea and wheat.<p> Additionally, the effect of long-term storage on the growth promoting traits of the PGPR strains selected; namely, Pseudomonas cepacia R55 and R85, P. aeruginosa R75, P. putida R105, and P. fluorescence R111 were investigated. These bacterial strains previously had been identified as PGPR, but had since undergone approximately twenty years of storage at -80¢ªC; thus, it was necessary to confirm that these strains had retained their plant growth promoting characteristics. Apparently, long-term storage had no significant adverse effect on the PGPR strains as all strains increased the total biomass of wheat significantly and demonstrated antagonism against fungal pathogens.<p> The possibility that spore-associated bacteria (SAB) could influence AMF associations, thereby affecting glomalin production, and subsequent crop yield potential was assessed. This was achieved by first isolating bacteria from disinfested spores of the AMF species and determining their potential as PGPR for wheat. According to fatty acid methyl ester (FAME) profiles, four genera of bacteria were isolated from AMF spores namely; Arthrobacter, Bacillus, Micrococcus, and Paenibacillus, of which Bacillus species were the most common SAB. None of these isolates, however, showed growth promoting abilities on wheat.<p> Based on the preliminary findings, the combined effects of the three AMF species and the five PGPR strains were examined on plant growth and glomalin production under gnotobiotic conditions using pea as the host plant. Interactions between G. intraradices and R75, R85, or R105 resulted in increased BRSP concentration in the mycorrhizosphere of pea. Additionally, significant interactions were observed between the AMF species and PGPR strains on BRSP concentration in pea rhizosphere under non-sterile conditions. As observed under sterile conditions, the co-inoculation of pea with G. intraradices and R75 or R85 increased BRSP concentrations in the rhizosphere of pea grown in non-sterile soil, although interaction effects were not significantly different from the control or when G. intraradices was applied alone. Significant AMF and PGPR interactions were observed to affect AMF colonization; however, the combination of these organisms did not significantly affect pea growth, nutrient uptake, and C and N storage in the plant rhizosphere. No correlations were detected between glomalin-related soil protein (GRSP), pea growth, nutrient concentrations in the plant tissue, and soil organic C and N content. This study demonstrated that although the potential exists to manipulate certain AMF and PGPR to enhance glomalin production, co-inoculation of AMF and PGPR did not enhance plant growth or C and N storage beyond that achieved by inoculation of either organism.

There are no seeding rates established for organic production of field pea and lentil in Saskatchewan and organic producers must rely upon rates recommended for conventional production of these crops. These seeding rates may not be suitable for organic production as the two systems differ in the use of inputs and in pest management. The objectives of this study were to determine an optimal seeding rate for organic production of field pea and lentil in Saskatchewan considering a number of factors, including yield, weed suppression, soil nitrogen (N) and phosphorus (P) concentrations, soil water storage, colonization of crop roots by arbuscular mycorrhizal fungi (AMF), plant P uptake, and profitability. A field experiment was conducted to determine the optimal seeding rates of field pea and lentil. Field pea seeding rates were 10, 25, 62, 156 and 250 plants m-2 and lentil seeding rates were 15, 38, 94, 235 and 375 plants m-2. Sites were established at Vonda, Vanscoy and Delisle, SK using a randomized complete block designs with summerfallow and green manure treatments included for each crop. Seed yield increased with increasing seeding rate for both crops, up to 1725 kg ha-1 for field pea and 1290 kg ha-1 for lentil. Weed biomass at physiological maturity decreased with increasing seeding rate for both crops. In field pea, weeds were reduced in weight by 68%, while lentil reduced weed biomass by 59% between the lowest and highest seeding rates. <p>Post-harvest soil phosphate-P levels did not change consistently between treatments, indicating that there was no trend in soil P concentration with seeding rate. Post-harvest soil inorganic N, however, was higher for the summerfallow and green manure treatments than for the seeding rate treatments in both crops. Inorganic N was higher at some sites for the highest two seeding rates in field pea. Soil water storage following harvest was not affected by treatment.<p>Colonization of crop roots by AMF increased for lentil with increasing seeding rate, but the same trend was not observed in field pea. A growth chamber experiment to study the rate of colonization of field pea between 10 and 50 d after emergence did not show any differences in AMF colonization between seeding rates. Colonization levels were high (70 to 85%) for both crops in both the field and growth chamber. Arbuscular mycorrhizal fungi colonization and seeding rate had no effect on plant P concentration for either field pea or lentil. Both crops became increasingly profitable as seeding rate increased. Field pea reached a maximum return at 200 plants m-2 and lentil return increased to the highest seeding rate of 375 plants m-2. Organic farmers should increase seeding rates of these crops to increase returns and provide better weed suppression.

Arbuscular mycorrhizas (AM) are the plant root-fungus interactions that are most widespread mycorrhiza in nature. As classically defined, there are two major AM morphologies named after the plant genera in which they were first described: Arum- (intercellular hyphae with arbuscules mainly in inner root cortex), Paris- (extensive hyphal coils in outer root cortex), as well as intermediate morphotypes. In this study, dandelions and chives harvested in Saskatoon (SK, Canada) were examined for AM colonization and morphological types. A Multiple Quantitation Method (MQM) was used for assessing fungal colonization intensity using magnified epifluorescence images of lactofuchsin stained roots, plus details analyzed by high-resolution confocal fluorescence imaging. The results showed that host plants harbored diverse endorhizal fungi, including arbuscular mycorrhizal fungi (AMF), septate endophytes (SE) and fine endophytes (FE), with varying abundances. The soil properties were assessed with respect to P status, organic matter and pH, but there was no correlation with the fungal abundance in this study. Both dandelion and chive roots had Arum- and Paris-type AM. In order to assess the applicability of a current model, I studied quantitative relationship between the cell packing pattern and AM morphotype. Cross sections of host roots were analyzed with Image J software to calculate the proportion of air spaces. The abundance of arbuscules (Arum-type) and hyphal coils (Paris-type) were significantly different in chive and dandelion roots. However, there was no difference in the proportion of air spaces in the inner or outer cortex. Therefore, host root cell packing does not appear to influence AM morphotype at least in the samples in this study. AM fungal diversity was preliminarily investigated by nested PCR with group specific primers, showing multiple PCR bands within root samples, and indicating the potential complexity of AMF groups. Further work to sequence the PCR products is needed to elucidate the AMF groups present.

There is accumulating evidence that arbuscular mycorrhizal fungi (AMF) produce a glycoprotein called glomalin, which has the potential to increase soil carbon (C) and nitrogen (N) storage, thereby reducing soil emissions of carbon dioxide (CO2) and nitrous oxide (N2O) into the atmosphere. However, other soil microorganisms such as plant growth-promoting rhizobacteria (PGPR) that interact with AMF could indirectly influence glomalin production. The objectives of this study were to determine the effects of AMF and PGPR interactions on glomalin production and identify possible combinations of these organisms that could enhance C and N storage in the rhizosphere. The effects of AMF and PGPR interactions on pea (Pisum sativum L.) growth and correlations between glomalin production and plant growth also were assessed.<p> A series of growth chamber and laboratory experiments were conducted to examine the effect of fungal and host plant species on glomalin production by comparing the amounts of glomalin produced by Glomus clarum, G. intraradices, and G. mosseae in association with corn (Zea mays L.), in addition to examining differences in the ability of corn, pea, and wheat (Triticum aestivum L.) to support glomalin production by G. intraradices. There were no significant differences in glomalin production [measured in the rhizosphere as Bradford-reactive soil protein (BRSP)] by the three AMF species, whereas host plant significantly affected glomalin production. Specifically, higher BRSP concentrations were found in the rhizosphere of corn as compared to pea and wheat.<p> Additionally, the effect of long-term storage on the growth promoting traits of the PGPR strains selected; namely, Pseudomonas cepacia R55 and R85, P. aeruginosa R75, P. putida R105, and P. fluorescence R111 were investigated. These bacterial strains previously had been identified as PGPR, but had since undergone approximately twenty years of storage at -80¢ªC; thus, it was necessary to confirm that these strains had retained their plant growth promoting characteristics. Apparently, long-term storage had no significant adverse effect on the PGPR strains as all strains increased the total biomass of wheat significantly and demonstrated antagonism against fungal pathogens.<p> The possibility that spore-associated bacteria (SAB) could influence AMF associations, thereby affecting glomalin production, and subsequent crop yield potential was assessed. This was achieved by first isolating bacteria from disinfested spores of the AMF species and determining their potential as PGPR for wheat. According to fatty acid methyl ester (FAME) profiles, four genera of bacteria were isolated from AMF spores namely; Arthrobacter, Bacillus, Micrococcus, and Paenibacillus, of which Bacillus species were the most common SAB. None of these isolates, however, showed growth promoting abilities on wheat.<p> Based on the preliminary findings, the combined effects of the three AMF species and the five PGPR strains were examined on plant growth and glomalin production under gnotobiotic conditions using pea as the host plant. Interactions between G. intraradices and R75, R85, or R105 resulted in increased BRSP concentration in the mycorrhizosphere of pea. Additionally, significant interactions were observed between the AMF species and PGPR strains on BRSP concentration in pea rhizosphere under non-sterile conditions. As observed under sterile conditions, the co-inoculation of pea with G. intraradices and R75 or R85 increased BRSP concentrations in the rhizosphere of pea grown in non-sterile soil, although interaction effects were not significantly different from the control or when G. intraradices was applied alone. Significant AMF and PGPR interactions were observed to affect AMF colonization; however, the combination of these organisms did not significantly affect pea growth, nutrient uptake, and C and N storage in the plant rhizosphere. No correlations were detected between glomalin-related soil protein (GRSP), pea growth, nutrient concentrations in the plant tissue, and soil organic C and N content. This study demonstrated that although the potential exists to manipulate certain AMF and PGPR to enhance glomalin production, co-inoculation of AMF and PGPR did not enhance plant growth or C and N storage beyond that achieved by inoculation of either organism.

Mémoire numérisé par la Division de la gestion de documents et des archives de l'Université de Montréal

Arbuscular mycorrhizal fungi (AMF) represent an ancient and critical symbiotic partner with the majority of land plants, understood to promote ecosystem productivity and biodiversity and are also important to ecologically sound land management practices. The fungus is thought to be over 400 million years old, and due to a lack of an observable sexual cycle, has been placed into a select group of eukaryotes called 'ancient asexuals', which seemingly defy evolutionary theory by persisting for an extended period of time in the absence of sexual reproduction. Recently however, molecular evidence has accumulated which may suggest AMF harbour a cryptic sexual cycle. In the first chapter of this thesis, entitled "Searching for clues of sexual reproduction in the genomes of arbuscular mycorrhizal fungi", I review evidence supporting this notion of a cryptic sexual cycle in AMF which includes: the presence of recombinational events, meiosis-specific genes as well as mating-specific transcription factors called SexM and SexP of the MATA_HMG protein family which are otherwise found only in the genomes of sexual fungi. In the second chapter, I present the main research of my MSc work where I used bioinformatic, population genetic, molecular and experimental approaches to build on this existing evidence of sexuality in AMF. These findings include the presence of a dramatically expanded family of MAT-HMG genes which are present in several isolates of the Rhizophagus irregularis and also harbour significant allele variation amongst these isolates, some of which resembles variation expected at MAT-genes in other fungi. Q-RT-PCR procedures revealed that at least some of these genes tend to increase in expression during crosses of R. irregularis isolates. We also uncovered the presence of a unique genomic region where at least three of these genes are located in tandem. Finally, several tests of recombination support the presence of intraisolate as well as interisolate recombination events occurring between these MAT-HMG genes.

A seca é um dos principais problemas que afetam a produção do feijoeiro. A despeito da importância de caracteres fenotípicos radiculares, muitos dos esforços de melhoramento genético da cultura tem focado na seleção de cultivares com maior produção de grãos. A simbiose estabelecida entre plantas e FMA aumentam o potencial de captação de água no solo através das extensas redes formadas pelas hifas e alteram vias metabólicas vitais para a manutenção das relações hídricas da planta. O modelo de interação feijoeiro (BAT 477) colonizado por uma mistura de FMA (Glomus clarum, Acaulospora scrobiculata e Gigaspora rosea) foi submetido a um déficit hídrico de 96 h durante o pré-florescimento. O transcritoma global de raízes inoculadas e não-inoculadas, sujeitas ou não à seca, foi comparado por RNA-Seq. Um conjunto de 71 transcritos foram induzidos por FMA durante a seca. Comparando-se os tratamentos estresse e controle, 12.086 unigenes foram regulados em plantas inoculadas e 11.938 em não-inoculadas, refletindo o alto potencial de tolerância da linhagem BAT 477 e indicando que a presença de FMA produz uma regulação fina no perfil de expressão de genes regularmente envolvidos na resposta da planta ao estresse. Foram selecionados 15 fatores de transcrição e seus perfis de expressão foram caracterizados por RT-qPCR tomando-se três períodos, 48, 72 e 96 h de déficit hídrico. Plantas inoculadas ativaram a expressão destes genes mais tardiamente (após 72 h), refletindo melhorias nas condições hídricas da planta que adiam a percepção do estresse. Adicionalmente, a expressão de 23 transcritos foi avaliada em três amostras teciduais diferentes obtidas por microscopia de microdissecção a laser. Glucan 1,3 ?-Glucosidase e PIP2,3, foram detectados somente em células do córtex radicular contendo arbúsculos indicando uma possível indução tecido específica dependente da presença dos fungos. Análises complementares apontaram a regulação de 171 unigenes envolvidos na resposta das FMA ao estresse. Estes resultados validam a hipótese inicial de que a inoculação com FMA altera os perfis de expressão de genes vitais para a resposta da planta ao déficit hídrico / Drought is one of the main problems that affect common bean\'s production. Despite the importance of root fenological characters, breeding efforts for the culture have focused on the selection of cultivars for grain yield. The symbiosis stablished between AMF and plants enhances the potential of water absorption from the soil through an extensive net formed by hyphae and alters vital metabolic pathways involved in the maintenance of the water relations in plants. The interaction model common bean (BAT 477) colonized by a mixture of AMF (Glomus clarum, Acaulospora scrobiculata and Gigaspora rosea) was exposed to a water deficit regime of 96 h during pre-flowering. Global transcriptome from inoculated and non-inoculated roots, exposed or not to drought, were compared through RNA-Seq. A set of 71 transcripts was induced by AMF during drought. Comparing both stress and control treatments, 12,086 unigenes were regulated in inoculated plants, and 11,938 in non-inoculated, reflecting the great tolerance potencial of the lineage BAT 477 and indicating that the presence of AMF produces a fine tune regulation on the expression of genes regularly involved on the drought response of the plant. It was selected 15 transcription factors and their expression profiles were characterized through RT-qPCR taking 3 periods, 48, 72 and 96 h of water deficit. AM plants activated earlier (after 72 h) the expression of these genes, reflecting improvements on the water conditions of the plant that delay the stress perception. Additionally, the expression of 23 transcripts was evaluated on three different tissue samples obtained through laser microdissection microscopy. Glucan 1,3 ?-Glucosidase and PIP2,3, were detected only in cortical cells containing arbuscules, pointing to a possible tissue specific induction dependent of the presence of the fungus. Additional analysis point to the regulation of 171 unigenes involved on the response of the AMF to drought. These results corroborate the initial hypothesis that the inoculation with AMF alters the gene expression profiles of genes that are vital for water deficit response in plants

Maize has great economical importance, while Brazil is the third largest producer, but does not stand out among the countries with the highest level of productivity, due to the large number of small producers who cultivate this cereal with low or no technological level. In this sense, given the importance of maize in the national scene, action research must be performed with maize cultivars used by small producers, especially cultivars of maize landraces, which have not gone through the process of genetic improvement in research institutions. Thus, there is a need to study the Land varieties of corn, about the efficiency in the development and establishment of colonization symbiotic mycorrhizal fungi, as this improves absorption of nutrients, and tolerance to abiotic stresses by the plant in order to minimize costs production mainly related to the use of phosphate fertilizers. The objective of this study was to evaluate the influence of phosphorus and mycorrhizal inoculation on initial Land varieties of corn, preserved by the Association of Seed Guardians Creole, the city of Ibarama, located in the Central Sierra, Rio Grande do Sul. The experiment was performed in a factorial 8x4x2, eight cultivars of maize landraces (Amarelão, Brancão, Cabo Roxo, Cateto Amarelo, Colorido, Cunha, Ferro and Palha Roxa), four phosphate 0 (control), 50 , 100 and 200 mg.kg-1 soil and a species of arbuscular mycorrhizal fungus (Glomus clarum Nicol. & Schenck) and treatment without inoculation, which evaluated the percentage of mycorrhizal colonization, shoot dry matter and plant height . The experiment was conducted in a greenhouse at the Federal University of Santa Maria and the data were subjected to analysis of variance, mean Scott Knott test (at 5% probability of error) and regression analysis. The results showed that significant difference occurs for percentage of mycorrhizal colonization in cultivars Brancão, Cabo Roxo, Cateto Amarelo, Colorido, Ferro and Palha Roxa, due to different levels of phosphorus in the soil, which does not occur in cultivars and Amarelão Cunha and also that the use of Glomus clarum inoculated into maize was effective in increasing dry matter of shoots and plant height in doses of phosphorus that stimulate mycorrhizal colonization. / A cultura do milho possui grande importância econômica mundial, sendo que o Brasil é o terceiro maior produtor, mas não se destaca entre os países com maior nível de produtividade, devido ao grande número de pequenos produtores que cultivam esse cereal com baixo ou nenhum nível tecnológico. Neste sentido, devido à importância do milho no cenário nacional, ações de pesquisa necessitam ser realizadas com cultivares de milho utilizadas por pequenos produtores, em especial as cultivares crioulas de milho, que não passaram por processo de melhoramento genético em instituições de pesquisa. Desta forma, surge a necessidade de estudar as cultivares crioulas de milho, quanto à eficiência no desenvolvimento e estabelecimento de colonização simbiótica com fungos micorrízicos arbusculares, pois esta melhora a absorção de nutrientes, e tolerância a estresses abióticos pela planta, de forma a minimizar custos de produção relacionados, principalmente ao uso de fertilizantes fosfatados. Assim, o objetivo deste trabalho foi avaliar a influência de doses de fósforo e da inoculação micorrízica, no desenvolvimento inicial de cultivares crioulas de milho, preservadas pela Associação dos Guardiões de Sementes Crioulas, do município de Ibarama, localizado na região Centro-Serra do Rio Grande do Sul. O experimento foi executado em esquema fatorial 8x4x2, sendo oito cultivares crioulas de milho (Amarelão, Brancão, Cabo Roxo, Cateto Amarelo, Colorido, Cunha, Ferro e Palha Roxa), quatro doses de fósforo 0 (testemunha), 50, 100 e 200 mg.kg-1 de solo e uma espécie de fungo micorrízico arbuscular (Glomus clarum Nicol. & Schenck) e o tratamento sem inoculação, onde avaliouse a percentagem de colonização micorrízica, matéria seca da parte aérea e altura de planta. O experimento foi conduzido em casa de vegetação da Universidade Federal de Santa Maria e os dados obtidos foram submetidos à análise de variância, teste de média Scott Knott (ao nível de 5% de probabilidade de erro) e análise de regressão. Os resultados obtidos mostraram que ocorre diferença significativa para percentagem de colonização micorrízica nas cultivares Brancão, Cabo Roxo, Cateto Amarelo, Colorido, Ferro e Palha Roxa, em função de doses de fósforo distintas existente no solo, o que não ocorre para as cultivares Amarelão e Cunha e, também, que o uso de Glomus clarum inoculado em milho mostrou-se eficiente no aumento da matéria seca da parte aérea e altura de planta em doses de fósforo que estimulam a colonização micorrízica.

Orientador: Ana Maria Rodrigues Cassiolato / Resumo: O solo degradado em estudo originou-se pela retirada da sua camada superficial, que compreende a vegetação, a matéria orgânica, os nutrientes e os microrganismos, cujos processos são essenciais para o solo ser considerado produtivo. Sua recuperação é importante para manter o equilíbrio da biodiversidade de flora e fauna e protege-lo contra a erosão, entre outros fatores. O objetivo deste trabalho foi o condicionamento do subsolo pela adição de cinza de biomassa de cana-de-açúcar, da inoculação de fungo micorrízico arbuscular (FMA – Glomus clarum) e da inoculação de fungos e de bactéria solubilizadores de fosfato, buscando uma técnica para a recuperação de solos degradados. A bananeira (Musa sp. cv. Mysore) foi utilizada como planta indicadora para as modificações que ocorreram no solo. O delineamento experimental foi em blocos casualizados, em esquema fatorial 2 x 2 x 3 + 2, com 4 repetições por tratamento, totalizando 12 tratamentos, com 4 repetições (bloco). O primeiro fator foi com e sem inoculação de FMA, o segundo foi com e sem adição de cinza proveniente da queima do bagaço de cana-de-açúcar, o terceiro foi com e sem inoculação de fungos e de bactéria solubilizadores de fosfato, e dois tratamentos adicionais, área conservada de Cerrado (SCRC) e área degradada sem intervenção (ADSI). Após 67 e 274 dias do plantio foram avaliados os atributos químicos e microbiológicos do solo na camada de 0,0 a 0,10 m, além da altura, do diâmetro, do índice relativo de clorofila e do P t... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The degraded soil under study originated by the removal of the superficial layer, which includes vegetation, organic matter, nutrients and microorganisms, whose processes are essential for the soil to be considered productive. Its recovery is important to maintain the balance of biodiversity of flora and fauna and protect it against erosion, among other factors. The objective of this work was the subsoil conditioning by the addition of sugarcane biomass ash, the inoculation of arbuscular mycorrhizal fungi (AMF - Glomus clarum) and the inoculation of fungi and bacteria phosphate solubilizers, searching for a technique for recovery of degraded soils. The banana plants (Musa sp. cv. Mysore) was used as an indicator plant for the changes that occurred in the soil. The experimental design was a randomized complete block design, with a 2 x 2 x 3 +2 factorial scheme, with 4 replicates per treatment, in a total of 12 treatments, with 4 replicates (block). The first factor was with and without AMF inoculation, the second was with and without ash added from the burning of sugarcane bagasse, the third was with and without inoculation of fungi and bacteria phosphate solubilizers and two controls area, Cerrado conservation area (CCAR) and degraded area without intervention (DAWI). After 67 and 274 days of planting, the soil chemical and microbiological attributes were evaluated in the 0.0 to 0.10 m layer, as well as the height, diameter, relative chlorophyll index, and total leaf P. The a... (Complete abstract click electronic access below) / Mestre

Submitted by Claudia Rocha (claudia.rocha@udesc.br) on 2018-03-06T15:24:22Z No. of bitstreams: 1 PGCS16MA139.pdf: 1337909 bytes, checksum: ef910e602df5926b0cb8296178289f55 (MD5) / Made available in DSpace on 2018-03-06T15:24:22Z (GMT). No. of bitstreams: 1 PGCS16MA139.pdf: 1337909 bytes, checksum: ef910e602df5926b0cb8296178289f55 (MD5) Previous issue date: 2016-02-16 / Capes / Promop / The arbuscular mycorrhizal fungi (AMF) have broad occurrence, ecological relevance for health and development of plants and edaphoclimatic factors variation sensitivity. These characteristics makes of AMF key group on environmental risk assessment of pollutants in the soil. The aims of this study was: (1) Selecting isolate AMF originated from tropical and subtropical soils, determining suitable incubation conditions to ecotoxicological tests with spores; (2) validating ISO/TS 10832:2009 protocol, basing on germination sensitivity to metals and pesticides and (3) determining if germ tube size (GT) and total length of hyphae (HP), parameters of pre-symbiotic growth, could be used as sensitive indicators of AMF isolates. In Experiment one, was evaluated spore germination of ten isolates of AMF: Acaulospora colombiana SCT115A, A. koskei SCT049C and SCT048A, A. morrowiae SCT400B, SCT056A and SCT063A, Claroideoglomus etunicatum SCT101A, Gigaspora albida SCT200A, Gi. margarita SCT077A and Rizhophagus clarus SCT720A in sand without contaminants. Five were chosen for Experiment two, where germination was evaluated in different times (14 and 28 days), temperatures (24 and 28°C) and substrates uncontaminated (sand, OECD soil (OECD nº 207, 1984) and Tropical Artificial Soil (TAS). In Experiment three, germination, GT and HP parameters of Gi.albida andGi. margarita were evaluated in two substrates (sand and TAS) with two contaminants (heavy metal cadmium and pesticide 18 19 chlorpyrifos). Gi.albida SCT200A, Gi. Margarita SCT077A and R. clarus RJ102A had high germination rate (>75%). Results showed that 14th days (according with ISO protocol) and 28 °C incubations increase germination of the isolates. Sand had huge variation, been inappropriate as control substrate, while TAS was viable and efficient, improving test performance. When exposed to contaminants, GT and HP parameters of Gi. albida SCT200A and Gi. margarita SCT077A had insensitive and variable behavior for doses tested. To both species germination,sensitive to contaminants was low (IC50 >20 mg kg-1 of Cd and IC50 >20 mg a.i. kg-1 of Chlorpyrifos). Additional tests with different doses, as well as other isolates could improving a sensitivity curve of species to contaminants / Os fungos micorrízicos arbusculares (FMAs) tem ampla ocorrência, relevância ecológica para a saúde e desenvolvimento das plantas e sensibilidade a variações nos fatores edafoclimáticos. Estas características tornam os FMAs em um grupo chave na análise de risco de contaminantes do solo. Os objetivos deste estudo foram: (1) selecionar isolados de FMAs originários de solos tropicais e subtropicais em condições adequadas de incubação para ensaios ecotoxicológicos com esporos; (2) validar o Protocolo ISO/TS 10832: 2009, baseando-se na sensibilidade de germinação à metais e agrotóxicos e (3) determinar se o tamanho do tubo germinativo (TG) e comprimento total de hifas (HF), parâmetros de crescimento pré-simbiótico, podem ser utilizados como indicadores de sensibilidade dos isolados de FMAs. No experimento um, avaliou-se a germinação de esporos de dez isolados de FMA: Acaulospora colombiana SCT115A, A. koskei SCT049C e SCT048A, A. morrowiae SCT400B, SCT056A e SCT063A, Claroideoglomus etunicatum SCT101A, Gigaspora albida SCT200A, Gi. margarita SCT077A e Rizhophagus clarus SCT720A, em areia sem contaminantes. Destes, cinco foram escolhidos para o experimento dois, onde a germinação foi avaliada em diferentes tempos (14 e 28 dias), temperaturas (24 e 28ºC) e substratos não contaminados (areia, solo OCDE (OECD nº 207, 1984) e Solo Artificial Tropical (SAT). No experimento três, os parâmetros germinação, TG e HF de Gi. albida e Gi. 14 15 margarita foram avaliados em dois substratos (areia e SAT) com dois contaminantes (cádmio e clorpirifós). Gi. albida SCT200A, Gi. margarita SCT077A e R. clarus RJ102A tiveram altas taxa de germinação ( >75% ). Resultados mostraram que 14 dias (de acordo com o protocolo ISO) e 28ºC de incubação aumentam a germinação dos isolados. A areia teve alta variação e considerada inapropriada como substrato controle, enquanto SAT foi viável e eficiente, melhorando o desempenho dos testes. Quando expostos aos contaminantes, os parâmetros TG e HF de Gi. albida SCT200A e Gi. margarita SCT077A se mostraram pouco sensíveis e com comportamento variável para as doses de contaminantes testadas. Para a germinação, ambas a espécies mostraram pouca sensibilidade aos contaminantes (CI50>20 mg kg-1 de Cd e CI50>20 mg i.a. kg-1 de clorpirifós). Ensaios adicionais com diferentes doses, bem como com outros isolados são indicados para poder melhorar a curva de sensibilidade das espécies aos contaminantes