Certain crop plants are susceptible to pathogens or unable to develop efficient microbial symbioses. These crops adversely impact soil biological quality with consequences on plant health and productivity of cropping systems. Chickpea is a rotational pulse crop with two types: kabuli and desi, and several cultivars. Cultivation of chickpea has inconsistent effects on soil microbial communities and subsequent wheat crops. I conducted field studies and used high throughput molecular analyses to explore the variations among chickpeas to identify cultivars developing fungal communities that are conducive to plant health and productivity. I also carried out greenhouse studies and used biochemical analyses to investigate the response of chickpea cultivars to arbuscular mycorrhizal (AM) fungi and non-AM fungal endophytes and identify the influence of root and root metabolites on the endophytic and pathogenic fungi. Cultivars and types of chickpeas and environmental conditions promoted different fungal communities in the root endosphere. Funneliformis and Claroideoglomus were the dominant AM fungal genera and Fusarium and Alternaria were the dominant non-AM fungal genera in the roots of chickpea. The roots of cultivars CDC Corrine, CDC Cory and CDC Anna hosted the most diverse fungal communities in contrast to CDC Alma and CDC Xena roots which hosted the least diverse communities. Plant response to AM and non-AM fungal endophytes varied with genotype and type of chickpea. The root symbiosis effectively promoted plant growth in CDC Cory, CDC Anna and CDC Frontier and stimulated nitrogen fixation in CDC Corrine. Cultivars of chickpea responded differently to dual inoculation of the AM and non-AM fungal endophytes. Co-inoculation with AM and non-AM fungal endophytes had additive effects on CDC Corrine, CDC Anna and CDC Cory but non-AM fungal endophytes reduced the positive effect of AM fungi in Amit and CDC Vanguard. Desi chickpea appeared to form more efficient symbioses with soil fungal resources than kabuli chickpea. Protein(s) up-regulated in the mycorrhizal roots of the desi chickpea CDC Anna suppressed the growth of the fungal endophytes Trichoderma harzianum and Geomyces vinaceus and of the pathogens Fusarium oxysporum and Rhizoctonia sp. The formation of AM symbiosis decreased the production of root bioactive metabolites soluble in 25% methanol. Some of the root metabolites stimulated the growth of Trichoderma harzianum and Geomyces vinaceus, and a few inhibited Rhizoctonia sp. and Fusarium oxysporum. A few metabolites with contrasting effects on the different fungal species were detected. The non-protein phytochemicals had selective effects on the endophytes and pathogens whereas the antifungal proteins of mycorrhizal roots were non-selective. Overall the study reveals a "genotype effect" of chickpea on the soil microbiota suggesting the possibility to improve the performance of this crop through the selection of genotypes improving the communities of root associated fungi, by associating and responding to beneficial fungi and repressing the pathogens.

No description available.

South Africa is the world's second largest exporter of fresh citrus and is ranked 14th in citrus production. Fungal pathogens such as Phytophthora and Pythium cause economic losses as a result of root rot and brown rot. Mycorrhizal fungi are specialized members of the fungal community forming a mutualistic relationship with plant roots. Mycorrhizal fungal structures are known to associate with other soil microorganisms and these may contribute to improved plant growth. A diverse group of bacteria that interact with the mycorrhizal fungi are known as Mycorrhizal Helper Bacteria (MHB). The aim of this study was to investigate the role of arbuscular mycorrhiza and associated bacteria isolated from spores and determine whether they had any plant growth promoting potential. A total of 19 bacteria were isolated from arbuscular mycorrhizal spores and were molecularly identified as belonging to several Bacillus, Micrococcus, Onchrobactrum and Staphylococcus sp. All bacterial isolates were tested for plant growth promotion abilities. One Bacillus isolate was able to solubilise phosphate. Four isolates Micrococcus sp, Micrococcus leteus, Ochrobacterum sp and Ochrobacterum antropi were able to produce Indole Acetic Acid and three isolates showed potential to reduce growth of Phytophthora nicotianae, P. citrocola and P. citrophthora in in vitro plate cultures. Further tests using culture supernatants of the Bacillus sp, Micrococcus sp and Bacillus cereus confirmed their ability to inhibit or reduce growth of the three Phytophthora species in a 96 well bioassay. Bacillus sp and Bacillus cereus were able to inhibit Phytophthora spp by 95 to 100 % and Micrococcus spp was able to decrease pathogen growth by 60 to 94 %. These bacterial isolates were further evaluated for plant growth promoting abilities on citrus rough lemon seedlings alone or in combination with arbuscular mycorrhizal inoculum. Bacterial and mycorrhizal inoculants influence the increase in shoot and root biomass. Bacillus cereus in combination with mycorrhizal inoculum significantly increased seedling shoot to root ratio while root biomass was significantly increased with mycorrhizal inoculation. Due to the short duration of the trial mycorrhizal colonisation could not be assessed. It is evident that selected combinations of bacteria and mycorrhizal fungi could promote citrus seedling growth and potentially improve seedling health. Further studies under nursery conditions are recommended.

Made available in DSpace on 2014-06-11T19:35:00Z (GMT). No. of bitstreams: 0 Previous issue date: 2006-05-25Bitstream added on 2014-06-13T19:05:01Z : No. of bitstreams: 1 russomanno_omr_dr_botfca.pdf: 951502 bytes, checksum: 65ab95dd8d5f795650c7b83e6aa5b10f (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Secretaria Agricultura / O objetivo do presente trabalho visou avaliar a influência dos FMA Glomus etunicatum Becker & Gerd. e Glomus clarum Nicol. & Schenck, no desenvolvimento de plantas de alecrim e manjericão, bem como verificar a capacidade destas plantas micorrizadas em superar os danos causados pelo fungo Fusarium oxysporum Schecht. Plantas de alecrim (Rosmarinus officinalis L.) e de manjericão (Ocimum basilicum L.) foram inoculadas, separadamente, com G. etunicatum e G. clarum, em casa de vegetação, com temperatura de 26 l 20C e luminosidade de 3000 Lux. Utilizou-se substrato autoclavado composto por uma parte de areia e uma de terra; o inóculo constou de esporos [500 esporos de G.etunicatum (50 mL-1) de solo e 700 esporos de G.clarum (50 mL-1) de solo] e ainda fragmentos de raízes infectadas e micélio. Em cada tipo de planta inoculada foram avaliadas as seguintes variáveis: altura das plantas (AP), peso da matéria seca da parte aérea (MSPA), peso da matéria fresca das raízes (MFR), esporulação (E), colonização radicular (CR) e teor de macro e micronutrientes no substrato e nas plantas (TMm). Foi avaliada também a influência de G. etunicatum e G. clarum no controle de F. oxysporum em plantas de alecrim e manjericão. A inoculação do patógeno (concentração de 5 x 103 esporos mL-1) foi realizada, separadamente, em plantas de alecrim e de manjericão com 90 dias de micorrização (5 vasos com G. etunicatum e 5 vasos com G. clarum) e ainda plantas testemunhas, não micorrizadas (5 vasos). No alecrim, G. clarum mostrou-se significativamente mais eficiente do que G. etunicatum em AP, MSPA e E; por outro lado, G. clarum apresentou CR menor do que G. etunicatum. Em relação às plantas testemunha, G. clarum diferiu significativamente destas em 2 todos as variáveis analisadas, porém G. etunicatum não diferiu estatisticamente das plantas testemunha em AP e MSPA. No manjericão, em relação... . / The purpose of the present work had the objective of evaluating the influence of AMF Glomus etunicatum Becker & Gerd. and Glomus clarum Nicol. & Schenck on the rosemary and basil plants development and also concerning the capacity of the mycorrhizal plants in resisting the wilt caused by Fusarium oxysporum Schecht. For that, rosemary (Rosmarinus officinalis L.) and basil (Ocimum basilicum L.) plants were previously inoculated with G. etunicatum and G. clarum in greenhouse under the temperature of 26 l 20C and luminosity of 3000 Lux. The soil was sterilized and composed by one part of sand and one part of earth. The inoculum was composed by the fungi spores [500 spores of G. etunicatum in (50 mL-1) soil and 700 spores of G. clarum in (50 mL-1) soil] and micelium and roots fragments infected by the AMF. In each plant inoculated the following variables were evaluated: plant height (PH), plant dry weigt (PDW), roots fresh weight (RFW), sporulation rate (SR), root mycorrhizal percentage (RMP) and macro and micronutrients level (MmL) present in the plants and in the soil. The G. clarum and G. etunicatum influence in the control of F. oxysporum wilt in both plants was also evaluated. The inoculation of the pathogen (5x103 spores mL-1) was separatety realized in the rosemary and basil plants after 90 days of the mycorrhization (5 pots of G. clarum and 5 pots of G. etunicatum); the control was also composed by 5 pots without mycorrhization. In the rosemary, G. clarum was significantely more efficient than G. etunicatum in the variables PH, RFW and SR; although G. clarum 4 presented RMP smaller than G. etunicatum. In relation to the control, G. clarum was significably better to the plants in all the variables, although G. etunicatum do not differed statistically for the control plants in PH and PDW. For basil in all the analised variables G. clarum differed statistically from G. etunicatum and was similar to the control treatment in all the variables.

Made available in DSpace on 2015-03-26T13:51:44Z (GMT). No. of bitstreams: 1 01 - capa_abstract.pdf: 58664 bytes, checksum: 66cb47131cc46bcd2d5d263065f54c20 (MD5) Previous issue date: 2008-07-29 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / The objective of this work was to evaluate the effects of the arbuscular mycorrhizal fungus (AMF), Glomus clarum, multiplied monoxenically in transformed carrot roots, on the growth, survival, mycorrhizal colonization and root development of micropropagated plantlets in different stages of rooting. Inoculation was performed in vitro and ex vitro conditions. Micropropagated plantlets with 0, 14 or 21 days maintained in rooting medium, supplemented or not with 1 mg L-1 of Indolbutiric acid (IBA), were transferred to a substrate composed by sand:soil:vermiculite (1:1/2:1). The in vitro system allowed the establishing mycorrhizal association, but no stimulatory effect on the development of shoot or root by mycorrhizal inoculation was observed. The period of time and addition of IBA did not affect plant growth. In ex vitro system, the stimulatory effects of arbuscular mycorrhizal association were evident in all evaluated characteristics, except to plant height. Plants that were not submitted to rooting showed growth similar or superior to those that were maintained in the rooting media, independent of IBA addition. Inoculated plants were shown to be more efficient at nutrient absorption, especially to phosphate. The arbuscular mycorrhizal association promotes beneficial effects when inoculation is performed ex vitro and the best period for inoculation of physic nut seedlings is in the beginning of acclimatization, without in vitro rooting phase. Inoculation of physic nut with arbuscular mycorrhizal fungi showed to be an important tool for micropropagated seedlings production. / O objetivo deste trabalho foi avaliar o efeito da inoculação do fungo micorrízico arbuscular (FMA), Glomus clarum, multiplicado monoxenicamente em raízes de cenoura transformadas, sobre o crescimento, a sobrevivência, a colonização micorrízica e o desenvolvimento radicular de plantas micropropagadas de pinhão-manso em diferentes estádios de enraizamento. A inoculação foi efetuada em condições in vitro e ex vitro. Plantas micropropagadas de pinhão-manso com 0, 14 ou 21 dias de permanência em meio de enraizamento, suplementado ou não com 1 mg L-1 de ácido indol-butírico (AIB), foram transferidas para substrato areia lavada:solo:vermiculita (1:1/2:1). O sistema in vitro utilizado permitiu o estabelecimento da associação micorrízica, mas não foi possível observar efeitos estimuladores da inoculação sobre o crescimento e desenvolvimento radicular das plantas. O tempo de permanência em meio de enraizamento e a adição de AIB também não afetaram o crescimento das plantas. No sistema ex vitro os efeitos estimuladores das micorrizas arbusculares foram evidenciados em todas as características relacionadas ao crescimento, com exceção da altura das plantas. As plantas que não foram submetidas à etapa de enraizamento mostraram crescimento semelhante ou superiores àquelas submetidas à etapa de enraizamento, independente da adição do AIB. As plantas inoculadas com G. clarum mostraram-se também eficientes na absorção de nutrientes, principalmente de fósforo. Pode-se concluir que os efeitos estimuladores das micorrizas arbusculares foram evidenciados quando foi realizada a inoculação ex vitro dessas plantas e que a melhor época de inoculação de J. curcas corresponde à fase de início da aclimatização, sem necessidade de passar pela fase de enraizamento in vitro. A inoculação com FMAs pode representar importante ferramenta para a produção de mudas micropropagadas de qualidade.

Le chêne-liège (Quercus suber) est une essence forestière d’une grande importance écologique et socio-économique pour les habitants de Méditerranée. Cependant, au cours des dernières décennies, ces subéraies et particulièrement les subéraies marocaines ont été soumises à de fortes contraintes climatiques, environnementales et humaines entrainant une accélération des processus de dégradation. La conservation de ces écosystèmes est fortement dépendante de notre capacité à prédire les changements induits par ces différentes pressions ainsi que du développement d’approches durables pour leur réhabilitation. Dans ce contexte, l’identification d’indicateurs biologiques de l’état de santé des subéraies et l'intensification des processus de facilitation entre plantes (arbres/arbustes) apparaissent comme des stratégies écologiques prometteuses. Le succès de ces approches est cependant assujetti à notre compréhension des interactions entre les communautés végétales et les champignons du sol, notamment les champignons mycorhiziens, éléments clés du fonctionnement des écosystèmes forestiers. Ce travail a visé le décryptage des réseaux fongiques, notamment mycorhiziens associés au chêne-liège et la végétation du sous-bois dans trois subéraies marocaines (Maâmora, Benslimane, Chefchaoun) caractérisées par différents niveaux de dégradation. La diversité fongique associée aux racines du chêne-liège et à plusieurs plantes arbustives représentatives des subéraies (Cistus salviifolius, Cistus monpeliensis et Lavandula stoechas) a été étudiée en combinant les méthodes traditionnelles basées sur l’aspect morphologique des mycorhizes et les nouvelles technologies de séquençage haut-débit par identification moléculaire des communautés fongiques.Les résultats obtenus représentent la plus vaste enquête de la diversité fongique du sol, notamment mycorhizienne, au sein des subéraies marocaines. Différents niveaux de structuration des communautés de champignons du sol ont été révélés, fonction de l’habitat, du type de plantes et de l'état de dégradation. Une large gamme d’indicateurs fongiques de l’état de dégradation de la subéraie, en lien avec la plantes hôte, ont pu être mise en évidence au sein des différents habitats, soulignant l’importance de plusieurs champignons ectomycorhiziens (notamment Cenococcum, Russula, Terfezia et Tomentella) mais aussi des champignons mycorhiziens éricoïdes (Cladophialophora, Oidiodendron) et à arbuscules (Rhizophagus, Redeckera, Racocetra, Paraglomus). Ce travail a permis d’établir une base de données majeure sur l’écologie des champignons du sol dans les subéraies marocaines, et de proposer un nouvel éclairage sur leur potentiel pour le suivi de l’état de santé des subéraies, ainsi que pour la mise en place de programmes de conservation adaptés tenant compte aussi des champignons associés. L’application des approches proposées à une plus large diversité d’écosystèmes forestiers devrait constituer un atout important pour la meilleure compréhension du fonctionnement biologique des écosystèmes forestiers et leur sauvegarde face à l’aggravation des pressions humaines et climatiques au niveau mondial. / The Cork oak (Quercus suber) forests play an important role in terms of ecological services and socio-economic development for the Mediterranean populations. However, the cork oak forests, notably in the Southern Mediterranean basin are highly threatened by increasing human and climate pressures, which accelerates desertication. The conservation of this ecosytem is strongly dependent of our ability to predict the environmental changes induced by these pressures as well as to develop sustainable approach for their restoration. In this context, the identification of biological indicators of cork oak health and the intensification of plant-plant facilitation processes appears as promisising ecological strategies. Their success is however subjected to our understanding of plant-fungal interactions, notably with fungal mycorrhiza, key factors of forest ecosystem functionning. The current work aimed at deciphering plant-fungal networks, notably mycorrhizal networks with cork oak and its understory shrub vegetation in three Moroccan cork oak habitats (Maâmora, Benslimane, Chefchaoun) characterized by different degradation levels. The root-fungal diversity associated to cork oak and major components of its understory shrub vegetation (Cistus salviifolius, Cistus monpeliensis et Lavandula stoechas) has been analysed by combining traditional methods based on morphological identification, and new generation high- throughput DNA sequencing methods to characterize communities at the molecular level.The study represents the most extensive survey of soil fungal diversity, notably mycorrhizal diversity, in Moroccan cork oak ecosystems. Different fungal community structures were revealed, depending on habitat, plant host type, and degradation forest status. A wide range of fungal indicators of plant type × forest status has been identified, highlighting the importance of several ectomycorrhizal fungi (notably Cenococcum, Russula, Terfezia and Tomentella) as well as ericoid mycorrhizal fungi (Cladophialophora, Oidiodendron) and arbuscular mycorrhizal fungi (Rhizophagus, Redeckera, Racocetra, and Paraglomus). The current work provides an extensive database on the ecology of soil fungi related to the Moroccan cork oak forest, offers new insights into the potential of soil fungi for monitoring the health of the cork oak forest, and for the developement of efficient conservation programs of this ecosystem by taking into account the soil fungal communties associated. The use of proposed appoaches to a larger diversity of forest ecosystems are promising to better understand the biological fonctionning of forest ecosystem and their conservation in response to the worsening of worldwide human and climate pressures.

Made available in DSpace on 2016-04-26T19:39:32Z (GMT). No. of bitstreams: 1 2007 - Michele de oliveira macedo.pdf: 1438139 bytes, checksum: 65185c2bcc2d92ca55054aa48e76edd3 (MD5) Previous issue date: 2007-07-31 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico / This study aimed to evaluate carbon and nitrogen stocks restoration, and soil fertility status in land reclamation with nodulated and mycorrhizal legume trees and fallow length on high productivity shifting cultivation. Evaluations were made of soil C and N stocks, concentration of free and occluded light fraction and soil available Ca, Mg, P, K, C and N, and amounts of mass and nutrients of litter in the dry and rainy season. One of the areas located in Angra dos Reis, RJ is under reclamation for 13 years. As reference area it was used a forest fragment and deforested area nearby. A second area is located in Serop?dica, RJ, and it is under reclamation for 17 years with the species Acacia auriculiformis and Mimosa caesalpiniifolia, and with Albizia guachapele for 13 years. As reference area it was used a secondary forest fragment and a pasture area. Both areas under recuperation in Angra dos Reis and in Serop?dica had been used as source of topsoil, having lost part of the upper soil horizons. The slash-and-burn area is located in Bom Jardim, Nova Friburgo, RJ. For this study, it was utilized a chronossequence of soil gradient: area cultivated for one year; area cultivated with crop rotation for three years; area cultivated with bananas for 18 years; area under fallowing for five years; and others two fallowing areas for 18 and 33 years. The usage of legume trees in land reclamation was able to increase stocks of C and N, from 0.04 to 0.15 Mg ha-1 year-1 for N, and of 0.13 to 1.73 Mg ha-1 year-1 for C. In the slash-and-burn agriculture system, the lowest soil C stock was found in the one and three years cultivated areas. The highest stock was observed under fallowing and in the area planted with bananas, and they were similar between areas. The highest soil N stock was found in fallow with five years. In Angra dos Reis area the concentration of free and occluded light fraction was similar between areas of forest and under reclamation. In Serop?dica, only the free light fraction of organic matter was re-established. There was no significant difference between the concentrations of free and occluded light fraction in Bom Jardim areas; however, the area with five years fallow and three years cultivation showed the same values of the free light fraction. In Angra dos Reis areas, the litter stocks were similar between the forest and the area under reclamation. In Serop?dica, except for the area under A.auriculiformis with greatest litter stock, all other areas showed similar litter stocks. In Bom Jardim, the litter stocks were similar among the fallowing areas. The nutrients contained in the litter, in general, were similar among the areas, and they were dependent on the litter mass. The soil fertility in the areas under reclamation was slightly higher than in the reference ones. In the slash-and-burn area, the soil fertility analyses indicated a residual effect of liming in the fallow with five years. Considering the attributes soil fertility and soil C and N stocks, the best fallowing period would be of five years. / Neste estudo foi avaliado o potencial do uso de leguminosas arb?reas fixadoras de nitrog?nio, e da utiliza??o de pousio em um sistema de agricultura itinerante de alta produtividade, em restabelecer a ciclagem de nutrientes e incrementar os estoques de C e N. Foram avaliados os estoques de C e N do solo, as concentra??es de fra??o leve livre e leve oclusa da mat?ria org?nica, os teores de Ca, Mg, P, K, C e N no solo, e o estoque e a quantidade de nutrientes presentes na serapilheira nas esta??es seca e chuvosa. Uma das ?reas avaliadas, no munic?pio de Angra dos Reis (RJ), est? em recupera??o h? 13 anos. Para fins de compara??o, utilizou-se como refer?ncia uma ?rea de mata e outra, desmatada, adjacente ? ?rea em recupera??o. As demais ?reas estudadas localizam-se em Serop?dica (RJ), e est?o em recupera??o h? 17 anos com as esp?cies Mimosa caesalpiniifolia e Acacia auriculiformis, e h? 13 anos, com a esp?cie Albizia guachapele. Como ?rea refer?ncia foi utilizada uma ?rea de capoeira e outra de pastagem. Tanto a ?rea em recupera??o localizada em Angra dos Reis quanto as ?reas em recupera??o em Serop?dica haviam sido utilizadas como ?reas de empr?stimo, tendo perdido parte dos horizontes superficiais do solo. A ?rea de agricultura itinerante situa-se em Bom Jardim, regi?o serrana do Rio de Janeiro. Foi avaliado um gradiente de uso do solo: ?rea cultivada h? um ano; ?rea h? tr?s anos com rota??o cultivos; ?rea h? 18 anos cultivada com bananas; ?rea em pousio h? cinco anos; e outras duas ?reas em pousio h? 18 e 33 anos. O uso de leguminosas incrementou os estoques de C e N do solo nas ?reas em recupera??o. As taxas de aumento variaram de 0,04 a 0,15 Mg ha-1 ano-1 para N, e de 0,13 a 1,73 Mg ha-1 ano-1 de C. No sistema de agricultura itinerante, os menores estoques de C foram observados nas ?reas cultivadas h? um e h? tr?s anos. Nas ?reas em pousio e no bananal os estoques foram similares entre si e superiores aos das ?reas cultivadas. O maior estoque de N foi observado na ?rea com cinco anos de pousio. As concentra??es de fra??o leve livre e leve oclusa foram similares entre as ?reas de mata e em recupera??o em Angra dos Reis. Nas ?reas em recupera??o em Serop?dica, somente a fra??o leve livre foi restabelecida. J? nas ?reas de Bom Jardim, n?o houve diferen?a significativa entre as concentra??es de fra??o leve livre e leve oclusa. No entanto, o pousio de cinco anos e o cultivo de tr?s anos apresentaram os menores valores de fra??o leve livre. Os estoques de serapilheira nas ?reas de mata e em recupera??o de Angra dos Reis foram similares. As ?reas em recupera??o no munic?pio de Serop?dica apresentaram estoques de serapilheira similares, com exce??o da esp?cie A. auriculiformis que mostrou os maiores estoques. Nas ?reas em pousio do sistema de agricultura itinerante, os estoques de serapilheira foram similares. O estoque de nutrientes da serapilheira, em geral, variou pouco, e foi dependente da massa de serapilheira. A fertilidade dos solos das ?reas em recupera??o foi ligeiramente maior que das ?reas refer?ncia. Na agricultura itinerante, pode-se observar efeito residual da calagem usada pelo agricultor no pousio de cinco anos. De acordo com os resultados de estoque de C, N e fertilidade do solo, o per?odo de pousio mais indicado para o corte e queima foi o de cinco anos.

Algumas actinobactérias habitantes da rizosfera são produtoras de substâncias capazes de combater micro-organismos patogênicos às plantas, o que as torna potenciais agentes de controle biológico, passíveis de serem utilizadas como princípio ativo de inoculantes de sementes e mudas. O presente trabalho teve como objetivo isolar e avaliar o potencial de isolados de actinobactérias no controle de doenças causadas por fungos nas espécies arbóreas Araucaria angustifolia (Araucária) e Pinus elliottii (Pinus). Além disso, foi iniciado o estudo do processo de elaboração de inoculante a base de actinobactérias antagônicas. Foram isoladas 215 estirpes de actinobactérias do rizoplano da Araucária, das quais 13 apresentaram, em testes in vitro, potencial como antagonistas contra os fungos fitopatogênicos Fusarium oxysporum, Cylindrocladium candelabrum, C. pteridis e Armillaria sp., e apenas os isolados Ac136 e Ac202 apresentaram os maiores valores de inibição nos testes com os quatro patógenos. Na avaliação de produção de algumas substâncias antimicrobianas, apenas celulases, quitinases e sideróforos foram produzidas pelos isolados, sendo este último o mais frequente. Nos testes de interação com organismos foi verificado que, embora os isolados de actinobactéria tenham inibido a germinação de esporos do fungo micorrízico arbuscular (FMA) Gigaspora rosea em teste in vitro, no experimento in vivo, em que foi utilizado o milho como planta hospedeira para inoculação com o FMA e os isolados A43, Ac136 e Ac202, os isolados Ac136 e Ac202 estimularam a colonização das raízes pelo FMA. Estes mesmos isolados também estimularam a germinação de sementes e desenvolvimento inicial de plântulas de Pinus, porém prejudicaram a germinação e desenvolvimento inicial de Araucária. Estes mesmos isolados foram capazes de reduzir a mortalidade de plântulas de Pinus em cerca de 25%, e esta diminuição foi atribuída à inibição de Fusarium sp.. No teste de viabilidade dos isolados em diferentes veículos o isolado que mostrou maior sobrevivência foi o Ac202, mantendo um número de propágulos viáveis correspondente a 5,48 log UFC mL-1, enquanto que o veículo mais apropriado para a elaboração de inoculante com actinobactérias foi a glicerina. A análise molecular mostrou que os isolados mais promissores apresentaram maior similaridade com S. kasugaensis. Dentre todos isolados Ac202 (S. kasugaensis) foi o mais promissor obtido neste trabalho para o uso como agente de biocontrole de doenças causadas por fungos, apresentando forte antagonismo contra os quatro patógenos testados, promovendo a germinação e desenvolvimento inicial de Pinus e aumentando a sobrevivência de plântulas contaminadas com fungos fitopatogênicos. / Some actinobacteria that inhabits the rhizosphere are producers of substances that are capable of combating plant-pathogenic microorganisms, what makes them potential biological control agents, which can be used as the active ingredient of seeds and seedlings inoculants. This study aimed to isolate and evaluate the potential of actinobacteria isolates in controlling diseases caused by fungi in Araucaria angustifolia and Pinus elliottii. In addition, the study initiated the process of elaboration of an actinobacteria-based inoculant. 215 actinobacterial strains were isolated from the Araucarias rhizoplane, and 13 of them showed potential as antagonists against the phytopathogenic fungi Fusarium oxysporum, Cylindrocladium candelabrum, C. pteridis and Armillaria sp. in in vitro tests, and only the Ac136 and Ac202 strains showed the highest inhibition in the tests against the four pathogens. Among the antimicrobial substances tested, only cellulases, chitinases and siderophores were produced, with the latter being the most frequent. In the interaction tests with other organisms it was found that although the actinobacterial strains have inhibited the germination of the arbuscular mycorrhizal fungi (AMF) Gigaspora rosea spores on the in vitro test, the in vivo experiment, with maize as host plant, inoculated with the AMF and A43, Ac136 and Ac202 actinobacterial strains, Ac136 and Ac202 stimulated root colonization by AMF. These strains also stimulated Pinus seed germination and seedling early development, but hindered the germination and early development of Araucaria. In addition, the same strains were able to reduce the mortality of pine seedlings by about 25%, and this decrease was attributed to the inhibition of Fusarium sp.. In the viability test of the strains in different vehicles the strain that showed the greatest shelf-life was Ac202, with 5.48 log CFU mL-1, and the most appropriate vehicle for the actinobacteriabased inoculant development was glycerin. Molecular analysis showed that the most promising isolates showed the greatest similarity with S. kasugaensis. Among all strains Ac202 (S. kasugaensis) was the most promising one for the use as biocontrol agent of fungal diseases, exhibiting a strong antagonism against the four pathogens tested, promoting germination and early development of Pinus and increasing survival of seedlings infected with pathogenic fungi.

Les plantes vivent en association avec une grande diversité de microorganismes qui forment son microbiota. Ce microbiote fournit des fonctions clés qui influencent tous les aspects de la vie d'une plante, de l'établissement à la croissance jusqu'à la production. Cette thèse a pour intention de déterminer les règlent d'assemblage du microbiote et ses conséquences pour le phénotypel l'adaptation et l'évolution des plantes. Pour atteindre cet objectif nous avont utilisé différentes approches expérimentales comprenant des plantes clonales comme organismes modèles ainsi que des mésocosmes prairiaux pour analyses à l'échelle des communautés. Nos résultats ont démontré i) que les Champignons Mycohiziens à Arbuscules induisent d'important es variations phénotypiques pour les traits des plantes clonales impliqués dans l'exploration de l'espace et l'exploitation des ressources. Ces changements dépendent de l'identité des symbiontes et altèrent les capacités des plantes à développer des réponses plastiques à l'hétérogénéité environnementale. ii) Les plantes ont évolué un méchanisme permettant la transmission d'une partie de leur microbiote a leur descendance, assurant la qualité de leur habitat. iii) Le contexte spécifique des communautés de plantes est un facteur majeur structurant l'assemblage du microbiota des plantes à échelle locale. L'abondance de certaines espèces de plante dans le voisinage d'une plante cible augmente ou diminue la diversité de son microbiote, déterminant in fine ses performances. De manière générale, cette thèse démontre l'importance des organismes symbiotiques dans la compréhension de l'adaptation et de l'évolution des plantes. / Plants live in association with a wide diversity of microorganisms forming the microbiota. The plant microbiota provides a variety of key functions that influence many aspects of plant's life comprising establishment, growth and reproduction. The present thesis aims at determining the assembly rules of the plant microbiota and its consequences for plant phenotype, adaptation and evolution. To fulfill this objective, we used different experimental approaches using either clonal plants as model organisms or grassland mesocosms for community-wide analyses. Our results demonstrated i) that Arbuscular Mycorrhizal Fungi induce important phenotypic variations in clonal plants traits involved in space exploration and resources exploitation. These changes depended on the identity of the symbionts and altered the plants ability to produce plastic responses to environmental heterogeneity. ii) Plants have evolved a mechanism allowing the transmission of a part of their microbiota to their progeny, ensuring thus their habitat quality. iii) The plant community context is a major factor structuring local plant microbiota assembly. Particular plant species identity in the neighborhood increase or decrease the microbiota diversity and ultimately determine the focal plant performance. This thesis overall demonstrates the importance of symbiotic microorganisms in the understanding of the plant adaptation and evolution. From the knowledges acquired we developed a novel understanding of symbiotic interactions in clonal plants by extending the holobiont theory to the meta-holobiont theory.

La phytoremédiaton assistée par les champignons mycorhiziens à arbuscules (CMA) compte parmi les méthodes émergentes de remédiation des sols pollués en raison de son adéquation avec le développement durable. Cette phytotechnologie présente un intêret à la fois dans la protection des plantes contre la phytotoxicité des polluants organiques tels que les hydrocarbures et dans leur dissipation. Cependant, son efficacité reste à prouver dans les sols historiquement multi-pollués. De plus, les mécanismes de dégradation et de tolérance mis en oeuvre par la symbiose mycorhizienne sont peu connus. Ainsi, ce travail de thèse avait pour premier objectif d'étudier chez les partenaires symbiotiques Medicago truncatula/Rhizophagus irregularis cultivés in vitro, l'expression de gènes potentiellement impliqués dans la tolérance au benzo[a]pyrène (B[a]P), un hydrocarbure aromatique polycyclique (HAP) de haut poids moléculaire fréquemment détecté dans les sols pollués. Pour cela, les expressions de gènes codant pour des enzymes antioxydantes, de détoxification des polluants et de réparation de l'ADN ont été mesurées par PCR quantitative en temps réel. Nos résultats ont montré une corrélation positive entre l'induction des systèmes antioxydants au niveau génique et enzymatique et la production d' H₂O₂ induite par l'accumulation de B[a]P dans les racines non mycorhizées et le CMA. En revanche, lorsque les racines sont colonisés par le CMA, celui-ci agirait comme une barrière physique en limitant l'accumulation de B[a]P et la production d'H₂O₂ dans les racines, et provoquerait la répression des systèmes antioxydants racinaires. Le second objectifs de cette thèse a consisté à évaluer l'apport d'un amendement mycorhizien dans la dégradation de deux types d'hydrocarbures : les HAP et les alcanes, présent dans un sol historiquement multi-pollué (site de l'Union). L'étude a été conduite en microcosmes en présence de blé (Triticul aestivum) inoculé ou non par R. irregularis. Une contribution positive de cette inoculation dans la biodégradation des hydrocarbures après 16 semaines de culture a été démontrée. Cette meilleure dégradation serait liée notamment à la stimulation de la microflore bactérienne du sol mais également aux capacités métaboliques de la plante. / Arbuscular mycorrhizal fungi (AMF)-assisted phytoremediation is one of innovative method for the remediation of polluted soils due to its relevance to sustainable development. This phytotechnology presents benefits both in the protection of plants against the phytotoxicity of organic pollutants such as hydrocarbons and in their dissipation. However, its efficiency remains to be proved in the aged multi-polluted soils and the mechanisms of degradation and tolerance implemented by the mycorrhizal symbiosis are poorly known. Thus, the thesis aims firstly to study in the symbiotic partners Medicago truncatula/Rhizophagus irregularis cultivated in vitro, the expression of genes potentially involved in the tolerance to benzo[a]pyrene (B[a]P), a high molecular weight polycyclic aromatic hydrocarbon (PAH) frequently detected in polluted soils. Expressions of genes encoding for antioxidant, pollutant, detoxification and DNA repairing enzymes were measured by real time PCR. Our results showed a positive correlation between the induction of antioxidant systems (genes and enzymes) and the production of H₂O₂ induced by the accumulation of B[a]P in non-mycorrhizal root and the AMF. In contrast, when the roots are colonized by the AMF, this one would act as a physical barrier limiting the accumulation of B[a]P and H₂O₂ production of in the roots and would cause the repression of root antioxidant systems. The second objective of the thesis consisted of evaluating the contribution of the arbuscular mycorrhizal amendment in the degradation of two types of hydrocarbons : the PAH and the alkanes, present in an aged multi-polluted soils (site of Union). The study was carried out in microcosms in the presence of non-inoculated or inoculated wheat (Triticul aestivum) by R. irregularis. A positive contribution of the mycorrhizal inoculation in the hydrocarbon biodegradation after 16 weeks of culture was demonstrated. This better degradation is related especially to the simulation of the bacterial microflora but also to the plant metabolic abilities.