CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / O Nordeste brasileiro reÃne condiÃÃes edafoclimÃticas favorÃveis ao cultivo da banana, que pode ser comprovado pela posiÃÃo de destaque dessa regiÃo no cenÃrio produtivo brasileiro. Sendo uma fruta de preferÃncia mundial, e uma das mais exploradas no mundo, assume importÃncia fundamental, pelo seu valor econÃmico, nutritivo e social. A salinidade dos solos à um importante fator de estresse, ocorrendo em regiÃes semiÃridas e Ãridas do Nordeste brasileiro, onde a bananeira à cultivada. Neste contexto, os fungos micorrizicos arbuscular (FMA) vÃm sendo pesquisados nos Ãltimos anos, com o objetivo de minimizar algum dos efeitos do estresse salino nas plantas. O presente trabalho teve como objetivo avaliar o efeito de nÃveis de sais da Ãgua de irrigaÃÃo sobre o acÃmulo de sais no solo, à colonizaÃÃo com FMA e ao desenvolvimento de mudas de bananeira Musa sp. colonizadas com fungos micorrizicos arbusculares nativos. Para isso foi instalado um experimento em casa de vegetaÃÃo do Departamento de CiÃncias do Solo da Universidade Federal do CearÃ, no Campus do Pici em Fortaleza. O delineamento experimental foi o inteiramente casualizado, em parcelas subdivididas, correspondendo a 4 perÃodos de colheita (40, 60, 80 e 100 DAT) e 5 nÃveis de salinidade (0,5; 1,5; 2,5; 3,5 e 4,5 dS m-1), com 4 repetiÃÃes totalizando 80 unidades experimentais. As mudas de bananeiras passaram por um perÃodo de aclimatizaÃÃo e micorrizaÃÃo de 40 dias apÃs o transplantio (DAT), sendo iniciada a aplicaÃÃo de Ãgua salina apÃs este perÃodo. A primeira coleta foi aos 40 DAT antes da aplicaÃÃo dos nÃveis de salinidade, sendo realizadas as outras coletas aos 60, 80 e 100 DAT, onde foram avaliados os parÃmetros: matÃria seca da parte aÃrea e altura das plantas, diÃmetro do XI pseudocaule, condutÃncia estomÃtica, taxa de transpiraÃÃo, taxa fotossintÃtica, pH do solo, condutividade elÃtrica do solo, colonizaÃÃo micorrÃzica radicular, densidade de esporos de FMA e respiraÃÃo basal do solo. O aumento da salinidade da Ãgua de irrigaÃÃo provocou acÃmulo de sais no solo, medido pela condutividade elÃtrica, porÃm nÃo influenciou significativamente o pH. O aumento nos nÃveis de salinidade nÃo ocasionou diferenÃa significativa na condutÃncia estomÃtica, taxa de transpiraÃÃo das plantas, altura das plantas e diÃmetro do pseudocaule, porÃm reduziu a taxa fotossintÃtica e a produÃÃo de matÃria seca das plantas, notadamente apÃs 80 DAT; o aumento da salinidade da Ãgua de irrigaÃÃo reduziu os teores de N e K, e aumento nos teores de Na, porÃm nÃo influenciou os teores de P nas plantas. O aumento da salinidade reduziu a colonizaÃÃo micorrÃzica radicular e a respiraÃÃo basal do solo, porÃm nÃo influenciou a densidade de esporos de FMA no solo. De modo geral, as respostas observadas no solo (quÃmicas e microbiolÃgicas) e na planta, em conseqÃÃncia da salinidade, foram influenciadas pelo tempo de exposiÃÃo do sistema solo/planta aos diferentes tratamentos de irrigaÃÃo. / The Northeast of Brazil region meets the soil-climate conditions for a favorable cultivation of the banana plant, which can be demonstrated by the prominent position of the region as a great banana producer in the country. The banana is a fruit consumed all over the world, so being intensively cropped everywhere; its importance involves economic, nutritional and social aspects. The soil salinity represents a fundamental stress factor on the banana yield and such soil salinity is very common in northeast of Brazil (semi-arid climate) soils where the crop is widely grown. Under this conditions, arbuscular mycorhizal fungi (AMF) have been investigated lately with the objective to evaluate the role of the fungus in decreasing the plant salinity stress effects. Thus, the present study had the objective to investigate the effects of different irrigation water salt levels on: (a) the soil salt accumulation, (b) on the fungus (AMF) colonization, and (c) on the banana Musa sp. seedling plant growth colonized with native AMF. An experiment was conducted under greenhouse conditions (belonging to Soil Science Department â Federal University of Cearà State â Campus PICI, Fortaleza city). The statistical design was an entirely randomized blocks, in subdivided plots, with a four harvest periods (40, 60, 80 and 100 days after planting), five level of soil salinity (0,5; 1,5; 2,5; 3,5 and 4,5 dS m-1) and four replicates, summing a total of 80 treatments. The banana seedling plants were subjected to a 40 days period of adaptation for fungus infection (mycorrization) and afterwards being irrigated with saline water. The first harvest was at the 40 days after the planting; the other harvests were done at the 60, 80, and 100 days after the planting. The following variables were evaluated: aerial plant dry matter, plant height, stem diameter, stomatal conductance, transpiration rate, photosynthetic rate, soil pH, soil electrical conductivity, root fungus colonization, estimates of AMF sporous, and soil basic respiration. Increase in the irrigation water salt level caused salt accumulation in the soil which was measured through soil electrical conductivity; the soil pH was not significantly affected. The increasing soil salinity did not cause significant differences in the following varables: stomatal conductance, plant transpiration rate, plant height, and stem diameter, otherwise, it reduced the photosynthetic rate and the plant aerial dry matter yield, notably at the 80th after planting. The increase in the salinity of the irrigation water caused a reduction of N and K plant concentrations, but increased the Na concentration an had no effect on the XIII P. The root fungus colonization and the basic soil respiration were both reduced by the salinity of the irrigation water; AMF fungus sporous formation in the soil was not affected. In general, the responses observed in both soil (chemical and microbiological) and plant, as results of the water salinity, were affected by the time exposure of the system soil/plant to the different salinity irrigation water levels.

La fertilisation phosphatée est très répandue dans les pratiques agricoles Nord-Américaines. Bien que généralement très efficace pour augmenter la production végétale, son utilisation peut engendrer certaines contaminations environnementales. Afin de diminuer ce problème, plusieurs pratiques de gestion sont envisagées. Parmi celles-ci, on retrouve l’intéressante possibilité de manipuler la flore microbienne car cette dernière est reconnue pour son implication dans bons nombres de processus fondamentaux liés à la fertilité du sol. Cette étude a démontré que lors d’essais en champs, la forme de fertilisant ajouté au sol ainsi que la dose de phosphore (P) appliquée avaient un impact sur la distribution des microorganismes dans les différentes parcelles. Une première expérience menée sur une culture de luzerne en prairie semi-aride a montré que les échantillons provenant de parcelles ayant reçu différentes doses de P présentaient des différences significatives dans leurs communautés bactériennes et fongiques. La communauté de CMA est restée similaire entre les différents traitements. Une deuxième expérience fut menée pendant trois saisons consécutives afin de déterminer l’effet de différentes formes de fertilisation organiques et minérale ajustées selon une dose unique de P sur les populations bactériennes et fongiques d’une culture intensive de maïs en rotation avec du soja. Les résultats des analyses ont montrés que les populations varient selon le type de fertilisation reçu et que les changements sont indépendants du type de végétaux cultivé. Par contre, les populations microbiennes subissent une variation plus marquée au cours de la saison de culture. La technique de DGGE a permis d’observer les changements frappant la diversité microbienne du sol mais n’a permis d’identifier qu’une faible proportion des organismes en cause. Parallèlement à cette deuxième étude, une seconde expérience au même site fut menée sur la communauté de champignons mycorhiziens à arbuscules (CMA) puisqu’il s’agit d’organismes vivant en symbiose mutualiste avec la majorité des plantes et favorisant la nutrition de même que l’augmentation de la résistance aux stress de l’hôte. Ceci permit d’identifier et de comparer les différents CMA présents dans des échantillons de sol et de racines de maïs et soja. Contrairement aux bactéries et aux champignons en général, les CMA présentaient une diversité très stable lors des différents traitements. Par contre, au cours des trois années expérimentales, il a été noté que certains ribotypes étaient significativement plus liés au sol ou aux racines. Finalement, l’ensemble de l’étude a démontré que la fertilisation phosphatée affecte la structure des communautés microbiennes du sol dans les systèmes évalués. Cependant, lors de chaque expérience, la date d’échantillonnage jouait également un rôle prépondérant sur la distribution des organismes. Plusieurs paramètres du sol furent aussi mesurés et ils présentaient aussi une variation au cours de la saison. L’ensemble des interactions possibles entre ces différents paramètres qui, dans certains cas, variaient selon le traitement appliqué, aurait alors probablement plus d’impact sur la biodiversité microbienne que la seule fertilisation. / Phosphorus fertilization is a widespread practice in North American agriculture. Although it is generally efficient to increase yields, its use can also induce some environmental contaminations. Several management practices are considered in order to decrease this problem. Among these possibilities there is the challenging one of manipulating microbial flora, which is well known for its implication in many processes related to soil fertility. We have demonstrated in field trials that both the form of fertilizer added to soil and the applied P amounts impact microbial distribution in plots. A first experiment performed on alfalfa monocultures in semi-arid prairie conditions demonstrated that samples coming from plots that had received different doses of P fertilizer presented significant differences on their bacterial and fungal communities. AMF population remained stable between treatments. A second experiment was conducted over three growing season of an intensive maize/soybean rotation cropping system. It aimed to determine the effect of different organic and mineral fertilizers containing equal P amount on bacterial and fungal populations. It was demonstrated that these communities varied according to the fertilizer type applied. Changes are independent from the grown crop. However, microbial populations have undergone greater variation within each growing season. DGGE approach allowed to observe changes occurring in soil microbial diversity but have only permit to identify a small proportion of organisms. A second experiment in the latter study was performed on the same site and focused on arbuscular mycorrhizal fungi (AMF) as they are organisms living in a mutualistic symbiosis with most land plants and increasing host nutrition and resistance to stresses. It led to the identification and comparison of the different AMF found in maize and soybean soil and root samples. In opposition to what was observed with bacteria and fungi previously, AMF presented a very stable diversity between the different treatments. However, some ribotypes were significantly more present in soil or roots during each growing season. Finally, our whole project demonstrated that P fertilization affected microbial community structure on studied sites. Nevertheless, in each experiment, sampling time also played a substantial role in the organism distribution. Many soil parameters were also monitored and presented a seasonal variation. The sum of possible interactions between these parameters, which in some cases varied according to treatment, would thus have more impact on microbial diversity that the sole fertilization.

La fertilisation phosphatée est très répandue dans les pratiques agricoles Nord-Américaines. Bien que généralement très efficace pour augmenter la production végétale, son utilisation peut engendrer certaines contaminations environnementales. Afin de diminuer ce problème, plusieurs pratiques de gestion sont envisagées. Parmi celles-ci, on retrouve l’intéressante possibilité de manipuler la flore microbienne car cette dernière est reconnue pour son implication dans bons nombres de processus fondamentaux liés à la fertilité du sol. Cette étude a démontré que lors d’essais en champs, la forme de fertilisant ajouté au sol ainsi que la dose de phosphore (P) appliquée avaient un impact sur la distribution des microorganismes dans les différentes parcelles. Une première expérience menée sur une culture de luzerne en prairie semi-aride a montré que les échantillons provenant de parcelles ayant reçu différentes doses de P présentaient des différences significatives dans leurs communautés bactériennes et fongiques. La communauté de CMA est restée similaire entre les différents traitements. Une deuxième expérience fut menée pendant trois saisons consécutives afin de déterminer l’effet de différentes formes de fertilisation organiques et minérale ajustées selon une dose unique de P sur les populations bactériennes et fongiques d’une culture intensive de maïs en rotation avec du soja. Les résultats des analyses ont montrés que les populations varient selon le type de fertilisation reçu et que les changements sont indépendants du type de végétaux cultivé. Par contre, les populations microbiennes subissent une variation plus marquée au cours de la saison de culture. La technique de DGGE a permis d’observer les changements frappant la diversité microbienne du sol mais n’a permis d’identifier qu’une faible proportion des organismes en cause. Parallèlement à cette deuxième étude, une seconde expérience au même site fut menée sur la communauté de champignons mycorhiziens à arbuscules (CMA) puisqu’il s’agit d’organismes vivant en symbiose mutualiste avec la majorité des plantes et favorisant la nutrition de même que l’augmentation de la résistance aux stress de l’hôte. Ceci permit d’identifier et de comparer les différents CMA présents dans des échantillons de sol et de racines de maïs et soja. Contrairement aux bactéries et aux champignons en général, les CMA présentaient une diversité très stable lors des différents traitements. Par contre, au cours des trois années expérimentales, il a été noté que certains ribotypes étaient significativement plus liés au sol ou aux racines. Finalement, l’ensemble de l’étude a démontré que la fertilisation phosphatée affecte la structure des communautés microbiennes du sol dans les systèmes évalués. Cependant, lors de chaque expérience, la date d’échantillonnage jouait également un rôle prépondérant sur la distribution des organismes. Plusieurs paramètres du sol furent aussi mesurés et ils présentaient aussi une variation au cours de la saison. L’ensemble des interactions possibles entre ces différents paramètres qui, dans certains cas, variaient selon le traitement appliqué, aurait alors probablement plus d’impact sur la biodiversité microbienne que la seule fertilisation. / Phosphorus fertilization is a widespread practice in North American agriculture. Although it is generally efficient to increase yields, its use can also induce some environmental contaminations. Several management practices are considered in order to decrease this problem. Among these possibilities there is the challenging one of manipulating microbial flora, which is well known for its implication in many processes related to soil fertility. We have demonstrated in field trials that both the form of fertilizer added to soil and the applied P amounts impact microbial distribution in plots. A first experiment performed on alfalfa monocultures in semi-arid prairie conditions demonstrated that samples coming from plots that had received different doses of P fertilizer presented significant differences on their bacterial and fungal communities. AMF population remained stable between treatments. A second experiment was conducted over three growing season of an intensive maize/soybean rotation cropping system. It aimed to determine the effect of different organic and mineral fertilizers containing equal P amount on bacterial and fungal populations. It was demonstrated that these communities varied according to the fertilizer type applied. Changes are independent from the grown crop. However, microbial populations have undergone greater variation within each growing season. DGGE approach allowed to observe changes occurring in soil microbial diversity but have only permit to identify a small proportion of organisms. A second experiment in the latter study was performed on the same site and focused on arbuscular mycorrhizal fungi (AMF) as they are organisms living in a mutualistic symbiosis with most land plants and increasing host nutrition and resistance to stresses. It led to the identification and comparison of the different AMF found in maize and soybean soil and root samples. In opposition to what was observed with bacteria and fungi previously, AMF presented a very stable diversity between the different treatments. However, some ribotypes were significantly more present in soil or roots during each growing season. Finally, our whole project demonstrated that P fertilization affected microbial community structure on studied sites. Nevertheless, in each experiment, sampling time also played a substantial role in the organism distribution. Many soil parameters were also monitored and presented a seasonal variation. The sum of possible interactions between these parameters, which in some cases varied according to treatment, would thus have more impact on microbial diversity that the sole fertilization.

Dans cette étude, nous avons isolé et cultivé des bactéries intimement liées aux spores du champignon mycorhizien Glomus irregulare prélevées dans la rhizosphère de plants d’Agrostis stolonifera L. récoltés dans un sol naturel. Le séquençage des 29 morphotypes isolés a révélé la présence de seulement sept taxons bactériens (Variovorax paradoxus, Microbacterium ginsengiosoli, Sphingomonas sp., Bacillus megaterium, B. simplex, B. cereus et Kocuria rhizophila). Des isolats de chacun de ces sept taxons ont ensuite été cultivés in vitro sur le mycélium de G. irregulare afin d’observer par microscopie leur capacité à croitre et à s’attacher au mycélium en absence d’éléments nutritifs autres que ceux fournis par le champignon. Tous les isolats, sauf B. cereus, ont été capables de bien croitre dans le système expérimental et de s’attacher au mycélium en formant des structures ressemblant à des biofilms sur la surface du champignon. Toutefois, B. simplex formait ces structures plus rapidement, soit en 15 jours, alors que les autres isolats les ont formés après 30 jours (K. rhizophila et B. megaterium) ou 45 jours (V. paradoxus, M. ginsengiosoli et Sphingomonas sp.). D’autre part, la technique PCR-DGGE a permis d’analyser la diversité bactérienne associée aux spores. La diversité des taxons associés aux spores de G. irregulare qu’il a été possible d’isoler et de cultiver in vitro a été nettement moindre que celle qui était présente sur la surface des spores, alors que la biodiversité bactérienne totale du sol a été encore beaucoup plus élevée. Les bactéries associées aux champignons mycorhiziens jouent probablement un rôle important dans la capacité des plantes à résister aux stress biotiques et abiotiques auxquels elles sont soumises. / In this study, we isolated and cultivated bacterial cells intimately associated with Glomus irregulare spores in a natural soil Agrostis stolonifera rhizosphere. Sequencing of the 29 morphotypes isolated revealed the presence of only seven bacterial taxa (Variovorax paradoxus, Microbacterium ginsengiosoli, Sphingomonas sp., Bacillus megaterium, B. simplex, B. cereus and Kocuria rhizophila). These seven isolates were cultivated in vitro on the mycelium of G. irregulare to allow microscopic observation of growth and attachment to the mycelium in absence of nutritive sources other than those derived from the fungal mycelium. All isolates but B. cereus were able to grow on the experimental system and to attach to the mycelium to form biofilm-like structures on their surface. However, B. simplex formed these structures more quickly, in 15 days, than the remaining isolates that have formed them only after 30 days (K. rhizophila and B. megaterium) or 45 days (V. paradoxus, M. ginsengiosoli and Sphingomonas sp.). In addition, PCR-DGGE was used to compare bacterial diversity. The bacterial biodiversity associated with spores of G. irregulare that were isolated and cultured in vitro was significantly lower than that present on the spore surface, while total soil bacterial diversity was much higher. The bacteria associated with mycorrhizal fungi probably have an important role in the ability of plants to withstand biotic and abiotic stresses to which they are submitted.

Dans cette étude, nous avons isolé et cultivé des bactéries intimement liées aux spores du champignon mycorhizien Glomus irregulare prélevées dans la rhizosphère de plants d’Agrostis stolonifera L. récoltés dans un sol naturel. Le séquençage des 29 morphotypes isolés a révélé la présence de seulement sept taxons bactériens (Variovorax paradoxus, Microbacterium ginsengiosoli, Sphingomonas sp., Bacillus megaterium, B. simplex, B. cereus et Kocuria rhizophila). Des isolats de chacun de ces sept taxons ont ensuite été cultivés in vitro sur le mycélium de G. irregulare afin d’observer par microscopie leur capacité à croitre et à s’attacher au mycélium en absence d’éléments nutritifs autres que ceux fournis par le champignon. Tous les isolats, sauf B. cereus, ont été capables de bien croitre dans le système expérimental et de s’attacher au mycélium en formant des structures ressemblant à des biofilms sur la surface du champignon. Toutefois, B. simplex formait ces structures plus rapidement, soit en 15 jours, alors que les autres isolats les ont formés après 30 jours (K. rhizophila et B. megaterium) ou 45 jours (V. paradoxus, M. ginsengiosoli et Sphingomonas sp.). D’autre part, la technique PCR-DGGE a permis d’analyser la diversité bactérienne associée aux spores. La diversité des taxons associés aux spores de G. irregulare qu’il a été possible d’isoler et de cultiver in vitro a été nettement moindre que celle qui était présente sur la surface des spores, alors que la biodiversité bactérienne totale du sol a été encore beaucoup plus élevée. Les bactéries associées aux champignons mycorhiziens jouent probablement un rôle important dans la capacité des plantes à résister aux stress biotiques et abiotiques auxquels elles sont soumises. / In this study, we isolated and cultivated bacterial cells intimately associated with Glomus irregulare spores in a natural soil Agrostis stolonifera rhizosphere. Sequencing of the 29 morphotypes isolated revealed the presence of only seven bacterial taxa (Variovorax paradoxus, Microbacterium ginsengiosoli, Sphingomonas sp., Bacillus megaterium, B. simplex, B. cereus and Kocuria rhizophila). These seven isolates were cultivated in vitro on the mycelium of G. irregulare to allow microscopic observation of growth and attachment to the mycelium in absence of nutritive sources other than those derived from the fungal mycelium. All isolates but B. cereus were able to grow on the experimental system and to attach to the mycelium to form biofilm-like structures on their surface. However, B. simplex formed these structures more quickly, in 15 days, than the remaining isolates that have formed them only after 30 days (K. rhizophila and B. megaterium) or 45 days (V. paradoxus, M. ginsengiosoli and Sphingomonas sp.). In addition, PCR-DGGE was used to compare bacterial diversity. The bacterial biodiversity associated with spores of G. irregulare that were isolated and cultured in vitro was significantly lower than that present on the spore surface, while total soil bacterial diversity was much higher. The bacteria associated with mycorrhizal fungi probably have an important role in the ability of plants to withstand biotic and abiotic stresses to which they are submitted.

Initially mycoheterotrophic plants have recently been declining in the wild, even without apparent causes. They are affected by a number of biotic and abiotic factors. The aim of this work is to investigate how selected factors, such as nitrate or symbiotic fungi, may influence their distribution in nature. In particular, the work applies in vitro experiments, molecular determination of fungal symbionts and stable isotope analyses. It describes both the effects of abiotic factor, specifically nitrate, and biotic interactions of initially mycoheterotrophic plants with fungal symbionts. The inhibition of germination by extremely low concentrations of nitrate in asymbiotic in vitro cultures was observed in several orchid species. The degree of sensitivity of each species to nitrate corresponds with the nitrate content of the soil and the nutrient availability requirements of the species according to Ellenberg indicator values. The inhibitory effect of nitrate on orchid germination was also observed in symbiotic in vitro cultures. Out of five tested fungal strains, only one Ceratobasidium was capable of eliminating the inhibitory effect of nitrate. Furthermore, the work reveals that green mixotrophic orchids use photosynthates to nourish the aboveground parts, whereas the belowground parts are...