Diss. (sammanfattning) Umeå : Sveriges lantbruksuniv., 2003. / Härtill 4 uppsatser.

Diss. (sammanfattning) Umeå : Sveriges lantbruksuniv., 2006. / Härtill 4 uppsatser.

Diss. (sammanfattning) Uppsala : Sveriges lantbruksuniversitet,2008. / Härtill 4 uppsatser.

La phytoremédiation s’impose depuis peu comme une alternative de décontamination des sols accessible et soucieuse de l’environnement. Le projet présenté ici a permis de tester, en serre, un traitement sur des sols modérément contaminés aux éléments traces (ET) (Cd, Cu, Pb et Zn), et comportant deux niveaux de contamination organique. L’étude, imbriquée au projet Génorem, a été réalisée à l’aide de plants de Salix purpurea et de mycorhizes Glomus irregulare. Les objectifs étaient 1) d’étudier l’effet de la présence des mycorhizes introduites sur le processus d’absorption des ET, ainsi que l’efficacité générale du traitement ; 2) de déterminer l’influence des processus de transfert et de mise en solution des ET au sein de la rhizosphère sur leur taux d’accumulation par les plantes, et 3) de mesurer l’impact des caractéristiques des sols et de la contamination organique sur la phytoextraction. La présence des mycorhizes inoculées n’a été perceptible ni sur l’accumulation et le transfert des ET, ni sur la production de biomasse. Seules les concentrations solubles de Cu et de Pb se sont avérées moins importantes dans la rhizosphère des sols inoculés que dans la matrice de sol. La performance des plants quant à leur niveau d’accumulation s’est avérée intéressante pour le Cd (moyennes de 2,80 à 10,1 mg/kg) et le Zn (de 54,1 à 410 mg/kg). Ces éléments se sont accumulés surtout dans les parties aériennes des plantes, alors que le Cu et le Pb ont plutôt été stockés dans les racines. Les concentrations solubles avaient pour la plupart augmenté entre le début et la fin de l’expérience, et les différences entre ces concentrations et les concentrations totales, dans la rhizosphère et la matrice, ont démontré que le temps de réponse du sol pour fournir aux plantes les éléments biodisponibles n’était pas un facteur limitant du système dans la plupart des cas. Les concentrations initiales totales dans les sols ont été corrélées à celles dans les parties des plantes uniquement pour le Cu dans les tiges (R2 : 0,54) et les racines (R2 : 0,58) des saules. Les facteurs de fertilité ont permis d’expliquer une part de la variance des concentrations (10 % du total de 25 % pour le Cd, 49 % du total de 36 % pour le Cu-aérien, et 28 % du total de 71 % pour le Cu-souterrain) et des quantités (15 % du total de 30 % pour le Cd) accumulées. La contamination mixte (organique et inorganique) n’a pas entravé l’extraction des ET, ni la dégradation des composés organiques. / Phytoremediation is a relatively new, low cost and environmentally friendly alternative method to decontaminating soils. This research, which involved Salix purpurea plants and Glomus irregulare mycorrhizae, was conducted through the Génorem project in a greenhouse. We tested a treatment on soils, which were contaminated with moderate amounts of trace elements (TE) (Cd, Cu, Pb and Zn), as well as with organic compounds (on two levels of concentration). The objectives were 1) to study the effect of the introduced mycorrhizae on TE’s absorption process, as well as the general treatment efficiency; 2) to determine the influence of TE transfer and solubilization processes at work within the rhizosphere on their accumulation rates by the plants, and 3) to measure the impact of soil characteristics as well as the organic contamination on phytoextraction. The presence of inoculate mycorrhizae was not noticeable on the accumulation or transfer of TE, nor was it on the biomass production. However, soluble concentrations of Cu and Pb were less important in the soil rhizosphere of the inoculated soils than in the soil matrix. Plants performance in regard to their accumulation was considerable for Cd (average of 2,80 to 10,1 mg / kg) and Zn (from 54.1 to 410 mg / kg). These elements were mainly accumulated in aboveground tissues, while Cu and Pb were instead stored in the roots. Additionally, most of the soluble concentrations had increased between the beginning and the end of the experiment. This, combined with the differences between the soluble and total-recoverable concentrations in the rhizosphere and the matrix, showed that the soil’s response in terms of time to provide bioavailable TE to plants was not a limiting factor of the system in most cases. Total initial soil concentrations were only correlated to concentrations plant tissues in the case of Cu within stems (R2 : 0,54), and within roots (R2 : 0,58). Fertility and availability factors allowed to explain parts of the variance in concentrations (10% of the total 25% for Cd, and 49% of the total 36% for Cu-aboveground, and 28% of the total 71% for Cu-aboveground), and content (15% of the total 30% for Cd) accumulated. Mixed contamination (organic and inorganic) did not significantly impede the TE extraction, neither the organic compounds degradation.

Os hormônios vegetais etileno e ABA são possíveis reguladores chave do desenvolvimento das micorrizas arbusculares (MAs). O primeiro objetivo deste trabalho foi avaliar o desenvolvimento da MA em mutantes superprodutores e insensíveis ao etileno (epinastic e Never ripe, respectivamente) e verificar se enzimas relacionadas ao sistema de defesa vegetal são reguladas diferencialmente pelo etileno durante o desenvolvimento da simbiose A colonização de raízes de epinastic (epi) por G. clarum foi significativamente reduzida quando comparada com Micro-Tom (MT), além disso, a funcionalidade da simbiose, demonstrada pelo acúmulo de transcritos de LePT4, foi fortemente reduzida quando comparada com MT. Por outro lado, o padrão de colonização e funcionalidade da simbiose no mutante Nr não diferiu de MT. Indução no acúmulo de transcritos de genes que codificam quitinases e enzimas relacionadas ao estresse oxidativo foi observada no mutante epi, enquanto o acúmulo de transcritos de uma -1,3-glucanase foi fortemente induzido no mutante Nr. O segundo objetivo foi verificar se a colonização reduzida observada em mutantes deficientes em ABA é dependente da produção do etileno. Para tanto, mutantes que diferem na deficiência de ABA e produzem diferentes quantidades de etileno (notabilis e sitiens) e o mutante epi foram inoculados com G. clarum e tratados com aminoethoxivinilglicina (AVG), um inibidor de etileno. A colonização intrarradicular de G. clarum nos mutantes epi e notabilis (not) foi significativamente reduzida quando comparada com MT, sendo que o mutante sitiens (sit) foi o menos susceptível à colonização em relação aos demais genótipos. A aplicação de AVG a 10 µM restaurou completamente a colonização em epi, mas não nos mutantes deficientes em ABA. O acúmulo de transcritos de LePT4 confirmou os resultados de colonização. Os genes que codificam enzimas chave na biossíntese do etileno, LeACS2 e LeACO4, foram regulados positivamente pela inoculação com G. clarum, e a restauração do desenvolvimento da MA observada em epi que recebeu AVG, provavelmente envolveu ACC oxidase (LeACO4). O acúmulo de transcritos dos genes que codificam ACC oxidase (LeACO1 e LeACO4) foi induzido significativamente em sit, mostrando que a superprodução de etileno observada neste mutante pode envolver a regulação da ACC oxidase. Por fim, indução no acúmulo de transcritos do gene LeNCED foi observada em todos os genótipos micorrizados e em raízes de epi que receberam AVG, demonstrando que LeNCED é regulado positivamente pela inoculação com G. clarum e que o etileno regula a expressão deste gene. Os resultados demonstram que etileno e ABA estão envolvidos na regulação do desenvolvimento de MA e que etileno modula a expressão de genes relacionados ao sistema de defesa vegetal permitindo o estabelecimento de uma simbiose funcional. / The plant hormones ethylene and ABA are probably key developmental regulators of arbuscular mycorrhizae (AM). The first objective of this work was to evaluate the development of MA in overproducing and insensitive mutants to ethylene (epinastic and Never ripe, respectively) and verify if enzymes related to plant defense system are differentially regulated by ethylene during the symbiosis development. The colonization of epinastic (epi) roots by G. clarum was significantly reduced when compared with Micro-Tom (MT), moreover, the functionality of the symbiosis as demonstrated by transcripts accumulation of LePT4, was strongly reduced when compared with MT. Still, the pattern of colonization and functionality of the symbiosis in the Nr mutant did not differ from MT. Induction in the transcripts accumulation of genes encoding chitinase and enzymes related to oxidative stress was observed in epi mutant, whereas transcripts accumulation of -1, 3-glucanase was strongly induced in the Nr mutant. The second objective was to determine whether the reduced colonization observed in mutants deficient in ABA is dependent on ethylene production. For this purpose, mutants that differ in ABA deficiency and produce different amounts of ethylene (notabilis and sitiens) and the epi mutant were inoculated with G. clarum and treated with an ethylene inhibitor called aminoethoxivinilglicin (AVG). The intraradical colonization of G. clarum in epi and notabilis (not) mutants was significantly reduced when compared with MT and sitiens mutant was less susceptible to colonization in comparison with other genotypes. The application of AVG to 10 µM completely restored the colonization in the epi but not in mutants deficient in ABA. The transcripts accumulation of LePT4 confirmed the results of colonization. The genes that encode key enzymes in the biosynthesis of ethylene, LeACS2 and LeACO4, were positively regulated by inoculation with G. clarum and the restoration in the development of MA observed in epi which received AVG probably involved ACC oxidase (LeACO4). Transcripts accumulation of genes that encode ACC oxidase (LeACO1 and LeACO4) were induced significantly in sit showing that overproduction of ethylene in this mutant can involve ACC oxidase. Finally, the induction in the accumulation of gene transcripts LeNCED was observed in all genotypes and mycorrhizal roots of epi which received AVG demonstrating that LeNCED is regulated positively by the inoculation with G. clarum and that ethylene regulates the expression of this gene. Results show that ethylene and ABA are involved in regulating of MA development and ethylene modulates the expression of genes related to the plant defense system allowing the establishment and functionality of the symbiosis.

Os hormônios vegetais etileno e ABA são possíveis reguladores chave do desenvolvimento das micorrizas arbusculares (MAs). O primeiro objetivo deste trabalho foi avaliar o desenvolvimento da MA em mutantes superprodutores e insensíveis ao etileno (epinastic e Never ripe, respectivamente) e verificar se enzimas relacionadas ao sistema de defesa vegetal são reguladas diferencialmente pelo etileno durante o desenvolvimento da simbiose A colonização de raízes de epinastic (epi) por G. clarum foi significativamente reduzida quando comparada com Micro-Tom (MT), além disso, a funcionalidade da simbiose, demonstrada pelo acúmulo de transcritos de LePT4, foi fortemente reduzida quando comparada com MT. Por outro lado, o padrão de colonização e funcionalidade da simbiose no mutante Nr não diferiu de MT. Indução no acúmulo de transcritos de genes que codificam quitinases e enzimas relacionadas ao estresse oxidativo foi observada no mutante epi, enquanto o acúmulo de transcritos de uma -1,3-glucanase foi fortemente induzido no mutante Nr. O segundo objetivo foi verificar se a colonização reduzida observada em mutantes deficientes em ABA é dependente da produção do etileno. Para tanto, mutantes que diferem na deficiência de ABA e produzem diferentes quantidades de etileno (notabilis e sitiens) e o mutante epi foram inoculados com G. clarum e tratados com aminoethoxivinilglicina (AVG), um inibidor de etileno. A colonização intrarradicular de G. clarum nos mutantes epi e notabilis (not) foi significativamente reduzida quando comparada com MT, sendo que o mutante sitiens (sit) foi o menos susceptível à colonização em relação aos demais genótipos. A aplicação de AVG a 10 µM restaurou completamente a colonização em epi, mas não nos mutantes deficientes em ABA. O acúmulo de transcritos de LePT4 confirmou os resultados de colonização. Os genes que codificam enzimas chave na biossíntese do etileno, LeACS2 e LeACO4, foram regulados positivamente pela inoculação com G. clarum, e a restauração do desenvolvimento da MA observada em epi que recebeu AVG, provavelmente envolveu ACC oxidase (LeACO4). O acúmulo de transcritos dos genes que codificam ACC oxidase (LeACO1 e LeACO4) foi induzido significativamente em sit, mostrando que a superprodução de etileno observada neste mutante pode envolver a regulação da ACC oxidase. Por fim, indução no acúmulo de transcritos do gene LeNCED foi observada em todos os genótipos micorrizados e em raízes de epi que receberam AVG, demonstrando que LeNCED é regulado positivamente pela inoculação com G. clarum e que o etileno regula a expressão deste gene. Os resultados demonstram que etileno e ABA estão envolvidos na regulação do desenvolvimento de MA e que etileno modula a expressão de genes relacionados ao sistema de defesa vegetal permitindo o estabelecimento de uma simbiose funcional. / The plant hormones ethylene and ABA are probably key developmental regulators of arbuscular mycorrhizae (AM). The first objective of this work was to evaluate the development of MA in overproducing and insensitive mutants to ethylene (epinastic and Never ripe, respectively) and verify if enzymes related to plant defense system are differentially regulated by ethylene during the symbiosis development. The colonization of epinastic (epi) roots by G. clarum was significantly reduced when compared with Micro-Tom (MT), moreover, the functionality of the symbiosis as demonstrated by transcripts accumulation of LePT4, was strongly reduced when compared with MT. Still, the pattern of colonization and functionality of the symbiosis in the Nr mutant did not differ from MT. Induction in the transcripts accumulation of genes encoding chitinase and enzymes related to oxidative stress was observed in epi mutant, whereas transcripts accumulation of -1, 3-glucanase was strongly induced in the Nr mutant. The second objective was to determine whether the reduced colonization observed in mutants deficient in ABA is dependent on ethylene production. For this purpose, mutants that differ in ABA deficiency and produce different amounts of ethylene (notabilis and sitiens) and the epi mutant were inoculated with G. clarum and treated with an ethylene inhibitor called aminoethoxivinilglicin (AVG). The intraradical colonization of G. clarum in epi and notabilis (not) mutants was significantly reduced when compared with MT and sitiens mutant was less susceptible to colonization in comparison with other genotypes. The application of AVG to 10 µM completely restored the colonization in the epi but not in mutants deficient in ABA. The transcripts accumulation of LePT4 confirmed the results of colonization. The genes that encode key enzymes in the biosynthesis of ethylene, LeACS2 and LeACO4, were positively regulated by inoculation with G. clarum and the restoration in the development of MA observed in epi which received AVG probably involved ACC oxidase (LeACO4). Transcripts accumulation of genes that encode ACC oxidase (LeACO1 and LeACO4) were induced significantly in sit showing that overproduction of ethylene in this mutant can involve ACC oxidase. Finally, the induction in the accumulation of gene transcripts LeNCED was observed in all genotypes and mycorrhizal roots of epi which received AVG demonstrating that LeNCED is regulated positively by the inoculation with G. clarum and that ethylene regulates the expression of this gene. Results show that ethylene and ABA are involved in regulating of MA development and ethylene modulates the expression of genes related to the plant defense system allowing the establishment and functionality of the symbiosis.

As plantações brasileiras de eucaliptos apresentam um dos maiores valores da produção primária bruta (PPB) entre os diversos ecossistemas do mundo. Nos ecossistemas florestais, o fluxo total de carbono no solo é constituído em grande parte pela produção e mortalidade das raízes finas (diâmetro <= 2 mm), podendo representar 20-60% da PPB. Esse estudo teve como objetivo principal avaliar a dinâmica e o prazo de vida (PV) das raízes finas, através do método não-destrutivo de minirhizotrons (MR), em plantio de E. grandis (2-4 anos de idade) cultivados em latossolos. Os objetivos específicos foram divididos em três capítulos: i-) avaliar a influência da textura (20 e 40% de argila) na dinâmica das raízes finas, em camadas superficiais do solo (0-30 cm); ii-) estudar a dinâmica das raízes no solo arenoso até 6 m de profundidade; iii-) investigar as associações simbióticas das raízes finas de eucaliptos (2 anos do plantio) com fungos micorrízicos arbusculares (FMA) na superfície de solos com texturas contrastantes, e fungos ectomicorrízicos (ECM) em camadas profundas do solo arenoso (4 anos do plantio). As imagens da interface solo-MR foram produzidas quinzenalmente, através de um scanner portátil no interior dos tubos MR em um período de dois anos. O software WinRHIZO Tron foi utilizado para medir o comprimento e diâmetro das raízes finas que apareceram durante o estudo. O conteúdo de água no solo foi monitorado, até 10 m de profundidade com auxilio de sensores CS615 (Campbell), continuamente durante todo o período de estudo. As avaliações de FMA e ECM foram realizadas através de amostragens destrutivas do solo. Os resultados observados com os MR mostraram que a textura do solo teve influência direta no comprimento radicular na camada de 0-30 cm, onde o solo arenoso apresentou valores superiores em relação ao solo argiloso durante todo o estudo. Ao final de dois anos, o solo arenoso teve o dobro de comprimento total acumulado em relação ao solo argiloso, com 30 m m-2 imagem. Na mesma profundidade, as raízes finas provenientes do solo arenoso tiveram uma maior colonização por FMA em relação ao solo argiloso, com médias de 38,5 e 10,5%, respectivamente. Uma fraca correlação entre umidade do solo e a dinâmica de raízes para textura e profundidade do solo foi observada. As médias de elongação diária das raízes finas foram de 0,10 e 0,22 cm dia-1 na camada de 0-2 e 2-6 m de profundidade, respectivamente. A elongação diária máxima no perfil do solo foi de 3,5 cm dia-1 na camada de 5-6 m. Através de análises moleculares, foi identificada uma espécie de ECM (Pisolithus) na profundidade de 2-3 m. No geral, o PV e a taxa de ciclagem das raízes finas de eucaliptos foram em torno de 500 dias e 0,70 ano-1, respectivamente, não sofrendo influência significativa da textura e profundidade do solo. Quando as raízes finas foram analisadas por classe de diâmetro (< 0,03, 0,3-0,5 e 0,5-2,0 mm) e micorrização, observou-se uma diferença significativa na sua longevidade. As árvores de eucaliptos apresentaram uma alta capacidade de exploração do solo / Brazilian eucalyptus plantations are among the forest ecosystems in the world with the highest gross primary productivity (GPP). The total belowground carbon allocation, with mainly production and mortality of fine roots (diameter <= 2 mm), can account for 20-60% of GPP in forest ecosystems. This study aimed to evaluate the dynamic and lifespan of fine roots in E. grandis plantations (2-4 years old) established in Oxisol soils, using the non-destructive method of minirhizotrons (MR). The specific objectives of this study were divided into three chapters: i-) to evaluate the influence of soil texture (20 and 40% clay) in the dynamics of fine roots in the topsoil (0-30 cm); ii-) to study the dynamics of the fine roots in a sandy soil up to 6 m deep; iii-) to investigate the symbiotic associations of Eucalyptus fine roots (2 years old after planting) with arbuscular mycorrhizal fungi (AMF) in the upper soil layers for two contrasting soil textures, and ectomycorrhizal fungi (ECM) in deep layers of a sandy soil (4 years old after planting). Images at the interface soil-MR were taken fortnightly through a portable scanner within the MR tube over a period of two years. The WinRhizo Tron software was used to measure the length and diameter of the fine roots that appeared throughout the study period. Soil water contents were continuously monitored down to a depth of 10 m using CS615 sensors (Campbell). AMF and ECM were studied sampling soil cores. The MR technique showed that the soil texture direct influenced fine root growth in the 0-30 cm layer, with higher values in the sandy soil than in the clayey soil throughout the study period. After two years, the sandy soil reached a total cumulative length of 30 m m-2 image, which was twice as high as the clayey soil. At the same depth, fine roots had a greater colonization by AMF in the sandy soil compared to the clayey soil, with means of 38,5 and 10,5%, respectively. A weak correlation between relative extractable water and dynamics of fine roots was observed, whatever the soil texture and depth. The means of daily elongation of fine roots were 0,10 cm day-1 in the 0-2 m soil layers and 0,22 cm day-1 in the 2-6 m soil layer. The maximum daily root elongation throughout the soil profiles reached 3,5 cm day-1 at a depth of 5-6 m. A specie of ECM (Pisolithus) was identified through molecular analyzes at a depth 2-3 m. In general, the lifespan and the turnover rates of Eucalyptus fine roots were about 500 days and 0.70 yr-1, respectively, and the influences of soil texture and soil depth were not significant. The fine root longevity was significantly affected by the diameter class (< 0,3, 0,3-0,5 and 0,5-2,0 mm) and the mycorrhizal status, there was a significant difference in their longevity. The Eucalyptus trees exhibited a remarkably high capacity of soil exploration in the Oxisol studied

As plantações brasileiras de eucaliptos apresentam um dos maiores valores da produção primária bruta (PPB) entre os diversos ecossistemas do mundo. Nos ecossistemas florestais, o fluxo total de carbono no solo é constituído em grande parte pela produção e mortalidade das raízes finas (diâmetro <= 2 mm), podendo representar 20-60% da PPB. Esse estudo teve como objetivo principal avaliar a dinâmica e o prazo de vida (PV) das raízes finas, através do método não-destrutivo de minirhizotrons (MR), em plantio de E. grandis (2-4 anos de idade) cultivados em latossolos. Os objetivos específicos foram divididos em três capítulos: i-) avaliar a influência da textura (20 e 40% de argila) na dinâmica das raízes finas, em camadas superficiais do solo (0-30 cm); ii-) estudar a dinâmica das raízes no solo arenoso até 6 m de profundidade; iii-) investigar as associações simbióticas das raízes finas de eucaliptos (2 anos do plantio) com fungos micorrízicos arbusculares (FMA) na superfície de solos com texturas contrastantes, e fungos ectomicorrízicos (ECM) em camadas profundas do solo arenoso (4 anos do plantio). As imagens da interface solo-MR foram produzidas quinzenalmente, através de um scanner portátil no interior dos tubos MR em um período de dois anos. O software WinRHIZO Tron foi utilizado para medir o comprimento e diâmetro das raízes finas que apareceram durante o estudo. O conteúdo de água no solo foi monitorado, até 10 m de profundidade com auxilio de sensores CS615 (Campbell), continuamente durante todo o período de estudo. As avaliações de FMA e ECM foram realizadas através de amostragens destrutivas do solo. Os resultados observados com os MR mostraram que a textura do solo teve influência direta no comprimento radicular na camada de 0-30 cm, onde o solo arenoso apresentou valores superiores em relação ao solo argiloso durante todo o estudo. Ao final de dois anos, o solo arenoso teve o dobro de comprimento total acumulado em relação ao solo argiloso, com 30 m m-2 imagem. Na mesma profundidade, as raízes finas provenientes do solo arenoso tiveram uma maior colonização por FMA em relação ao solo argiloso, com médias de 38,5 e 10,5%, respectivamente. Uma fraca correlação entre umidade do solo e a dinâmica de raízes para textura e profundidade do solo foi observada. As médias de elongação diária das raízes finas foram de 0,10 e 0,22 cm dia-1 na camada de 0-2 e 2-6 m de profundidade, respectivamente. A elongação diária máxima no perfil do solo foi de 3,5 cm dia-1 na camada de 5-6 m. Através de análises moleculares, foi identificada uma espécie de ECM (Pisolithus) na profundidade de 2-3 m. No geral, o PV e a taxa de ciclagem das raízes finas de eucaliptos foram em torno de 500 dias e 0,70 ano-1, respectivamente, não sofrendo influência significativa da textura e profundidade do solo. Quando as raízes finas foram analisadas por classe de diâmetro (< 0,03, 0,3-0,5 e 0,5-2,0 mm) e micorrização, observou-se uma diferença significativa na sua longevidade. As árvores de eucaliptos apresentaram uma alta capacidade de exploração do solo / Brazilian eucalyptus plantations are among the forest ecosystems in the world with the highest gross primary productivity (GPP). The total belowground carbon allocation, with mainly production and mortality of fine roots (diameter <= 2 mm), can account for 20-60% of GPP in forest ecosystems. This study aimed to evaluate the dynamic and lifespan of fine roots in E. grandis plantations (2-4 years old) established in Oxisol soils, using the non-destructive method of minirhizotrons (MR). The specific objectives of this study were divided into three chapters: i-) to evaluate the influence of soil texture (20 and 40% clay) in the dynamics of fine roots in the topsoil (0-30 cm); ii-) to study the dynamics of the fine roots in a sandy soil up to 6 m deep; iii-) to investigate the symbiotic associations of Eucalyptus fine roots (2 years old after planting) with arbuscular mycorrhizal fungi (AMF) in the upper soil layers for two contrasting soil textures, and ectomycorrhizal fungi (ECM) in deep layers of a sandy soil (4 years old after planting). Images at the interface soil-MR were taken fortnightly through a portable scanner within the MR tube over a period of two years. The WinRhizo Tron software was used to measure the length and diameter of the fine roots that appeared throughout the study period. Soil water contents were continuously monitored down to a depth of 10 m using CS615 sensors (Campbell). AMF and ECM were studied sampling soil cores. The MR technique showed that the soil texture direct influenced fine root growth in the 0-30 cm layer, with higher values in the sandy soil than in the clayey soil throughout the study period. After two years, the sandy soil reached a total cumulative length of 30 m m-2 image, which was twice as high as the clayey soil. At the same depth, fine roots had a greater colonization by AMF in the sandy soil compared to the clayey soil, with means of 38,5 and 10,5%, respectively. A weak correlation between relative extractable water and dynamics of fine roots was observed, whatever the soil texture and depth. The means of daily elongation of fine roots were 0,10 cm day-1 in the 0-2 m soil layers and 0,22 cm day-1 in the 2-6 m soil layer. The maximum daily root elongation throughout the soil profiles reached 3,5 cm day-1 at a depth of 5-6 m. A specie of ECM (Pisolithus) was identified through molecular analyzes at a depth 2-3 m. In general, the lifespan and the turnover rates of Eucalyptus fine roots were about 500 days and 0.70 yr-1, respectively, and the influences of soil texture and soil depth were not significant. The fine root longevity was significantly affected by the diameter class (< 0,3, 0,3-0,5 and 0,5-2,0 mm) and the mycorrhizal status, there was a significant difference in their longevity. The Eucalyptus trees exhibited a remarkably high capacity of soil exploration in the Oxisol studied

La symbiose mycorhizienne à arbuscule (MA) est une association mutualiste s’établissant entre les racines des plantes et des champignons du sol appartenant à l’embranchement des Gloméromycètes. Dans cette association, le champignon agit comme un fertilisant naturel, fournissant à la plante divers minéraux (phosphore, mais aussi azote et soufre …) en échange de sources de carbone indispensables à son développement. Une caractéristique originale de ces champignons est leur très large spectre d’hôte : de l’ordre de 80% des espèces végétales ont l’aptitude à former cette symbiose, et certains espèces de champignons MA ne semblent pas avoir de limitation de spectre d’hôte. Les champignons MA possèderaient-ils des « clés universelles » de compatibilité cellulaire avec leur hôte, ou de contournement de l’immunité végétale ? Pour aborder cette problématique, nous avons entrepris l’étude du sécrétome du champignon MA Rhizophagus irregularis dont plusieurs données génomiques étaient disponibles. Les microorganismes eucaryotes sécrètent en effet dans leur environnement des protéines agissant sur leurs structures exogènes (paroi cellulaire), leur environnement, et pouvant agir sur l’immunité des cellules hôtes. Ces protéines sécrétées (SPs) sont dans ce dernier cas appelées « effecteurs ». Sur la base de deux assemblages différents, un catalogue de 872 SPs de R. irregularis (RiSPs) a été défini pour lesquelles les profils d’expression dans trois plantes hôtes ont été comparés. Nous avons également comparé ces SPs à celles que nous avons définies sur une autre espèce de champignon MA, Gigaspora rosea. Après enrichissement du catalogue de RiSPs avec des séquences de petite taille identifiées sur un assemblage transcriptomique propre, puis sélection des candidats dont les cadres de lecture sont robustes et présentant un niveau d’expression élevé (FC>10) dans les 3 hôtes testés, un jeu de 33 RiSPs d’intérêt a été défini, dont 18 ont été sélectionnées pour effectuer des analyses fonctionnelles. En absence de protocole de transformation de ces champignons, l’analyse fonctionnelle a porté sur la cytolocalisation de protéines de fusion RiSP:citrine dans les cellules végétales par surexpression dans des feuilles de tabac et des racines de luzerne tronquée. Différents compartiments cellulaires sont ciblés par ces RiSPs, très souvent le compartiment vacuolaire. Des approches par surexpression in planta de plusieurs candidats RiSP ont permis d’identifier une activité stimulatrice de 3 RiSPs sur l’établissement de la symbiose. Parallèlement, des essais de stimulation de la symbiose MA par apport exogène de différents SPs sur plantules en cours de mycorhization en chambre ont été initiés. Ils devront être poursuivis sur les 3 candidats issus du crible de surexpression. En perspective, l’évaluation de la spécificité d’action de ces SPs sur la symbiose MA comparativement à d’autres interactions plante-champignon ouvrira la voie à des essais d’application au champ. / Arbuscular mycorrhizal (AM) symbiosis is a mutualistic association established between plant roots and soil fungi belonging to the phylum Glomeromycota. In this association, the fungus acts as a natural fertilizer, supplying the plant with various minerals (phosphorus, but also nitrogen and sulfur) in exchange to carbon sources essential for its development. An original feature of these fungi is their very broad host spectrum: c.a. 80% of plant species have the ability to form this symbiosis, and some species of AM fungi do not seem to have a restrictive host spectrum. Would MA fungi possess "universal keys" for cell compatibility with their host, or to by-pass plant immunity? To address this problem, we studied the secretome of the AM fungus Rhizophagus irregularis from which several genomic data were available. Eukaryotic microorganisms indeed secrete in their environment proteins acting on their exogenous structures (cell wall), on their environment (nutrient recruitment), and even on host plant cell immunity. These last secreted proteins (SPs) are defined as effectors. Based on two different assemblies, a repertoire of 872 SPs of R. irregularis was defined for which transcriptional expression profiles obtained in three hosts were compared, as well with SPs from another species of AM fungus, Gigaspora rosea. After adding sequences of small size identified from an in-house transcriptomic assembly, screening unambiguous open reading frame, and selecting strongly expressed candidates (FC> 10) in the 3 plant hosts analyzed, a set of 33 RiSPs of interest was defined, of which 18 were selected for functional analysis. As genetic transformation protocol is unavailable for AM fungi, RiSP:citrine fusion proteins were overexpressed in tobacco leaves and barrel medic roots for plant cell localization. Different cell compartments were targeted by these RiSPs, and often localised in the vacuolar compartment. In planta overexpression of several candidates allowed identifying 3 RiSPs that stimulate the establishment of the symbiosis. In the same time, attempts to enhance MA symbiosis by addition of exogenous RiSPs on seedlings during mycorrhizal establishment were initiated. Such assays should be pursued on the 3 active candidates revealed by overexpression assays. Evaluating the specificity of action of these RiSPs on AM symbiosis compared to other plant-fungus interactions will open the way to field trials.

Striga, a genus of obligate parasitic weeds in the family Orobanchaceae, has been identified as the most important biological factor limiting agricultural productivity in sub-Saharan Africa. Germination of Striga seeds is triggered by strigolactone root exudates from host plants. Strigolactones also induce hyphal branching in arbuscular mycorrhizal (AM) fungi, which are important for plant uptake of phosphorus in low phosphorus soils. Mechanisms of Striga resistance based on reduced strigolactone production may also convey resistance to AM fungi which would require higher inputs of phosphorus fertilizer to attain optimal crop growth. There is evidence for genetic differences in mycorrhizal responsiveness in other grain crops; therefore it is beneficial for breeders to be aware of these differences when developing Striga-resistant sorghum cultivars. This research aims to determine phosphorus and mycorrhizal responsiveness of sorghum genotypes important for or developed by breeders working on Striga resistance. Phosphorus response curves were determined for twelve sorghum genotypes using pasteurized low phosphorus soil amended to achieve four different phosphorus levels. Simple linear regression was performed on root and shoot dry weight data. Results indicate variability in phosphorus responsiveness within Striga resistant and susceptible genotypes. Seven of these genotypes were selected for continued research, which analyzed responsiveness to phosphorous and differences in mycorrhizal responsiveness in relation to reported mechanisms of Striga resistance. Treatments included three levels of phosphorus amendments and the addition of Funneliformis mosseae inoculum. All genotypes were strongly responsive to P amendment when grown without AM fungi and showed a decrease in responsiveness to P when inoculated with F. mosseae. Trends for all genotypes indicate a greater uptake of P, Zn, and Mg by mycorrhizal plants as compared to nonmycorrhizal plants. All seven genotypes were responsive to mycorrhizae, with a significant increase in biomass for all genotypes, especially at the lowest phosphorus level. The responsiveness to the mycorrhizal fungus does not appear to be directly related to the susceptibility of genotypes to the parasitic weed Striga. / Graduation date: 2012