Made available in DSpace on 2014-06-11T19:29:42Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-08-10Bitstream added on 2014-06-13T20:19:53Z : No. of bitstreams: 1 faria_tm_me_ilha.pdf: 2501450 bytes, checksum: 35741ba09de4fee809d4e55df75c6446 (MD5) / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / A competição entre as plantas é uma concorrência constante entre as espécies que vivem em comunidades, porém a capacidade de certas espécies interferirem na germinação de sementes e desenvolvimento de plantas por meio dos efeitos alelopáticos provenientes de substâncias que liberam na atmosfera ou, quase sempre, no solo, ainda é pouco compreendida no meio agronômico. Muitas destas espécies vegetais podem influenciar a vegetação de um local, sucessão de plantas, indução de dormência, preservação e germinação de sementes, atividade microbiana, produtividade de culturas, entre outros fatores. Visando melhor entender possíveis interferências de restos vegetais nos cultivos subseqüentes, este trabalho tem como objetivo avaliar os possíveis efeitos alelopáticos de sorgo (Sorghum bicolor L. Moench) milheto (Pennisetum glaucum (L.) R. Brown) e soja (Glycine max L.), como cobertura vegetal, sobre a emergência, micorrização, atividade microbiana e crescimento de milho (Zea mays L.), soja (G. max L.) e feijão (Phaseolus vulgaris L.). Os experimentos foram conduzidos nas dependências da fazenda experimental da UNESP/Campus de Ilha Solteira. Foram avaliadas a emergência de plântulas e sua taxa de crescimento, população final da cultura, ocorrência, identificação e quantificação de plantas daninhas nas áreas, produção de grãos, colonização micorrízica e número de esporos, carbono de CO2 liberado. Os compostos alelopáticos liberados na decomposição das coberturas de milheto, sorgo e soja, interferirem positivamente na COL micorrízica em milho e negativamente em feijão e soja. As coberturas vegetais testadas liberam aleloquímicos que desestabilizam o meio, proporcionado um aumento na esporulação micorrízica nas culturas testadas. Os agentes aleloquímicos liberados pelas coberturas vegetais... / The competition between plants species that lives in the same community is constant. However some species can interfere in the seed germination and plant development through allelophatic effects that occurs because plants release substances in atmosphere or in the soil. This capacity that certain species have is not well understood in the agricultural area. Many of those species can influence the local vegetation, ensuing plantations, dormancy, preservation and seed germination of seeds, microbial activity, cultures productivity and other factors. To a better understanding of possible interferences of vegetable rests in ensuing croop, this work aims to evaluate possible allelophatic effects of sorghum (Sorghum bicolor L. Moench), millet (Pennisetum glaucum (L.) R. Brown) and soy (Glycine max L.) when used as vegetable covers under mycorrhization, microbial activity and the growth of bean (Phaseolus vulgaris L.). The experiments were carried out in the experimental farm of UNESP/Campus de Ilha Solteira and could evaluate the seedling emergence and its growth rate, final population of croop; occurrence, identification and quantification of weed plants in the area, seed production, mycorrhizal colonization, spore number and carbon of CO2 released. In the decomposition of the covers millet, sorghum and soy some allelophatic compounds were released and interfered positively in the mycorrhizal harvest of corn but interfered negatively for bean and soy. The vegetable covers that were tested released allelochemics that destabilized the environment providing an increase of mycorrhizal fungi in tested cultures. The allelochemic agents released by the vegetable covers interfered negatively in the carbon of CO2 released. When used as a dead cover sorghum interferes negatively... (Complete abstract click electronic access below)

I have studied arbuscular mycorrhizal symbiosis of 44 herbaceous invasive neophytes occuring in the Czech Republic. My results show that about 70% of the examined species are capable to form symbiotic association with arbuscular mycorrhizal fungi (AMF) in the field. To my knowledge, mycorrhizal status of 23 invasive species is reported here for the first time. I predicted, based on the hypothesis of Urcelay & Díaz (2003), that the intensity of AMF in the roots of invasive species will be correlated with that of the native dominant species of invaded plant community, but collected data did not support this hypothesis. In addition, the effects of habitat and community characteristics on the intensity of AMF colonization of the invasive species{\crq} roots were tested. My results show that, at the within-species level, plants in the habitats with higher light and temperature conditions have less colonized roots whereas intensity of mycorrhizal colonization increases with habitat moisture. At the among-species level, invasive species occurring in the habitats with elevated nitrogen availability have higher mycorrhizal colonization of their roots. The effect of nitrogen availability is revealed at the among-species level and stays significant even after phylogenetic correction, suggesting this is an evolutionary adaptation rather than a phenotypic plasticity.

As plantas apresentam diversas adaptações fisiológicas à baixa disponibilidade de fósforo (Pi) do solo. Este trabalho discute os custos fisiológicos e energéticos associados com essas estratégias, focado nas respostas da cana-de-açúcar (Saccharum spp.) à disponibilidade de Pi durante a simbiose com micorrizas arbusculares (Glomus clarum). Esses custos são importantes componentes para a adaptação a solos com baixo Pi, afetando a aquisição e conteúdo de fósforo; o crescimento e concentração de açúcares em tecidos vegetais. Plantas de cana-de-açúcar foram cultivadas em vasos com ou sem micorrizas (Glomus clarum), e sob a disponibilidade de baixo (20 mg kg-1) ou alto (202 mg kg-1) fósforo. Raízes e parte-aérea foram coletadas para as análises após 14, 30, 44 e 58 dias pós-inoculação (dpi) com Glomus clarum . A condição de BP causou a deficiência de Pi nas plantas, micorrízicas ou não. As plantas sob AP continham um teor foliar de Pi adequado, e partir dos 44 dpi acumularam pelo menos 6 vezes mais Pi parte-aérea, do que as cultivadas sob BP, efeito mais evidente nas micorrízicas. A eficiência de absorção, indicada pelo acúmulo de Pi na parte-aérea a uma dada biomassa da raiz, foi igual para todos os tratamentos, sugerindo que as eficiências radicular e micorrízica da absorção de Pi foram similares, independentemente da doses de Pi. A disponibilidade de fósforo não afetou a biomassa total das plantas, sendo as cultivadas sob BP mais eficientes na utilização deste nutriente. Por outro lado, as plantas micorrízicas suplementadas com BP apresentaram maior crescimento da raiz e redução na parte-aérea, resultando no aumento da proporção raiz:parte-aérea. Aos 58 dpi, a glicose, frutose e sacarose presente nas folhas de plantas micorrízicas foi 3,8, 2,3 e 2,4 vezes respectivamente mais concentrada do que nas não micorrízicas. Esses resultados sugerem que, nestas condições experimentais, o estabelecimento da simbiose não foi uma associação mutualística típica, afetando o perfil de crescimento e a alometria da cana cultivada com BP. As concentrações de fotoassimilados na folhas de planta micorrízicas indicam que houve aumentos nas taxas fotossintéticas, mas isso não resultou no maior crescimento do macrosimbionte. A tecnologia de amplificação quantitativa de transcritos reversos (RT-PCR) se tornou uma opção para a validação funcional de genes, com alta sensibilidade, acurada quantificação e eficácia. A quantificação relativa da expressão gênica é conseqüentemente fácil e determina a expressão de um gene em relação a outro expresso e relativamente constante. Neste trabalho foi analisada a variabilidade de expressão dos genes de cana-de-açúcar codificando a actina (Actina), gliceraldeido fosfato desidrogenase (GAPDH), tubulina (Tubulina), e ubiquitinas (UbiQ1 e UbiQ2) em diversos tecidos, e comparou-se a variabilidade obtida utilizando os programas Genorm e NormFinder. Em seguida, foram realizadas análises de expressão gênica utilizando o programa REST para a validação estatística da expressão de genes de cana-de-açúcar. O gene UbiQ1 foi mais estável nos tecidos ou órgãos testados: meristema, inflorescência, folha, colmo e raízes tratadas com alto e baixo fósforo. Tendo o gene UbiQ1 como referência, a expressão relativa dos genes transportadores de fosfato de alta afinidade de cana-de-açúcar PT7 e PT8 foi avaliada em amostras de raiz fertilizadas com alto ou baixo Pi, inoculadas ou não com fungo micorrízico e coletadas aos 58 dpi. Esses dois genes pertencem à família Pht1 de transportadores de fosfato e são similares aos ortólogos de arroz ORYsa;tPht1;7 e ORYsa;Pht1;8. Sob a deficiência de Pi, o transportador de fosfato PT7 foi induzido em raízes não colonizadas; já nas micorrízicas foi pouco expresso, sendo que altas taxas de colonização radicular suprimiram a expressão do PT7. O gene PT8 pouco variou sua expressão, sendo sutilmente mais expresso em plantas micorrízicas do que nas não micorrízicas sob o suprimento de BP. Estes resultados indicam que o PT7 é induzido em raízes sob estresse por fósforo e provavelmente associado à absorção radicular de Pi. Enquanto o gene PT8 possui uma modulação pouco variável provavelmente envolvido na manutenção do fluxo ou homeostase de Pi, possivelmente associado com a absorção radicular e micorrízica de fosfato. O PT7 e o PT8 foram expressos em tratamentos de médio/longo prazo, apresentando expressão ou indução em resposta a privação por Pi, o que é consistente com a função proposta de aquisição e mobilização de Pi para esta família de transportadores. A cana-de-açúcar micorrízica mostrou alta plasticidade de resposta ao BP / Plants display a wide array of physiological adaptations to low soil phosphorus (Pi) availability. This work discussed physiological and energetic costs associated with these strategies, focusing on sugarcane responses to Pi availability during the development of arbuscular mycorrhizal (AM) symbiosis. Such costs are important components of adaptation to low phosphorus soils affecting phosphorus acquisition and leve, growth and soluble sugars concentration in plant tissues. Sugarcane plants were grown in pots, with or without AM (Glomus clarum), and with low (20 mg kg-1) or high (200 mg kg-1) phosphorus supply. Roots and shoots were harvest for analysis after 14, 30, 44 and 58 days post-inoculation (dpi) with the fungus Glomus clarum,. The low Pi supply caused Pi deficiency in mycorrhizedl or non-mycorrhizedl plants. The efficiency of Pi absorption, indicated by shoot Pi accumulation in correlation to root biomass, suggested that root and mycorrhizal Pi absorption were similar, regardless of the Pi doses. Phosphorus availability did not affect the whole-plant biomass, and plants under low Pi supply, used more efficiently this nutrient. On the other hand, mycorrhized plants supplemented with low Pi presented the highest root growth and shoot reduction, resulting in high root:shoot ratio. At 58 dpi, glucose, fructose and sucrose concentrations in leaves of mycorrhized plants were 3.8, 2.3 and 2.4-fold higher respectively, than in non-mycorrhizedl plants. These results suggested that, under these experimental conditions, mycorrhizal symbiosis establishment was not a typical mutualistic association affecting sugarcane growth profile and allometry, when cultivated under low Pi. The photosynthate levels of leaves from mycorrhizd plants indicated an increase in photossynthetic rate but withut resulting in higher macrobiont growth. The quantitative amplification of reversed transcripts (RT-PCR) technology has become a method of choice for functional gene validation, with sensitiveness, accurate quantification and high-throughput. The relative quantification is easier determined by relative expression in comparison to a constitutively expressed refernce gene. Here, the expression variability of a sugarcane actin gene (Actin) glyceraldehydo phosphate dehydrogenase (GAPDH), tubulin (Tubulina), and ubiquitins (UbiQ1 and UbiQ2) from various tissues were analysed and compared based on the ranking list from the Genorm and NormFinder softwares. Expression analysis based on the REST software gave the proper statistic validation. UbiQ1 was the most stable gene among the candidate gen-references along the various tissues or organs tested: meristem, inflorescence, leaf, stem and roots treated with high and low phosphorus. Considering UbiQ1 as the reference gene, the relative expression of the sugarcane high-affinity phosphate transporters genes PT7 and PT8 were assessed from roots fertilized with low Pi or high Pi , inoculated or not with the mycorrhizal fungus, harvest 58 dpi. Both genes belong to the Pht1 Pi transporter and share similarity with the rice orthologs ORYsat;Pht1;7 and ORYsat;Pht1;8. Under Pi deficiency, the phosphate transporter PT7 was induced in non-colonized roots, but less expressed in mycorrhizedl ones, with high root colonization rate suppressing PT7 expression. PT8 showed low variability in expression, slightly more expressed in mycorrhized plants than in non-mycorhized plants under low Pi supply. These results indicated that PT7 was induced in Pi stressed roots, and possibly associated with the root Pi uptake, while PT8 had limited modulation in expression and probably involved on Pi fluxes or homeostasis, likely associated with both root and mycorrhizal phosphate uptake pathways. PT7 and PT8 were induced during medium/long-term treatments, showing induction or constant expression in both acquisition and mobilization of Pi in response to Pi deprivation, which is consistent with the proposed role of this tranporter family. Mycorrhizedl sugarcane showed a highly plastic response to low Pi

Submitted by Marco Antônio de Ramos Chagas (mchagas@ufv.br) on 2018-06-05T12:51:35Z No. of bitstreams: 1 texto completo.pdf: 1056418 bytes, checksum: a4f83dbd4ba9d8c5874aa1cc1d9000d5 (MD5) / Made available in DSpace on 2018-06-05T12:51:35Z (GMT). No. of bitstreams: 1 texto completo.pdf: 1056418 bytes, checksum: a4f83dbd4ba9d8c5874aa1cc1d9000d5 (MD5) Previous issue date: 2018-02-23 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / A obtenção de mudas de qualidade é fundamental para o sucesso da implantação de um projeto florestal, pois o crescimento e a sobrevivência das plantas estarão atrelados à sua qualidade. A nutrição mineral e a seleção de microrganismos simbiontes a serem adicionados ao substrato de cultivo são de grande importância e influenciam a qualidade de uma muda. Contudo, para grande parte das espécies arbóreas nativas do Cerrado, esse conhecimento ainda não foi obtido, podendo dificultar ou inviabilizar a produção das mudas. O presente estudo teve como objetivo avaliar o crescimento inicial e a qualidade de mudas de Hancornia speciosa (mangabeira) e Brosimum gaudichaudii (mama-cadela) em resposta a doses de P e inoculação de microrganismos simbiontes provenientes do solo próximo às plantas matrizes e inóculo de fungos micorrízicos arbusculares (FMA). Foram avaliados dois experimentos, sendo ambos conduzidos no delineamento inteiramente casualizado (DIC). O experimento um foi composto por cinco doses de P (0, 100, 200, 300 e 400 mg dm -3 ), com seis repetições, totalizando 30 unidades experimentais por espécie, sendo o substrato uma mistura do solo rizosférico de plantas matrizes (SRPM) das espécies em estudo + horizonte subsuperficial de um Latossolo Vermelho-Amarelo distrófico autoclavado (LVAd-Aut) + substrato comercial Tropstrato Florestal® autoclavado (1:1:1,v:v:v). O experimento dois foi composto por quatro formulações de substrato contendo SRPM, LVAd-Aut e substrato comercial autoclavados (SC-Aut), onde esses receberam ou não a adição de inóculo de FMA (Mix), composto pelas espécies Gigaspora decipiens, Rhizophagus clarus e Scutellospora heterogama. Os tratamentos deste experimento consistiram de: T1 (LVAd-Aut + SC-Aut), T2 (LVAd-Aut + SC-Aut + Mix), T3 (LVAd-Aut + SC-Aut + SRPM) e T4 (LVAd-Aut + SC-Aut + SRPM + Mix), com seis repetições, totalizando 24 unidades experimentais por espécie. Ao final de cada experimento, realizou-se a determinação da taxa de colonização micorrízica (CM), a abundância de esporos presentes no substrato (AE) e as medições da altura total da parte aérea (H), diâmetro do coleto (DC), volume do sistema radicular (VR), massa de matéria seca da parte aérea (MSPA), raízes (MSR) e total (MST); relações H/DC, H/MSPA, MSPA/MSR, Índice de Qualidade de Dickson (IQD) e, para o experimento dois, foi quantificada, também, a dependência micorrízica (DM). No experimento um, plantas de mangabeira sofreram redução da CM em resposta aos níveis de P testados e tiveram resposta linear positiva à aplicação de P ao analisar as relações H/MSPA e MSPA/MSR, enquanto a relação H/DC não sofreu efeito do P. As demais variáveis em mudas de H. speciosa foram influenciadas negativamente pela aplicação de P. Em relação ao experimento dois, as espécies em estudo tiveram DM variando de moderada a alta quando o SRPM foi adicionado ao substrato de cultivo. A adição do SRPM gerou incremento na H, DC, VR, MSPA, MSR, MST e aumento da qualidade de mudas de H. speciosa e B. gaudichaudii, em comparação às plantas do tratamento controle. Para a produção de mudas de qualidade de H. speciosa e B. gaudichaudii, ao usar o substrato utilizado neste ensaio, a adubação fosfatada é desnecessária, sendo recomendada a utilização de solo rizosférico da planta matriz e, ou inoculação com FMA para garantir a produção de mudas de mangabeira e mama-cadela. / Obtaining seedlings of quality is essential to the successful implementation of a forestry project, because the growth and survival of plants will be tied to its quality. Mineral nutrition and selection of symbionts microorganisms to be added to the substrate of seedling production are of great importance and influence the quality of the seedlings. However, for most of the native tree species of Cerrado, this knowledge has not yet been obtained, and may hinder or derail the production of seedlings. The present study aimed to evaluate the initial growth and the quality of seedlings of Hancornia speciosa (mangabeira) and Brosimum gaudichaudii (mama-cadela) in response to P and inoculation doses of microorganisms symbionts from the soil around the plants and inoculum of arbuscular mycorrhizal fungi (AMF). Two experiments were evaluated, both being conducted in completely randomized design (CRD). The experiment was composed of five doses of P (0, 100, 200, 300 and 400 mg dm -3 ), with six repetitions, totalizing 30 experimental units per species, being the substrate a mixture of rhizospheric soil coming from the mother plant species (RSMP) + subsurface horizon of a Latossolo Vermelho-Amarelo distrófico (Oxisol) autoclaved (LVAd-Aut) + commercial Forest substrate autoclaved (SC-Aut) (1:1:1, v:v:v). The experiment two was composed of four substrate formulations containing RSMP, LVAd-Aut and SC-Aut, where these had or had not received the addition of AMF inoculum (Mix), composed by the species Gigaspora decipiens, Rhizophagus clarus and Scutellospora heterogama. The treatments evaluated in experiment two were T1 (LVAd-Aut + SC-Aut), T2 (LVAd-Aut + SC-Aut + Mix), T3 (LVAd-Aut + SC-Aut + RSMP) and T4 (LVAd-Aut + SC-Aut + RSMP + Mix), with six repetitions, totalizing 24 experimental units per species. At the end of each experiment, the determination of the rate of mycorrhizal colonization (CM), the abundance of spores present in the substrate (AE) and measurements of the total height of shot (H), stalk diameter (DC), volume of the root system (VR), mass of shoot dry matter (MSPA), mass of roots dry matter (MSR), and total dry matter (MST),beyond their relationships, and the Dickson’s quality index (IQD) and, for the experiment two, was quantified also the mycorrhizal dependence (DM). In one experiment, plants of mangabeira suffered reduction of CM in response to P levels tested and had positive linear response to P application to analyze relations H/MSPA and MSPA/MSR, while the ratio H/DC suffered no effect of P. The other variables in seedlings of H. speciosa were influenced negatively by applying P. compared to experiment two, the studied species had DM ranging from moderate to high when the RSMP was added to the substrate. The addition of the RSMP generated increase in H, DC, VR, MSPA, MSR, and MST, and increased the quality of seedlings of H. speciosa and B. gaudichaudii, as compared to the control treatment seedlings. For the production of quality seedlings of H. speciosa and B. gaudichaudii, when using the substrate used in this test, the phosphate fertilization it is recommended to use of rhizospheric soil of the mother plant and, or inoculation with AMF for guarantee the seedling production of mangabeira and mama- cadela.

The study of arbuscular mycorrhizal symbiosis (AMS) - the mutualist relationship between the most of land plants and evolutionary old fungal group Glomeromycota - is becoming a prestigious topic. The prevalence of and extent of physiological action of AMS on plants is very interesting for the plant biology itself, but its importance grows, notably in time of global climate change, frequent soil degradation and ascending exhaustion of mineral fertilizer reserves. Although the flows in AMS of some minerals, like of phosphorus was enlightened, carbon exchange between the symbionts is still poorly understood. In this experimental work, I utilized the boom of molecular and bioinformatic methods in the quest for completely unexplained carbon flows. The organisms used include barrel medic (Medicago truncatula), the model legume for symbiotic relationships, biotic, and abiotic stresses; Rhizophagus irregularis, the widely used fungus for such experimental studies of AMS; and Sinorhizobium meliloti, the nodulating nitrogen-fixing bacterium compatible with the barrel medic. Two variants - mycorrhizal (M+) and non-mycorrhizal (NM) plants were subjected to several levels of analysis. I have checked the variants, did the measurements of phosphorus and nitrogen contents, as well as I probed the plants with...

The symbiotic unit of an arbuscular mycorrhizal fungus and its host is able toachieve and maintain far higher inflow of nutrients than non-mycorrhizal roots. The colonization strategy of the mycobiont within the plant is intrinsic to the symbiosis with respect to both structural adaptations and nutrient exchange. An investigation into the effect of Glomus etunicatum colonization on the structure and phloem transport in Eragrostis curvula (Umgeni) allowed for greater insight into the dynamic of the symbiosis. The combined use of stains (such as Trypan Blue, Chlorazol Black, Safranin and Fast Green), and techniques, (such as freeze-microtome transverse sectioning and permanent slide preparations) contributed to a successful general observation of an intermediate colonization strategy using light microscopy methods. However, clarity into structural detail of mycorrhizal forms required electron microscopy studies. The SEM method used with freeze fracture was a relatively quick and simple method allowing for the observation of surface and internal features. The TEM method allowed for highresolution images providing insight into the variations in the apoplasmic compartmental form, and how this may relate to the function of the symbiosis with regard to fungal coils or arbuscules. The apoplasmic nature of mycorrhizas was substantiated and no symplasmic connections were found between symbionts. Fluorescence studies demonstrated that 5,6-carboxyfluorescein was transported through the phloem into the roots of E. curvula, but remained predominantly in the root phloem. Unloading only occurred in optimal nutrient exchange areas of meristimatic lateral or apical growth regions. It was not possible, using fluorescence techniques and related equipment available, to conclusively establish if there were symplasmic connections between the mycobiont and its host or if bidirectional transfer of nutrients occurred at the same interface.

No description available.

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

La plupart des espèces végétales terrestres vivent en symbiose avec les champignons mycorhiziens à arbuscules (MA). Il s’agit d’une symbiose très ancienne datant de plus de 400 millions d’années. Les champignons MA sont des champignons du sol qui appartiennent aux Gloméromycètes. Ils sont présents dans la plupart des écosystèmes terrestres. Ainsi, ils peuvent être considérés comme une composante intégrale des racines des plantes. Ils forment dans les cellules racinaires corticales des structures fonctionnelles essentielles appelées arbuscules où ils apportent à la plante des minéraux nutritifs en échange de sucres. L’auxine est une phytohormone impliquée dans de nombreux processus de développement des plantes, y compris la dominance apicale, les tropismes, la structuration vasculaire et la formation de racines latérales. Le principal objectif de notre travail était d’étudier de manière approfondie le rôle de l’auxine dans le processus de développement des mycorhizes. On sait déjà que la symbiose MA stimule la formation de racines latérales dans les plantes hôtes, ce qui pourrait être due à une modification du métabolisme de l’auxine, de son transport ou de sa perception. Les microARNs (miARNs) sont des molécules d’ARN non codantes de ~ 21 nucléotides capables de réprimer l’expression de gènes en ciblant et clivant spécifiquement leur ARNm correspondant. Plusieurs miARNs interagissent avec la signalisation de l’auxine et parmi eux miR393 qui cible les récepteurs à l’auxine. Nous avons étudié le rôle de miR393 dans la colonisation mycorhizienne. Nous mettons en évidence que chez Solanum lycopersicum (Solanacées), Medicago truncatula (Fabaceae) et Oryza sativa (Poaceae), l’expression des précurseurs de miR393 diminue lors de la mycorhization. En outre nous montrons que DR5-GUS, un gène rapporteur de réponse à l’auxine, est préférentiellement exprimé dans les cellules de la racine contenant les arbuscules. En sur-exprimant miR393 dans les racines et donc en régulant négativement l’expression des gènes de récepteurs à l’auxine, nous montrons également que les arbuscules ne se développent pas normalement. En tant que composantes des complexes récepteurs d’auxine, les protéines Aux/IAA jouent un rôle majeur dans la voie de signalisation de l’auxine en réprimant l’activité des facteurs de transcription de type ARF. Nous avons vérifié dans des racines de tomate mycorhizées l’expression de 25 gènes AUX/IAA. Nous nous sommes concentrés sur IAA27 dont l’expression est induite lors des premiers stades de la symbiose MA. Nous observons qu’une répression par ARNi de l’expression de IAA27 dans des plants de tomate conduit à une forte diminution de la colonisation MA et du nombre des arbuscules. Puis nous montrons par des approches différentes que la régulation positive de la mycorhization par IAA27 est liée à la biosynthèse des strigolactones. Globalement, ces résultats appuient fortement l’hypothèse selon laquelle la signalisation de l’auxine joue un rôle important aussi bien dans le stade précoce de la mycorhization que dans la formation des arbuscules. / Most land plant species live in symbiosis with arbuscular mycorrhizal (AM) fungi. This is a very ancient symbiosis dating back to 450 million years. AM fungi are soil fungi that belong to the Glomeromycota. They are present in most terrestrial ecosystems. Thus they can be considered as an integral root component of plants. They form essential functional structures called arbuscules in root cortical cells at which mineral nutrients are released to the plant in exchange of sugars. The phytohormone auxin is involved in many developmental processes in plants, including apical dominance, tropisms, vascular patterning and lateral root formation. The main objective of our work was to investigate further the role of auxin in the mycorrhizal developmental process. We already know that AM symbiosis stimulates the lateral root formation in host plants, which could be due to modification of auxin metabolism, transport or perception. The microRNAs (miRNAs) are ~21-nucleotides noncoding RNAs that target corresponding mRNA transcripts for cleavage and transcriptional repression. Several miRNAs interact with auxin signaling and among them miR393 that targets auxin receptors. We investigated the role of miR393 in AM root colonization. In Solanum lycopersicum (Solanaceae), Medicago truncatula (Fabaceae) and Oryza sativa (Poaceae), expression of the precursors of the miR393 was down-regulated during mycorrhization. In addition DR5-GUS, a reporter for auxin response, was found to be preferentially expressed in root cells containing arbuscules. By over-expressing miR393 in roots and therefore down-regulating auxin receptor genes, arbuscules could not develop normally. As components of auxin receptor complexes, Aux/IAA proteins play a major role in auxin signaling pathway by repressing the activity of ARF type transcription factors. We checked the expression of 25 AUX/IAA genes in AM roots. Among them, we focused on IAA27 that was significantly up-regulated during the early stages of AM symbiosis. IAA27 down-regulation in plants led to a strong decrease of AM colonization and arbuscule abundance. We showed by different approaches that the positive regulation of mycorrhization by IAA27 was linked to strigolactone biosynthesis. Overall these results strongly support the hypothesis that auxin signaling plays an important role both in the early stage of mycorrhization and in the arbuscule formation.

Thesis (M.Sc. (Horticulture)) -- University of Limpopo, 2017 / Refer to document / National Research Foundation of South Africa, and Agricultural Research Council-Universities Collaboration Centreꞌ for scholarship and research