Orientador: Angelo Luiz Cortelazzo / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-11T20:45:08Z (GMT). No. of bitstreams: 1 Vilhalva_DivinaAparecidaAnunciacao_D.pdf: 8473615 bytes, checksum: 16013933ce12bdce13be2d0f84d9563f (MD5) Previous issue date: 2008 / Resumo: O acúmulo de metais pesados no solo provoca grandes danos ambientais, em especial o do cádmio, devido ao seu alto nível de toxicidade e efeitos acumulativos nos organismos em geral. Poucas pesquisas envolvendo a resposta de diferentes vegetais a metais pesados foram realizadas com espécies nativas de cerrado. O objetivo deste trabalho foi avaliar, em casa de vegetação, o comportamento de duas espécies do cerrado: Campulocliniumchlorolepis (Asteraceae) e Galianthe grandifolia (Rubiaceae) em relação ao excesso de cádmio (Cd) no solo. Ambas foram irrigadas com soluções aquosas contendo 0, 0,3 e 3 mmol L-1 de cádmio. Os resultados demonstraram que as espécies são diferentemente tolerantes às concentrações de cádmio a que foram submetidas. Campuloclinium chlorolepis suportou por 14 dias as concentrações de 0,3 e 3 mmol L-1 de Cádmio e, para estas mesmas concentrações, as plantas de Galianthe grandifolia, toleraram até 28 dias. Os principais sintomas de toxicidade do Cd foram: clorose, mancha e murchamento das folhas e inibição do crescimento em Campuloclinium chlorolepis; epinastia e manchas nas folhas em Galianthe grandifolia. O Cd não inibiu o crescimento de Galianthe grandifolia, havendo estímulo no crescimento das plantas até mesmo na maior concentração (3 mmol L-1 de Cd). A análise do teor de Cd mostrou que Galianthe grandifolia é mais tolerante e hiperacumuladora desse metal, concentrando em média 120,37 mg Kg¿1 nos tecidos da parte aérea e 298,97 mg Kg-1 de Cd na parte subterrânea. Campuloclinium chlorolepis apresentou concentrações maiores na parte aérea (22,07 mg Kg-1 de Cd) que nas raízes (7,97 mg Kg-1), mas inferiores às concentrações observadas na outra espécie. Neste trabalho foram também investigadas particularidades relevantes de ambas as espécies. Em Campuloclinium chlorolepis destaca-se a ocorrência de frutanos do tipo inulina, em forma de glóbulos e localizados em todos os tecidos de suas raízes tuberosas, com exceção da epiderme. No sistema subterrâneo de Galianthe grandifolia constatou-se a ocorrência de coléteres protegendo as gemas caulinares originadas em seu xilopódio. Ressalta-se também que as duas espécies possuem associação com fungos micorrízicos arbusculares e fungos endofíticos ¿Dark Septate¿, os quais são co-ocorrentes nas raízes tuberosas e finas de Campuloclinium chlorolepis, enquanto colonizaram apenas as raízes adventícias de Galianthe grandifolia. / Abstract: The accumulation of toxic metals in the soil brings forth great environmental damages, especially cadmium, due to its high toxicity and cumulative effects on the organisms in general. There are a few studies regarding heavy metals and Cerrado native species. The aim of this work was to assess the behavior of two Cerrado species cultivated in the greenhouse: Campuloclinium chlorolepis (Asteraceae) and Galianthe grandifolia (Rubiaceae), as affected by cadmium (Cd) excess in the soil. Both species were watered with aqueous solutions of 0, 0.3, and 3 mmol L-1 of Cd. The results showed that the species have different tolerance levels to the Cadmium concentrations to which they were submitted. Campuloclinium chlorolepis resisted during fourteen days, subjected to the concentrations of 0.3 and 3 mmol L-1 of Cadmium and, at these same concentrations, Galianthe grandifolia remained alive up to 28 days. The main symptoms of Cd fitotoxicity were: clorosis, stained and withered leaves and growth inhibition in Campuloclinium chlorolepis; epinasty and stained leaves in Galianthe grandifolia. There was no growth inhibition in Galianthe grandifolia, but rather growth stimulation, even in the highest concentration (3 mmol L-1 of Cd). The Cd content analysis revealed that Galianthe grandifolia is more tolerant and a hyperaccumulator of this metal, with a mean accumulation of 120.37 mg Kg-1 in the tissues of shoots, and 298.97 mg Kg-1 of Cd in the underground system. Campuloclinium chlorolepis showed greater concentrations of Cd in the shoots (22.07 mg Kg-1) rather than in the roots (7.97 mg Kg-1). In this work, peculiarities of both species were also analyzed. Globular inulin-type fructans were detected in all tissues of the tuberous roots of Campuloclinium chlorolepis, except for the epidermis. In the underground system of Galianthe grandifolia, the shoot buds sprouting from the xylopodium are protected by colleters. In addition, the two species showed an association with both arbuscular mycorrhizal fungi and dark septate endophytic fungi, which co-occur in the tuberous and the thin roots of Campuloclinium chlorolepis, but colonize only the adventitious roots of Galianthe grandifolia. / Doutorado / Doutor em Biologia Vegetal

Abstract Most plants grow in association with arbuscular mycorrhizal (AM) fungi in their roots forming the so-called AM symbiosis. AM symbiosis is usually beneficial to the host as it improves plant survival and performance. However, AM symbiosis also entails a cost to the plant in terms of the carbon allocated to the fungus. In sexually dimorphic plants, more than one type of individual can be recognised with regard to their sexual expression or gender. The cost of reproduction in these plants will differ in relation to the relative investment in male versus female function, as the female and the male sexual functions incur different costs. This different cost of reproduction may be translated into differences in other plant functions between the sexes as all functions are connected through trade-offs. Therefore, since sexes differ in resource needs and allocation patterns, and AM mediate resource acquisition and allocation patterns through imposing both costs and benefits to the plant, the sexes of dimorphic plant species may possess, at least theoretically, a different relationship with their AM roots symbionts. In this thesis, I have investigated whether the sexes in sexually dimorphic plant species differ in their mycorrhizal relationship, and if so, in which ways. Several plant life history traits were studied in the dioecious species Antennaria dioica and also in the gynodioecious Geranium sylvaticum using greenhouse, common-garden and field experiments. Resource acquisition, resource allocation, and both plant and fungal benefits from AM symbiosis were considered. Mainly beneficial effects of AM symbiosis were observed in both sexes of the two dimorphic plant species for most of the studied plant life history traits. Overall, both partners benefited from the AM association. However, several sex-specific benefits were detected which were not uniformly present in all experiments for any given trait. Moreover, the responses observed in certain life history traits were dependent on both the AM fungal and plant species involved in the symbiosis. Remarkably, plants gained sex-specific benefits from the same species of AM fungi and the fungal benefit differed depending on the sex of the host plant. In addition, mycorrhizal benefits were lost under certain environmental conditions. To summarise, the results obtained in this study highlight the complexity of AM interactions. My results suggest that the plant-mycorrhizal fungus relationship may differ depending on the sex of the host plant. Through sex-specific effects on survival, growth and reproduction of the hosts, AM fungi may play a role in the evolution of the life histories in the studied species. In addition, sex-specific relationships between plants and their mycorrhizal symbionts may have potential important consequences for the population dynamics of the sexual morphs and the coevolution of the mycorrhizal relationship.

La symbiose mycorhizienne à arbuscules (SMA) est le résultat de l'interaction entre les racines de plus de 80% des familles de plantes terrestres et les champignons MA. Divers types de membranes jouent un rôle crucial dans la mise en place et le fonctionnement de la SMA chez l’hôte végétal. Si l’électrophorèse bidimensionnelle (2-DE) reste la méthode la plus couramment utilisée pour des analyses protéomiques quantitatives dans la SMA, elle résout difficilement les protéines membranaires en raison de leur hydrophobicité, leur précipitation au point isoélectrique (pI) et leur faible abondance comparativement aux protéines cytosoliques. Donc peu nombreuses sont les protéines membranaires identifiées comme étant régulées en réponse à la symbiose. Afin d’avoir accès à cette catégorie de protéines et contourner les défauts de la 2-DE, l’application de nouvelles méthodes permet de réaliser des analyses quantitatives avec marquage chimique (comme l’iTRAQ) ou non (label-free). Dans ce contexte, deux méthodes de protéomique quantitative, iTRAQ-OFFGEL-CL-SM/SM et « label-free » 1-DE-CL-SM/SM, sont adoptées dans ce travail visant à identifier et quantifier les variations d’accumulation des protéines microsomales de racines de Medicago truncatula inoculées par Rhizophagus irregularis, préalable indispensable à l’analyse de leur rôle fonctionnel dans la SMA. Un protocole d’extraction donnant accès à des fractions radiculaires enrichies en protéines microsomales nécessaires pour les analyses ultérieures est décrit dans cette étude. En plus de l’analyse quantitative du protéome membranaire en réponse à la SMA, une approche méthodologique a été mise en place afin d’étudier l’impact du marquage iTRAQ sur le pI des peptides. / Arbuscular mycorrhizas (AM) are widespread symbiotic associations between plant roots and AM fungi. Deep membrane alterations are the foremost morphological changes occurring in the host plant in response to AM symbiosis. Two-dimensional gel electrophoresis (2-DE) is the workhorse method in AM proteomics. Membrane proteins are under-represented in 2-DE because of their hydrophobicity, low abundance, and precipitation at their isoelectric point, thereby few are the identified membrane proteins involved in sustaining the AM symbiosis. Membrane proteomics is still challenging due to 2-DE related shortcomings, however latest trends and advancements in mass spectrometry (MS)-based quantitative proteomics offer enormous potential to monitor membrane protein change in abundance in large scale experiments. In the current work microsomal proteins of Medicago truncatula roots inoculated with Rhizophagus irregularis were, for the first time, scrutinised by state-of-the-art MS-based proteomic approaches iTRAQ-OFFGEL-LC-MS/MS and label-free 1-DE-LC-MS/MS. The applied workflows combine two novel proteomic procedures, label-based and -free, targeting an insight view on the membrane proteome changes in AM symbiosis. A subcellular fractionation method is herein described to access the total membrane-associated proteins with sufficient recovery and purity for their subsequent in-depth analysis. In addition to the biological gain by shedding the light on candidate AM-related membrane proteins, a methodological approach was carried out in the present work in order to elucidate the iTRAQ labelling impact on peptide isoelectric points.

Fundamental processes underpinning the biotransformation of coal by fungal biocatalysts have been intensively investigated, however, limited large-scale industrial applications using such systems have been reported. The un-anticipated sporadic growth of Cynodon dactylon on the surface of un-rehabilitated discard coal dumps has been noted and this was found to be coupled with the breakdown of coal into a humic soil-like material in the top 1.5 metres of the dumps. Extensive fungal growth was observed to be associated with the Cynodon dactylon root system and examination of plant roots indicated the presence of mycorrhizal fungi. Analysis of the Cynodon dactylon plant roots around which coal biotransformation was occurring confirmed the presence of arbuscular mycorrhizal colonisation with the species Glomus clarum, Paraglomus occultum, Gigaspora gigantea and Glomus mosseae identified to be associated with the plants. Further molecular characterisation of non-mycorrhizal rhizospheric fungi showed the presence of fungal species with coal-degrading capabilities that most likely played a role in the coal biotransformation observed. The discard coal dump environment was simulated in pot and column studies and coal biotransformation was reproduced, with this process enhanced by the addition of mycorrhizal and non-mycorrhizal rhizospheric fungal inocula to the environment. Mycorrhizal and non-mycorrhizal species in the inoculum were re-isolated from the simulated environment fulfilling a number of Koch’s postulates and indicating a causal role in the biotransformation of coal. An inversion of conventional mycorrhizal colonisation was demonstrated in this system with reduction in extraradicular presence and an increase in intracellular colonisation compared to soil controls. A descriptive model was formulated suggesting a two-part fungal system involving organic carbon and nutrient exchange between the plant, mycorrhizal fungi and non-mycorrhizal coal-degrading rhizospheric fungi ultimately resulting in the biotransformation of coal. The biotransformation observed was comparable to reports of “rock-eating fungi”. Results suggest that the biological degradation of coal in situ with the production of a soil-like substrate could provide a feasible method of discard coal dump rehabilitation as well as provide a humic-rich substrate that can be utilised in further industrial applications.

Soybeans (Glycine max L. Merr.) are a symbiont of two beneficial associations: biological nitrogen fixation (BNF) with Bradyrhizobium japonicum, and arbuscular mycorrhizal fungi (AMF). Within the Northern Great Plains of the USA, iron deficiency chlorosis (IDC) of soybean is a yield-limiting factor. The effects of IDC on BNF and AMF are not well defined. This study was conducted to determine the effects of IDC on BNF and AMF. A laboratory study was performed to compare three methods of measuring ureide-N, a product of BNF in soybeans. Field studies in soybean were performed at three locations at eastern N011h Dakota. The experimental design was a factorial combination of three cultivars and three treatments. The three cultivars, in order of decreasing chlorosis susceptibility, were NuTech NT-0886, Roughrider Genetics RG 607, and Syngenta S01-C9 RR. The three treatments were control, Sorghum bicolor L. companion crop planted with the soybean seed, and FeEDDHA applied with the soybean seed. Chlorosis severity was the greatest and least for the NuTech and Syngenta cultivars, respectively. The FeEDDHA treatment decreased chlorosis severity. Ureide levels were abnormally high in plants severely stunted by JDC. The excess accumulation of ureides in IDC-stunted plants suggests that plant growth was reduced more than the rate of nitrogen fixation. The AMF population \vas at an adequate level at all locations and not affected by cultivar or treatment, in general. In the laboratory study, the Patterson et al. method had greater ureide concentrations due to the non-specific measuring of ammonium compounds compared to the Vogels and Van der Drift and Goos methods. / North Dakota Soybean Council

No description available.

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

The plant genus Tolpis (Asteraceae) is a predominantly insular plant lineage. It inhabits four of the five archipelagoes that comprise the Atlantic region of Macaronesia and also occurs in Mediterranean Europe and North Africa. Twelve species are currently recognized in Tolpis, of which ten are insular and two continental. The majority of the insular species inhabit the five western Canarian islands, where they constitute endemics to specific ecological habitats. A comprehensive molecular phylogeny of Tolpis is generated via DNA sequences of one nuclear ribosomal and two low-copy nuclear DNA markers. Considerable phylogenetic uncertainty among inferred tree topologies is detected, and incongruence between these topologies is resolved via statistical hypotheses testing. The extant diversity of the genus is identified to be the result of two independent colonization pathways and adaptive radiations on several islands. Moreover, potential hybridization is detected between species that inhabit different islands and archipelagoes, indicating a more widespread historical distribution of the genus. Details of the biogeographic history of Tolpis are inferred via ancestral area reconstructions under parsimony and likelihood optimality criteria. The hypothesis that Tolpis may have undergone a back-dispersal from an island to a continental habitat is also tested. Uncertainty in taxon cladograms owing to the presence of hybrid or allopolyploid taxa is characterized and a potential adjustment strategy evaluated. Averaging reconstruction results over all optimal phylogenetic trees and the manual pruning of cloned DNA sequences are found potential adjustment strategies against the impact of topological uncertainty owning to hybrid or allopolyploid taxa. Adjusted ancestral area reconstructions in Tolpis do not support the scenario that the genus has undergone a reverse colonization of the continent. In addition to the phylogenetic and biogeographic history of the genus, the diversity of symbiotic mycorrhizal fungi associated with Tolpis is characterized. A molecular survey using two nuclear ribosomal DNA markers and 454 pyrosequencing is performed. Particular emphasis is placed on the quality filtering of resulting fungal DNA sequences, the generation of operational taxonomic units, and their taxonomic assignment via similarity searches against DNA sequence databases. Numerous potentially novel fungal genotypes are identified. / text

Roots of silver maple (Acer saccharinum L.) trees from downtown Montreal and the municipal nursery in Terrebonne were sampled to determine their vesicular-arbuscular mycorrhizal (VAM) colonization levels. Soil was analyzed to determine the concentration of various soil nutrients, pH, and bulk density. It was found that trees from downtown Montreal grow in soils with lower phosphorus and magnesium, and higher copper, zinc, sodium, pH and bulk density than nursery trees. Downtown tree health, in terms of % dieback, was positively correlated with soil phosphorus, potassium, calcium, the phosphorus:copper ratio and negatively correlated with soil copper, zinc, bulk density and tree age. It is suggested that tree health and survival are associated with tree location. Older and healthier trees grow on streets that provide larger soil volumes. Downtown trees had significantly higher VAM colonization levels (44.9%) than those from the nursery (36.1%). The colonization level in downtown trees was positively correlated with the phosphorus:copper ratio and tree age, and negatively correlated with soil potassium. It is hypothesized that downtown trees living under the most stress will be those with the highest VAM colonization. Under controlled conditions 81.5% of the variability of % VAM infection of silver maple seedlings could be explained by soil zinc, phosphorus, copper, sodium, and potassium. Zinc, phosphorus and copper negatively affected VAM colonization, while a slight positive effect of sodium and potassium was found. This suggests that VAM colonization in downtown trees may be affected by soil factors, but the effects may be masked by other conditions, such as water stress, light, or size of root systems.

Thesis (MScFor (Forest and Wood Science))--University of Stellenbosch, 2007. / The study was undertaken to determine the potential physiological benefits to plants provided by the double colonisation of host plant roots by endomycorrhizal (AM) and ectomycorrhizal (ECM) fungi, when growing under normal and under salt stress conditions. Plants of the Eucalyptus grandis x E. camaldulensis clone were grown in a sterile soil with 0 and 75 mM NaCl and with or without infection with the fungi Glomus etunicatum (an AM fungus) and Pisolithus tinctorius (an ECM fungus). The Eucalyptus clone formed both ECM and AM in single and double inoculation. The mycorrhizal symbiosis did not provide any nutritional benefits to the hosts. The double colonisation had no effect on plant growth under normal growth conditions while single colonisations of AM and ECM reduced growth. Double colonisation reduced host plant specific leaf mass by 12% and increased total leaf area by 43% compared with the control under these growth conditions. This colonisation also reduced photosynthesis per leaf area by 29% compared with the control. The reduced photosynthesis of the double colonisation did not result in reduced plant growth because these plants may have had a high total plant photosynthesis because of their large total leaf area. The double symbiosis however did not reduce salt stress when host plants were exposed to 75 mM NaCl, while the AM fungus increased plant dry weight by 13% compared to the control. AM and ECM colonisation in the double colonised roots under salt stress was decreased by 18 and 43% compared to that in plants under normal growth. The reduced colonisation may have reduced the fungi’s abilities to be beneficial to the host plant. The double symbiosis is recommended based on the documented positive effects of this symbiosis to plant growth and the considered possible long-term benefits to host plants growing in saline soils.