A mesofauna edáfica, compreendendo ácaros e colêmbolos, é sensível a alterações no solo, sendo considerada um indicador de qualidade, uma vez que realiza a fragmentação de matéria orgânica, controle populacional de determinados grupos da biota do solo e ingestão de microrganismos. A colonização de plantas por micro-organismos, como a de leguminosas por rizobios, pode levar a mudanças na rizosfera vegetal e influenciar a mesofauna. Este estudo teve por objetivo avaliar o efeito da inoculação de plantas de trevo vermelho com rizóbios sobre os ácaros e colêmbolos edáficos e avaliar se colêmbolos alimentados com rizóbios poderiam inocular plantas hospedeiras. Para isso, plantas de trevo vermelho, inoculadas com as estirpes SEMIA 2081, SEMIA 2082 e SEMIA 222, e plantas controle sem inoculação foram plantadas em tubos-armadilha e inseridas em campo. A seguir, a mesofauna atraída foi extraída e identificada. Outro experimento em laboratório utilizou plantas de trevo vermelho que foram colocadas em contato com colêmbolos da família Hypogastruridae e dos gêneros Proisotoma, Onychiurus e Orthonychiurus, os quais haviam sido alimentados com um substrato impregnado com o rizóbio da estirpe SEMIA 2081 As plantas inoculadas com as três estirpes de rizóbios atrairam ácaros edáficos mas não colêmbolos, quando comparadas com as plantas que não foram inoculadas nem receberam fonte de nitrogênio. Considerando os números totais de ácaros e colêmbolos, somente as plantas inoculadas com a estirpe SEMIA 222 apresentaram atratividade quando comparadas ao controle sem inoculação e sem fonte de nitrogênio. A atratividade de ácaros e colêmbolos observada no tratamento controle com nitrogênio foi similar aos tratamentos inoculados. No experimento de laboratório, os colêmbolos vivos e o macerado de colêmbolos da família Hypogastruridae, e gêneros Proisotoma e Orthonychiurus foram capazes de transferir os rizóbios que receberam pela alimentação às plantas, formando nódulos radiculares. As plantas que receberam o macerado de colêmbolos da família Hypogastruridae apresentaram maior número de nódulos comparadas às plantas que receberam os mesmos colêmbolos vivos. / The soil mesofauna comprising mites and springtails, is sensitive to changes in soil, and it is considered a quality indicator since it performs fragmentation of organic matter, population control of certain soil biota groups and ingestion of microorganisms. The plants colonized by microorganisms, such as legumes by rhizobia can lead to changes in plant rhizosphere and influence mesofauna. This study aimed to evaluate the effect of inoculation of red clover plants with rhizobia on edaphic mites and springtails and assess if springtails fed with rhizobia could inoculate host plants. For this, red clover plants inoculated with strains SEMIA 2081, SEMIA 2082 and SEMIA 222, and control plants without inoculation were planted in trap-tubes and inserted on field. Then, the attracted mesofauna was submitted to extraction and identification. Another laboratory experiment used red clover plants put in contact with springtails. These springtails of Hypogastruridae family and Proisotoma, Onychiurus and Orthonychiurus genera, had been fed with a substrate impregnated with rhizobia strain SEMIA 2081. Plants inoculated with three Rhizobia strains attracted edaphic mites, but not Collembola, when compared to uninoculated plants with no nitrogen source Considering the total numbers of mites and springtails, only plants inoculated with SEMIA 222 strain showed attractiveness when compared to the control without inoculation and no nitrogen source. The attractiveness of mites and springtails observed in the control treatment with nitrogen was similar to the inoculated treatments. In the laboratory experiment, the living springtails and the macerate of springtails of Hypogastruridae family, and Proisotoma and Orthonychiurus genera were able to transfer the rhizobia that they received by feeding to the plants, forming root nodules. The plants that received the macerate of springtails of the Hypogastruridae family had a higher number of nodules compared to the plants that received the same living springtails.

Cloning and molecular characterization of new genes constitutes a useful approach in studying the symbiotic interactions between the model plant Medicago truncatula and Synorhizobium meliloti. Large numbers of expressed sequence tags (ESTs) available for Medicago truncatula, along with numerous cDNA, oligonucleotides, and Affimetrix DNA microarray chips, represent useful tools for gene discovery. In an attempt to identify a new gene that might be involved in the process of nodulation in Medicago truncatula, preliminary data reported by Fedorova et al. (2002), who identified 340 putative gene products or tentative consensus sequences (TCs) expressed only in nodules, was used. This research was focused on TC33166 (TC103185), which has 3 ESTs in the TC, and whose strongest BLASTX hit of TC103185 is the incomplete root hair elongation (IRE) protein kinase-like protein (NP_192429) from Arabidopsis thaliana. The Arabidopsis IRE gene is required for normal root hair growth, and a role in apical growth was suggested (Oyama et al., 2002). Infection thread growth can be looked at as an inward growth of the root hair. Thus, TC103185 was a good candidate for identifying a gene that may be involved in early events of nodulation. MtIRE (GenBank accession AC122727) is organized in 17 exons and 16 introns, similarly to the Arabidopsis IRE gene. MtIRE is a new member of the IRE family and it is a putative Ser/Thr protein kinase. MtIRE is a nodule- and flower-specific gene, suggesting that nodulation may have recruited it from other developmental processes. MtIRE is likely to be involved in the invasion process, or in the maturation of the symbiosome, or of the cells that contain rhizobia, rather than infection thread initiation and elongation or in nitrogen fixation. Nodule invasion precedes the onset of MtIRE expression and the expression pattern changes in time within the nodule. RNA interference results support MtIRE expression data and suggest a possible role in preventing extensive defense responses. Our study demonstrates the existence of an Arabidopsis IRE homolog in Medicago truncatula root nodules with an entirely new function and regulation.

In recent years, the rhizobial taxonomy changed significantly with the discovery of novel symbiotic associations between legumes and nodulating bacteria. This was aided by the focus shift from studying only agricultural crops to legumes indigenous to certain regions, ultimately to discover new inoculant strains and to uncover the secrets of the rhizobium¬legume symbiosis. In previous studies on the diversity of South African rhizobia, it has become clear that our country has a wealth of rhizobia. Cyclopia is a legume genus, which belongs to the fynbos biome of South Africa. Honeybush tea is a herbal infusion manufactured from the leaves and stems of certain Cyclopia spp. Commercial cultivation of this potentially new agricultural crop is now developed to protect the natural Cyclopia spp. populations from harvesting and ultimately extinction. Superior inoculant strains are necessary for these commercial seedlings. The diversity of root-nodulating strains isolated from 14 Cyclopia spp. was determined using 16S-23S IGS-RFLP and partial 16S rDNA base sequencing. Based on 16S-23S IGS-RFLP and partial 16S rDNA base sequencing most of the isolates, with the exception of seven strains, were found to belong to the genus Burkholderia. More extensive phylogenetic, symbiotic and phenotypic studies of selected strains were performed using near full-length 168 rDNA base sequencing, nodA base sequencing and substrate utilisation analysis. In the genus Burkholderia, the isolates belonged to the novel root-nodulating species Burkholderia tuberum and several novel, undescribed Burkholderia genotypes. However, no new Burkholderia species could formally be proposed, since DNA-DNA hybridisation analysis, which is a prerequisite for the description of new species could not be performed in our laboratory. The seven strains not affiliated with the Burkholderia genus belonged to two Bradyrhizobium genospecies, R tropici and a possibly new genus in the a-Proteobacteria. The nodA sequences of all the Cyclopia isolates corresponded to a large extent, indicating that different chromosomal genotypes harbour the same symbiotic genotype. All the isolates of the Cyclopia genus appear to be acid-tolerant, which is in agreement with the acidic nature of the soil from which the strains were isolated. / Thesis (PhD(Microbiology))--University of Pretoria, 2004. / Microbiology and Plant Pathology / unrestricted

The Rhizobiaceae bacteria are known for its varied nitrogen metabolism: especially its nitrogen fixation, so called diazotrofia, its capability of symbiosis with plants, or on the contrary plant pathogenesis (genus Agrobacterium). Rhizobium radiobacter R89 (stored in Czech Collection of Microorganism, in Brno as R89-1) is bacteria strain capable of codeine and morphine biotransformation to whole spectrum of pharmacologically significant derivates. This work is focused on purification and characterization of two selected enzymes that catalyze oxidation-reduction reactions, the crucial beginning of morphine skelets biodegradation. At the first, the biotransformational potential of the strain was evaluated and biochemical, molecular biological and bioinformatical approaches were employed to purify the responsible enzymes. Considering unsuccessful purification from Rhizobium radiobacter R89-1, the proteins structural genes were cloned and heterologously expressed in bacterial system and afterward characterized. Found protein sequence and enzyme characterization (basic kinetic measurements, substrate specificity and thermo- stability) have revealed different origin of the enzymes and clarify reasons why the original purification procedure was not successful.

Rhizobia are Gram-negative, rod-shaped, soil bacteria with the ability to fix atmospheric nitrogen into ammonia as symbiont bacteroids within nodules of leguminous plant roots. Here, resident Rhizobium plasmids were studied as possible sources of components for the construction of a cloning vector for Rhizobium species.

The Rhizobia are a group of free-living soil bacteria known for their ability to symbiotically infect the roots of specific host plants as well as to produce siderophores in order to compete with other microorganisms for the limited availability of iron in the rhizosphere. In this study, Rhizobium leguminosarum ATCC 14479, which preferentially infects the red clover Trifolium pratense, was found to produce the trihydroxamate siderophore vicibactin (C 33H55N6O15) under iron restricted conditions. In addition, two other iron-binding, siderophore-like compounds: C20H36N4O10, C31H 55N6O15, were isolated and purified from the culture media. Due to the structural similarity of the latter compounds to vicibactin based on electrospray-mass spectrometry and nuclear magnetic resonance data, these heretofore unreported molecules are thought to be either modified or degraded products of vicibactin. Although vicibactin has previously been found to be commonly produced by other rhizobial strains, this is the first time it has been chemically characterized from a clover infecting strain of R. leguminosarum.

The Rhizobia are a group of free-living soil bacteria known for their ability to symbiotically infect the roots of specific host plants as well as to produce siderophores in order to compete with other microorganisms for the limited availability of iron in the rhizosphere. In this study, Rhizobium leguminosarum ATCC 14479, which preferentially infects the red clover Trifolium pratense, was found to produce the trihydroxamate siderophore vicibactin (C 33H55N6O15) under iron restricted conditions. In addition, two other iron-binding, siderophore-like compounds: C20H36N4O10, C31H 55N6O15, were isolated and purified from the culture media. Due to the structural similarity of the latter compounds to vicibactin based on electrospray-mass spectrometry and nuclear magnetic resonance data, these heretofore unreported molecules are thought to be either modified or degraded products of vicibactin. Although vicibactin has previously been found to be commonly produced by other rhizobial strains, this is the first time it has been chemically characterized from a clover infecting strain of R. leguminosarum.

No description available.

No description available.

No description available.