<p>C₄-dicarboxylic acids appear to be metabolized via the TCA cycle in N₂-fixing bacteria (bacteroids) within legume nodules. In <em>Sinorhizobium meliloti</em> bacteroids from alfalfa, NAD⁺-malic enzyme (DME) is required for symbiotic N₂-fixation and this activity is thought to be required for the anaplerotic synthesis of pyruvate. In contrast, in the pea symbiont <em>Rhizobium leguminosarum</em> pyruvate synthesis can occur via either the DME pathway or a pathway catalyzed by phosphoenolpyruvate carboxykinase (PCK), pyruvate kinase (PYK), and pyruvate dehydrogenase. Here we report that <em>dme</em> mutants of <em>Sin</em>or<em>hizobium sp</em>. NGR234 formed root nodules on a broad range of plants and that the level of N₂-fixation varied from 90% to 20% of wild type depending on the host plants inoculated. NGR234 bacteroids had significant PCK activity and while single <em>pckA</em> and single <em>dme</em> mutants fixed N₂ on <em>Macroptilium atropurpureum</em> and <em>Leucaena leucocephala</em> (albeit at a reduced rate), a <em>pckA</em> <em>dme</em> double mutant had no N₂-fixing activity (Fix^-). Thus, NGR234 bacteroids appear to synthesize pyruvate from TCA cycle intermediates via DME or PCK pathways. These NGR234 data, together with other reports, suggested that the completely Fix^- phenotype of <em>S. meliloti dme </em>mutants may be specific to the alfalfa-<em>S. meliloti </em>symbiosis. We therefore examined the ME-like genes <em>azc3656 </em>and <em>azc0119 </em>from <em>Azorhizobium caulinodans</em>, as <em>azc3656 </em>mutants were previously shown to form Fix^- nodules on the tropical legume <em>Sesbania rostrata</em>. We found that purified AZC3656 protein is an NAD (P)⁺-malic enzyme whose activity is inhibited by acetyl-coenzyme A (acetyl-CoA) and stimulated by succinate and fumarate. Thus, whereas DME is required for symbiotic N₂ fixation in <em>A. caulinodans </em>and <em>S. meliloti</em>, in other rhizobia this activity can be bypassed via another pathway(s).</p> <p>In <em>S. meliloti</em> both malic enzymes DME and TME share similar apparent <em>K_m</em>s for substrate and cofactors, but differ in their responses to TCA cycle intermediates, with DME activity inhibited by acetyl-CoA and induced by succinate and fumarate. Previous results in our laboratory indicated that DME is essential for symbiotic N₂ fixation, while TME fails to functionally replace DME. One possible reason for it is that a high ratio of NADPH/NADP⁺in<em> S. meliloti </em>bacteroids prevents TME from functioning in nodules. We sought to lower the<em> </em>NADPH/NADP⁺ratio by overexpressing a soluble pyridine nucleotide transhydrogenase (STH). However, metabolite measurements indicated that overproducing STH failed to lower the ratio of NADPH/NADP⁺ in<em> S. meliloti</em>.</p> <p>Previous studies assumed that DME and TME might play different roles in central carbon metabolism. To gain insight of their physiological functions, genome-wide microarray analysis was conducted in <em>S. meliloti</em> single<em> dme and</em> <em>tme</em> mutants grown on glucose or succinate. The most striking changes of gene expression were observed in <em>S. meliloti</em> <em>dme</em> mutants grown on succinate. The functions of upregulated genes suggested that DME might play an important role in regulating TCA cycle intermediates, important for the maintenance of metabolic flux through TCA cycle during C₄-dicarboxylate oxidation. However, changes of gene expression found in <em>tme </em>mutants were not significant enough to predict the physiological functions of TME protein in central carbon metabolism.</p> / Doctor of Philosophy (PhD)

En raison de leur capacité à former une symbiose avec des bactéries du sol appelées Rhizobium, ces bactéries fixent l’azote atmosphérique et leurs plantes-hôtes n’ont donc pas besoin de fertilisant azoté, les légumineuses jouent un rôle important dans l'agriculture. En outre, certaines souches de Rhizobium ont la capacité de solubiliser le phosphate, fournissant ainsi du phosphore assimilable aux plantes hôtes. Une aptitude à lutter contre certains agents pathogènes a aussi été démontrée dans plusieurs études. La présente étude évalue l’efficacité symbiotique de rhizobiums tunisiens, leur capacité à solubiliser le phosphate, et leur aptitude à lutter contre la verticilliose ainsi que d’autres champignons pathogènes chez Medicago truncatula. Trente-six isolats de rhizobiums prélevés sur des nodules racinaires de M. truncatula provenant de différentes régions de Tunisie ont été obtenus pour ce travail. Environ 60% de ces isolats étaient capables de solubiliser le phosphate in vitro. Dans une seconde étape, trois rhizobiums solubilisant le phosphate et un isolat incapable de solubiliser le phosphate in vitro ont été utilisés pour des essais en phytotron afin de voir l’effet de la présence des rhizobiums sur les paramètres de croissance des plantes en présence de phosphate inorganique sous forme de CaHPO4 et rocheux sous forme brute. Les résultats de l’essai montrent que les plantes de la lignée A17 traitées au CaHPO4, ont tendance à produire plus de nodules et de biomasse aérienne que la lignée F83005.5 et que la forme du phosphate utilisé, soluble ou non soluble, affecte les paramètres étudiés. L'inoculation de quatre lignées de M. truncatula avec 16 isolats de rhizobium sélectionnés auparavant a montré une interaction significative entre les isolats et les lignées pour la symbiose visible par la formation de nodules. Tous les isolats de rhizobium testés ont augmenté la biomasse aérienne des plantes, réduit la biomasse racinaire et entraîné une teneur plus élevée en azote mais l’effet dépendait de l’isolat de rhizobium et de la lignée de M. truncatula utilisés. Enfin, ces isolats ont été testés pour leur capacité à protéger des plantes de M. truncatula contre une maladie racinaire, la verticilliose. Des activités antagonistes in vitro contre divers champignons pathogènes dont Verticillium et Fusarium ont également été recherchées permettant d’identifier un isolat efficace pour la lutte biologique. Les résultats de cette étude suggèrent que des isolats de rhizobium sélectionnés pourraient être utilisés comme biofertilisants dans les sols pauvres pour réduire l'utilisation d'engrais chimiques azotés et phosphorés mais pas pour lutter contre la verticilliose. / Because of their ability to form a symbiosis with soil bacteria called Rhizobium, legumes play an important role in agriculture. These bacteria fix atmospheric nitrogen; hence their host plants do not need nitrogen fertilizers. In addition, some strains of Rhizobium have the ability to solubilize phosphate, thus providing phosphorus to host plants. An ability to control certain pathogens has also been demonstrated in several studies. The present study evaluates the symbiotic efficiency of Tunisian rhizobia, their ability to solubilize phosphate, and their ability to control Verticillium wilt and other pathogenic fungi in Medicago truncatula. Thirty-six rhizobial isolates were obtained from root nodules of M. truncatula from different parts of Tunisia were used in this work. About 60% of these isolates were able to solubilize phosphate in vitro. In a second step, three phosphate solubilizing rhizobia and one isolate unable to solubilize phosphate in vitro were used for phytotron assays to see the effect of the presence of rhizobia on plant growth parameters in the presence of soluble and insoluble forms of phosphate. The results of the experiment show that A17 plants treated with CaHPO4, tend to produce more nodules and shoot biomass than F83005.5 and that the phosphate form used, soluble or non-soluble, affects parameters studied. Inoculation of four M. truncatula lines with 16 previously selected rhizobial isolates showed significant interaction between isolates and lines for symbiotic abilities as visualised by nodule formation. All rhizobial isolates tested increased above-ground biomass, reduced root biomass, and increased nitrogen content with strains effects of plant genotype and bacterial isolate. Finallly, these isolates were tested for their ability to protect M. truncatula plants against Verticillium wilt, and to inhibit the growth of pathogenic fungi such as Verticillium and Fusarium in vitro. However, no isolate could be identified as effective for biological control. The results of this study suggest that selected rhizobial isolates could be used as biofertilizers in poor soils to reduce the use of nitrogen and phosphorus fertilizers but not to control Verticillium wilt.

Contrary to the predictions of numerous theoretical models, variation in partner quality continues to persist in mutualisms, including in the symbiosis between legumes and rhizobia. One potential explanation for the maintenance of this genetic diversity is genotype × genotype interactions, however it is unknown which genetic regions might underlie these interactions. To investigate this question, I performed a quantitative trait loci mapping experiment with two different rhizobium strains to locate potential regions of the genome influencing genotype × genotype interactions between the legume Medicago truncatula and its symbiont Sinorhizobium meliloti. I found no evidence for genotype × genotype or QTL × rhizobium interactions, however some of the QTLs colocalized with genes involved in the symbiosis signaling pathway, suggesting variation in these genes could potentially affect plant performance and fitness traits. These findings have important implications for the evolutionary interactions between legumes and rhizobia, and the genetic architecture of Medicago truncatula.

Contrary to the predictions of numerous theoretical models, variation in partner quality continues to persist in mutualisms, including in the symbiosis between legumes and rhizobia. One potential explanation for the maintenance of this genetic diversity is genotype × genotype interactions, however it is unknown which genetic regions might underlie these interactions. To investigate this question, I performed a quantitative trait loci mapping experiment with two different rhizobium strains to locate potential regions of the genome influencing genotype × genotype interactions between the legume Medicago truncatula and its symbiont Sinorhizobium meliloti. I found no evidence for genotype × genotype or QTL × rhizobium interactions, however some of the QTLs colocalized with genes involved in the symbiosis signaling pathway, suggesting variation in these genes could potentially affect plant performance and fitness traits. These findings have important implications for the evolutionary interactions between legumes and rhizobia, and the genetic architecture of Medicago truncatula.

CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / FundaÃÃo Cearense de Apoio ao Desenvolvimento Cientifico e TecnolÃgico / Banco do Nordeste do Brasil / A regeneraÃÃo natural da vegetaÃÃo à o procedimento mais barato para reabilitar Ãreas degradadas, porÃm, à lento. O plantio de mudas de arbÃreas nativas pode ser uma soluÃÃo. Nesse contexto, conduziu-se o presente trabalho em casa de vegetaÃÃo por 120 dias, testando 5 doses de P (0, 75, 150, 300 e 600 mg L-1) com fosfato de rocha natural na colonizaÃÃo por FMA e por rizÃbio nativos, e no desenvolvimento de mudas de embaÃba (Cecropia pachystachya TrÃcul) e de sabià (Mimosa caesalpiniaefolia Benth), cultivadas em solo natural (SN), solo natural com 50 % esterilizado (SND) e solo natural adicionado de 25 % de material orgÃnico (SNM). Analisou-se o comprimento, massa da matÃria seca, teores de N, P, K da parte aÃrea e o diÃmetro do colo das plantas, nÃmero de esporos e colonizaÃÃo micorrÃzica, massa da matÃria seca dos nÃdulos de rizÃbios, carbono da biomassa microbiana, respiraÃÃo basal e quociente metabÃlico do solo. O crescimento em comprimento, o diÃmetro do colo e a produÃÃo de massa da matÃria seca da parte aÃrea das plantas de embaÃba nÃo sofreram efeito dos tratamentos. Os substratos SN e SND proporcionaram a maior esporulaÃÃo e colonizaÃÃo de FMA, favorecendo o aporte de P na parte aÃrea dessas plantas. Essa atividade foi reduzida nas maiores doses de P. A RespiraÃÃo Basal e o carbono da biomassa microbiana do solo (CBMS) foram menores no SND e nÃo sofreram influÃncia das doses de P. O substrato SND proporcionou maior aporte de N na parte aÃrea das plantas, mas sem influÃncia das doses de P. Enquanto que o SNM proporcionou maior aporte de P e K na parte aÃrea das plantas. Os FMA favoreceram o aporte de P na parte aÃrea das plantas de embaÃba, mas com reduÃÃo nas maiores doses de P. O crescimento em comprimento das plantas de sabià nÃo sofreu efeito dos tratamentos, porÃm o diÃmetro do colo e a massa da matÃria seca da parte aÃrea das plantas apresentaram melhores resultados no substrato SNM, mas sem influÃncia das doses de P. O SNM apresentou o menor nÃmero de esporos, maior colonizaÃÃo de FMA na dose de 75 mg de P, todavia com reduÃÃo nas maiores doses, e maior CBMS, mas sem influÃncia das doses de P. O SND e o rizÃbio nativo proporcionaram maior aporte de N na parte aÃrea e maior massa nodular nas plantas de sabiÃ, sendo influenciados pelas doses de P. O SNM proporcionou maior aporte de P e K na parte aÃrea das plantas de sabiÃ, tendo a dose de 600 mg de P favorecido os maiores valores desses nutrientes. Os FMA proporcionaram um discreto incremento no teor de P na parte aÃrea das plantas de sabiÃ. / Natural regeneration of vegetation is the procedure cheaper to rehabilitate degraded areas, however, is slow. The planting of native tree seedlings may be a solution. In this context, an experiment was carried out in a greenhouse for 120 days, testing 5 P rates (0, 75, 150, 300 and 600 mg L-1) with rock phosphate colonization by AMF and native rhizobia in development of seedlings embaÃba (Cecropia pachystachya TrÃcul) and sabià (Mimosa caesalpiniaefolia Benth), grown in natural soil (NS), natural soil sterilized with 50% (NSS) and natural soil added with 25% organic compost (NSC). We analyzed the length, dry matter weight, N, P, K aerial part and the stem diameter of the plant, number of spores and mycorrhizal colonization, dry matter weight of nodules rhizobia, microbial biomass carbon, basal respiration and metabolic quotient soil. The growth in length, stem diameter and dry matter weight of embaÃba not affected by the treatments. The substrates NS and NSS provided the greatest colonization and sporulation of AMF, favoring the input of P in shoots of these plants. This activity was reduced at the highest levels of P. The soil basal respiration and microbial biomass carbon (MBC) were lower in the NSS and not influenced by P fertilization. The substrate NSS provided major input of N in shoots, not influenced by P fertilization. Whereas the NSC provided major input of P and K in shoots of embaÃba plants. The AMF favored the input of P in shoots of embaÃba plants, but this activity was reduced at the highest levels of P. The growth of sabià was not affected by treatments, but stem diameter and dry mass weight showed better results in substrate NSC. However, were not affected by the rates of P. The NSC had the lowest number of spores, the highest colonization of AMF at rate of 75 mg of P, however with reduction in higher rates, and higher MBC, but without influence of P rates. The NSS and native rhizobia provided major input of N in shoots and increased nodular mass in sabià and levels of phosphorus influenced these parameters. The NSC provided major input of P and K in shoots of sabiÃ, in rate of 600 mg P, favored higher values of these nutrients. AMF provided a mild increase the levels of P in the shoots of sabià plants.

Submitted by (lucia.rodrigues@ufrpe.br) on 2016-08-10T13:22:32Z No. of bitstreams: 1 Maria de Fatima da Silva.pdf: 619107 bytes, checksum: 566cd7dafe2052de47a3a66eb87eda27 (MD5) / Made available in DSpace on 2016-08-10T13:22:32Z (GMT). No. of bitstreams: 1 Maria de Fatima da Silva.pdf: 619107 bytes, checksum: 566cd7dafe2052de47a3a66eb87eda27 (MD5) Previous issue date: 2011-02-28 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / Cowpea (Vigna unguiculata (L.) Walp) in Brazil is grown in the most different climatic conditions. By the adaptability to tropical conditions and recognized as a culture of high socioeconomic value, this legume has an important role for the capitalization of small farmers in the Northeast region. Cowpea may obtain adequate quantities of N by the process of biological nitrogen fixation (FBN) when associated with specific and effective bacteria. However, in Brazil the practice of inoculation is not widely processed due to the low agricultural technology and inconsistent responses using the seed inoculation process. The main goal of this study was to evaluate the effect of different rates of inoculum on effectivity and competitiveness of Bradyrhizobium (strain BR 3267) on cowpea grown in greenhouse conditions. The first experiment was carried out with five different concentration of strain BR 3267 (rhizobia cells/mL of inoculant), N mineral fertilizer treatment without inoculation, and a control without inoculation and N mineral fertilizer. The second experiment conducted in Leonard jars with inoculant produced with BR 3267 strain, 15 native rhizobia (isolated from the used soil), and a control treatment without inoculation and no N mineral fertilizer applied. The application of the low recommended rate increased number and dry biomass of nodules, shoot dry biomass and shoot N total uptake. In the Leonard jars experiment the rhizobia strain BR 3267 was more effective on nitrogen fixation and showed greater competitiveness compared to the native rhizobia isolates. The BRS Pujante cultivar was benefit by the FBN process when inoculated with strain BR 3267 proportional to the inoculum rate. / O feijão - caupi (Vigna unguiculata (L.) Walp.) é cultivado, no Brasil, nas mais diferentes condições edafoclimáticas. Devido a sua boa adaptabilidade às condições tropicais e por ser uma cultura de alto valor sócio-econômico, tem um papel importante para a capitalização de pequenos agricultores na região Nordeste. Por ser uma leguminosa, o feijão-caupi pode adquirir N em quantidades adequadas para suprir suas necessidades através do processo de fixação biológica do nitrogênio (FBN) quando associado com bactérias específicas e efetivas. Entretanto a prática da inoculação não é muito utilizada devido ao baixo aporte tecnológico nas aéreas de cultivo e respostas inconsistentes com os inoculantes utilizados. O objetivo do presente estudo foi de avaliar o efeito da aplicação de diferentes taxas de inóculo, habilidade competitiva e eficiência para a FBN da estirpe de Bradyrhizobium sp. BR 3267 na cultura do feijão-caupi em casa de vegetação. No primeiro experimento, conduzido em vasos com solo foram usados sete tratamentos, compreendendo cinco concentrações de células da estirpe BR 3267 (células/mL de inoculante), um tratamento sem inoculação e com adição de fertilizante nitrogenado, e um controle sem inoculação e sem adição de fertilizante nitrogenado. O segundo experimento constituiu de 17 tratamentos, compostos por 15 estirpes nativas, a estirpe BR 3267 e um tratamento sem inoculação e sem adição de nitrogênio, em vasos de Leonard com substrato estéril. A aplicação da dose mínima recomendada incrementou número e biomassa seca dos nódulos, biomassa seca da parte aérea e o acúmulo de nitrogênio na parte aérea. Em substrato estéril a população rizobiana nativa não foi mais eficiente quando comparada à estirpe BR 3267. A estirpe BR 3267 foi mais competitiva e eficiência em relação à população rizobiana nativa. A cultivar BRS Pujante foi mais beneficiada pela FBN quando inoculada com a estirpe BR 3267, proporcionalmente à taxa de inóculo.

FundaÃÃo Cearense de Apoio ao Desenvolvimento Cientifico e TecnolÃgico / Os diferentes solos e manejos culturais afetam o equilÃbrio entre solo e organismos endÃgenos, os quais, por sua vez afetam a sustentabilidade do solo. Desse modo acredita-se que a diversidade dos organismos do solo tenha uma relaÃÃo estreita com a diversidade de outros organismos, tanto na superfÃcie, quanto no prÃprio solo e que as interaÃÃes dessa diversidade microbiana possam levar a uma alteraÃÃo de funÃÃo reduzindo ou ampliando a sustentabilidade dos ecossistemas. InteraÃÃes mutualÃsticas sÃo muito comuns na natureza e desempenham importante papel em muitos processos de diversos ecossistemas. Desse modo, a identificaÃÃo dos padrÃes da estrutura espacial e abundÃncia de microrganismos à um elemento importante e, necessÃrio para identificar esse processo.AssociaÃÃes mutualÃsticas entre plantas e organismos do solo sÃo essenciais para a sobrevivÃncia e crescimento das plantas na maioria dos ecossistemas terrestres. Assim, o uso combinado de leguminosas e microrganismos na reabilitaÃÃo de solos deteriorados à um processo efetivo na reestabilizaÃÃo dos ciclos de nutrientes nesse sistema, pois a estrutura alimentar do solo pode afetar o desenvolvimento da vegetaÃÃo. O mutualismo entre rizÃbios e leguminosas à possÃvel de manipulaÃÃo experimental. Diferente de alguns mutualistas, rizÃbios podem crescer e ser cultivados em meios seletivos. AlÃm disso, seu comportamento mutualista dentro dos nÃdulos pode ser manipulado e monitorado de modo nÃo invasivo. objetivo deste trabalho foi avaliar a diversidade de estirpes nativas de rizÃbio e a relaÃÃo com algumas espÃcies de leguminosas arbÃreas nativas ocorrentes na Reserva Particular do PatrimÃnio Natural (RPPN) Serra das Almas (05 00â a 05 20â S e 40 48 a 41 12â W) no estado do Cearà (Brasil),em uma Ãrea de caatinga no municÃpio de CrateÃs-Ce, dista 390 Km de Fortaleza, entre cotas de 300 a 350 m de altitude, e que caracteriza-se pro apresentar clima semi Ãrido e pluviosidade mÃdia de 881 mm anuais distribuÃda de Janeiro a Abril. Foram identificadas oito espÃcies de leguminosas arbÃreas, que apresentaramassociaÃÃes com rizÃbios: Anadenanthera colubrina var. cebil (Griseb)Altschu (Angico), Bauhinia cheilantha (Bong.) Stend (MororÃ), Poincianella pyramidalis (Tul.) L.P. Queiroz (Catingueira), Erythrina velutina Willd. (Mulungu), Mimosa caesalpiniifolia Benth (SabiÃ), Minosa acustistipula (Mart.) Benth (Juremabranca), Mimosa tenuiflora (Willd.) Poir (Jurema-preta), Amburana Cearensis (AllemÃo) A.C. Smith (Emburana). Foram coletados nÃdulos e solo rizosfÃrico para a identificaÃÃo de bactÃrias diazotrÃficas, em dois perÃodos, na estaÃÃo chuvosa e na seca. Foi realizado o cultivo destes rizÃbios nas plantas-isca, Macropitillium atropurpureum (DC) Urban, Vigna unguiculata (L., Walp.), Cajanus cajan var. flavus DC e Mimosa pudica L, bem como a caracterizaÃÃo cultural caracterizaÃÃo cultural de estirpes de rizÃbio isolados, testes de tolerÃncia a nÃveis crescentes de NaCl e a altas temperaturas.Verificou-se que 92,42% dos isolados apresentara crescimento rÃpido e 52,24% acidificaram o meio 79. Um total de 84,93% isolados possuem tolerÃncia a altas temperaturas (45 C), e 90,75% isolados apresentaram tolerÃncia Ãs concentraÃÃes salinas a 5%.Os resultados obtidos demonstraram que hà relaÃÃo entre a tolerÃncia à salinidade e à temperatura quando avaliado in vitro para os isolados testados

Symbioses between microbes and multicellular eukaryotes are found in all biomes, and encompass a spectrum of symbiotic lifestyles that includes parasitism and disease, commensalism, and mutually beneficial interdependent host-microbe relationships. Regardless of outcome, these symbiotic lifestyles are governed by a complex molecular "courtship" between microbe and potential host. This courtship is the primary determinant of the host range of a given microsymbiont. Host immunity poses a formidable barrier to the establishment of host-microbe relationships, and the majority of microbial suitors will be thwarted by it. Only by successfully "wooing" the host cell's immune defenses with the appropriate molecular signals can a microsymbiont successfully colonize its host. A strategy common to microsymbionts across the spectrum of symbiotic lifestyles and host organisms is the delivery of microbial-encoded effector proteins into the cytoplasm of host cells to manipulate the host cell's molecular machinery for the purposes of subverting host immunity. Bacteria, in particular, have adapted a number of secretion systems for this purpose. The most well-characterized of these is the type III secretion system (T3SS), a molecular apparatus that specializes in injecting type III effector (T3Es) proteins directly into host cells. The work in this thesis focuses on T3Es of plant-associated bacteria, with particular emphasis on mutualistic bacteria. We present evidence that collections of T3Es from Sinorhizobium fredii and Bradyrhizobium japonicum are, in stark contrast to those of phytopathogenic bacteria, in a co-evolutionary equilibrium with their hosts. This equilibrium is characterized by highly conserved T3E collections consisting of many "core" T3Es with little variation in nucleotide sequence. The T3Es of Mesorhizobium loti MAFF303099 suggest a completely different picture of the evolution of T3Es. MAFF303099 recently acquired its T3SS locus, and the work in this thesis provides an evolutionary snapshot of a mutualist that is innovating a T3E collection primarily through horizontal gene transfer. Collectively, this work represents the first comprehensive catalog of T3Es of rhizobia and, in the case of Sinorhizobium and Bradyrhizobium, the first evidence of purifying selection for T3Es. / Graduation date: 2012

Herbicides have become a key component in modern agricultural production. Meanwhile, there is a concern that some herbicides persist past the growing season of the treated crop, and negatively influence the production of the subsequently planted crops. Amongst various herbicides used in western Canada, acetohydroxyacid synthase (AHAS)-inhibiting herbicides warrant special attention given their residual properties and acute plant toxicity at low concentrations in soil. Soil residual AHAS inhibitors have the potential to influence both leguminous host plants and their bacterial symbiotic partners; consequently, the use of an AHAS inhibitor in a given year can negatively influence the inoculation success and grain yield of legumes cropped in the following year. <p>The present thesis project focused on one of the AHAS inhibiting herbicides (flucarbazone) and studied its potential for carryover injury and negative influence on the success of inoculation in field pea. A series of growth chamber and field experiments were conducted to test the following null hypothesis: the presence of residual flucarbazone in soil does not affect nodulation of field pea by inoculum rhizobia. <p>A growth chamber experiment clearly demonstrated the susceptibility of field pea to the presence of flucarbazone in soil where the lowest concentration of flucarbazone amendment (5 ìg kg1) significantly reduced the crop growth. In contrast, a field study failed to reveal any negative effects of flucarbazone use on crop growth and N2 fixation. <p>It was concluded that if the weather and soil conditions favour decomposition of flucarbazone as described in the present study, flucarbazone applied at the recommended field rate will not persist into the following season at high enough concentrations to negatively influence field pea growth, grain yields, and inoculation success. To ensure safety of rotational crops, it is important to strictly adhere to the herbicide application guidelines. Additionally, producers are cautioned to be particularly aware of the environmental and soil conditions that may reduce the rate of herbicide degradation.

Herbicides have become a key component in modern agricultural production. Meanwhile, there is a concern that some herbicides persist past the growing season of the treated crop, and negatively influence the production of the subsequently planted crops. Amongst various herbicides used in western Canada, acetohydroxyacid synthase (AHAS)-inhibiting herbicides warrant special attention given their residual properties and acute plant toxicity at low concentrations in soil. Soil residual AHAS inhibitors have the potential to influence both leguminous host plants and their bacterial symbiotic partners; consequently, the use of an AHAS inhibitor in a given year can negatively influence the inoculation success and grain yield of legumes cropped in the following year. <p>The present thesis project focused on one of the AHAS inhibiting herbicides (flucarbazone) and studied its potential for carryover injury and negative influence on the success of inoculation in field pea. A series of growth chamber and field experiments were conducted to test the following null hypothesis: the presence of residual flucarbazone in soil does not affect nodulation of field pea by inoculum rhizobia. <p>A growth chamber experiment clearly demonstrated the susceptibility of field pea to the presence of flucarbazone in soil where the lowest concentration of flucarbazone amendment (5 ìg kg1) significantly reduced the crop growth. In contrast, a field study failed to reveal any negative effects of flucarbazone use on crop growth and N2 fixation. <p>It was concluded that if the weather and soil conditions favour decomposition of flucarbazone as described in the present study, flucarbazone applied at the recommended field rate will not persist into the following season at high enough concentrations to negatively influence field pea growth, grain yields, and inoculation success. To ensure safety of rotational crops, it is important to strictly adhere to the herbicide application guidelines. Additionally, producers are cautioned to be particularly aware of the environmental and soil conditions that may reduce the rate of herbicide degradation.