Submitted by Sandra Pereira (srpereira@ufrrj.br) on 2017-01-09T10:11:25Z No. of bitstreams: 1 2014 - Joel Quintino de Oliveira Junior.pdf: 986333 bytes, checksum: 680ec4322bce52cb9979155c6978ea36 (MD5) / Made available in DSpace on 2017-01-09T10:11:25Z (GMT). No. of bitstreams: 1 2014 - Joel Quintino de Oliveira Junior.pdf: 986333 bytes, checksum: 680ec4322bce52cb9979155c6978ea36 (MD5) Previous issue date: 2014-02-28 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Leguminosae comprises a wide variety of species that form symbiosis with mycorrhizal fungi (AMF), and mostly with nitrogen fixing bacteria. Within this frame fits the group Pitadenia encompassing genres: Mimosa , Piptadenia , Parapiptadenia , Anadenathera , Pseudopiptadenia , Microlobius , Stryphnodendron and Adenopia , comprising over 500 species, of which most are found in the New World , especially in Brazil . Their habits vary from tree and herbaceous species to thrive in flooded, dry, high altitude grasslands , savanna ecosystems among other areas . Are well known and used in the rehabilitation of degraded soils by their aggressiveness and ability to grow under adverse soil conditions. Results indicate that in some species of this group only nodulated with the presence of mycorrhizal fungi. Legumes use for the realization of these two symbioses very similar molecular mechanisms and some of leguminous species from the subfamily Mimosoideae show atypical and high potential exploration of evolutionary symbiosis between the two skills. The objective of this work is to investigate and confirm this dependence of the presence of arbuscular mycorrhizal fungi for nodulation and nitrogen fixation in species group Piptadenia nodulated by Burkholderia sp . under controlled conditions / A fam?lia Leguminosae compreende um imensa variedade de esp?cies que formam simbiose com fungos micorr?zicos arbusculares (FMAs) e, em sua grande maioria, com bact?rias fixadoras de nitrog?nio. Dentro desse quadro se encaixa o grupo de Pitadenia que engloba os g?neros: Mimosa, Piptadenia, Parapiptadenia, Anadenathera, Pseudopiptadenia, Microlobius, Stryphnodendron e Adenopia, compreendendo mais de 500 esp?cies, sendo que a maioria delas ? encontrada no novo mundo, em especial no Brasil. Seus h?bitos variam desde esp?cies herb?ceas at? arb?reas e se desenvolvem em ?reas alagadas, secas, campos de altitude, cerrado entre outros ecossistemas. S?o bem conhecidas e utilizadas na reabilita??o de solos degradados por sua agressividade e capacidade de se desenvolver nas mais adversas condi??es ed?ficas. Resultados indicam que em algumas esp?cies desse grupo s? nodulariam com a presen?a de fungos micorr?zicos. As leguminosas usam para a realiza??o dessas duas simbioses mecanismos moleculares muito similares e algumas dessas esp?cies de leguminosas nativas da subfam?lia Mimosoideae mostram habilidades at?picas e com alto potencial de explora??o evolucion?ria entre as duas simbioses. O objetivo deste trabalho ? investigar e confirmar essa depend?ncia da presen?a de fungos micorr?zicos arbusculares para a nodula??o e fixa??o biol?gica de nitrog?nio em esp?cies do grupo de Piptadenia noduladas por Burkholderia sp. sob condi??es controladas

Genotypic differences in growth and yield of two common bean (Phaseolus vulgaris L) cultivars to Rhizobium inoculation and management were investigated. In 2003-04, the two bean cultivars (Scylla and T-49) were combined with three inoculant treatments (strains CC 511 and RCR 3644, and a control of no inoculation), two fertiliser levels (0 and 150 kg N ha⁻¹) and two irrigation treatments (irrigated and rainfed). There was no nodulation on either cultivar. To further investigate the symbiotic relationship, 16 rhizobial isolates, including the two used in the first field experiment, were combined with the cultivar Scylla and evaluated in a greenhouse. Subsequently, five Rhizobium isolates were chosen for further field evaluation, based on signs of early nodulation in the greenhouse trial. The second field experiment in 2004-05 combined the five inoculant strains (RCR 3644, UK 2, H 20, PRF 81, PhP 17 and a control) with two bean cultivars (Scylla and T-49). In the greenhouse, nodule number varied from 7 (UK 2) to 347 (H 441) nodules plant⁻¹ at 51 DAS and from 13 (UK 1) to 335 (CIAT 899) nodules plant⁻¹ at 85 DAS. In 2004-05, in the field, nodulation was also variable, ranging between 1 and approximately 70 nodules plant⁻¹, with higher nodules numbers plant⁻¹ being found on cultivar T-49. Of the isolates used in the field, strains H 20, PRF 81 and PhP 17 produced 70, 25 and 12 nodules plant⁻¹ at 70, 40 and 54 DAS respectively. Nodules formed were of various sizes and more than 80 % were pink to dark red in colour denoting the presence of leghaemoglobin and active N fixation. The remaining nodules were either green or white. The importance of selecting an appropriate cultivar for the growing conditions was highlighted in these experiments. Leaf area index, leaf area duration intercepted radiation and final utilisation efficiency were significantly affected by cultivar. In both seasons cv. T-49 reached maturity (dry seed) before Scylla, while unirrigated plants reached green pod maturity seven days before irrigated plants. Plants of cv. Scylla gave a final TDM of 730 g m⁻²; compared to the 530 g m⁻² produced by T-49. The average growth rate was 7.0 and 5.2 g m⁻² day⁻¹ for Scylla and T-49 respectively (2003-04). Plants receiving 150 kg N ha⁻¹ produced 665 g m⁻² TDM which was 12 % more than was produced by unfertilised plants. The application of 150 kg N ha⁻¹ gave an average growth rate of 6.4 g m⁻² day⁻¹ compared to 5.7 g m⁻² day⁻¹ from plants with no N. Inoculation in the field had no significant effect on TDM in both seasons. Temperature affected growth and DM accumulation. Accumulated DM was highly dependent on cumulative intercepted PAR. Air temperatures below the base temperature (10 °C) affected growth in 2004-05, resulting in plants accumulating just 0.24 g DM MJ⁻¹ PAR during early growth. This increased to 2.26 g DM MJ⁻¹ PAR when the temperature was increased above the base temperature. There was a strong relationship between LAI and intercepted PAR. A LAI of 4.0-4.5 was required to intercept 90-95 % of incident solar radiation. Cultivar significantly (p < 0.001) affected radiation use efficiency (RUE). Scylla had a RUE of 1.02 g DM MJ⁻¹ PAR compared to T-49 at 1.18 g DM MJ⁻¹ PAR. Seed yield was significantly (p < 0.001) affected by cultivar and fertiliser application. Cultivar Scylla produced 467 g m⁻² which was 76 % more than T-49, while a 12 % increase in seed yield was observed in N fertilised plants over unfertilised plants. Only cultivar significantly affected HI, while the yield components that had the greatest effect on seed yield were hundred seed weight and pods plant⁻¹. Inoculation significantly (p< 0.05) affected 100 seed weight (2004-05). Plants inoculated with strain H 20 had the highest 100 seed weight at 25.2 g with cv. Scylla producing larger seeds than T-49. The belief that local environmental conditions play a major role on field survival of bacteria, led to the use of PCR methods to identify field nodulating organisms. Amplification of genomic DNA from parent isolates using primers fC and rD generated a single band for each isolate. Isolates were identified to the species level as either Rhizobium or Agrobacterium, using the highly conserved internally transcribed spacer (ITS) region and are known to nodulate common bean. The DNA extracted from the isolates recovered from nodules of field grown beans gave multiple bands with primers fC and rD. Five distinct banding patterns were observed. All of these were different from those of parent isolates. Sequencing of the 16S rRNA demonstrated that nodules of field grown beans in Canterbury were inhabited by Pseudomonads either alone or in association with other root nodulating organisms. The inability to identify the inoculant strains in nodules of field grown beans does not rule out their infection and nodulating function in the cultivars used. The results suggest the possibility of both Rhizobium and Pseudomonads cohabiting in the nodules of field grown beans. The aggressive nature of Pseudomonads on artificial media, possibly out competing the inoculant rhizobia is proposed, leading to the inability to identify the inoculant strain from the nodules of the field grown beans by PCR methods. The need to identify the nodule forming or nodule inhabiting bacteria in the nodules is necessary to classify the importance of these organisms and their economic benefit to agricultural production. This study also underlines the importance of using PCR methods to gain valuable insights into the ecological behaviour of Rhizobium inoculants and nodule inhabiting organisms.

Made available in DSpace on 2015-04-20T12:31:32Z (GMT). No. of bitstreams: 1 Aloisio Freitas Chagas Junior.pdf: 1425510 bytes, checksum: 861fcee7c2ea43864c650307852db610 (MD5) Previous issue date: 2007-04-13 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / To increase the rhizobia contribution to plants nutrition and development is necessary to develop researches for selecting more efficient nitrogen fixation strains. This research work had as objectives, to accomplish morphologic characterization of rhizobia isolated from Amazonian soils; laboratory evaluations about the tolerance to acidity and toxic Al, the capacity to solubilize calcium (P-Ca) and aluminum (P-Al) phosphates, the indole-acetic acid production (AIA); the ability of the isolates to induce nodulation and to fix nitrogen under the acidity and low fertility conditions of the regional soils in greenhouse experiment; to genetically characterize the isolated ones selected using the PCR-RFLP technique. The infectivity test showed that there was significant increase in the cowpea biomass. With relationship to the phenotypic characterization, the 200 isolates showed a great diversity. Among these, 165 and 127 presented high tolerance within the media with pH 4,5 and pH 4,5 + Al, respectively. From the 200 isolates, 68 solubilized P-Ca and 47 P-Al, and 32 isolates solubilized both P-Ca and P-Al. The AIA production significantly increased (p <0,01) with tryptophan quantities added in the medium for 19 of the 92 isolates tested in the experiment. It was observed production of dry matter and nodule number and mass superior simbiotic to the control treatment and similar to the fertilized treatment for various rhizobia isolates. The 20 selected isolates evaluated presented high genotypic diversity, with the derived dendrogram PCR-RFLP profiles showing that they can be divided inside two main genders, Rhizobium and Bradyrhizobium. Associating the results of simbiotic efficiency and nodulation, it was observed that the isolates of rhizobia from Amazonian soils presented good results, and they can be recommended for tests of agronomic efficiency for cowpea bean. / Para aumentar a contribuição de rizóbia na nutrição e desenvolvimento de plantas é necessário desenvolver pesquisas de seleção de estirpes mais eficientes quanto à fixação do N2. Este trabalho teve como objetivos, realizar a caracterização morfológica de isolados de rizóbio oriundos de solos amazônicos; avaliar em laboratório, a tolerância à acidez e Al tóxico, a capacidade de solubilização de fosfato de cálcio (P-Ca) e alumínio (P-Al), a produção de acido indol-acético (AIA); a capacidade de isolados em induzirem nodulação e fixarem nitrogênio sob as condições de acidez e baixa fertilidade dos solos regionais em casa-de-vegetação; caracterizar geneticamente os isolados selecionados usando a técnica de PCR-RFLP. Os testes de infectividade mostraram que houve aumento significativo na biomassa de caupi. Quanto à caracterização fenotípica, os 200 isolados mostraram uma grande diversidade. Dentre estes, 165 e 127 isolados apresentaram elevada tolerância nos meios de cultura com pH 4,5 e pH 4,5 + Al, respectivamente. Dos 200 isolados testados, 68 solubilizaram P-Ca e 47 P-Al, sendo que 32 solubilizaram tanto o P Ca quanto o P Al. A produção de AIA aumentou significativamente com as doses de triptofano adicionadas ao meio (p<0,01) para 19 dos 92 isolados testados no experimento. Observou-se produção de matéria seca, número e massa seca dos nódulos e acúmulo de nitrogênio superiores ao tratamento testemunha e similares ao tratamento adubado para vários isolados de rizóbia. Os 20 isolados selecionados e avaliados apresentaram alta diversidade genotípica, onde o dendrograma derivado de perfis de PCR-RFLP mostrou que podem ser divididos dentro de dois gêneros principais, Rhizobium e Bradyrhizobium. Associando-se os resultados de eficiência simbiótica e nodulação, observou-se que os isolados de rizóbio de solos da Amazônia apresentaram bom desempenho e podem ser recomendados para testes de eficiência agronômica em feijão caupi.