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Jucimara Anunciação De Jesus Sousa INTERAÇÕES ECOLÓGICAS E POTENCIAL BIOFERTILIZANTE DE ACTINOBACTÉRIAS E BACTÉRIAS DIAZOTRÓFICAS NA AGRICULTURA JUCIMARA ANUNCIAÇÃO DE JESUS SOUSA UNIVERSIDADE ESTADUAL DO NORTE FLUMINENSE - UENF CAMPOS DOS GOYTACAZES - RJ FEVEREIRO, 2017 INTERAÇÕES ECOLÓGICAS E POTENCIAL BIOFERTILIZANTE DE ACTINOBACTÉRIAS E BACTÉRIAS DIAZOTRÓFICAS NA AGRICULTURA JUCIMARA ANUNCIAÇÃO DE JESUS SOUSA “Tese apresentada ao Centro de Biociências e Biotecnologia da Universidade Estadual do Norte Fluminense Darcy Ribeiro como parte das exigências para obtenção do título de Doutora em Biociências e Biotecnologia” Orientador: Prof. D. Sc. Fabio Lopes Olivares UNIVERSIDADE ESTADUAL DO NORTE FLUMINENSE - UENF CAMPOS DOS GOYTACAZES - RJ FEVEREIRO, 2017 INTERAÇÕES ECOLÓGICAS E POTENCIAL BIOFERTILIZANTE DE ACTINOBACTÉRIAS E BACTÉRIAS DIAZOTRÓFICAS NA AGRICULTURA JUCIMARA ANUNCIAÇÃO DE JESUS “Tese apresentada ao Centro de Biociências e Biotecnologia da Universidade Estadual do Norte Fluminense Darcy Ribeiro como parte das exigências para obtenção do título de Doutora em Biociências e Biotecnologia” Aprovada em 27 de outubro de 2016 Comissão Examinadora: Jean Luiz Simões de Araújo (D.Sc., Ciências Biológicas) - EMBRAPA Agrobiologia Alessandro Coutinho Ramos (D.Sc., Produção Vegetal) - UENF Solange Silva Samarão (D.Sc., Biociências e Biotecnologia) - UENF Tatiane Sanches Soares (D.Sc., Ciências Biológicas) - UENF (Revisora) Fabio Lopes Olivares (D.Sc., Agronomia) - UENF (Orientador) iii DEDICATÓRIA Esta tese é dedicada às pessoas que mais amo no mundo, minha família. Em especial, meu amado esposo Rony e minha mãe Juçara. iv AGRADECIMENTOS AGRADECIMENTOS A Deus, autor e consumador da minha fé. Fonte de força, sabedoria e perseverança; À minha amada mãe e amiga, Juçara, pelo amor e conforto com sábias palavras; À minha família pelo apoio e alegria; A Rony, meu esposo, meu amor, companheiro sempre presente nessa luta. Por tolerar minhas oscilações de humor, enxugar minhas lágrimas e me fazer rir de tudo no final; Ao professor Fabio Lopes Olivares pelo aprendizado; Aos membros da banca: D.Sc. Jean Luiz Simões, D.Sc. Alessandro Coutinho Ramos, D.Sc. Solange Silva Samarão e D.Sc. Tatiane Sanches Soares pelas valorosas contribuições; A todos os amigos e companheiros de trabalho, pelas conversas, orientações e momentos de descontração; A todos os demais professores, técnicos e funcionários da UENF; A todos que, de algum modo, contribuíram para eu concluir esta etapa; À FAPERJ e UENF pela concessão da bolsa de doutorado; À UENF e ao Programa de Pós-Graduação em Biociências e Biotecnologia pela oportunidade de realização do doutorado. v “A menos que modifiquemos nossa maneira de pensar, não seremos capazes de resolver os problemas causados pela forma como nos acostumamos a ver o mundo”. (Albert Einstein) vi SUMÁRIO SUMÁRIO RESUMO GERAL .................................................................................................................. x ABSTRACT ...........................................................................................................................xi INTRODUÇÃO ...................................................................................................................... 1 CAPÍTULO 1 .......................................................................................................................... 3 PLANT GROWTH PROMOTION BY STREPTOMYCETES: ECOPHYSIOLOGY, MECHANISMS AND APPLICATIONS ................................................................................... 3 ABSTRACT ........................................................................................................................ 4 INTRODUCTION ................................................................................................................ 5 REVIEW ............................................................................................................................. 6 Historical and taxonomic aspects .................................................................................... 6 Morphological differentiation and physiology ................................................................... 7 Ecology of Streptomycesplant host interaction ............................................................ 12 Plant growth promotion by streptomycetes ................................................................... 15 Biotechnological application of PGPS in agriculture ...................................................... 19 Concluding remarks and perspectives .......................................................................... 21 REFERENCES ................................................................................................................. 23 CAPÍTULO 2 .......................................................................................................................... 1 MICROBIOLOGICAL AND AGRO-BIOTECHNOLOGICAL CHARACTERIZATION OF HUMIC ACID USING-STREPTOMYCETES ISOLATED FROM MATURED VERMICOMPOST. ........ 1 ABSRACT ........................................................................................................................ 38 INTRODUCTION .............................................................................................................. 39 MATERIAL AND METHODS ............................................................................................ 40 Actinobacteria isolation procedures in vermicompost material ...................................... 40 Molecular characterization ............................................................................................ 41 Use of different carbon and nitrogen sources ................................................................ 42 Production of extracellular enzymes ............................................................................. 43 Calcium phosphate and zinc oxide solubilization .......................................................... 43 Production of related indole compounds ....................................................................... 44 Salinity and pH tolerance .............................................................................................. 44 DR5::GUS reporter Micro-Tom tomato assay ............................................................... 45 Assessment of maize growth promotion ....................................................................... 45 Statistical analysis ........................................................................................................ 46 RESULTS ........................................................................................................................ 47 Isolation of actinobacteria associated with matured vermicompost ............................... 47 vii Identification of actinobacteria isolates .......................................................................... 48 Carbon and nitrogen nutritional requirements ............................................................... 48 Other Physiological Features ........................................................................................ 52 Micro-Tom DR5::GUS reporter response to Streptomyces secreted metabolites .......... 52 Gnobiotic studies for maize growth promotion response inoculated with the Streptomyces strains .................................................................................................... 56 DISCUSSION ................................................................................................................... 57 CONCLUSIONS ............................................................................................................... 62 REFERENCES ................................................................................................................. 63 CAPÍTULO 3 ........................................................................................................................ 69 INTERAÇÃO ENTRE A BACTÉRIA PROMOTORA DO CRESCIMENTO VEGETAL Herbaspirillum seropedicae estirpe HRC54 E ESTREPTOMICETOS ISOLADOS DE VERMICOMPOSTOS EM PLÂNTULAS DE TOMATE. ........................................................ 69 RESUMO ......................................................................................................................... 69 ABSTRACT ...................................................................................................................... 70 INTRODUÇÃO ................................................................................................................. 71 MATERIAL E MÉTODOS ................................................................................................. 73 Bactéria diazotrófica e isolados de estreptomicetos: condições de cultivo .................... 73 Ensaio de compatibilidade ............................................................................................ 74 Potencial de interações para promoção de crescimento de plântulas de tomate .......... 74 Interação entre H. seropedicae estirpe HRC54 e S. bellus isolado AC06, um produtor de amilases ....................................................................................................................... 75 Flutuação populacional de H. seropedicae estirpe HRC54 em co-cultura com S. bellus isolado AC06 ............................................................................................................. 75 Avaliação do sistema tripartite H. seropedicae estirpe HRC54, S. bellus isolado AC06 e plântulas de tomate ...............................................................................................
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