Submitted by (lucia.rodrigues@ufrpe.br) on 2016-06-29T14:32:08Z No. of bitstreams: 1 Carla Regine Reges Silva Franca.pdf: 474566 bytes, checksum: b95c31472908cbb435a486697979ad3b (MD5) / Made available in DSpace on 2016-06-29T14:32:08Z (GMT). No. of bitstreams: 1 Carla Regine Reges Silva Franca.pdf: 474566 bytes, checksum: b95c31472908cbb435a486697979ad3b (MD5) Previous issue date: 2009-05-27 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / This study had for objective to evaluate the growth kinetics and characteristics morphophysiological and symbiotic strains of rhizobia of four, front the conditioning different liquids. The experiment was composed of four strains (two of the strains of Rhizobium tropici (SEMIA 4077-SEMIA 4080) and two of Bradyrhizobium japonicum (SEMIA 5079-SEMIA 5080), respectively for bean (Phaseolus vulgaris) and soybean (Glycine max), three conditioning (NaCl 0,1%, glicerol 20% and carboximetilcelulose 0,2%) more fresh bacteria, that you/they were grown close to each evaluation period. After the growth period in liquid middle YM 0,5 mL of the bacterial broth of each strains was transferred for tubes with capacity for 2 mL, containing 1,5 mL of each conditioning liquid and maintained in room temperature by 180 days. After the accomplishment of the different formulations for the conservation of the strains, it was made the counting of UFC that contained 108 cells of the rhizobia culture approximately using serial dilutions and the method of the drop. During the experiment, readings were accomplished in the time 0, 1, 7, 15, 21 days after the conservation, following for monthly samples 30, 60, 90, 120, 150, 180 days. The efficiency symbiotic was evaluated in the periods of 15, 60 and 120 days after the conservation in bean plants and soy. The results allow to observe that the concentration of viable cells varied among the preservantes. The preservante NaCl 0,1% maintained the concentration of viable cells around 107 and 108 UFC mL-1 for the strains SEMIA 4077, 5080 and 5079 for up to 180 days of conservation and it nodulation to the 15 and 120 days of conservation for the strains SEMIA 4077 and SEMIA 4080, when inoculated in bean seeds. The conditioning Glicerol 20%, didn't present desirable characteristics for a good conservation method with the strains SEMIA 4080, SEMIA 5079 and SEMIA 5080. The carboximetilcelulose was promising for the conservation of the ancestries SEMIA 4077 and SEMIA 5079 in room temperature. Nodulation was influenced by the preservative used. In all of the preservantes there was decrease in the number of nodules during the conservation period. With base in the results the conditioning liquids NaCl 0,1% and Carboximeticelulose 0,2% were the one that got maintain the cells of the strains for a period of short period, besides preserving the characteristics morfofisiológicas of the strains of Rhizobium and Bradyrhizobium. / Este estudo visou avaliar a cinética de crescimento e características morfofisiológicas e simbiótica de quatro estirpes de rizóbio, frente a diferentes condicionadores líquidos. O experimento foi composto de quatro estirpes (sendo duas de Rhizobium tropici (SEMIA 4077 – SEMIA 4080) e duas de Bradyrhizobium japonicum (SEMIA 5079 – SEMIA 5080), recomendadas respectivamente para feijão (Phaseolus vulgaris) e soja (Glycine max), três condicioadores líquidos (NaCl 0,1 % , glicerol 20 % e carboximetilcelulose 0,2 %). Após cultivo em meio líquido YM 0,5 mL do caldo bacteriano de cada estirpe foi transferido para tubos com capacidade para 2 mL, contendo 1,5 mL de cada condicionador líquido e mantidas em temperatura ambiente por 180 dias. Em seguida, fez-se a contagem utilizando diluições seriadas e o método da gota das UFC que foi de aproximadamente 108 células. Durante o experimento, foram realizadas determinações da população bacteriana aos 0, 1, 7, 15, 21 dias após a conservação, seguido por amostras aos 30, 60, 90, 120, 150, 180 dias. A eficiência simbiótica foi avaliada nos períodos de 15, 60 e 120 dias após a conservação em plantas de feijão e soja. Os resultados permitem observar que a concentração de células viáveis variou entre os preservantes. O preservante NaCl 0,1% manteve a concentração de células viáveis entre 107 e 108 UFC mL-1 para as estirpes SEMIA 4077, 5080 e 5079 por até 180 dias de conservação e produziu efetiva nodulação aos 15 e 120 dias de conservação para as estirpes SEMIA 4077 e SEMIA 4080, quando inoculados em sementes de feijão. O preservante Glicerol 20%, não apresentou características desejáveis para um bom método de preservação com as estirpes SEMIA 4080, SEMIA 5079 e SEMIA 5080. O carboximetilcelulose foi promissor para a conservação das estirpes SEMIA 4077 e SEMIA 5079 em temperatura ambiente. A nodulação foi influenciada pelo preservante utilizado. Em todos os preservantes houve diminuição no número de nódulos durante o período de conservação. Com base nos resultados osxii condicionadores líquidos NaCl 0,1 % e Carboximeticelulose 0,2 % foram os que conseguiram manter as células das estirpes por um período de curto prazo, além de preservar as características morfofisiológicas das estirpes de Rhizobium e Bradyrhizobium.

Submitted by Celso Magalhaes (celsomagalhaes@ufrrj.br) on 2018-09-13T17:03:37Z No. of bitstreams: 1 2017 - Gustavo Feitosa de Matos.pdf: 1844017 bytes, checksum: 0ed6966082e21d15123a12cd0e5777ef (MD5) / Made available in DSpace on 2018-09-13T17:03:38Z (GMT). No. of bitstreams: 1 2017 - Gustavo Feitosa de Matos.pdf: 1844017 bytes, checksum: 0ed6966082e21d15123a12cd0e5777ef (MD5) Previous issue date: 2017-02-23 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico - CNPQ / Recent molecular biology studies have indicated that bacteria of the genus Bradyrhizobium and Rhizobium may also play a role in the biological nitrogen fixation process (BNF) when associated with sugarcane. The use of Vigna unguiculata trap plants allowed obtaining a diverse collection of endophytic Bradyrhizobium spp. from sugarcane roots. The present work aimed to characterize a group of bacteria of the genus Bradyrhizobium from sugarcane roots, as well as to evaluate the ability of a representative strain of this group and an isolate of the genus Rhizobium to promote growth in this crop. To define the taxonomic position of the isolates of the group, phylogenetic analyzes were performed with the individual 16S rRNA, ITS, nodC and recA genes and multilocus sequence analysis (MLSA) involving four housekeeping genes (recA, dnaK, glnII and atpD), in four representative isolates (BR 10280T, BR 10266, BR 10555 and BR 10556). In addition, biochemical and morphophysiological tests were performed. Experiments in which the isolates P9-20 (BR 10280) (Bradyrhizobium sp.) and P5-2 (Rhizobium sp.) were inoculated in sugarcane seedlings were conducted in a greenhouse to evaluate the growth promoting effect of these isolates. Two harvests were performed at 30 and 75 days after transplanting (DAT). Among the analyzed variables were the budding speed index (BSI), dry mass and total nitrogen (N). Phylogenetic analyzes positioned the isolates in the superclade of B. japonicum in an independent branch close to B. huanghuaihaiense, a soybean micro-symbiont. Despite the proximity of the group under study to B. huanghuaihaiense, these isolates did not induce nodulation in Glycine max. In addition, unlike B. huanghuaihaiense, the new isolates induced nodule formation in Phaseolus vulgaris. In morphophysiological studies, significant differences were demonstrated between the representative isolates from this study and B. huanghuaihaiense. In the greenhouse experiment, an increase on BSI and root dry mass of the inoculated treatment with the P5-2 isolate was observed in the first harvest. In the second harvest, increments were found in the root dry mass and root volume, as well as in the total N of the roots and aerial part, in the treatment inoculated with P5-2, although no statistical difference was detected. These results of the characterization indicate that the sugarcane isolates of the genus Bradyrhizobium represent a new species of this genus. In relation to the greenhouse experiment, the isolate of the genus Rhizobium sp. presented potential as a growth promoter in the sugarcane crop. / Estudos recentes de biologia molecular indicaram que bact?rias do g?nero Bradyrhizobium e Rhizobium tamb?m podem ter um papel no processo de fixa??o biol?gica de nitrog?nio (FBN) quando associadas ? cana-de-a??car. O uso de plantas de Vigna unguiculata como ?isca? possibilitou a obten??o de uma cole??o diversa de Bradyrhizobium spp. endof?ticos a partir de ra?zes de cana-de-a??car. O presente trabalho teve como objetivo caracterizar um grupo de bact?rias do g?nero Bradyrhizobium proveniente de ra?zes de cana-de-a??car, assim como avaliar a capacidade de uma estirpe representante desse grupo e de um isolado do g?nero Rhizobium em promover efeito de crescimento nessa cultura. Para definir o posicionamento taxon?mico dos isolados do grupo, an?lises filogen?ticas foram realizadas com os genes individuais 16S rRNA, ITS, nodC e recA e an?lise de sequ?ncia de multilocus (MLSA) envolvendo quatro genes ?housekeeping? (recA, dnaK, glnII e atpD), em quatro isolados representantes (BR 10280T, BR 10266, BR10555 e BR 10556). Al?m disso, foram realizados testes bioqu?micos e morfofisiol?gicos. Experimento onde os isolados P9-20 (BR 10280) (Bradyrhizobium sp.) e P5-2 (Rhizobium sp.) foram inoculados em minitoletes de cana-de-a??car foi conduzido em casa de vegeta??o. Foram realizadas coletas aos 30 e 75 dias ap?s transplantio (DAT). Entre as vari?veis analisadas est?o o ?ndice de velocidade de brotamento (IVB), massa seca e Nitrog?nio (N) total. An?lises filogen?ticas posicionaram os isolados no super clado de B. japonicum em um ramo independente pr?ximo a B. huanghuaihaiense, um microssimbionte de soja. Apesar da proximidade do grupo em estudo com B. huanghuaihaiense, estes isolados n?o induziram nodula??o em Glycine max. Al?m disso, diferente de B. huanghuaihaiense, os novos isolados induziram a forma??o de n?dulos em Phaseolus vulgaris. Em estudos morfofisiol?gicos foram demostradas diferen?as significativas entre os isolados representantes do grupo em estudo e B. huanghuaihaiense. Em experimento em casa de vegeta??o, observou-se na primeira coleta um incremento sobre o IVB e massa seca da raiz no tratamento inoculado com o isolado P5-2, embora n?o tenham sido observadas diferen?as estat?sticas. Na segunda coleta, incrementos foram encontrados na massa seca de raiz e volume de raiz, assim como, no N total das ra?zes e parte a?rea, no tratamento incoculado com P5-2, embora n?o tenham apresentado diferen?a estat?stica. Massa seca da parte a?rea do tratamento inoculado com P5-2 apresentou incremento na primeira e segunda coleta, em rela??o aos controles, embora n?o tenha sido detectada diferen?a estat?stica. Os resultados da caracteriza??o do grupo indicam que os isolados de cana-de-a??car do g?nero Bradyrhizobium representam uma nova esp?cie desse g?nero. Em rela??o ao experimento em casa de vegeta??o, o isolado do g?nero Rhizobium sp. apresentou potencial como promotor de crescimento na cultura de cana-de-a??car

Das Bakterium Bradyrhizobium japonicum ist wie andere Rhizobien in der Lage mit Pflanzen der Familie Fabales (Leguminosen) eine Symbiose einzugehen. Im symbiontischen Zustand fixieren die Mikrosymbionten atmosphärischen, molekularen Stickstoff und stellen diesen den Pflanzen in verwertbarer Form zur Verfügung. Im Gegenzug erhalten die Bakterien von den Pflanzen verschiedene Verbindungen als Kohlenstoff- und Energiequelle. Dem geht ein komplexer Signalaustausch voraus um die gegenseitige Erkennung der Partner und die Spezies-spezifische Symbiose zu ermöglichen. Auf Seite der Bakterien erfolgt die Reaktion auf die Gegenwart pflanzlicher Signalmoleküle. In B. japonicum induziert das Flavonoid Genistein die Transkription einer Reihe von Genen. Durch die Expression der nod-Gene erfolgt eine Synthese von Lipochitooligosacchariden. Diese sogenannten Nodulationsfaktoren rufen wiederum eine Reaktion in der Wirtspflanze hervor. Um die zugrunde liegende Genexpression und deren Regulationsmechanismen zu erforschen, standen bis dato nur Methoden für die Untersuchung einzelner Gene zur Verfügung. Die erstmals 1995 publizierte Microarraytechnologie eröffnete die Möglichkeit zu einem Zeitpunkt die Gentranskription eines gesamten Organismus zu untersuchen (Schena et al. 1995). Um mit dieser Technologie die Gentranskription bei B. japonicum zu untersuchen, wurde ein auf PCR-Produkten basierender Microarray hergestellt. Grundlage für die Synthese genspezifischer PCR-Produkte war die symbiontische Region von B. japonicum und andere zu diesem Zeitpunkt bekannte Gensequenzen (Göttfert et al. 2001). Nach der 2002 erfolgten Veröffentlichung des Genoms von B. japonicum erfolgte ein Abgleich der bisher verwendeten Sequenzen mit der vollständigen Sequenzinformation (Kaneko et al. 2002a). Nach Synthese der PCR-Produkte und deren Kontrolle durch Sequenzierung erfolgte die Herstellung des Microarrays unter Einsatz eines Microarrayspotters. Geeignete Techniken der cDNA-Markierung und die Microarray-Hybridisierung wurden mit Total-RNA von B. japonicum-Kulturen ausgetestet und etabliert. Eine differentielle Expremierung war eine Stunde nach Genistein- bzw. Methanolgabe zum Kulturmedium nicht nachweisbar. Zum darauffolgenden Zeitpunkt konnte die bekannte Induktion der nod-Gene durch dass Flavonoid Genistein beobachtet werden. Diese Induktion fiel in den verbleibenden zwei Zeitpunkten stark ab. Dieser Abfall der Induktionsrate ließ sich nicht mit dem Genisteingehalt im Kulturmedium erklären, da dieser Zeitraum konstant blieb. Vier Stunden nach Genisteingabe war die Induktion der Gene nolA und nodD2, deren Produkte sind an der negativen Regulation der nod-Gene beteiligt, nachweisbar. Dies wäre eine mögliche Erklärung für den Abfall der Induktion der nod-Gene. Im gleichen untersuchten Zeitraum wurde die Transkription verschiedener Gene der Stickstofffixierung und Denitrifikation durch das Flavonoid Genistein induziert. Auch für die bekannten Regulatoren dieser Gene war eine Induktion nachweisbar. Nach bisherigen Arbeiten war die Expression dieser Gene auf mikroaerobe und anaerobe Zustände, wie z.B. dem symbiontischen Stadium, beschränkt. Eine Induktion durch Flavonoide ist bisher nicht beobachtet worden. Um die Verwendbarkeit des Microarrays auch für den symbiontischen Zustand von B. japonicum zu testen, wurden Versuche mit Wurzelknöllchen von Sojabohne (Glycine max) durchgeführt. Dafür wurden Keimlinge der Sojabohne (G. max) mit B. japonicum inokuliert und die Total-RNA der entstehenden Wurzelknöllchen isoliert. Jedoch war eine Kreuzhybridisierung mit pflanzlicher Total-RNA zu beobachten. Der auf PCR-Produkten basierende Microarray ist für die Untersuchung des symbiontischen Stadiums von B. japonicum nicht einsetzbar. Im Vergleich dazu wurde ein auf Oligomeren basierender, kommerzieller Microarray für diesen Versuch getestet. Die Kreuzhybridisierung mit pflanzlicher Total-RNA war auf die Oligomere für die 16S rDNA und 23S rDNA reduziert.

Les Légumineuses ont développé une interaction symbiotique avec des bactéries du sol, les rhizobia, qui fixent l’azote atmosphérique et le transfèrent à la plante sous forme assimilable.Cette interaction a lieu, au sein des nodosités, des organes racinaires où les bactéries intracellulaires se différencient en bactéroïdes. Chez Medicago truncatula, ces bactéroïdes correspondent à un stade de différentiation terminale corrélée à une endoréplication de leur génome, une augmentation de la taille des cellules, une modification des membranes et une faible capacité à se propager. Cette différentiation est induite par des facteurs de la plante appelés NCR (Nodule-specific Cysteine Rich). Les peptides NCRs ressemblent à des défensines, des peptides antimicrobiens ayant une activité antimicrobienne in vitro, tuant des bactéries. Ainsi, un élément clef dans la différenciation des bactéroïdes est la protéine bactérienne BacA, un transporteur membranaire qui confère une résistance contre l’activité antimicrobienne des peptides. Dans le cadre de ce travail de thèse, j’ai montré que l'expression des NCR est soumise à une régulation stricte et qu’ils sont activés dans trois vagues dans les cellules symbiotiques polyploïdes.Les mécanismes de contrôle par la plante sur les rhizobia intracellulaires demeurent à ce jourpeu connus et le seul modèle étudié, au début de ce travail de thèse, restait l'interaction entre M. truncatula et S. meliloti. Je me suis donc intéressée à la symbiose de certaines Légumineuses tropicales du genre Aeschynomene appartenant au clade des Dalbergoïdes où jemontre qu’ils utilisent une classe différente de peptides riches en cystéine (NCR-like) pour induire la différenciation des bactéroïdes. Ce mécanisme est analogue à celui décrit précédemment chez Medicago qui était jusqu'à présent supposé être limitée aux légumineuses appartenant au clade des IRLC. J’ai également montré que Bradyrhizobium, symbionte d’Aeschynomene possèdent un transporteur de type ABC homologues à BacA de Sinorhizobium nommé BclA. Ce gène permet l'importation d'une variété de peptides comprenant des peptides NCR. En l'absence de ce transporteur, les rhizobiums sont incapables de se différencier et de fixer l'azote.Cette étude a permis d'élargir nos connaissances sur l'évolution de la symbiose en montrant qu’au cours de l’évolution, deux clades de Légumineuses relativement éloignés (IRLC et Dalbergoïdes) aient convergé vers l’utilisation de peptides de l’immunité innée afin de contrôler leur symbionte bactérien et d’en tirer un bénéfice maximal au cours de l’interaction symbiotique. / The ability of legumes to acquire sufficient nitrogen from the symbiosis with Rhizobium relies on the intimate contact between the endosymbiotic, intracellular rhizobia, called bacteroids, and their host cells, the symbiotic nodule cells. A well-studied example is the symbiotic nitrogen fixing bacterium Sinorhizobium meliloti, which nodulates the legume Medicago truncatula. Nodules of M. truncatula produce an enormous diversity of peptides called NCRs which are similar to antimicrobial peptides (AMPs) of innate immune systems. These NCRs are involved in maintaining the homeostasis between the host cells in the nodules and the large bacterial population they contain. Although many NCRs are genuine AMPs which kill microbes in vitro, in nodule cells they do not kill the bacteria but induce them into the terminally differentiated bacteroid state involving cell elongation, genome amplification, membrane fragilization and loss of cell division capacity. Protection against the antimicrobial action of NCRs by the bacterial BacA protein is critical for bacteroid survival in the symbiotic cells and thus for symbiosis. As a part of my PhD thesis, I have shown that the differentiation of the symbiotic cells in M. truncatula is associated with a tremendous transcriptional reprogramming involving hundreds of genes, mainly NCR genes, which are only expressed in these cells. Although the extensive work on the model M. truncatula/S. meliloti, little is known how the plant controls its intracellular population and imposes its differentiation into a functional form, the bacteroids in other symbiotic systems.In my PhD work, I provide several independent pieces of evidence to show that tropical legumes of the Aeschynomene genus which belong to the Dalbergoid legume clade use a different class of cysteine rich peptides (NCR-like) to govern bacteroid differentiation. This mechanism is similar to the one previously described in Medicago which was up to now assumed to be restricted to the advanced IRLC legume clade, to which it belongs. I have also shown that the Bradyrhizobium symbionts of Aeschynomene legumes possess a multidrug transporter, named BclA, which mediates the import of a diversity of peptides including NCR peptides. In the absence of this transporter, the rhizobia do not differentiate and do not fix nitrogen. BclA has a transmembrane domain of the same family as the transmembrane domain of the BacA transporter of Rhizobium and Sinorhizobium species which is known to be required in these rhizobia to respond to the NCR peptides of IRLC legumes. Again this is a mechanism which is analogous to the one described in S. meliloti the symbiont of Medicago.This study broaden our knowledge on the evolution of symbiosis by showing that the modus operandi involving peptides derived from innate immunity used by some legumes to keep their intracellular bacterial population under control is more widespread and ancient than previously thought and has been invented by evolution several times.

Acacia spirorbis est une légumineuse endémique de Nouvelle-Calédonie se développant sur des sols calcaires, métallifères et volcano-sédimentaires, établissant des symbioses avec des bactéries fixatrices d’azote. Pour comprendre la contribution de la symbiose dans l’adaptation de la plante à des milieux contrastés et parfois extrêmes, nous avons évalué la fixation d’azote en conditions naturelles, caractérisé les rhizobia associés à cette plante et analysé la réponse adaptative de la plante aux éléments traces métalliques dans ses tissus racinaires, notamment au niveau des nodules.Nous avons mis en évidence que la symbiose rhizobienne fournissait plus de 80% de l’azote total chez des populations naturelles d’A. spirorbis se développant sur des sites d’études présentant des sols calcaires, métallifères et volcano-sédimentaires. Cette valeur est remarquable puisque chez A. mangium, A. melanoxylon et A. mucronata, les valeurs moyennes sont respectivement de 50%, 43% et 58%. Les rhizobia symbiotiques associés à A. spirorbis appartiennent aux alpha- et bêta-protéobactéries, genres Bradyrhizobium et Paraburkholderia, révélant ainsi une très large gamme de symbiontes et une faible sélectivité de partenaire. De manière remarquable, la taxonomie et la phénotypie de ces souches sont structurées et adaptées aux conditions édaphiques. Enfin, les signatures chimiques des tissus internes des nodules reflètent les propriétés chimiques des sols dans lesquels ils se sont développés, indiquant une potentielle gestion des éléments traces métalliques dans ces tissus.Tous ces éléments suggèrent que la symbiose fixatrice d’azote contribue de manière significative dans l’adaptation d’Acacia spirorbis à des sols contrastés et pouvant présenter une toxicité polymétallique extrême. / Acacia spirorbis is a leguminous tree from New Caledonia naturally found on a wide range of soils (calcareous, ultramafic and volcano-sedimentary) and able to establish symbioses with soil microorganisms, including nitrogen-fixing bacteria. The contribution of this symbiosis in the plant adaptation to contrasted edaphic environments, sometimes extremes, has been investigated according to i) the nitrogen-fixing potential of A. spirorbis in its natural ecosystems, ii) the characterization of its rhizobia and iii) the plant adaptive response to heavy metals inside its roots tissues, especially in nodules.Therefore, we revealed that the nitrogen-fixing symbiosis provided more than 80% of the plant total nitrogen in natural population naturally occurring on calcareous, ultramafic and volcano-sedimentary soils. This value is top notch among other Acacia species, where A. mangium, A. melanoxylon and A. mucronata presented mean values of 50%, 43% and 58%, respectively. Acacia spirorbis established nitrogen-fixing symbioses with alpha- and bêta-proteobacteria, genus Bradyrhizobium and Paraburkholderia, respectively, thus revealing a wide range of symbiotic partner and a low selectivity. Noteworthy, the taxonomy and phenotypes of these symbionts are structured and adapted to edaphic parameters. Finally, chemical signatures of internal nodules tissues presented similarities with soils chemical properties, thus indicating a potential management of heavy metals inside these tissues.Altogether, these data suggest that the nitrogen-fixing symbiosis might significantly contributes to Acacia spirorbis adaptation towards contrasted soils with strong edaphic conditions as an extreme polymetallic toxicity.

Submitted by Maria Beatriz Vieira (mbeatriz.vieira@gmail.com) on 2017-04-03T12:43:52Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) dissertacao_adhyvan_karmo_dalmolin.pdf: 1510521 bytes, checksum: 38b309e7f6b36a685bcf42f2e4c0bfea (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2017-04-07T18:49:11Z (GMT) No. of bitstreams: 2 dissertacao_adhyvan_karmo_dalmolin.pdf: 1510521 bytes, checksum: 38b309e7f6b36a685bcf42f2e4c0bfea (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2017-04-07T18:50:28Z (GMT) No. of bitstreams: 2 dissertacao_adhyvan_karmo_dalmolin.pdf: 1510521 bytes, checksum: 38b309e7f6b36a685bcf42f2e4c0bfea (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2017-04-07T18:50:50Z (GMT). No. of bitstreams: 2 dissertacao_adhyvan_karmo_dalmolin.pdf: 1510521 bytes, checksum: 38b309e7f6b36a685bcf42f2e4c0bfea (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2015-08-18 / Sem bolsa / O presente trabalho teve como objetivo avaliar a aplicação foliar de produto comercial contendo molibdênio e cobalto na cultura da soja, visando aumento nos componentes de rendimento e qualidade fisiológica das sementes produzidas. O experimento foi realizado na fazenda Sucupira, município de Sorriso, Estado do Mato Grosso. O delineamento experimental adotado foi de blocos casualizados com quatro repetições e seis tratamentos com diferentes doses. O produto usado na aplicação foi o Néctar (cobalto e molibdênio) da empresa Microquímica, aplicado com um pulverizador costal na vazão de 160 L.ha-1. A pulverização ocorreu quando a soja estava no estádio de desenvolvimento V5. Foram conduzidos seis tratamentos (0; 100; 200; 300; 400 e 500 mL.ha-1 do produto comercial). Foi analisado os componentes de rendimento (peso de mil sementes; rendimento de grãos, número de vagens por planta e número de sementes por vagem) a qualidade fisiológica das sementes (primeira contagem da germinação, germinação, envelhecimento acelerado, emergência em campo e tetrazólio) e análise de sanidade (fungos e bactéria). A aplicação foliar de produto comercial contendo cobalto e molibdênio na dose de até 240 mL.ha-1 afeta de forma positiva o número de sementes por vagem da cultura, não houve incrementos significativos na produtividade da soja. As doses de molibdênio e cobalto tem efeito positivo na qualidade fisiológica. / This study aimed to evaluate the effect of molybdenum and cobalt different doses applied by foliar fertilization in the soybean crop, aiming at increasing the yield components and physiological seed quality. The essay was carried out at Sucupira farm, city of Sorriso, in Mato Grosso State. The experimental design was a randomized block with four replications and six treatments with different doses. Product used in the application was the Nectar (cobalt and molybdenum) of Microquímica Company, applied with a backpack sprayer at a flow rate of 160 L.ha-1. Spraying occurred in the V5 growth soybean stage. Six treatments were conducted (0, 100, 200, 300, 400 and 500 commercial product mL.ha-1). It was analyzed the yield components (thousand seed weight, grain yield, number of pods per plant and number of seeds per pod) Physiologic seed quality (first count of germination, standard germination test, accelerated aging, field seedlings emergence and tetrazolium test) and health test (fungi and bacteria). Foliar spraying of cobalt and molybdenum up to 240 mL.ha-1 influences positively the number of seeds per pod culture, there was no significant increase in soybean yield. The doses of molybdenum and cobalt has a positive effect on physiological quality.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / New technologies are intended to assist the farmer in the inoculation process, as is the case of antecipated inoculation (pre-inoculation) provided by using osmoprotectors. In search of higher yields, the kind of inoculation and management with nitrogen fertilizer also need to be clarified. In this study, two experiments were conducted to evaluate the agronomic performance of two inoculation strategies with Bradyrhizobium alone (conventional inoculation) or combined with Azsopirillum brasilense (co-inoculation). The first experiment, we evaluated the pre-inoculation (0, 7 and 10 days before seeding). The second experiment evaluated the response to supply urea (0, 75 and 150 kg N ha-1) split into two periods of three soybeans cultivars development stage (BMX Ativa RR, TEC 6029 IPRO and BMX Potência RR). We evaluated the interaction of nodulation, of morphophysiological components and grain yield at conditions of Santa Maria. Co-inoculation and conventional inoculation, both plus an osmoprotectors, showed increases in grain yield relative to treatment not inoculated 31 and 25%, respectively. With seven days before to sowing, use of osmoprotectors for both types of inoculation, did not differ statistically from the other treatments inoculated the same day of sowing. Co-inoculation provided increment in grain yield of 240 kg ha-1 compared to conventional inoculation. BMX Ativa, TEC 6029 IPRO and BMX Potência when co-inoculated showed increases in grain yield 6, 4 e 12%, respectively. The addition of 150 kg N ha-1 for the two indeterminate cultivars had an increases, but without economic return. The cultivars tested when inoculated conventionally not respond positively to the application of urea. Therefore, these results confirm the effectiveness of the use osmoprotectores and co-inoculation ond the seeds and low economic returns provided by the use of nitrogen fertilizer. It emphasizes the use of activation and protection substances on rhizobial inoculants for the formulation, as well as more studies concerning the efficiency of co-inoculation with rhizobacteria promote plant growth to obtain higher yields. / Novas tecnologias têm surgido para auxiliar o agricultor no processo de inoculação, como é o caso da pré-incoulação (inoculação antecipada) possibilitada pelo uso de osmoprotetores. Na busca por produtividades superiores, o tipo de inoculação e o manejo com fertilizante nitrogenado também precisam ser melhor esclarecidos. No presente trabalho, foram conduzidos dois experimentos para avaliar o desempenho agronômico de duas estratégias de inoculação com bactérias do gênero Bradyrhizobium isoladamente (inoculação convencional) ou combinadas com Azospirillum brasilense (co-inoculação). O primeiro experimento, testou a pré-inoculação (0, 7 e 10 dias anteriores à semeadura). O segundo experimento, testou à resposta ao fornecimento de ureia (0, 75 e 150 kg de N ha-1) fracionado em dois momentos do estádio de desenvolvimento de três cultivares de soja (BMX Ativa RR, TEC 6029 IPRO e BMX Potência RR). Avaliou-se os componentes da nodulação, morfofisiológicos e da produtividade de grãos nas condições edafoclimáticas de Santa Maria. A co-inoculação e a inoculação convencional, ambas acrescidas do osmoprotetor, apresentaram acréscimos na produção de grãos em relação ao tratamento não inoculado de 31 e 25%, respectivamente. Com sete dias anteriores à semeadura, o uso do osmoprotetor para os dois tipos de inoculação, não diferiu estatisticamente dos demais tratamentos inoculados no mesmo dia da semeadura. A co-inoculação proporcionou incremento na produtividade de grãos de 240 kg ha-1 quando comparado à inoculação convencional. Os cultivares BMX Ativa, TEC 6029 e BMX Potência quando co-inoculadas apresentaram incrementos na produtividade de 6, 4 e 12% respectivamente. Com a adição de 150 kg de N ha-1, os dois cultivares indeterminados apresentaram incremento na produtividade de 300 kg ha-1, porém, sem retorno econômico. Os cultivares testados quando inoculados convencionalmente não respondem positivamente à aplicação de ureia. Portanto, esses resultados confirmam a eficácia do uso de osmoprotetores e co-inoculação de sementes e o baixo retorno econômico proporcionado pelo uso de fertilizante nitrogenado. Enfatiza-se o uso de substâncias de ativação e proteção para os rizóbios na formulação de inoculantes, assim como, mais estudos quanto a eficiência da co-inoculação com rizobactérias promotoras de crescimento de plantas para obtenção de maiores produtividades.

Master of Science / Department of Agronomy / Kraig Roozeboom / Nitrogen fixation by Bradyrhizobium japonicum in soybean [Glycine max] is highly beneficial in soybean crop production. Nodulation issues have been encountered on fields new to growing soybeans in recent years in Kansas. The purpose of this research was to evaluate soybean nodulation performance under various situations and seed handling practices in order to educate producers on how to achieve reliable nodulation consistency in the field. The objectives of the study were to: 1) compare inoculant products using single and double rates and in combination with one another on fields with varying soybean history; 2) determine if there was a negative interaction between inoculant products and common seed treatments; and 3) discover the influence of inoculated seed storage conditions before planting on the rhizobia’s ability to successfully nodulate soybean roots. Field experiments were conducted on diverse Kansas sites in 2011 and 2012. Inoculant treatment and seed treatment interaction trials had ten and seven experimental sites respectively. Inoculated seed storage conditions were evaluated in a greenhouse experiment during the spring of 2013. All studies used a randomized complete block design with four replications. The Novozymes inoculant products generally provided superior nodulation performance over other company products in the study where soybean had not been in recent rotation with an average increase of 167% in nodule number verses the control. The combination of dry and liquid inoculant products provided a significant increase in root nodule number at five of the environments out of recent rotation with a 76% increase over single inoculant rates. Although there were early season nodulation differences between treatments in new soybean ground, these did not carry through to seed yield differences in the majority of research sites. Hot and dry summer conditions reduced yields, making detection of treatment differences difficult. There were no negative effects on nodulation performance with any of the seed treatments. Although soybean seed yield was 634 kg ha[superscript]-[superscript]1 greater for the Novozyme combination treatment compared to the check at one location in 2011, the control yielded as well or better than all other treatment/inoculant combinations, implying that yield differences were likely not related to inoculant treatments. At other sites, yield was not influenced by seed treatment and inoculant combinations. Results indicate that seed treatment formulations did not significantly impact bacterial inoculant product performance, soybean nodulation, or yield. Storage conditions had no effect on nodulation performance in the greenhouse study, likely due to survival of Bradyrhizobium japonicum in the heat-treated growth medium.

Orientador: Sandra Cristina de Oliveira / Resumo: Está dissertação teve como objetivo principal, averiguar os efeitos da inoculação de sementes de amendoim, com as bactérias Bradyrhizobium sp. e Azospirillum brasilense de forma isolada e em conjunto (coinoculação). Para tanto foram realizadas analises bioquímicas das folhas, afim de analisar os teores de compostos fotossintetizantes, açucares solúveis totais, ureídeos, feofitinas totais e compostos nitrogenados e também foi foram feitas avaliações referentes a parâmetros fitotécnicos. Primeiramente foi realizada uma revisão bibliográfica por meio de levantamentos de dados sobre o panorama da produção de amendoim a nível mundial, nacional e algumas cidades localizadas no Oeste paulista, região a qual é destaque na produção de amendoim. Além do levantamento bibliográfico, foi realizado um experimento à campo e em laboratório, a fim de investigar os efeitos da inoculação e coinoculação em sementes de amendoim com bactérias promotoras do crescimento de plantas. Na parte experimental utilizou-se o delineamento experimental de blocos ao acaso com quatro repetições e sete tratamentos, sendo: Controle (sem inoculação); Az 2 mL kg-¹ (Azospirillum brasilense na dose de 2 mL para 1 kg de sementes); Az 4 mL kg-¹ (Azospirillum brasilense na dose de 4 mL para 1 kg de sementes); Bd 2 mL kg-¹ (Bradyrhizobium sp. na dose de 2 mL para 1 kg de sementes); Bd 4 mL kg-¹ (Bradyrhizobium sp. na dose de 4 mL kg-¹ para 1 kg de sementes); Az + Bd 2 mL kg-¹ (Azospirillum brasilense + Bradyrhizobium sp.... (Resumo completo, clicar acesso eletrônico abaixo) / Mestre

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