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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
| Identifer | oai:union.ndltd.org:IBICT/oai:localhost:jspui/2436 |
| Date | 23 February 2017 |
| Creators | Matos, Gustavo Feitosa de |
| Contributors | Baldani, Jos? Ivo, Rouws, Luc Felicianus Marie, Zilli, Jerri ?dson, Medici, Leonardo Oliveira |
| Publisher | Universidade Federal Rural do Rio de Janeiro, Programa de P?s-Gradua??o em Fitotecnia, UFRRJ, Brasil, Instituto de Agronomia |
| Source Sets | IBICT Brazilian ETDs |
| Language | Portuguese |
| Detected Language | English |
| Type | info:eu-repo/semantics/publishedVersion, info:eu-repo/semantics/masterThesis |
| Format | application/pdf |
| Source | reponame:Biblioteca Digital de Teses e Dissertações da UFRRJ, instname:Universidade Federal Rural do Rio de Janeiro, instacron:UFRRJ |
| Rights | info:eu-repo/semantics/openAccess |
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