Caracterização de rizóbios indicados para produção de inoculantes por meio de sequenciamento parcial do 16S rRNA

Toledo, Bethânia Figueiredo Barbosa de; Marcondes, Jackson; Lemos, Eliana Gertrudes de Macedo

doi:10.1590/s0100-204x2009000400008

Cited by 6 publications

(6 citation statements)

References 12 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…These last three genus were, respectively, the taxons presenting the highest frequencies regarding the number of clones from the library (Figure 3). Partial sequencing of the 16S rRNA gene is a reliable technique, and has been used frequently for the identification of microorganisms, these being from cultures (Toledo et al, 2009), or available from environmental samples (Nercessian et al, 2008 On the other hand, the curve with grouping criterion that presented 90 % of similarity attained its plateau when coming close to 20 taxonomic units. These results reveal that the number of 16S rDNA clones sequenced was close to the total number of individuals which composed the bacterial community sampled.…”

Section: Resultsmentioning

confidence: 99%

“…The identification of microorganisms that use hydrocarbons as a C source and that present tolerance to contaminants has been reported in areas with a history of contamination by petroleum hydrocarbons and in many different types of soil (Richard & Vogel, 1999;Van Beilen et al, 2002;Widada et al 2002). The predominance of the Pseudomonas genus was expected, based on the number of published studies that describe the frequency of these in contaminated soils (Wunsche et al, 1995;Pirôllo, 2008;Higashioka et al 2009), as well as their efficiency in the degradation of hydrocarbons (Whyte et al, 1997, Obuekwe et al, 2008, Bishnoi et al, 2009.…”

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Molecular analysis of the bacterial diversity in a specialized consortium for diesel oil degradation

Paixão

Dimitrov

Pereira

et al. 2010

Rev. Bras. Ciênc. Solo

Self Cite

View full text Add to dashboard Cite

SUMMARYDiesel oil is a compound derived from petroleum, consisting primarily of hydrocarbons. Poor conditions in transportation and storage of this product can contribute significantly to accidental spills causing serious ecological problems in soil and water and affecting the diversity of the microbial environment. The cloning and sequencing of the 16S rRNA gene is one of the molecular techniques that allows estimation and comparison of the microbial diversity in different environmental samples. The aim of this work was to estimate the diversity of microorganisms from the Bacteria domain in a consortium specialized in diesel oil degradation through partial sequencing of the 16S rRNA gene. After the extraction of DNA metagenomics, the material was amplified by PCR reaction using specific oligonucleotide primers for the 16S rRNA gene. The PCR products were cloned into a pGEM-T-Easy vector (Promega), and Escherichia coli was used as the host cell for recombinant DNAs. The partial clone sequencing was obtained using universal oligonucleotide primers from the vector. The genetic library obtained generated 431 clones. All the sequenced clones presented similarity to phylum Proteobacteria, with Gammaproteobacteria the most present group (49.8 % of the clones), followed by Alphaproteobacteira (44.8 %) and Betaproteobacteria (5.4 %). The Pseudomonas genus was the most abundant in the metagenomic library, followed by the Parvibaculum and the Sphingobium genus, respectively. After partial sequencing of the 16S rRNA, the diversity of the bacterial consortium was estimated using DOTUR software. When comparing these sequences to the database from the National Center for Biotechnology Information (NCBI), a strong correlation was found between the data generated by the software used and the data deposited in NCBI. PCR, clonagem, 16S rRNA, metagenoma, solo, biosurfactantes.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Molecular analysis of the bacterial diversity in a specialized consortium for diesel oil degradation

Paixão

Dimitrov

Pereira

et al. 2010

Rev. Bras. Ciênc. Solo

Self Cite

View full text Add to dashboard Cite

show abstract

“…This strain belongs to the collection of BFNN of the Laboratory of Soil Microbiology of UFLA/MG, efficient in establishing symbiosis with common bean and remains in the test phase. However, the CIAT 899 strain (SEMIA 4077) of Rhizobium tropici is authorized by the Ministry of Livestock and Supply (Ministério da Agricultura Pecuária e Abastecimento -MAPA) for the production of commercial bean inoculants (TOLEDO et al, 2009;BARBOSA;GONZAGA, 2012). In addition to the two strains, treatment without inoculation was used.…”

Section: Methodsmentioning

confidence: 99%

Evaluation of Rhizobia strains UFLA 02 100 and CIAT 899 in line LEP 02 11 of Phaseolus vulgaris L.

et al. 2019

View full text Add to dashboard Cite

The growing quest for sustainability in agricultural production systems has been pushing researchers to develop new technologies under different soil and climatic conditions. Based on this, and knowing the socioeconomic importance of the bean, the use of biological nitrogen fixation (BNF) is explored. Thus, the quest for quantitative knowledge of BNF at the phenological stages in a new bean line using promising and commercial strains becomes necessary. The objective of this study was to evaluate the strains UFLA 02 100 and CIAT 899 in the different phenological stages of the LEP 02 11 common bean strain using the biological nitrogen fixation. The experiment was conducted in a greenhouse on the Maringá-PR campus of the State University of Maringá. The design was a randomized complete block design in a 6x4 factorial scheme with four replications. The factor A comprised six nitrogen sources: Control, strain UFLA 02 100, strain CIAT 899, Nitrogen 60 kg ha-1, strain UFLA 02 100 + Nitrogen 30 kg ha-1, strain CIAT 899 + Nitrogen 30 kg ha-1 and the B factor is the phenological stages V4, R5, R6 and R8. The analyzed variables were: dry mass of nodules (DMN, grams/plant), dry mass of shoot (DMS, grams/plant), nitrogen content in shoot (NCS, %) and accumulation of nitrogen in shoot (ANS, grams/plant). For the statistical analysis of the data, the means test was used for the variables in the stages. The CIAT 899 strain statistically outperformed UFLA 02 100 for most variables and it was found that nitrogen fertilization negatively affected nodulation.

show abstract

“…In addition, functional differentiation between rRNA operons has allowed the differential expression of them in response to environmental changes and resource availability (Klappenbachet al, 2000). Toledo et al (2009) used 16S rDNA for the characterization of rhizobia suitable for the production of inoculants and stated that the molecular characterization of strains collections on the basis of conserved genes is essential to compare and identify soil rhizobia. Comparative studies based on 16S rDNA analysis are suitable not only among bacteria but also with 16S rDNA sequences from archeobacteria and the 18S rDNA from eukaryotes (Claridge III, 2004).…”

Section: Qpcr Data and 16s Rdna Quantificationmentioning

confidence: 99%

Detection and Enumeration of Bradyrhizobia Cells by Real-Time PCR Quantification

2015

JAES

View full text Add to dashboard Cite

Bradyrhizobiumelkanii and Bradyrhizobiumjaponicum are nitrogen-fixing bacteria that establish symbiosis with soybean plants. The optimization of this symbiosis and the success of biological nitrogen fixation are reached by inoculating the seeds with strains of bradyrhizobia. The aim of this work consisted in the enumeration of bacterial cells of inoculants from different ages by real-time PCR quantification (qPCR) based on 16S rDNA and the evaluation of surviving cells rates under desiccation experiments. The results of counting by colony forming units (CFU) were correlated with those obtained by qPCR. All qPCR values were higher than those for CFU values in cell enumeration for all samples and conditions observed. Cell resistance to desiccation was shown until 24 hours onto soybean seeds, with considerable fall in all the inoculants after 48 hours under desiccation. This was probably due to biochemical and physiological changes in its metabolism, making use of defensive mechanism to the adverse conditions for its survival.Keywords: Cellular enumeration, soybean inoculants, biological nitrogen fixation, bradyrhizobia IntroductionThe main sources of nitrogen (N) for soybean crops provide from application of nitrogen fertilizers and biological nitrogen fixation (BNF) that uses atmospheric N2 through symbiotic association between legumes and rhizobia BNF is the process that involves the conversion of inert N2 gas into ammonia (NH3) through expending large amounts of energy to break triple chemical bonds N atoms (Oldroydet al., 2011). Bradyrhizobia are nitrogenfixing bacteria that possess a complex enzyme named nitrogenase that is able to realize N2 conversion to produced NH3, which in turn is assimilated into organic compounds by soybean plants able to establish symbiotic interactions with those microrganisms (Kuykendalet al., 1992). The maximization of the symbiosis events can be achieved by inoculation with selected strains of rhizobia performing the best and desirable characteristics, reducing the cost of soybean production. On the other hand, rhizobia inoculation is also a factor that contributes to reducing pollution in the environment by decreasing N fertilizers applications in the field (Abdel-Fattah I El-Shaarawiet al., 2011). Official recommendation of Bradyrhizobiumelkanii and/or Bradyrhizobiumjaponicum strains for soybean's inoculants production in Brazil is made by Ministry of Agriculture, Livestock and Food, Supply (MAPA). These bradyrhizobia are genetic and physiological different (Soares and Passaglia 2010) and can be used separately or in combination. MAPA requires a periodic control of inoculants quality and found that in some cases there is a discrepancy between the numbers of cells detected by the CFU (colony forming units) traditional method of counting performed by different laboratories. In this paper, the development and applying of a new method for inoculants cell enumeration was tested by real-time PCR quantification (qPCR) based on 16S rDNA. This technique was applied both to pure ...

show abstract

Caracterização de rizóbios indicados para produção de inoculantes por meio de sequenciamento parcial do 16S rRNA

Cited by 6 publications

References 12 publications

Molecular analysis of the bacterial diversity in a specialized consortium for diesel oil degradation

Molecular analysis of the bacterial diversity in a specialized consortium for diesel oil degradation

Evaluation of Rhizobia strains UFLA 02 100 and CIAT 899 in line LEP 02 11 of Phaseolus vulgaris L.

Detection and Enumeration of Bradyrhizobia Cells by Real-Time PCR Quantification

Contact Info

Product

Resources

About