Fast and efficient symbiotic gene-based duplex PCR approach for the preliminary selection of legume root nodule bacteria

Silva, Valéria Borges da; Silva, Aleksandro Ferreira da; Silva, Thaise Rosa da; Santos, Jonnathan Whiny Moraes dos; Silva, Jéssica Fernanda da; Souza, Adaílson Pereira de; Freitas, Ana Dolores Santiago de; Fernandes‐Júnior, Paulo Ivan

doi:10.1016/j.rhisph.2019.100144

Cited by 7 publications

(2 citation statements)

References 19 publications

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“…The preliminary amplification of nodC fragment is a promising strategy to increase the probability to select rhizobia rather than non-rhizobial isolates. This strategy as already applied to rhizobial culture collections with good results (Mothapo et al 2013;Hollowell et al 2016;Silva et al 2019) and along with the nifH primer was recently proposed as criterion for separation of rhizobia to non-rhizobia in culture collections (Silva et al 2019).…”

Section: Discussionmentioning

confidence: 99%

“…The DNA was extracted using the Wizard® genomic DNA purification kit (Promega, USA) following the manufacturer's instructions. First of all, we amplified the nodC gene as strategy to preliminary selection of putative rhizobia (Mothapo et al 2013;Silva et al 2019). The 16S-23S rRNA intergenic spacer (IGS) region was amplified and subjected to endonuclease digestion for the restriction length fragment polymorphism (RFLP) technique on the IGS fragment (IGS-RFLP) using HaeIII, AluI, and HindIII (Thermo Scientific, USA) restriction enzymes.…”

Section: Molecular Analysismentioning

confidence: 99%

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Molecular, Physiological, and Symbiotic Characterization of Cowpea Rhizobia from Soils Under Different Agricultural Systems in the Semiarid Region of Brazil

Sena

Nascimento

Lino

et al. 2020

J Soil Sci Plant Nutr

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The objectives of this study were to characterize the genetic diversity and evaluate the ability to tolerate stress as well as to assess the symbiotic efficiency of bacteria from cowpea nodules in agricultural soils with different uses in the semiarid region of Bahia state (Brazil). Soil samples were collected from six crop lands and one from the pristine Caatinga biome. After a trap-host experiment, the bacteria were isolated and culturally characterized. Isolates with typical characteristics of Bradyrhizobium were subjected to the nodC symbiotic gene amplification and those positive were evaluated by 16S-23S IGS-RFLP. Twenty-seven isolates belonging to different genetic clusters were selected for 16S-23S IGS sequencing. In additions, the selected bacteria were characterized biochemically and symbiotically. Among 420 characterized isolates, approximately 60% (251 isolates) displayed typical Bradyrhizobium cultural features. A total of 161, out of 251 isolates, showed positive amplification of the nodC gene fragment. The IGS-RFLP profiles analysis generated 33 groups and 27 were selected for further analysis. The fertility of the soils influenced the distribution of the isolates in the IGS-RFLP clusters. The bacteria were assigned to two genera, Bradyrhizobium and Microvirga, with 26 and 1 representative bacteria, respectively. Some isolates were able to tolerate NaCl as well as acidic and alkaline pH. In addition, isolates showed the abilities to produce biofilm under stress and to produce indole compounds, as well as efficient nodulation and nitrogen fixation. The isolates displayed great genetic, biochemical, and symbiotic variability with promising biotechnological potential.

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Section: Discussionmentioning

confidence: 99%

Section: Molecular Analysismentioning

confidence: 99%

Molecular, Physiological, and Symbiotic Characterization of Cowpea Rhizobia from Soils Under Different Agricultural Systems in the Semiarid Region of Brazil

Sena

Nascimento

Lino

et al. 2020

J Soil Sci Plant Nutr

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Phenotypic and genotypic diversity of root nodule bacteria from wild Lathyrus and Vicia species in Gaziantep, Turkey

Basbuga,

Can

et al. 2024

Folia Microbiol

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Are Cowpea-Nodulating Bradyrhizobial Communities Influenced by Biochar Amendments in Soils? Genetic Diversity and Symbiotic Effectiveness Assessment of Two Agricultural Soils of Brazilian Drylands

Oliveira

Sena

Nascimento

et al. 2019

J Soil Sci Plant Nutr

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The objective of this study was to evaluate the influence of biochar amendment on the bacterial genetic diversity and symbiosis in cowpea-nodulating rhizobial communities in two agricultural soils of Brazilian drylands. Vertisol and Ultisol surface layer soil samples were collected in the Brazilian semiarid region. The soils were mixed with biochar and cowpea seeds (BRS Marataoã) and sown in pots. Forty-five days after plant emergence, the nodule bacteria were isolated. The genetic variability of the nodule bacteria was evaluated by means of restriction fragment length polymorphism of 16S-23S rRNA intergenic spacer region (IGS-RFLP), and those that shared identical IGS-RFLP profiles were fingerprinted by BOX-PCR. By selecting of representative isolates in the genetic clusters, 27 bacteria were identified by 16S-23S IGS sequencing and symbiotically assessed. The IGS-RFLP dendrogram of the 73 isolates had 20 clusters; the multivariate correspondence analysis grouped both Vertisol treatments, and the Ultisol treatments were placed far away in the biplot. Ultisol showed higher bradyrhizobial diversity than the Vertisol. The fingerprinting indicated the presence of only three clonal colonies, showing high intraspecific diversity. The 16S-23S IGS sequences indicated the prevalence of bacteria related to Bradyrhizobium guangxiense in the Vertisol and Bradyrhizobium zhanjiangense in the Ultisol, in addition to the presence of three putative Microvirga spp. in the Ultisol. The symbiotic efficiency of 22 out of 27 bacteria was comparable with that of the inoculant strain Bradyrhizobium yuanmingense BR 3267. The genetic diversity of the cowpea-nodulating bradyrhizobia was not primarily affected by the biochar application in two agricultural soils of Brazilian drylands. Both soils harbored symbiotically efficient bacteria, without the influence of biochar amendment .

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Fast and efficient symbiotic gene-based duplex PCR approach for the preliminary selection of legume root nodule bacteria

Cited by 7 publications

References 19 publications

Molecular, Physiological, and Symbiotic Characterization of Cowpea Rhizobia from Soils Under Different Agricultural Systems in the Semiarid Region of Brazil

Molecular, Physiological, and Symbiotic Characterization of Cowpea Rhizobia from Soils Under Different Agricultural Systems in the Semiarid Region of Brazil

Phenotypic and genotypic diversity of root nodule bacteria from wild Lathyrus and Vicia species in Gaziantep, Turkey

Are Cowpea-Nodulating Bradyrhizobial Communities Influenced by Biochar Amendments in Soils? Genetic Diversity and Symbiotic Effectiveness Assessment of Two Agricultural Soils of Brazilian Drylands

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