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Isolation of Bacterial Endophytes from Phalaris arundinacea and their Potential in Diclofenac and Sulfamethoxazole Degradation

ANNA WĘGRZYN
ANNA WĘGRZYN
 y 
EWA FELIS
EWA FELIS
   | 04 sept 2018
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Article Category: original-paper
Publicado en línea: 04 sept 2018
Páginas: 321 - 331
Recibido: 30 oct 2017
Aceptado: 20 may 2018
DOI: https://doi.org/10.21307/pjm-2018-039
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© 2018 Anna Węgrzyn and Ewa Felis et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Fig. 1.

The scheme of a lab-scale constructed wetlands system, from which the plant samples were collected.
The scheme of a lab-scale constructed wetlands system, from which the plant samples were collected.

Fig. 2.

Workflow for isolation of cultivable endophytic bacteria from P. arundinacea plants non-exposed and exposed to DCF and SMX mixed pollution (three plants per condition from each column were harvested).
Workflow for isolation of cultivable endophytic bacteria from P. arundinacea plants non-exposed and exposed to DCF and SMX mixed pollution (three plants per condition from each column were harvested).

Fig. 3.

The number of bacteria (CFU/g FW) obtained from the surface of the sterilized reed canary grass root samples. Light bars – the number of bacterial endophytes grown on nutrient agar. Grey bars – the number of bacterial endophytes grown on nutrient medium enriched with DCF and SMX. N = 6. Significant differences have been marked using * (p < 0.05; t-test). Time regime of wastewater supply in brackets.
The number of bacteria (CFU/g FW) obtained from the surface of the sterilized reed canary grass root samples. Light bars – the number of bacterial endophytes grown on nutrient agar. Grey bars – the number of bacterial endophytes grown on nutrient medium enriched with DCF and SMX. N = 6. Significant differences have been marked using * (p < 0.05; t-test). Time regime of wastewater supply in brackets.

Fig. 4.

The removal of 2 mg/l DCF by MG7 strain in the adsorption control, without additional carbon source and with phenol at a concentration of 100 mg/l as the additional carbon source.
The removal of 2 mg/l DCF by MG7 strain in the adsorption control, without additional carbon source and with phenol at a concentration of 100 mg/l as the additional carbon source.

Fig. 5.

The removal of 2 mg/l SMX by MG7 strain in the adsorption control, without additional carbon source and with phenol at a concentration of 100 mg/l as the additional carbon source.
The removal of 2 mg/l SMX by MG7 strain in the adsorption control, without additional carbon source and with phenol at a concentration of 100 mg/l as the additional carbon source.

Fig. 6.

Changes in biomass of MG7 strain in the cultures that contained DCF, monitored as optical density at 600 nm, without additional carbon source and with phenol at a concentration of 100 mg/l as additional carbon source.
Changes in biomass of MG7 strain in the cultures that contained DCF, monitored as optical density at 600 nm, without additional carbon source and with phenol at a concentration of 100 mg/l as additional carbon source.

Fig. 7.

Changes in biomass of MG7 strain in the cultures that contained SMX, monitored as optical density at 600 nm, without additional carbon source and with phenol at a concentration of 100 mg/l as additional carbon source.
Changes in biomass of MG7 strain in the cultures that contained SMX, monitored as optical density at 600 nm, without additional carbon source and with phenol at a concentration of 100 mg/l as additional carbon source.

Fig. 8.

DGGE pattern of 16S rRNA gene fragments of 180 bp amplified from DNA obtained from the surface-sterilized root tissues of P. arundinacea.A1-3, C1-3 – the root samples non-exposed to DCF and SMX, B1-3, D1-3 – the root samples exposed to DCF and SMX. The frequency of wastewater supply: A1-3 and B1-3 – 0.35 l four times a day; C1-3 and D1-3 1.4 l – once a day. Description of the bands in the manuscript text.
DGGE pattern of 16S rRNA gene fragments of 180 bp amplified from DNA obtained from the surface-sterilized root tissues of P. arundinacea.A1-3, C1-3 – the root samples non-exposed to DCF and SMX, B1-3, D1-3 – the root samples exposed to DCF and SMX. The frequency of wastewater supply: A1-3 and B1-3 – 0.35 l four times a day; C1-3 and D1-3 1.4 l – once a day. Description of the bands in the manuscript text.

Fig. 9.

Shannon biodiversity index based on DGGE profiles.A1-3, C1-3 – the root samples non-exposed on DCF and SMX; B1-3, D1-3 – the root samples exposed on DCF and SMX. The frequency of wastewater supply: A1-3 and B1-3 – 0.35 l four times a day; C1-3 and D1-3 – 1.4 l once a day.
Shannon biodiversity index based on DGGE profiles.A1-3, C1-3 – the root samples non-exposed on DCF and SMX; B1-3, D1-3 – the root samples exposed on DCF and SMX. The frequency of wastewater supply: A1-3 and B1-3 – 0.35 l four times a day; C1-3 and D1-3 – 1.4 l once a day.

Identification of bacterial endophytes, based on the 16S rRNA gene sequence.

IsolateClostest matchSimilarity, %Sequence ID
MG2Variovorax boronicumulans96NR_114214.1
MG5Bacillus wiedmannii97NR_152692.1
MG7Microbacterium flavescens98NR_029350.1
MG11Agrobacterium tumefaciens97NR_041396.1
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