Valorization of Lignin as an Immobilizing Agent for Bioinoculant Production using Azospirillum brasilense as a Model Bacteria
Plant growth-promoting bacteria (PGPB) have been largely considered as beneficial in harsh and limiting environments given their effects on alleviating plant stress. For practical applications, most of the PGPB are prepared in immobilization matrices to improve the stability and benefits of bacteria...
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| Published in | Molecules (Basel, Switzerland) Vol. 24; no. 24; p. 4613 |
|---|---|
| Main Authors | , , , |
| Format | Journal Article |
| Language | English |
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Switzerland
MDPI AG
17.12.2019
MDPI |
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| Online Access | Get full text |
| ISSN | 1420-3049 1420-3049 |
| DOI | 10.3390/molecules24244613 |
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| Abstract | Plant growth-promoting bacteria (PGPB) have been largely considered as beneficial in harsh and limiting environments given their effects on alleviating plant stress. For practical applications, most of the PGPB are prepared in immobilization matrices to improve the stability and benefits of bacteria. Despite the long list of immobilizing agents/carriers tested to date, a long list of desired requirements is yet to be achieved. Here, lignin stands as a scarcely tested immobilizer for bioinoculants with great potential for this purpose. The aim of this work was to demonstrate the feasibility of lignin as a carrier of the nitrogen-fixing Azospirillum brasilense. These bacteria were cultured in liquid media with recovered organosolv lignin added for bacterial immobilization. Then, lignin was recovered and the immobilized biomass was quantified gravimetrically by DNA extraction and serial dilution plating. Fluorescent microscopy as well as Congo red agar plating showed the immobilization of the bacterial cells in the lignin matrix and crystal violet dyeing showed the biofilms formation in lignin particles. A high number of cells were counted per gram of dried lignin. Lignin can be readily used as low-cost, health-safe bioinoculant carrier to be used in soil and agricultural applications. |
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| AbstractList | Plant growth-promoting bacteria (PGPB) have been largely considered as beneficial in harsh and limiting environments given their effects on alleviating plant stress. For practical applications, most of the PGPB are prepared in immobilization matrices to improve the stability and benefits of bacteria. Despite the long list of immobilizing agents/carriers tested to date, a long list of desired requirements is yet to be achieved. Here, lignin stands as a scarcely tested immobilizer for bioinoculants with great potential for this purpose. The aim of this work was to demonstrate the feasibility of lignin as a carrier of the nitrogen-fixing
Azospirillum brasilense
. These bacteria were cultured in liquid media with recovered organosolv lignin added for bacterial immobilization. Then, lignin was recovered and the immobilized biomass was quantified gravimetrically by DNA extraction and serial dilution plating. Fluorescent microscopy as well as Congo red agar plating showed the immobilization of the bacterial cells in the lignin matrix and crystal violet dyeing showed the biofilms formation in lignin particles. A high number of cells were counted per gram of dried lignin. Lignin can be readily used as low-cost, health-safe bioinoculant carrier to be used in soil and agricultural applications. Plant growth-promoting bacteria (PGPB) have been largely considered as beneficial in harsh and limiting environments given their effects on alleviating plant stress. For practical applications, most of the PGPB are prepared in immobilization matrices to improve the stability and benefits of bacteria. Despite the long list of immobilizing agents/carriers tested to date, a long list of desired requirements is yet to be achieved. Here, lignin stands as a scarcely tested immobilizer for bioinoculants with great potential for this purpose. The aim of this work was to demonstrate the feasibility of lignin as a carrier of the nitrogen-fixing Azospirillum brasilense. These bacteria were cultured in liquid media with recovered organosolv lignin added for bacterial immobilization. Then, lignin was recovered and the immobilized biomass was quantified gravimetrically by DNA extraction and serial dilution plating. Fluorescent microscopy as well as Congo red agar plating showed the immobilization of the bacterial cells in the lignin matrix and crystal violet dyeing showed the biofilms formation in lignin particles. A high number of cells were counted per gram of dried lignin. Lignin can be readily used as low-cost, health-safe bioinoculant carrier to be used in soil and agricultural applications. Plant growth-promoting bacteria (PGPB) have been largely considered as beneficial in harsh and limiting environments given their effects on alleviating plant stress. For practical applications, most of the PGPB are prepared in immobilization matrices to improve the stability and benefits of bacteria. Despite the long list of immobilizing agents/carriers tested to date, a long list of desired requirements is yet to be achieved. Here, lignin stands as a scarcely tested immobilizer for bioinoculants with great potential for this purpose. The aim of this work was to demonstrate the feasibility of lignin as a carrier of the nitrogen-fixing Azospirillum brasilense. These bacteria were cultured in liquid media with recovered organosolv lignin added for bacterial immobilization. Then, lignin was recovered and the immobilized biomass was quantified gravimetrically by DNA extraction and serial dilution plating. Fluorescent microscopy as well as Congo red agar plating showed the immobilization of the bacterial cells in the lignin matrix and crystal violet dyeing showed the biofilms formation in lignin particles. A high number of cells were counted per gram of dried lignin. Lignin can be readily used as low-cost, health-safe bioinoculant carrier to be used in soil and agricultural applications.Plant growth-promoting bacteria (PGPB) have been largely considered as beneficial in harsh and limiting environments given their effects on alleviating plant stress. For practical applications, most of the PGPB are prepared in immobilization matrices to improve the stability and benefits of bacteria. Despite the long list of immobilizing agents/carriers tested to date, a long list of desired requirements is yet to be achieved. Here, lignin stands as a scarcely tested immobilizer for bioinoculants with great potential for this purpose. The aim of this work was to demonstrate the feasibility of lignin as a carrier of the nitrogen-fixing Azospirillum brasilense. These bacteria were cultured in liquid media with recovered organosolv lignin added for bacterial immobilization. Then, lignin was recovered and the immobilized biomass was quantified gravimetrically by DNA extraction and serial dilution plating. Fluorescent microscopy as well as Congo red agar plating showed the immobilization of the bacterial cells in the lignin matrix and crystal violet dyeing showed the biofilms formation in lignin particles. A high number of cells were counted per gram of dried lignin. Lignin can be readily used as low-cost, health-safe bioinoculant carrier to be used in soil and agricultural applications. In some cases, these PGPB are directly inoculated in crop seeds; however, these have a short viability, which make it necessary to develop formulations able to sustain microbial growth, to resist drying and rehydration, and, of course, to deliver living microorganisms to the soil [1]. [...]formulations, also called bioinoculants, are prepared in immobilizing agents or carriers. Apart from its stimulatory action in plant nutrient absorption, Azospirillum spp. enhance the root mass formation, providing physical stability to the plant against wind and water erosion and resistance to abiotic stress. [...]Azospirillum spp. have the ability to grow under extreme pH and heavy metal conditions [13]. [...]the peak at 1422–1430 cm−1 is an indicator of aromatic skeleton vibrations combined with C–H in-plane deformations [14]. Given the difficulties in cell enumeration on adhered living cells in the lignin surface, free bacterial cells were dry-weighed and correlated with extracted DNA. Since this method does not differentiate among living and dead cells, the serial dilution plating technique was applied and the viable cell number was also correlated with the extracted DNA to estimate the number of living cells in the lignin particles. Plant growth-promoting bacteria (PGPB) have been largely considered as beneficial in harsh and limiting environments given their effects on alleviating plant stress. For practical applications, most of the PGPB are prepared in immobilization matrices to improve the stability and benefits of bacteria. Despite the long list of immobilizing agents/carriers tested to date, a long list of desired requirements is yet to be achieved. Here, lignin stands as a scarcely tested immobilizer for bioinoculants with great potential for this purpose. The aim of this work was to demonstrate the feasibility of lignin as a carrier of the nitrogen-fixing . These bacteria were cultured in liquid media with recovered organosolv lignin added for bacterial immobilization. Then, lignin was recovered and the immobilized biomass was quantified gravimetrically by DNA extraction and serial dilution plating. Fluorescent microscopy as well as Congo red agar plating showed the immobilization of the bacterial cells in the lignin matrix and crystal violet dyeing showed the biofilms formation in lignin particles. A high number of cells were counted per gram of dried lignin. Lignin can be readily used as low-cost, health-safe bioinoculant carrier to be used in soil and agricultural applications. |
| Author | Tapia-Olivares, Victor Rogelio Escalante, Froylán M.E. Aguilar-Garnica, Efrén Vazquez-Bello, Eimy Alejandra |
| AuthorAffiliation | Department of Biotechnology and Environmental Sciences, Universidad Autónoma de Guadalajara, AC. Av. Patria 1201, Lomas del Valle, 45129 Zapopan, Mexico; vtaov4@gmail.com (V.R.T.-O.); eimy98@live.com.mx (E.A.V.-B.); efren.aguilar@edu.uag.mx (E.A.-G.) |
| AuthorAffiliation_xml | – name: Department of Biotechnology and Environmental Sciences, Universidad Autónoma de Guadalajara, AC. Av. Patria 1201, Lomas del Valle, 45129 Zapopan, Mexico; vtaov4@gmail.com (V.R.T.-O.); eimy98@live.com.mx (E.A.V.-B.); efren.aguilar@edu.uag.mx (E.A.-G.) |
| Author_xml | – sequence: 1 givenname: Victor Rogelio surname: Tapia-Olivares fullname: Tapia-Olivares, Victor Rogelio – sequence: 2 givenname: Eimy Alejandra surname: Vazquez-Bello fullname: Vazquez-Bello, Eimy Alejandra – sequence: 3 givenname: Efrén surname: Aguilar-Garnica fullname: Aguilar-Garnica, Efrén – sequence: 4 givenname: Froylán M.E. orcidid: 0000-0002-5788-9918 surname: Escalante fullname: Escalante, Froylán M.E. |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/31861075$$D View this record in MEDLINE/PubMed |
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| Keywords | biofertilizer organosolv nitrogen-fixing bacteria Azospirillum |
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| SubjectTerms | Advantages Azospirillum brasilense - growth & development Azospirillum brasilense - metabolism Bacteria Biofilms Biomass Ethanol Lignin Lignin - chemistry Microorganisms Nitrogen Nitrogen - metabolism Nitrogen Fixation Plant Development Spectroscopy, Fourier Transform Infrared Symbiosis |
| Title | Valorization of Lignin as an Immobilizing Agent for Bioinoculant Production using Azospirillum brasilense as a Model Bacteria |
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