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 inMolecules (Basel, Switzerland) Vol. 24; no. 24; p. 4613
Main Authors Tapia-Olivares, Victor Rogelio, Vazquez-Bello, Eimy Alejandra, Aguilar-Garnica, Efrén, Escalante, Froylán M.E.
Format Journal Article
LanguageEnglish
Published Switzerland MDPI AG 17.12.2019
MDPI
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ISSN1420-3049
1420-3049
DOI10.3390/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.
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.)
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CitedBy_id crossref_primary_10_1007_s42729_024_01947_y
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crossref_primary_10_1016_j_soilbio_2024_109578
crossref_primary_10_3390_agronomy13041032
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Keywords biofertilizer
organosolv
nitrogen-fixing bacteria
Azospirillum
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Snippet Plant growth-promoting bacteria (PGPB) have been largely considered as beneficial in harsh and limiting environments given their effects on alleviating plant...
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...
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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
URI https://www.ncbi.nlm.nih.gov/pubmed/31861075
https://www.proquest.com/docview/2333430960
https://www.proquest.com/docview/2329737635
https://pubmed.ncbi.nlm.nih.gov/PMC6943589
Volume 24
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