Abstract
Using microalgal growth-promoting bacteria (MGPB) to improve the cultured microalga metabolism during biotechnological processes is one of the most promising strategies to enhance their benefits. Nonetheless, the culture condition effect used during the biotechnological process on MGPB growth and metabolism is key to ensure the expected positive bacterium growth and metabolism of microalgae. In this sense, the present research study investigated the effect of the synthetic biogas atmosphere (75% CH4–25% CO2) on metabolic and physiological adaptations of the MGPB Azospirillum brasilense by a microarray-based transcriptome approach. A total of 394 A. brasilense differentially expressed genes (DEGs) were found: 201 DEGs (34 upregulated and 167 downregulated) at 24 h and 193 DEGs (140 upregulated and 53 downregulated) under the same conditions at 72 h. The results showed a series of A. brasilense genes regulating processes that could be essential for its adaptation to the early stressful condition generated by biogas. Evidence of energy production is shown by nitrate/nitrite reduction and activation of the hypothetical first steps of hydrogenotrophic methanogenesis; signal molecule modulation is observed: indole-3-acetic acid (IAA), riboflavin, and vitamin B6, activation of Type VI secretion system responding to IAA exposure, as well as polyhydroxybutyrate (PHB) biosynthesis and accumulation. Moreover, an overexpression of ipdC, ribB, and phaC genes, encoding the key enzymes for the production of the signal molecule IAA, vitamin riboflavin, and PHB production of 2, 1.5 and 11 folds, respectively, was observed at the first 24 h of incubation under biogas atmosphere Overall, the ability of A. brasilense to metabolically adapt to a biogas atmosphere is demonstrated, which allows its implementation for generating biogas with high calorific values and the use of renewable energies through microalga biotechnologies.
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Data availability
The datasets generated and/or analyzed during the current study are available from the corresponding author on reasonable request.
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Acknowledgements
Francisco J. Choix acknowledges the Consejo Nacional de Humanidades, Ciencia y Tecnología (CONAHCYT, Mexico) for the support under the Program-Project 90 Cátedras CONAHCYT, as well as CONAHCYT-Frontiers of Science 2019 Project 15769; CIBNOR technical support of Julio Antonio Hernandez; Lorena Chávez González, Simón Guzmán León, and Jorge Ramírez for technical assistance in the microarray determinations; Diana Fischer for English edition.
Funding
This research was supported by CONACYT-Frontiers of Science 2019 Project 15769.
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All the authors contributed to the study conception and design. Material preparation, data collection and the first draft were performed by CG-M; FJC contributed to financial support and the design of the experiment; CAC contributed to data acquisition and analyses; LEdeB contributed to data analyses and discussion of the final draft; GAG-A contributed to experimental and microarray designs; OAP contributed to the experimental design, analysis of results, discussion and wrote the final draft. All the authors read and approved the final manuscript.
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This is an observational study. This article does not contain any studies with human participants or animals performed by any of the authors.
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Communicated by PANKAJ BHATT.
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Originality-Significance Statement: This research study provides novel and significant evidence that microalgal growth-promoting bacteria A. brasilense can adapt to biogas atmosphere and maintain plant/microalgal growth-promotion traits. Thus, the possibility of using this bacterium opens an environmentally sustainable biotechnology strategy to enhance CO2 capture from biogas by biological mechanisms.
Supplementary Information
Below is the link to the electronic supplementary material.
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Online resource 1. Modified Kitasato flask to test the effect of biogas and air atmospheres in liquid cultures. (XLSX 10 KB)
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Online resource 2. List of primers used for qRT‑PCR validation of differentially expressed mRNAs in the current study. (XLSX 30 KB)
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Online resource 3. List of differentially expressed Azospirillum brasilense genes growing under biogas at two different time points compared to an air atmosphere. Differential expression testing was performed using genArise software. Genes were considered significantly differentially expressed between biogas and air atmosphere conditions when they showed a z-score ≥ 1.5 or ≤ -1.5. (XLSX 32 KB)
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Online resource 4. List of enriched KEGG pathways in Azospirillum brasilense growing under biogas at two different time points compared to an air atmosphere. Where input number is the number of differentially expressed genes (DEGs) mapped to the indicated pathway, background number is the total number of genes in a specific pathway, and the rich factor is the ratio of the number of DEGs to the total gene number in a specific pathway. Pathways were considered significantly enriched with FDR-corrected P-value < 0.05. (PNG 1102 KB)
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Online resource 5. Phenotypic variation in Azospirillum brasilense. The parental strain Cd (A) growing under an air atmosphere (hydrocarbon-free air; control) was exposed to a biogas atmosphere (75% CH4−25% CO2; treatment) for 72 h (B), after which a white phenotypic variant that lacked the ability to produce carotenoids was observed. (PNG 31 KB)
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Online resource 6. Relative expression of ipdC, phaC, and ribB transcripts in Azospirillum brasilense growing under biogas at 24 h (grey) and 72 h (black) compared to an air atmosphere (=1). Data points represent the mean, and whisker lines represent standard error (SE), n = 3. Different lower letters between interval times in the same gene and different capital letters between genes in the same time indicate significant differences in relative expression after analysis of variance (ANOVA) and Fisher's LSD post hoc test (P < 0.05). (JPG 158 KB)
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Garciglia-Mercado, C., Contreras, C.A., Choix, F.J. et al. Metabolic and physiological adaptations of microalgal growth-promoting bacterium Azospirillum brasilense growing under biogas atmosphere: a microarray-based transcriptome analysis. Arch Microbiol 206, 173 (2024). https://doi.org/10.1007/s00203-024-03890-z
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DOI: https://doi.org/10.1007/s00203-024-03890-z