Effect of subtilisin, a protease from Bacillus sp., on soil biochemical parameters and microbial biodiversity

Caballero, Pablo; Macías-Benítez, Sandra; Revilla, Elisa; Tejada, Manuel; Parrado, Juan; Castaño, Angélica

doi:10.1016/j.ejsobi.2020.103244

Cited by 8 publications

(5 citation statements)

References 44 publications

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“…Recent results in our group have revealed that the exogenous application of naturallyproduced-in-soil microbial enzymes yielded interesting results not only at the biochemical level, stimulating soil microbial enzymes, but also over the microbial biodiversity [15]. Thus, by applying subtilisin from Bacillus licheniformis, we found an interesting stimulation of PGPB.…”

Section: Introductionmentioning

confidence: 71%

“…The experimental design was stablished according to previous studies [15]. Thus, microcosms of 250 g of soil were preincubated at 30-40% of their water holding capacity for 7 days.…”

Section: Design Of the Soil Biostimulation Studymentioning

confidence: 99%

“…The Rubrobacteriaceae family was negatively affected along the experiment in all of the soil groups including the control. We have previously described that after applying subtilisin, a soil extracellular endopeptodase from Bacillus sp., relative abundance of Rubrobacter decrease in soil and the effect was more pronounced when subtilisin was applied in combination with keratins [15]. In general, the Rubrobacter strains are associated with extreme environments (e.g., high temperature environments) such as the fumarole heated stream, soil adjacent to volcanic caldera, deteriorated monuments, or even halophilic species [47], and show low ecological value.…”

Section: Changes In Soil At Microbiological Levelmentioning

confidence: 99%

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Biochemical and Microbiological Soil Effects of a Biostimulant Based on Bacillus licheniformis-Fermented Sludge

et al. 2022

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Biostimulants are substances and/or microorganisms that are applied to plants or to the rhizosphere in order to enhance the natural process improving the absorption of nutrients and the quality of crops as well as the tolerance to abiotic stresses. A new biostimulant was developed from sewage sludge through its fermentation with Bacillus licheniformis as a plant growth-promoting bacteria (PGPB). The fermented product includes three classes of biostimulant components: the B. licheniformis biomass; the enzymatic secretion of said microorganism, which are mainly peptidases and amidases related to nitrogen metabolism and glucanases, related to carbohydrate metabolism; and finally, the hydrolyzed sludge organic matter, with a high content of protein hydrolysates. The biostimulant was evaluated in soil at the biochemical (enzymatic activities) and microbiological levels (metabarcoding analysis). Metabarcoding analysis revealed that the biostimulant complex, mainly the soluble fraction containing the Bacillus multienzyme complex and protein hydrolysate, induced PGPB soil bacteria, and it was detected that the inoculation in the soil of B. licheniformis remained active throughout the study. These results show the fermentation process with B. licheniformis as an interesting option for the total valorization of activated sewage sludge aimed at obtaining products of agronomic/environmental interest.

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Section: Introductionmentioning

confidence: 71%

“…The experimental design was stablished according to previous studies [15]. Thus, microcosms of 250 g of soil were preincubated at 30-40% of their water holding capacity for 7 days.…”

Section: Design Of the Soil Biostimulation Studymentioning

confidence: 99%

Section: Changes In Soil At Microbiological Levelmentioning

confidence: 99%

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Biochemical and Microbiological Soil Effects of a Biostimulant Based on Bacillus licheniformis-Fermented Sludge

et al. 2022

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“…Plant microbiome members, unlike other microbiota, develop an intimate relationship with plant metabolism and influence it in harmony to alleviate various biotic and abiotic stress conditions. Among all, the firmicutes are well known for their abundance on plants, and the genus Bacillus has a special place with extensive studies on agroecology, including nutrient cycling [ 16 – 18 ], disease suppression [ 19 – 21 ], plant growth regulation [ 22 , 23 ], diversity enhancement [ 24 , 25 ] and phytoremediation [ 26 , 27 ]. With this relevance, the pearl millet microbiome is under exploration of late, and associated microbes from the root, leaf, and stem regions of pearl millet are attributed in many studies to its drought stress tolerance ability [ 28 ].…”

Section: Introductionmentioning

confidence: 99%

Comprehensive genomic analysis of Bacillus subtilis and Bacillus paralicheniformis associated with the pearl millet panicle reveals their antimicrobial potential against important plant pathogens

Ashajyothi,

Mahadevakumar,

Venkatesh

et al. 2024

BMC Plant Biol

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Background Plant microbiome confers versatile functional roles to enhance survival fitness as well as productivity. In the present study two pearl millet panicle microbiome member species Bacillus subtilis PBs 12 and Bacillus paralicheniformis PBl 36 found to have beneficial traits including plant growth promotion and broad-spectrum antifungal activity towards taxonomically diverse plant pathogens. Understanding the genomes will assist in devising a bioformulation for crop protection while exploiting their beneficial functional roles. Results Two potential firmicute species were isolated from pearl millet panicles. Morphological, biochemical, and molecular characterization revealed their identities as Bacillus subtilis PBs 12 and Bacillus paralicheniformis PBl 36. The seed priming assays revealed the ability of both species to enhance plant growth promotion and seedling vigour index. Invitro assays with PBs 12 and PBl 36 showed the antibiosis effect against taxonomically diverse plant pathogens (Magnaporthe grisea; Sclerotium rolfsii; Fusarium solani; Alternaria alternata; Ganoderma sp.) of crops and multipurpose tree species. The whole genome sequence analysis was performed to unveil the genetic potential of these bacteria for plant protection. The complete genomes of PBs 12 and PBl 36 consist of a single circular chromosome with a size of 4.02 and 4.33 Mb and 4,171 and 4,606 genes, with a G + C content of 43.68 and 45.83%, respectively. Comparative Average Nucleotide Identity (ANI) analysis revealed a close similarity of PBs 12 and PBl 36 with other beneficial strains of B. subtilis and B. paralicheniformis and found distant from B. altitudinis, B. amyloliquefaciens, and B. thuringiensis. Functional annotation revealed a majority of pathway classes of PBs 12 (30) and PBl 36 (29) involved in the biosynthesis of secondary metabolites, polyketides, and non-ribosomal peptides, followed by xenobiotic biodegradation and metabolism (21). Furthermore, 14 genomic regions of PBs 12 and 15 of PBl 36 associated with the synthesis of RiPP (Ribosomally synthesized and post-translationally modified peptides), terpenes, cyclic dipeptides (CDPs), type III polyketide synthases (T3PKSs), sactipeptides, lanthipeptides, siderophores, NRPS (Non-Ribosomal Peptide Synthetase), NRP-metallophone, etc. It was discovered that these areas contain between 25,458 and 33,000 secondary metabolite-coding MiBiG clusters which code for a wide range of products, such as antibiotics. The PCR-based screening for the presence of antimicrobial peptide (cyclic lipopeptide) genes in PBs 12 and 36 confirmed their broad-spectrum antifungal potential with the presence of spoVG, bacA, and srfAA AMP genes, which encode antimicrobial compounds such as subtilin, bacylisin, and surfactin. Conclusion The combined in vitro studies and genome analysis highlighted the antifungal potential of pearl millet panicle-associated Bacillus subtilis PBs12 and Bacillus paralicheniformis PBl36. The genetic ability to synthesize several antimicrobial compounds indicated the industrial value of PBs 12 and PBl 36, which shed light on further studies to establish their action as a biostimulant for crop protection.

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“…can secrete many endopeptidases, such as S1, S8, M4, and C40, and exopeptidases such as M24, S13, S11, and S26 to synergistically degrade macromolecular organic nitrogen. A recent study showed that proteases from the genus Bacillus stimulated microbial activity, which in turn promotes the mineralization of organic matter and phosphorus ( Caballero et al, 2020 ). Therefore, adding an appropriate number of low temperature Bacillus to the plateau environment could promote the conversion of organic nitrogen to available nitrogen.…”

Section: Discussionmentioning

confidence: 99%

A functional metagenomics study of soil carbon and nitrogen degradation networks and limiting factors on the Tibetan plateau

et al. 2023

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IntroductionThe Three-River Source Nature Reserve is located in the core area of the Qinghai-Tibetan Plateau, with the alpine swamp, meadow and steppe as the main ecosystem types. However, the microbial communities in these alpine ecosystems, and their carbon and nitrogen degrading metabolic networks and limiting factors remain unclear.MethodsWe sequenced the diversity of bacteria and fungi in alpine swamps, meadows, steppes, and their degraded and artificially restored ecosystems and analyzed soil environmental conditions.ResultsThe results indicated that moisture content had a greater influence on soil microbial community structure compared to degradation and restoration. Proteobacteria dominated in high moisture alpine swamps and alpine meadows, while Actinobacteria dominated in low moisture alpine steppes and artificial grasslands. A metabolic network analysis of carbon and nitrogen degradation and transformation using metagenomic sequencing revealed that plateau microorganisms lacked comprehensive and efficient enzyme systems to degrade organic carbon, nitrogen, and other biological macromolecules, so that the short-term degradation of alpine vegetation had no effect on the basic composition of soil microbial community. Correlation analysis found that nitrogen fixation was strong in meadows with high moisture content, and their key nitrogen-fixing enzymes were significantly related to Sphingomonas. Denitrification metabolism was enhanced in water-deficient habitats, and the key enzyme, nitrous oxide reductase, was significantly related to Phycicoccus and accelerated the loss of nitrogen. Furthermore, Bacillus contained a large number of amylases (GH13 and GH15) and proteases (S8, S11, S26, and M24) which may promote the efficient degradation of organic carbon and nitrogen in artificially restored grasslands.DiscussionThis study illustrated the irrecoverability of meadow degradation and offered fundamental information for altering microbial communities to restore alpine ecosystems.

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Effect of subtilisin, a protease from Bacillus sp., on soil biochemical parameters and microbial biodiversity

Cited by 8 publications

References 44 publications

Biochemical and Microbiological Soil Effects of a Biostimulant Based on Bacillus licheniformis-Fermented Sludge

Biochemical and Microbiological Soil Effects of a Biostimulant Based on Bacillus licheniformis-Fermented Sludge

Comprehensive genomic analysis of Bacillus subtilis and Bacillus paralicheniformis associated with the pearl millet panicle reveals their antimicrobial potential against important plant pathogens

A functional metagenomics study of soil carbon and nitrogen degradation networks and limiting factors on the Tibetan plateau

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