Construction of lignocellulose-degrading compound microbial inoculum and its effects on green waste composting.

The high content of recalcitrant lignocellulose in green waste (GW) makes composting and degradation challenging. Conventional GW composting typically employs single-strain microbial inoculants (MIs) with limited enzyme production capabilities, resulting in low composting efficiency and suboptimal compost product quality. In this study, Bacillus amyloliquefaciens (J1), Clonostachys rogersoniana (B2), and Streptomyces thermoviolaceus (J3) was utilized to optimize cultivation conditions and strain ratios based on enzyme activity indicators. The aim was to develop a potent three-strain lignocellulose-degrading MIs and test the hypothesis that its performance is superior to that of single-strain and two-strain MIs in terms of lignocellulose degradation and compost maturation. The results indicated that, the optimal treatment was T7, which was inoculated with a three-strain MIs composed of the spore suspensions of J1, B2, and J3 with a volume ratio of 3:3:2. Specifically, compared to the control (without MI), T7 increased the content of particle size between 0.25 and 2.00 mm and humic acid by 17% and 291%, respectively. Furthermore, T7 enhanced the degradation rates of cellulose, hemicellulose, and lignin by 197%, 145%, and 113%, respectively, and increased the activities of laccase, manganese peroxidase, lignin peroxidase, and carboxymethyl cellulase by 605%, 269%, 180%, and 228%, respectively. Additionally, T7 increased the relative abundance of bacteria (e.g. Pseudomonas) and fungi (e.g. Parascedosporium) that facilitated lignocellulose degradation, enhanced the alpha diversity index and promoted the formation of a microbial community structure characterized by prominent dominant species and greater diversity. Remarkably, the inoculation with the three-strain MI yielded high-quality compost within 32 days.

Recent advances in the use of Trichoderma-containing multicomponent microbial inoculants for pathogen control and plant growth promotion.

Chemical pesticides and fertilizers are used in agricultural production worldwide to prevent damage from plant pathogenic microorganisms, insects, and nematodes, to minimize crop losses and to preserve crop quality. However, the use of chemical pesticides and fertilizers can severely pollute soil, water, and air, posing risks to the environment and human health. Consequently, developing new, alternative, environment-friendly microbial soil treatment interventions for plant protection and crop yield increase has become indispensable. Members of the filamentous fungal genus Trichoderma (Ascomycota, Sordariomycetes, Hypocreales) have long been known as efficient antagonists of plant pathogenic microorganisms based on various beneficial traits and abilities of these fungi. This minireview aims to discuss the advances in the field of Trichoderma-containing multicomponent microbiological inoculants based on recent experimental updates. Trichoderma strains can be combined with each other, with other fungi and/or with beneficial bacteria. The development and field performance of such inoculants will be addressed, focusing on the complementarity, synergy, and compatibility of their microbial components.

Microbial Catabolic Activity: Methods, Pertinence, and Potential Interest for Improving Microbial Inoculant Efficiency.

Microbial catabolic activity (MCA) defined as the degrading activity of microorganisms toward various organic compounds for their growth and energy is commonly used to assess soil microbial function potential. For its measure, several methods are available including multi-substrate-induced respiration (MSIR) measurement which allow to estimate functional diversity using selected carbon substrates targeting specific biochemical pathways. In this review, the techniques used to measure soil MCA are described and compared with respect to their accuracy and practical use. Particularly the efficiency of MSIR-based approaches as soil microbial function indicators was discussed by (i) showing their sensitivity to different agricultural practices including tillage, amendments, and cropping systems and (ii) by investigating their relationship with soil enzyme activities and some soil chemical properties (pH, soil organic carbon, cation exchange capacity). We highlighted the potential of these MSIR-based MCA measurements to improve microbial inoculant composition and to determine their potential effects on soil microbial functions. Finally, we have proposed ideas for improving MCA measurement notably through the use of molecular tools and stable isotope probing which can be combined with classic MSIR methods. Graphical abstract describing the interrelation between the different parts and the concepts developed in the review.

Characterization and in vitro evaluations of Baru pulp from almond production waste ensiled with different additives.

The silage process is an efficient way of storing nutrients for animal nutrition. Our hypothesis was that the Baru pulp can be preserved in the form of silage, regardless of the use of additives to aid the process. Silages of Baru pulp containing different additives were evaluated through in vitro analysis, fermentation parameters, and composition and thermal analysis including differential scanning calorimetry and thermogravimetry. The treatments consisted of (1) silage with Baru pulp (BP) in natura without additive; (2) BP in natura with acetic acid; (3) BP in natura with formic acids; and (4) BP in natura added with microbial inoculums in a randomized experimental design with three replications per treatment and analyzed in duplicate. The ensiled material was kept in anaerobic conditions for a period of 30 days. BP before and after the silage process presented averages of 67.31 and 66.24% for in vitro digestibility of DM (IVDMD). Microbial inoculant additive was the most effective in reducing pH, followed by acetic acid and formic acid. There were effects of additives on silages for all degradation parameters in ruminal liquid in vitro. It was observed that BP before ensiling had the highest A fraction (7.9 mL gas/100 mg DM), without differing from the silage treated with formic acid (7.1 mL gas/100 mg DM). Similar effects were observed on mass loss (TG) and heat flux (DSC) between the silages. Our findings suggest that Baru pulp with formic acid is more efficient in the conservation and preservation of fermentable carbohydrates as well as in silage production.

Characterization of plant growth-promoting rhizobacterial isolates associated with food plants in South Africa.

The region around the plant root referred to as the rhizosphere, is the zone where various microbial activity occurs. It performs crucial functions such as increasing the uptake of nutrients for plant development and preventing plant against plant pathogens. Keeping in mind the beneficial role performed by rhizospheric microorganisms, rhizobacterial species were isolated from the maize and soybean plant's rhizosphere. The isolated microorganisms were evaluated for their biochemical characteristics, plant growth-promoting potentials, tolerance to different environmental conditions, and their antifungal activity against Fusarium graminearum, a fungal pathogen that infects maize. The rhizobacterial isolates with multiple plant growth-promoting potentials were identified as Bacillus spp (80.77%), Rhodocyclaceae bacterium (3.85%), Enterococcus spp (3.85%). Massilia spp (3.85%. and Pseudomonas (7.69%) species based on their 16S rRNA molecular characterization. The bacterial isolates possessed antifungal activities against Fusarium graminearum, promote maize and soybeans seed under laboratory conditions, and exhibited different levels of tolerance to pH, temperature, salt, and heavy metal. Based on this, the whole genome sequencing of Bacillus sp. OA1, Pseudomonas rhizosphaerea OA2, and Pseudomonas sp. OA3 was performed using Miseq Illumina system to determine the functional genes and secondary metabolites responsible for their plant growth-promoting potential Thus, the result of this research revealed that the selected bacterial isolates possess plant growth-promoting potentials that can make them a potential candidate to be employed as microbial inoculants for protecting plants against phytopathogens, environmental stress and increasing plant growth and productivity.

Evaluation of biochars as carriers for Rhizobium leguminosarum.

Peat is the standard carrier material used for commercial microbial inoculants produced in Canada and the United States. Peat is a slowly renewable resource and its production is extremely vulnerable to variable weather conditions. Furthermore, it may not be widely available in all countries. We investigated the potential to develop biochar as a carrier material. Our goal was to evaluate if different biochars perform comparably in supporting rhizobial survival, and what characteristics contribute to their ability to support rhizobial survival. Evaluation included characterization of the biochars, assessment of biochar phytotoxicity, survival of Rhizobium on biochars, and growth chamber evaluation of two biochars as Rhizobium carriers for inoculating pea. Of the original nine biochars evaluated, six supported Rhizobium leguminosarum for 84 days at 4 °C; of this six, two supported numbers >1 × 106 cfu·(g biochar)-1. The only characteristics that correlated with survival were C/N ratio and percent C. The two biochars evaluated delivered R. leguminosarum to pea that initiated nodulation, biomass production, and biomass N at levels higher than a noninoculated control and heat-killed inoculated biochars. We demonstrate that there is considerable potential to develop biochar as a carrier for rhizobial inoculants.

Fermentative profile and lactic acid bacterial dynamics in non-wilted and wilted alfalfa silage in tropical conditions.

This study was conducted to evaluate the fermentative profile and microbial populations of wilted and non-wilted alfalfa silages ensiled with or without inoculant and the population dynamics of lactic acid bacteria (LAB) of wilted alfalfa plant and theirs silage. A 2 × 2 × 6 factorial arrangement was used, with the absence or presence of wilting (W), with and without bacterial inoculant (I) and six fermentation periods (P) (1, 3, 7, 14, 28 and 56 days), in a completely randomized design, with three replicates. The alfalfa was slightly wilted for 6 h and increased the dry matter content from 133.9 to 233.4 g/kg. It was performed the cultivation, followed by the isolation of LAB from samples of alfalfa forage before ensiling and its silage only in non-inoculated silages, after different fermentation periods. DNA was extracted from the isolated strains of LAB; the 16S rRNA gene sequences were amplified by PCR and the sequences were compared to those available from the GenBank database. Wilting provided silages with lower pH, ammonia nitrogen and acetic acid concentrations. The wilting process did not alter the amount of LAB; however, it affected the LAB diversity of the silages. The Lactobacillus plantarum was the predominant species in non-wilted and wilted silages.

Effect of microbial inoculants on fermentation quality and aerobic stability of sweet potato vine silage.

OBJECTIVE: This study was conducted to evaluate the effect of homo or hetero fermentative inoculants on fermentation quality and aerobic stability of sweet potato vine (SPV) silage containing Italian ryegrass hay as moisture absorbent. METHODS: The SPV was harvested at 15% dry matter, mixed with Italian ryegrass hay at 1:1 ratio on a fresh weight basis, and chopped to 3 to 5 cm length. After then, the chopped forage mixture was ensiled into 20-L mini silos in quadruplicate for 7, 48, and 100 days after application of microbial inoculants at 1.2×105 colony forming units (cfu)/g of forage following: no inoculant (CON), Lactobacillus plantarum as a homo fermentative (LP), Lactobacillus buchneri as a hetero fermentative (LB), and mixture of LP and LB at 1:1 ratio as a combo fermentative (MIX). RESULTS: The LP and MIX silages had lowest pH (p<0.001) on 7 and 48 days, while MIX and CON silages had greatest lactate concentrations (p<0.05) on 7 and 48 days, respectively. Acetate concentrations were highest (p<0.01) in LB and MIX silages on 7 days, and in LB silage on 48 days, while lactate to acetate ratios were lowest (p<0.001) in LB silages. The chemical compositions and nutrient digestibility of silage ensiled for 100 days was not affected by inoculants. On 100 days of ensiling, LB silage had lowest (p<0.01) lactate concentration and lactate to acetate ratio, but highest acetate concentration. Aerobic stability was highest (p<0.001) in LB silage followed in MIX silage. On contrast, LB silage had lowest (p<0.05) lactic acid bacteria and mold. CONCLUSION: The results indicated that application of LB solely had a better effect on aerobic stability than not only LP, but also MIX. However, LP application did not show beneficial effects from the viewpoints of fermentation quality and aerobic stability compared to CON.

Impact of plant growth-promoting rhizobacteria on root colonization potential and life cycle of Rhizophagus irregularis following co-entrapment into alginate beads.

AIMS: This study aimed at evaluating the impact of seven plant growth-promoting rhizobacteria (PGPR) on root colonization and life cycle of Rhizophagus irregularis MUCL 41833 when co-entrapped in alginate beads. METHODS AND RESULTS: Two in vitro experiments were conducted. The first consisted of the immobilization of R. irregularis and seven PGPR isolates into alginate beads to assess the effect of the bacteria on the pre-symbiotic growth of the fungus. In the second experiment, the best performing PGPR from experiment 1 was tested for its ability to promote the symbiotic development of the AMF in potato plantlets from three cultivars. Results showed that only one isolate identified as Pseudomonas plecoglossicida (R-67094) promoted germ tube elongation and hyphal branching of germinated spores during the pre-symbiotic phase of the fungus. This PGPR further promoted the symbiotic development of the AMF in potato plants. CONCLUSIONS: The co-entrapment of Ps. plecoglossicida R-67094 and R. irregularis MUCL 41833 in alginate beads improved root colonization by the AMF and its further life cycle under the experimental conditions. SIGNIFICANCE AND IMPACT OF THE STUDY: Co-entrapment of suitable AMF-PGPR combinations within alginate beads may represent an innovative technology that can be fine-tuned for the development of efficient consortia-based bioformulations.

Effects of Lactobacillus formosensis S215T and Lactobacillus buchneri on quality and in vitro ruminal biological activity of condensed tannins in sweet potato vines silage.

AIMS: This study investigated the influence of two microbial inoculants; Lactobacillus formosensis and Lactobacillus buchneri on fermentation quality, chemical composition, aerobic stability and in vitro ruminal biological activity of condensed tannins in sweet potato vines silage. METHODS AND RESULTS: Sweet potato vines were ensiled for 28 and 60 days; without inoculant (CON), with Lact. buchneri (LB) or with Lact. formosensis (LF), both inoculants applied to achieve 1 × 106  CFU g-1 fresh forage. Lactobacillus formosensis silage had lower pH and higher lactic acid than all treatments. Yeasts and moulds were not detected in LB silage after ensiling. Lactobacillus buchneri silage was more aerobically stable than all treatments, whereas LF was more stable than CON silage. In vitro ruminal biological activity of condensed tannins was lower in microbial-inoculated silages than CON after ensiling. CONCLUSION: Lactobacillus formosensis improved fermentability by reducing silage pH and improved aerobic stability by producing more propionate, which inhibited yeast activity. Lactobacillus buchneri improved aerobic stability of the silage by producing more acetate. Both strains effectively reduced the antinutritional effect of condensed tannins after ensiling. SIGNIFICANCE AND IMPACT OF THE STUDY: Lactobacillus formosensis has the potential to be used as a silage inoculant because of its ability to improve fermentability and aerobic stability in sweet potato vines silage.

Does nature make provision for backups in the modification of bacterial community structures?

Self-balancing is an inherent character in nature in response to community structure modification pressure and modern biotechnology has revolutionized the way such detections are made. Presented here is an overview of the forces and process interactions between released bacteria and indigenous microflora which encompass soil bacterial diversity, community structure, indigenous endorhizosphere micro-organisms, molecular detection methodologies, and transgenic plants and microbes. Issues of soil bacterial diversity and community structure as well as the interpretation of results from various findings are highlighted and discussed as inferred from research articles. An understanding of the factors influencing bio-inoculant modification of bacterial community structure in the colonization of the rhizosphere is essential for improved establishment of biocontrol agents, and is critically reviewed.

Integrative analysis of microbiome and metabolome revealed the effect of microbial inoculant on microbial community diversity and function in rhizospheric soil under tobacco monoculture.

Over-application of chemical fertilizers and continuous cropping obstacles seriously restrict the sustainable development of tobacco production. Localized fertilization of beneficial microbes has potential advantages in achieving higher productivity, but the underlying biological mechanisms of interactions between rhizospheric microorganisms and the related metabolic cycle remain poorly characterized. Here, an integrative analysis of microbiomes with non-targeted metabolomics was performed on 30 soil samples of rhizosphere, root surrounding, and bulk soils from flue-cured tobacco under continuous and non-continuous monocropping systems. The analysis was conducted using UPLC-MS/MS platforms and high-throughput amplicon sequencing targeting the bacterial 16S rRNA gene and fungal ITS gene. The microbial inoculant consisted of Bacillus subtilis, B. velezensis, and B. licheniformis at the ratio of 1:1:1 in effective microbial counts, improved the cured leaf yield and disease resistance of tobacco, and enhanced nicotine and nitrogen contents of tobacco leaves. The bacterial taxa Rhizobium, Pseudomonas, Sphingomonadaceae, and Burkholderiaceae of the phylum Proteobacteria accumulated in high relative abundance and were identified as biomarkers following the application of the microbial inoculant. Under continuous monocropping, metabolomics demonstrated that the application of the microbial inoculant significantly affected the soil metabolite spectrum, and the differential metabolites were significantly enriched to the synthesis and degradation of nicotine (nicotinate and nicotinamide metabolism and biosynthesis of alkaloids derived from nicotinic acid). In addition, microbes were closely related to the accumulation of metabolites through correlation analysis. The interactions between plant roots and rhizospheric microorganisms provide valuable information for understanding how these beneficial microbes affect complex biological processes and the adaption capacity of plants to environments.IMPORTANCEThis study elaborated on how the microbial fertilizer significantly changed overall community structures and metabolite spectrum of rhizospheric microbes, which provide insights into the process of rhizosphere microbial remolding in response to continuous monocropping. we verified the hypothesis that the application of the microbial inoculant in continuous cropping would lead to the selection of distinct microbiota communities by establishing models to correlate biomarkers. Through correlation analysis of the microbiome and metabolome, we proved that rhizospheric microbes were closely related to the accumulation of metabolites, including the synthesis and degradation of nicotine. The interactions between plant roots and rhizospheric microorganisms provide valuable information for understanding how these beneficial microbes affect complex biological processes and the adaption capacity of plants to environments.

Amendment of straw with decomposing inoculants benefits the ecosystem carbon budget and carbon footprint in a subtropical wheat cropping field.

The incorporation of straw with decomposing inoculants into soils has been widely recommended to sustain agricultural productivity. However, comprehensive analyses assessing the effects of straw combined with decomposing inoculants on greenhouse gas (GHG) emissions, net primary production (NPP), the net ecosystem carbon budget (NECB), and the carbon footprint (CF) in farmland ecosystems are scant. Here, we carried out a 2-year field study in a wheat cropping system with six treatments: rice straw (S), a straw-decomposing Bacillus subtilis inoculant (K), a straw-decomposing Aspergillus oryzae inoculant (Q), a combination of straw and Bacillus subtilis inoculant (SK), a combination of straw and Aspergillus oryzae inoculant (SQ), and a control with no rice straw or decomposing inoculant (Control). We found that all the treatments resulted in a positive NECB ranging between 838 and 5065 kg C ha-1. Relative to the Control, the S treatment increased CO2 emissions by 16%, while considerably enhancing the NECB by 349%. This difference might be attributed to the straw C input and an increase in plant productivity (NPP, 30%). More importantly, in comparison to that in S, the NECB in SK and SQ significantly increased by 27-35% due to the positive response of NPP to the decomposing inoculants. Although the combination of straw and decomposing inoculants yielded a 3% increase in indirect GHG emissions, it also exhibited the lowest CF (0.18 kg CO2-eq kg-1 of grain). This result was attributed to the synergistic effects of straw and decomposing inoculants, which reduced direct N2O emissions and increased wheat productivity. Overall, the findings of the present study suggested that the combined amendment of straw and decomposing inoculants is an environmentally sustainable management practice in wheat cropping systems that can generate win-win scenarios through improvements in soil C stock, crop productivity, and GHG mitigation.

Virome and metagenomic sequencing reveal the impact of microbial inoculants on suppressions of antibiotic resistome and viruses during co-composting.

Co-composting with exogenous microbial inoculant, presents an effective approach for the harmless utilization of livestock manure and agroforestry wastes. However, the impact of inoculant application on the variations of viral and antibiotic resistance genes (ARGs) remains poorly understood, particularly under varying manure quantity (low 10 % vs. high 20 % w/w). Thus, employing virome and metagenomic sequencing, we examined the influence of Streptomyces-Bacillus Inoculants (SBI) on viral communities, phytopathogen, ARGs, mobile genetic elements, and their interrelations. Our results indicate that SBI shifted dominant bacterial species from Phenylobacterium to thermotropic Bordetella, and the quantity of manure mediates the effect of SBI on whole bacterial community. Major ARGs and genetic elements experienced substantial changes with SBI addition. There was a higher ARGs elimination rate in the composts with low (∼76 %) than those with high manure (∼70 %) application. Virus emerged as a critical factor influencing ARG dynamics. We observed a significant variation in virus community, transitioning from Gemycircularvirus- (∼95 %) to Chlamydiamicrovirus-dominance. RDA analysis revealed that Gemycircularvirus was the most influential taxon in shaping ARGs, with its abundance decreased approximately 80 % after composting. Collectively, these findings underscore the role of microbial inoculants in modulating virus communities and ARGs during biowaste co-composting.

Adsorption mechanism of Cd2+ on microbial inoculant and its potential for remediation Cd-polluted farmland soils.

The adsorption characteristics and mechanism of Cd2+ on microbial inoculant (MI) mainly composed of Bacillus subtilis, Bacillus thuringiensis and Bacillus amyloliquefaciens, and its potential for remediation Cd polluted soils through batch adsorption and soil incubation experiments. It was found that the Freundlich isotherm model and the pseudo-second-order kinetics were more in line with the adsorption processes of Cd2+. The maximum adsorption capacity predicted by Langmuir isotherm model suggested that of MI was 57.38 mg g-1. Scanning electron microscopy and energy dispersive spectroscopy (SEM-EDS) images exhibited the surface structure of MI was damaged to varying degrees after adsorption, and Cd element was distributed on the surface of MI through ion exchange. X-ray diffraction (XRD) results showed that CdCO3 was formed on the surface of MI. Moreover, the functional groups (-OH, C-H, and -NH) involved in the adsorption of Cd2+ through fourier transform infrared spectroscopy (FTIR). After applying MI to Cd-contaminated soil, it was found that soil pH, conductivity (EC) and soil organic matter (SOM) increased by 0.84 %-2.43 %, 31.6 %-241.48 %, and 8.11 %-24.1 %, respectively, when compared with the control treatments. The content of DTPA-Cd in the soils was significantly (P < 0.05) reduced by 15.48 %-29.68 % in contrast with CK, and the Cd speciation was transformed into a more stable residual fraction. The activities of urease, phosphatase and sucrose were increased by 3.5 %-45.18 %, 57.00 %-134.18 % and 52.51 %-70.52 %, respectively, compared with CK. Therefore, MI could be used as an ecofriendly and sustainable material for bioremediation of Cd-contaminated soils.

Response of antioxidant activity, active constituent and rhizosphere microorganisms of Salvia miltiorrhiza to combined application of microbial inoculant, microalgae and biochar under Cu stress.

Salvia miltiorrhiza, a widely used medicinal herb renowned for its properties in promoting blood circulation, removing blood stasis and alleviating pain, is currently facing quality degradation due to excessive heavy metal levels, posing a threat to medication safety. In order to investigate the effects of microbial inoculant, microalgae and biochar on the growth of Salvia miltiorrhiza under copper (Cu) stress, as well as its Cu absorption, antioxidant activity, active component contents and rhizosphere microbial community, a pot experiment was conducted. Salvia miltiorrhiza plants were cultivated in the soil containing 400 mg/kg of Cu for six months and treated with microbial inoculant, microalgae and biochar, either individually or in combination. Almost all soil amendment treatments led to an increase in root biomass. Notably, co-application of microbial inoculant and microalgae had the optimal effect with a 63.07 % increase compared to the group treated solely with Cu. Moreover, when microbial inoculant was applied alone or in combination with microalgae, the Cu content in plant roots was reduced by 19.29 % and 25.37 %, respectively, whereas other treatments failed to show a decreasing trend. Intriguingly, Cu stress increased the active component contents in plant roots, and they could also be enhanced beyond non-stress levels when microbial inoculant and microalgae were applied together or in combination with biochar. Analyses of plant antioxidant activity, soil properties and rhizosphere microorganisms indicated that these amendments may alleviate Cu stress by enhancing peroxidase activity, facilitating plant nutrient absorption, and enriching beneficial microorganisms capable of promoting plant growth and mitigating heavy metal-induced damage. This study suggests that the combined application of microbial inoculant and microalgae can reduce Cu levels in Salvia miltiorrhiza while enhancing its quality under Cu stress.

From waste to protein: a new strategy of converting composted distilled grain wastes into animal feed.

Distilled grain waste (DGW) is rich in nutrients and can be a potential resource as animal feed. However, DGW contains as much as 14% lignin, dramatically reducing the feeding value. White-rot fungi such as Pleurotus ostreatus could preferentially degrade lignin with high efficiency. However, lignin derivatives generated during alcohol distillation inhibit P. ostreatus growth. Thus, finding a new strategy to adjust the DGW properties to facilitate P. ostreatus growth is critical for animal feed preparation and DGW recycling. In this study, three dominant indigenous bacteria, including Sphingobacterium thermophilum X1, Pseudoxanthomonas byssovorax X3, and Bacillus velezensis 15F were chosen to generate single and compound microbial inoculums for DGW composting to prepare substrates for P. ostreatus growth. Compared with non-inoculated control or single microbial inoculation, all composite inoculations, especially the three-microbial compound, led to faster organic metabolism, shorter composting process, and improved physicochemical properties of DGW. P. ostreatus growth assays showed the fastest mycelial colonization (20.43 µg·g-1 ergosterol) and extension (9 mm/d), the highest ligninolytic enzyme activities (Lac, 152.68 U·g-1; Lip, 15.56 U·g-1; MnP, 0.34 U·g-1; Xylanase, 10.98 U·g-1; FPase, 0.71 U·g-1), and the highest lignin degradation ratio (30.77%) in the DGW sample after 12 h of composting with the three-microbial compound inoculation when compared to other groups. This sample was relatively abundant in bacteria playing critical roles in amino acid, carbohydrate, energy metabolism, and xenobiotic biodegradation, as suggested by metagenomic analysis. The feed value analysis revealed that P. ostreatus mycelia full colonization in composted DGW led to high fiber content retention and decreased lignin content (final ratio of 5% lignin) but elevated protein concentrations (about 130 g·kg-1 DM). An additional daily weight gain of 0.4 kg/d was shown in cattle feeding experiments by replacing 60% of regular feed with it. These findings demonstrate that compound inoculant consisting of three indigenous microorganisms is efficient to compost DGW and facilitate P. ostreatus growth. P. ostreatus decreased the lignin content of composted DGW during its mycelial growth, improving the quality of DGW for feeding cattle.

Deciphering Microbial Community and Nitrogen Fixation in the Legume Rhizosphere.

Nitrogen is the most limiting factor in crop production. Legumes establish a symbiotic relationship with rhizobia and enhance nitrogen fixation. We analyzed 1,624 rhizosphere 16S rRNA gene samples and 113 rhizosphere metagenomic samples from three typical legumes and three non-legumes. The rhizosphere microbial community of the legumes had low diversity and was enriched with nitrogen-cycling bacteria (Sphingomonadaceae, Xanthobacteraceae, Rhizobiaceae, and Bacillaceae). Furthermore, the rhizosphere microbiota of legumes exhibited a high abundance of nitrogen-fixing genes, reflecting a stronger nitrogen-fixing potential, and Streptomycetaceae and Nocardioidaceae were the predominant nitrogen-fixing bacteria. We also identified helper bacteria and confirmed through metadata analysis and a pot experiment that the synthesis of riboflavin by helper bacteria is the key factor in promoting nitrogen fixation. Our study emphasizes that the construction of synthetic communities of nitrogen-fixing bacteria and helper bacteria is crucial for the development of efficient nitrogen-fixing microbial fertilizers.

Nutritive value, fermentation characteristics, and in situ disappearance kinetics of sorghum silage treated with inoculants.

Fibrolytic enzymes and microbial inoculants have the potential to improve the value of sorghum feedstuff and feedstock. An experiment was conducted to determine nutritive value, ensiling characteristics, and in situ disappearance kinetics of 4 sorghum (Sorghum bicolor L.) silage varieties: Dairy Master BMR (DBMR; brown midrib; Richardson Seed, Vega, TX), PS 747 (PS; photoperiod sensitive; Pogue Seed, Kenedy, TX), Silo 700D (S700D; conventional forage type; Richardson Seed), and MMR 381/73 (MMR; conventional forage type; Richardson Seed) pretreated with fibrolytic enzyme (xylanase plus cellulase, XC; 50:50 mixture of Cellulase Plus and Xylanase Plus; Dyadic, Juniper, FL) or microbial [Promote ASB (Lactobacillus buchneri and Lactobacillus plantarum); Cargill Animal Nutrition, Indianapolis, IN; PRO] inoculants. The greatest yield was for cultivar PS and the least for MMR. Neutral detergent fiber (NDF) concentration was least for XC-treated silage, and acid detergent fiber (ADF) concentration was least for XC- and PRO-treated silage. When silage was treated with XC, concentrations of NDF concentrations decreased, on average, 4.81% across all cultivars and ADF concentrations decreased, on average, 3.23% in all cultivars except MMR. Inoculant PRO reduced the NDF concentration of DBMR by 6.47%. The ADF concentrations of DBMR and PS treated with PRO were decreased by 3.25%. Treating sorghum silage with XC or PRO reduced the NDF and ADF fractions, which increased cell wall degradability. In vitro true digestibility was greatest for PRO-treated DBMR, whereas acid detergent lignin was least for PRO-treated DBMR. Aerobic stability was not improved by PRO; however, aerobic stability of XC-treated MMR was 63 h greater than that of the control. Acetate concentrations were greatest for XC-treated MMR, which explains the 63-h improvement in aerobic stability due to the inhibition of fungi. However, inoculant PRO did not improve yeast and mold counts or aerobic stability of sorghum silage compared with the control, which may be due to the lesser acetate concentrations, especially of PRO-treated S700D silage. Generally, in situ disappearance kinetics were improved with the application of XC and PRO, and XC had the greatest effect on silage with greater NDF and ADF concentrations.

Synthesis and application of a compound microbial inoculant for effective soil remediation.

Currently, there is a noticeable scarcity of applications that harness composite microbial inoculants to stimulate straw decomposition, nitrogen fixation, and crop growth. This study addresses this gap by selecting and coculturing three bacterial strains to create a composite microbial inoculant named HY-1. This innovative inoculant exhibits multifunctional capabilities, including nitrogen fixation, straw decomposition, and crop growth promotion. Furthermore, we aimed to explore its impact on soil microbial communities. The results showed that the optimal preparation conditions for the compound microbial inoculant HY-1 were 28.5 ± 0.6 °C, pH = 7.34 ± 0.40, and bacteriophage ratio 1:2:1 (Microbacterium: Streptomyces fasciatus: Bacillus amyloliquefaciens). Compared to single strains, the combination exhibited higher levels of cellulose-degrading and nitrogen-fixing enzyme activity, increased the straw degradation rate by 37.91% within 180 days, and significantly promoted the growth of corn seedlings. Under the condition of straw return, the compound bio-fungicide HY-1 effectively improved the soil microbial diversity. At that time, the soil had the highest number of unique bacterial operational taxonomic units (166), and the abundance of Proteobacteria in the soil increased by 7.24%, while that of Acidobacteriota decreased by 2.27%. The biosynthetic function of the cell wall/membrane/periplasm and the metabolic function of transporting inorganic ions were significantly enhanced. In this study, we discovered that employing coculturing techniques to produce the composite microbial inoculant HY-1 and applying it in the field effectively compensates for the limitations of single-strain inoculants, which often exhibit fewer functions and less pronounced effects. This approach demonstrates significant potential for enhancing the quality of agricultural soils.