Integrated Application of Salicylic Acid and PGPRs to Control Fusarium Wilt of Chickpea.

BACKGROUND: Fusarium wilt and Ascochyta blight are the most important diseases of chickpea. The current study was designed to investigate the individual and combined effect of salicylic acid (SA) with Pseudomonas stutzeri and Pseudomonas putida to suppress Fusarium wilt and promote growth of chickpea varieties: Thal-2006 and Punjab-2008. METHODS: At the time of sowing, inoculum of Fusarium oxysporum was applied to the soil and the incidence of Fusarium wilt was recorded after 60 days. The seeds were inoculated with Pseudomonas stutzeri and Pseudomonas putida prior to sowing. Chickpea plants were treated with salicylic acid at seedling stage. RESULTS: The combination of P. stutzeri and SA significantly increased root length (166% and 145%), shoot height (50% and 47%) and shoot biomass (300% and 233%) in cv. Thal-2006 and cv. Punjab-2008, respectively, in infected plants. Similarly, the combined treatment of P. putida + SA, also enhanced the plant growth parameters of chickpea varieties. Maximum reduction in disease severity was observed in both P. stutzeri + SA (90% and 84%) and P. putida + SA (79% and 77%) treatments in cv. Thal-2006 and Punjab-2008, respectively. Both P. putida + SA and P. stutzeri + SA treatments resulted in increased leaf relative water and total protein content, peroxidase, superoxide dismutase, phenylalanine ammonia-lyase and polyphenol oxidase activities in both resistant (cv. Thal-2006) and susceptible (cv. Punjab-2008) cultivars. Both treatments also significantly reduced malondialdehyde (MDA) and proline content in cv. Thal-2006 and Punjab-2008. Cultivar Thal-2006 was more effective than cv. Punjab-2008. CONCLUSIONS: The results suggested that, in combination, salicylic acid and P. stutzeri may play an important role in controlling Fusarium wilt diseases by inducing systemic resistance in chickpea.

Screening of potential phosphate solubilizing bacteria inoculants should consider the contrast in phosphorus bio-solubilization rate along with plant growth promotion and phosphorus use efficiency.

AIMS: Although phosphate solubilizing bacteria (PSB) have been globally reported to improve soil phosphorus (P) availability and plant growth, technical gaps such as the lack of an ideal screening approach, is yet to be addressed. The potential of non-halo-forming PSB remains underestimated because of the currently adopted screening protocols that exclusively consider halo-forming and PSB with high phosphorus solubilization (PS) capacities. Yet, caution should be taken to properly assess PSB with contrasting PS rates regardless of the presence or absence of the solubilization halo. METHODS AND RESULTS: This study sought to examine the PS rate and plant growth promotion ability of 12 PSB categorized as high PSB (H-PSB), medium PSB (M-PSB), and low PSB (L-PSB) based on their PS rates of rock phosphate (RP). The non-halo-forming PSB Arthrobacter pascens was categorized as H-PSB, which might have been eliminated during the classical screening process. In addition, induction of organic acids and phosphatase activity in rhizosphere soils by H-, M-, and L-PSB was proportional to increased wheat P content by 143.22, 154.21, and 77.76 mg P g-1 compared to uninoculated plants (18.1 mg P g-1). CONCLUSIONS: Isolates considered as M- and L-PSB could positively influence wheat above-ground physiology and root traits as high as H-PSB. In addition, non-halo-forming PSB revealed significant PS rates along with positive effects on plant growth as high as halo-forming PSB.

Mitigation of arsenic toxicity in rice by the co-inoculation of arsenate reducer yeast with multifunctional arsenite oxidizing bacteria.

The study aimed to explicate the role of microbial co-inoculants for the mitigation of arsenic (As) toxicity in rice. Arsenate (AsV) reducer yeast Debaryomyces hansenii NBRI-Sh2.11 (Sh2.11) with bacterial strains of different biotransformation potential was attempted to develop microbial co-inoculants. An experiment to test their efficacy (yeast and bacterial strains) on plant growth and As uptake was conducted under a stressed condition of 20 mg kg-1 of arsenite (AsIII). A combination of Sh2.11 with an As(III)-oxidizer, Citrobacter sp. NBRI-B5.12 (B5.12), resulted in ∼90% decrease in grain As content as compared to Sh2.11 alone (∼40%). Reduced As accumulation in rice roots under co-treated condition was validated with SEM-EDS analysis. Enhanced As expulsion in the selected combination under in vitro conditions was found to be correlated with higher As content in the soil during their interaction with plants. Selected co-inoculant mediated enhanced nutrient uptake in association with better production of indole acetic acid (IAA) and gibberellic acid (GA) in shoot, support microbial co-inoculant mediated better biomass under stressful condition. Boosted defense response in association with enhanced glutathione-S-transferase (GST) and glutathione reductase (GR), activities under in vitro and in vivo conditions were observed. These results indicated that the As(III) oxidizer-B5.12 accelerated the As detoxification property of the As(V) reducer-Sh2.11. Henceforth, the results confer that the coupled reduction-oxidation process of the co-inoculant reduces the accumulation of As in rice grain. These co-inoculants can be further developed for field trials to achieve higher biomass with alleviated As toxicity in rice.

Six complex microbial inoculants for removing ammonia nitrogen from waters.

To determine the effect of microbial inoculants on the removal of ammonia nitrogen (NH4 + -N), six different complex microbial inoculants were studied. In this study, their effectiveness on NH4 + -N removal was compared, and their microbial community composition was determined. High-throughput sequencing results showed that Proteobacteria and Firmicutes were the dominant phyla in six samples. Before the reaction, Bacillus, Cyanobacteria, and Mitochondria genera were the dominant genera. The dominant genera were significantly different after the reaction with the addition of bacterial agents. The six water samples were Massilia, Escherichia-Shigella, Brevibacillus, Mitsuaria, Bacillus, and Ralstonia. Among the six complex microbial inoculants, "Gandu nitrifying bacteria (NR4 )" have the best removal effect on NH4 + -N. In addition, the removal effect of six different bacterial agents on chemical oxygen demand (COD) was compared. The results showed that "Bilaiqing ammonia nitrogen removal bacteria agent (NR5 )" has the best removal effect on COD. Single-factor experiments suggested that the optimal conditions for NR4 bacteria were pH 7, 30°C, 1.0 g/L of bacterial agent dosage and a wide range of NH4 + -N from 30 to 300 mg/L. PRACTITIONER POINTS: The nitrogen removal effects of six different microbial agents were compared. High-throughput sequencing provides important insights into the study of ammonia nitrogen removal by microbial communities. Analysis of six different complex bacterial agents by high-throughput sequencing. The relative abundance of microorganisms is not proportional to the ability to remove NH4 + -N Good application effect in urban landscape water body.

An Insight into Microbial Inoculants for Bioconversion of Waste Biomass into Sustainable "Bio-Organic" Fertilizers: A Bibliometric Analysis and Systematic Literature Review.

The plant-microbe holobiont has garnered considerable attention in recent years, highlighting its importance as an ecological unit. Similarly, manipulation of the microbial entities involved in the rhizospheric microbiome for sustainable agriculture has also been in the limelight, generating several commercial bioformulations to enhance crop yield and pest resistance. These bioformulations were termed biofertilizers, with the consistent existence and evolution of different types. However, an emerging area of interest has recently focused on the application of these microorganisms for waste valorization and the production of "bio-organic" fertilizers as a result. In this study, we performed a bibliometric analysis and systematic review of the literature retrieved from Scopus and Web of Science to determine the type of microbial inoculants used for the bioconversion of waste into "bio-organic" fertilizers. The Bacillus, Acidothiobacillus species, cyanobacterial biomass species, Aspergillus sp. and Trichoderma sp. were identified to be consistently used for the recovery of nutrients and bioconversion of wastes used for the promotion of plant growth. Cyanobacterial strains were used predominantly for wastewater treatment, while Bacillus, Acidothiobacillus, and Aspergillus were used on a wide variety of wastes such as sawdust, agricultural waste, poultry bone meal, crustacean shell waste, food waste, and wastewater treatment plant (WWTP) sewage sludge ash. Several bioconversion strategies were observed such as submerged fermentation, solid-state fermentation, aerobic composting, granulation with microbiological activation, and biodegradation. Diverse groups of microorganisms (bacteria and fungi) with different enzymatic functionalities such as chitinolysis, lignocellulolytic, and proteolysis, in addition to their plant growth promoting properties being explored as a consortium for application as an inoculum waste bioconversion to fertilizers. Combining the efficiency of such functional and compatible microbial species for efficient bioconversion as well as higher plant growth and crop yield is an enticing opportunity for "bio-organic" fertilizer research.

Effects of Two Bacillus Velezensis Microbial Inoculants on the Growth and Rhizosphere Soil Environment of Prunus davidiana.

Microbial inoculants, as harmless, efficient, and environmentally friendly plant growth promoters and soil conditioners, are attracting increasing attention. In this study, the effects of Bacillus velezensis YH-18 and B. velezensis YH-20 on Prunus davidiana growth and rhizosphere soil bacterial community in continuously cropped soil were investigated by inoculation tests. The results showed that in a pot seedling experiment, inoculation with YH-18 and YH-20 resulted in a certain degree of increase in diameter growth, plant height, and leaf area at different time periods of 180 days compared with the control. Moreover, after 30 and 90 days of inoculation, the available nutrients in the soil were effectively improved, which protected the continuously cropped soil from acidification. In addition, high-throughput sequencing showed that inoculation with microbial inoculants effectively slowed the decrease in soil microbial richness and diversity over a one-month period. At the phylum level, Proteobacteria and Bacteroidetes were significantly enriched on the 30th day. At the genus level, Sphingomonas and Pseudomonas were significantly enriched at 15 and 30 days, respectively. These bacterial phyla and genera can effectively improve the soil nutrient utilization rate, antagonize plant pathogenic bacteria, and benefit the growth of plants. Furthermore, inoculation with YH-18 and inoculation with YH-20 resulted in similar changes in the rhizosphere microbiome. This study provides a basis for the short-term effect of microbial inoculants on the P. davidiana rhizosphere microbiome and has application value for promoting the cultivation and production of high-quality fruit trees.

Interactions between Bacterial Inoculants and Native Soil Bacterial Community: the Case of Spore-forming Bacillus spp.

Microbial diversity can restrict the invasion and impact of alien microbes into soils via resource competition. However, this theory has not been tested on various microbial invaders with different ecological traits, particularly spore-forming bacteria. Here we investigated the survival capacity of two introduced spore-forming bacteria, Bacillus mycoides (BM) and B. pumillus (BP) and their impact on the soil microbiome niches with low and high diversity. We hypothesized that higher soil bacterial diversity would better restrict Bacillus survival via resource competition, and the invasion would alter the resident bacterial communities' niches only if inoculants do not escape competition with the soil community (e.g. through sporulation). Our findings showed that BP could not survive as viable propagules and transiently impacted the bacterial communities' niche structure. This may be linked to its poor resource usage and low growth rate. Having better resource use capacities, BM better survived in soil, though its survival was weakly related to the remaining resources left for them by the soil community. BM strongly affected the community niche structure, ultimately in less diverse communities. These findings show that the inverse diversity-invasibility relationship can be valid for some spore-forming bacteria, but only when they have sufficient resource use capacity.

Microbiome of rehydrated corn and sorghum grain silages treated with microbial inoculants in different fermentation periods.

Due to the co-evolved intricate relationships and mutual influence between changes in the microbiome and silage fermentation quality, we explored the effects of Lactobacillus plantarum and Propionibacterium acidipropionici (Inoc1) or Lactobacillus buchneri (Inoc2) inoculants on the diversity and bacterial and fungal community succession of rehydrated corn (CG) and sorghum (SG) grains and their silages using Illumina Miseq sequencing after 0, 3, 7, 21, 90, and 360 days of fermentation. The effects of inoculants on bacterial and fungal succession differed among the grains. Lactobacillus and Weissella species were the main bacteria involved in the fermentation of rehydrated corn and sorghum grain silage. Aspergillus spp. mold was predominant in rehydrated CG fermentation, while the yeast Wickerhamomyces anomalus was the major fungus in rehydrated SG silages. The Inoc1 was more efficient than CTRL and Inoc2 in promoting the sharp growth of Lactobacillus spp. and maintaining the stability of the bacterial community during long periods of storage in both grain silages. However, the bacterial and fungal communities of rehydrated corn and sorghum grain silages did not remain stable after 360 days of storage.

Mixed consortium of microbial inoculants improves yield and essential oil profile of coriander.

Coriander (Coriandrum sativum L.), one of the oldest spice plants globally, has wide usage, mainly owing to its essential oil content. This study investigated the effects of rhizobacteria, mycorrhizae, and their combination on the yield and essential oil profile of coriander. The treatments resulted in statistically higher yield properties and essential oil values than the control. The effects of the microorganism treatments on the yield characteristics were not statistically different. However, the treatments significantly affected the essential oil content and yield. While the arbuscular mycorrhizal fungi and combined application of the microorganisms were statistically equal, arbuscular mycorrhizal fungi had a higher value in essential oil content and yield by 0.75% and 11.8 L ha-1, respectively. The combined application resulted in higher values of linalool (9.47%) and γ-terpinene (6.75%), the components with the highest rate in the essential oil composition. The principal component analysis highlighted the importance of the combined application on the crucial components. In the light of the results, beneficial microorganism treatments were considered significant for yield and essential oil enhancement. Therefore, the wide use of these treatments will highly contribute to coriander cultivation.

Bacillus velezensis strain Ag75 as a new multifunctional agent for biocontrol, phosphate solubilization and growth promotion in maize and soybean crops.

Soybean and maize are some of the main drivers of Brazilian agribusiness. However, biotic and abiotic factors are of great concern, causing huge grain yield and quality losses. Phosphorus (P) deficiency is important among the abiotic factors because most Brazilian soils have a highly P-fixing nature. Thus, large amounts of phosphate fertilizers are regularly applied to overcome the rapid precipitation of P. Searching for alternatives to improve the use of P by crops is essential to reduce the demand for P input. The use of multifunctional rhizobacteria can be considered one of these alternatives. In this sense, the objective of the present work was to select and validate bacterial strains with triple action (plant growth promoter, phosphate solubilizer, and biocontrol agent) in maize and soybean, aiming to develop a multifunctional microbial inoculant for Brazilian agriculture. Bacterial strains with high indole acetic acid (IAA) production, phosphate solubilization, and antifungal activity against soil pathogenic fungi (Rhizoctonia solani, Macrophomina phaseolina, and Fusarium solani) were selected from the maize rhizosphere. Then, they were evaluated as growth promoters in maize under greenhouse conditions. Based on this study, strain 03 (Ag75) was selected due to its high potential for increasing biomass (root and shoot) and shoot P content in maize. This strain was identified through genomic sequencing as Bacillus velezensis. In field experiments, the inoculation of this bacterium increased maize and soybean yields by 17.8 and 26.5%, respectively, compared to the control (25 kg P₂O₅). In addition, the inoculation results did not differ from the control with 84 kg P₂O₅, indicating that it is possible to reduce the application of phosphate in these crops. Thus, the Ag75 strain has great potential for developing a multifunctional microbial inoculant that combines the ability to solubilize phosphate, promote plant growth, and be a biocontrol agent for several phytopathogenic fungi.

Insights into enzyme activity and phosphorus conversion during kitchen waste composting utilizing phosphorus-solubilizing bacterial inoculation.

The main objective of this research was to investigate the effects of Phosphorus-Solubilizing Bacterial (PSB) inoculant on the bacterial structure and phosphorus transformation during kitchen waste composting. High throughput sequencing, topological roles, and multiple analysis methods were conducted to explain the links between phosphorus fractions, enzyme contents, and microbial community structure and function. The findings indicated that bacterial inoculant improved environmental parameters and increased the concentration of total phosphorus, Olsen phosphorus, citric acid phosphorus, OM decomposition, and bacterial diversity. Network analysis concluded that the inoculation treatment was more complex (nodes and edges) and contained more positive links than the control, implying the inoculation effect. The structural equation model also displayed that pH and enzyme activity directly enhanced the phosphorus conversion and bacterial structure. Overall, these results suggest that bacterial inoculation may considerably increase enzyme activity, thus improving biological phosphorus transformation and nutrient content in composting products.

Changes in Composition and Diversity of Epiphytic Microorganisms on Field Pea Seeds, Partial Crop Peas, and Whole Crop Peas during Maturation and Ensiling with or without Lactic Acid Bacteria Inoculant.

The present study was conducted under the hypothesis that, in field peas, type of plant material, stage of maturity, ensiling, silage additive, and aerobic stress affect the composition and diversity of epiphytic microbial communities. Epiphytic microbial composition and diversity of pea seeds, partial crop peas, and whole crop peas was analyzed at different stages of late maturity, before and after ensiling, and with or without the use of lactic acid bacteria (LAB) as inoculant. Suitable combinations among pea crop variants, maturity stages, and inoculant use for the production of stable silages with sufficient aerobic stability after opening and during feed-out were identified. Genomic DNA was extracted, and 16S and 18S rRNA gene amplicons were sequenced. To assess the quality of the various silages, nutrient concentration, pH value, concentration of lactic acid, short chain fatty acids, and alcohols, and aerobic stability were determined. Pea seeds were barely colonized by epiphytic microorganisms. In partial and whole crop peas, composition and α-diversity (Shannon index) of bacterial communities did not differ between crop variants but differed among maturity stages. Epiphytic eukaryotes were rarely found on partial and whole crop peas. Bacterial composition and α-diversity were affected by ensiling and subsequent aerobic storage. In partial and whole crop peas, plant maturation caused an increase of the relative abundance of naturally occurring LAB (Weissella, Pediococcus, and Lactobacillus spp.). As a possible result, natural LAB support stable ensiling conditions even without the use of inoculants beginning with a maturity of 78 on the BBCH scale. This corresponded with a dry matter (DM) concentration of 341 and 363 g/kg in partial and whole crop peas, respectively. Addition of LAB inoculants, however, reduced ammonia, acetic acid, and butanol concentrations, and supported aerobic stability. Earlier stages of plant maturity (BBCH 76 and 77, 300 g DM/kg or less) were more prone to microbial spoilage. Stable pea seed silages can be produced at a maturity between BBCH 78 (427 g DM/kg) and 79 (549 g DM/kg), but they undoubtedly require LAB inoculation or application of other ensiling agents. IMPORTANCE Field peas are important protein suppliers for human and animal nutrition. They can be grown in many areas of the world, which may reduce imports of protein plants and has beneficial economic and ecological effects. Ensiling is a method of preserving feed that can be implemented easily and cost-effectively at the farm. Peas harvested as seeds, partial crop, or whole crop at different maturities enable a wide range of applications. The study characterized epiphytic microbial communities on peas in terms of composition and diversity depending on the maturity of the plants and feed conservation by ensiling as they play an essential role for the production of silages. Even if this study did not consider year, site, or cultivar effects, the results would show which part of the plant is probably well suited for the production of stable and high-quality silages and at which stage of maturity.

Bacterial inoculants as effective agents in minimizing the non-target impact of azadirachtin pesticide and promoting plant growth of Vigna radiata.

Microbes regulate soil health by negating ecological disturbances, and improve plant productivity in a sustainable manner. Indiscriminate application of pesticides creates a detrimental impact on the rhizospheric microbiota, thereby affecting soil health. Azadirachtin, earlier believed to be an environment-friendly alternative to chemical pesticides, exhibits a non-target impact on microbial communities. This study aimed to employ potent bacteria to promote the growth of mungbean plant (Vigna radiata), and mitigate the non-target impact of azadirachtin. Bacterial strains were isolated by enrichment from mungbean rhizosphere. A plant growth experiment was performed with mungbean, amended with azadirachtin to assess the impact of bacterial bioinoculants on the rhizospheric microbiota. The impact of azadirachtin on rhizospheric bacterial community was analyzed qualitatively and quantitatively by 16S rRNA PCR-DGGE and qPCR of various markers, respectively. Residual concentration of azadirachtin in the soil was estimated by HPLC. The bacterial inoculants used in combination significantly promoted plant growth and enhanced the diversity and abundance of total bacterial community in the presence of azadirachtin. Further, the abundance of specific bacterial groups (α-Proteobacteria, ß-Proteobacteria, Actinobacteria, Acidobacteria, and Firmicutes) were significantly boosted. Compared to the control, the isolates significantly facilitated the reduction in residual concentration of azadirachtin in the mungbean rhizosphere. Bacterial inoculants can serve a tripartite role in reducing the stress imparted by botanical pesticides, together with promoting plant growth and enriching the rhizospheric bacterial community structure.

Microbial inoculants reshape structural distribution of complex components of humic acid based on spectroscopy during straw waste composting.

This study aimed to explore the mechanism of actinomycetes inoculation to promote humification based on spectroscopy during straw waste composting. Results showed that inoculating actinomycetes could significantly increase the humification index and humification ratio, which were 2.53% and 21.79% respectively (P < 0.05). A spectroscopic analysis suggested that actinomycetes promoted the relative content of complex components of humic acid and reshaped the structural distribution of two sub fluorescence peaks in it. Furthermore, variance partitioning analysis demonstrated that compared with the intensity, the high-quality uniform distribution of fluorescence peaks had a greater contribution to the improvement of humification. In addition, structural equation model further verified that actinomycetes inoculation promoted the transformation of fulvic acid to humic acid, and then promoted the formation of humic acid. Therefore, actinomycetes inoculation can promote the humification of straw compost by reshaping the complex components of humic acid.

Isolation and Characterization of Phosphate Solubilizing Bacteria from Paddy Field Soils in Japan.

Phosphorus (P) is abundant in soil and is essential for plant growth and development; however, it is easily rendered insoluble in complexes of different types of phosphates, which may lead to P deficiency. Therefore, increases in the amount of P released from phosphate minerals using microbial inoculants is an important aspect of agriculture. The present study used inorganic phosphate solubilizing bacteria (iPSB) in paddy field soils to develop microbial inoculants. Soils planted with rice were collected from different regions of Japan. Soil P was sequentially fractionated using the Hedley method. iPSB were isolated using selective media supplemented with tricalcium phosphate (Ca-P), aluminum phosphate (Al-P), or iron phosphate (Fe-P). Representative isolates were selected based on the P solubilization index and soil sampling site. Identification was performed using 16S rRNA and rpoB gene sequencing. Effectiveness was screened based on rice cultivar Koshihikari growth supplemented with Ca-P, Al-P, or Fe-P as the sole P source. Despite the relatively homogenous soil pH of paddy field sources, three sets of iPSB were isolated, suggesting the influence of fertilizer management and soil types. Most isolates were categorized as ß-Proteobacteria (43%). To the best of our knowledge, this is the first study to describe the genera Pleomorphomonas, Rhodanobacter, and Trinickia as iPSB. Acidovorax sp. JC5, Pseudomonas sp. JC11, Burkholderia sp. JA6 and JA10, Sphingomonas sp. JA11, Mycolicibacterium sp. JF5, and Variovorax sp. JF6 promoted plant growth in rice supplemented with an insoluble P source. The iPSBs obtained may be developed as microbial inoculants for various soil types with different P fixation capacities.

Micromanaging the nitrogen cycle in agroecosystems.

While large inputs of synthetic nitrogen fertilizers enable our current rate of crop production and feed a growing global population, these fertilizers come at a heavy environmental cost. Driven by microbial processes, excess applied nitrogen is lost from agroecosystems as nitrate and nitrous oxide (N2O) contaminating aquatic ecosystems and contributing to climate change. Interest in nitrogen-fixing microorganisms as an alternative to synthetic fertilizers is rapidly accelerating. Microbial inoculants offer the promise of a sustainable and affordable source of nitrogen, but the impact of inoculants on nitrogen dynamics at an ecosystem level is not fully understood. This review synthesizes recent studies on microbial inoculants as tools for nutrient management and considers the ramifications of inoculants for nitrogen transformations beyond fixation.

Cadmium phytoextraction through Brassica juncea L. under different consortia of plant growth-promoting bacteria from different ecological niches.

Combined bioaugmentation inoculants composed of two or more plant growth-promoting bacteria (PGPB) were more effective than single inoculants for plant growth and cadmium (Cd) removal in contaminated soils. However, the principles of consortia construction still need to be discovered. Here, a pot experiment with Cd natural polluted soil was conducted and PGPB consortia with different ecological niches from hyperaccumulator Sedum alfredii Hance were used to compare their effects and mechanisms on plant growth condition, Cd phytoextraction efficiency, soil enzymatic activities, and rhizospheric bacterial community of Brassica juncea L. The results showed that both rhizospheric and endophytic PGPB consortia inoculants promoted plant growth (6.9%-22.1%), facilitated Cd uptake (230.0%-350.0%) of oilseed rape, increased Cd phytoextraction efficiency (343.0%-441.0%), and enhanced soil Cd removal rates (92.0%-144.0%). PGPB consortia inoculants also enhanced soil microbial carbon by 22.2%-50.5%, activated the activities of soil urease and sucrase by 74.7%-158.4% and 8.4%-61.3%, respectively. Simultaneously, PGPB consortia inoculants increased the relative abundance of Flavobacterium, Rhodanobacter, Kosakonia, Pseudomonas and Paraburkholderia at the genus level, which may be beneficial to plant growth promotion and bacterial phytopathogen biocontrol. Although the four PGPB consortia inoculants promoted oilseed growth, amplified Cd phytoextraction, and changed bacterial community structure in rhizosphere soil, their original ecological niches were not a decisive factor for the efficiency of PGPB consortia. therefore, the results enriched the present knowledge regarding the significant roles of PGPB consortia as bioaugmentation agents and preliminarily explored construction principles of effective bioaugmentation inoculants, which will provide insights into the microbial responses to combined inoculation in the Cd-contaminated soils.

Effect of dry matter content on the microbial community and on the effectiveness of a microbial inoculant to improve the aerobic stability of corn silage.

Silage inoculants are commonly used as a tool to improve the fermentation and aerobic stability of corn silage fed to dairy cows. However, their effectiveness can be inconsistent. Our objective was to determine the effect of the dry matter (DM) content of freshly chopped whole-plant corn on its microbial community as affected by an inoculant containing Lentilactobacillus hilgardii, Lentilactobacillus buchneri, and Pediococcus pentosaceus on improving the aerobic stability of silage. Whole-plant corn was harvested at low (31.80%, LDM), medium (33.32%, MDM), or high (39.44%, HDM) DM content and treated with nothing (CTR) or an inoculant (INO) containing L. hilgardii CNCM I-4785 at 150,000 cfu/g fresh forage, L. buchneri NCIMB 40788 at 150,000 cfu/g fresh forage, P. pentosaceus NCIMB12455 at 100,000 cfu/g of fresh forage, ß-glucanase (5,750 IU/g), and xylanase (30,000 IU/g) and ensiled for 20 and 60 d. Data were analyzed as a completely randomized design in a 3-by-2 factorial arrangement of treatments. Fresh LDM forage had a higher concentration of reducing sugars, a less rich, diverse, and even bacterial community, and greater relative abundance of Saccharomycetales than MDM and HDM forages. Silages at 20 and 60 d, inoculated LDM had a more modest proliferation of culturable lactic acid bacteria than inoculated MDM. At 20 d, regardless of treatment, LDM had greater concentrations of lactic and acetic acids. Also at 20 d, LDM had lower numbers of culturable yeasts but greater relative abundance of Enterobacteriaceae than MDM and HDM. For silage at 20 d, HDM silage was more aerobically stable than LDM and MDM and inoculation improved aerobic stability 1.8-fold compared with CTR. For silage at 60 d, there was an interaction between DM content and inoculation. The improvements in stability by inoculation, compared with CTR, were greater in MDM (261 vs. 41 h) and HDM (320 vs. 66 h) silages than in LDM (85 vs. 46 h). The lower DM content and possible slower pH decline in LDM might have facilitated the development of undesirable bacteria and coupled with its greater concentration of reducing sugars and lactic and acetic acids, which are substrates for aerobic microorganisms, might explain the more modest improvements in aerobic stability from inoculation in LDM compared with MDM and HDM. Our findings suggest that the DM content of whole-plant corn affected its epiphytic microbial community and the effectiveness of the inoculant, which improved aerobic stability at all DM but to a greater extent in HDM and MDM than in LDM, especially after 60 d of ensiling.

Integrating 16S rRNA amplicon metagenomics and selective culture for developing thermophilic bacterial inoculants to enhance manure composting.

Composting is an important method for treating and recycling organic waste, and the use of microbial inoculants can increase the efficiency of composting. Herein, we illustrate an approach that integrate 16S rRNA amplicon metagenomics and selective culture of thermophilic bacteria for the development of inoculants to improve manure composting. The 16S rRNA amplicon sequencing analysis revealed that Firmicutes and Actinobacteria were dominant in the composting mixture, and that different microbial hubs succeeded during the thermophilic stage. All isolated thermophilic bacteria were affiliated with the order Bacillales, such as Geobacillus, Bacillus, and Aeribacillus. These isolated thermophilic bacteria were grouped into 11 phylotypes, which shared ＞99% sequence identity to 0.15% to 5.32% of 16S rRNA reads by the amplicon sequencing. Three of these phylotypes transiently enriched during the thermophilic stage. Six thermophilic bacteria were selected from the three phylotypes to obtain seven microbial inoculants. Five out of seven of the microbial inoculants enhanced the thermophilic stage of composting by 16.9% to 52.2%. Three-dimensional excitation emission matrix analysis further revealed that two inoculants (Thermoactinomyces intermedius and Ureibacillus thermophilus) stimulated humification. Additionally, the 16S rRNA amplicon sequencing analysis revealed that inoculation with thermophilic bacteria enhanced the succession of the microbial community during composting. In conclusion, 16S rRNA amplicon metagenomics is a useful tool for the development of microbial inoculants to enhance manure composting.

Meta-analysis of the effects of chemical and microbial preservatives on hay spoilage during storage.

A meta-analysis was performed to evaluate the effects of chemical (50 articles) and microbial (21 articles) additives on hay preservation during storage. Multilevel linear mixed-effects models were fit with response variables calculated as predicted differences (Δ) between treated and untreated samples. Chemical preservatives were classified into five groups such as propionic acid (PropA), buffered organic acids (BOA), other organic acids (OOA), urea, and anhydrous ammonia (AA). Moderators of the models included preservative class (PC), forage type (FT; grass, legumes, and mixed hay), moisture concentration (MC), and application rate (AR). Dry matter (DM) loss during storage was affected by PC × FT (P = 0.045), PC × AR (P < 0.001), and PC × MC (P = 0.009), relative to the overall effect of preservatives (-0.37%). DM loss in PropA-treated hay was numerically reduced to a greater extent in grasses (-16.2), followed by mixed hay (-1.76), but it increased (+2.2%) in legume hay. Increasing AR of PropA resulted in decrease in DM loss (slope = -1.34). Application of BOA, OOA, PropA, and AA decreased visual relative moldiness by -22.1, -29.4, -45.5, and -12.2 percentage points, respectively (PC; P < 0.001). Sugars were higher in treated grass hay (+1.9) and lower in treated legume hay (-0.8% of DM) relative to their untreated counterparts (P < 0.001). The application of all preservatives resulted in higher crude protein (CP) than untreated hay, particularly urea (+7.92) and AA (+5.66% of DM), but PropA, OOA, and BOA also increased CP by 2.37, 2.04, and 0.73 percentage points, respectively. Additionally, preservative application overall resulted in higher in vitro DM digestibility (+1.9% of DM) relative to the untreated hay (x¯=58.3%), which increased with higher AR (slope = 1.64) and decreased with higher MC (slope = -0.27). Microbial inoculants had small effects on hay spoilage because the overall DM loss effect size was -0.21%. Relative to untreated (x¯=4.63% DM), grass hay preserved more sugars (+1.47) than legumes (+0.33) when an inoculant was applied. In conclusion, organic acid-based preservatives prevent spoilage of hay during storage, but their effectiveness is affected by FT, MC, and AR. Microbial inoculants had minor effects on preservation that were impaired by increased MC. Moreover, legume hay was less responsive to the effects of preservatives than grass hay.

Storing hay that has not been properly dried to a moisture concentration (MC) below 20% can lead to the growth of undesirable microbes, such as molds, that decrease the nutritive value of the hay and compromise animal performance and health. Hay preservatives such as propionic acid (PropA), buffered organic acids (BOA), other organic acids (OOA), urea, anhydrous ammonia (AA), and microbial inoculants have been commonly applied to high moisture (>20%) hay to reduce microbial growth during hay storage. The present study compiled the results of 62 published articles on hay preservatives and performed a quantitative analysis (meta-analysis) to determine the effect of preservative treatment on dry matter (DM) loss, moldiness, bale heating, nutritional composition, and DM digestibility, and their interactions with forage type, application rate, and bale moisture. Organic acid-based preservatives (PropA, BOA, and OOA) were effective at reducing DM loss, moldiness, bale maximum temperature, and insoluble nitrogen, and at preserving sugars and DM digestibility during storage to different extents. Microbial inoculants had small effects on the prevention of hay spoilage, and were negatively affected by the increase of hay MC. Legume hay was less responsive to the effects of preservatives than grass hay during storage.