Use of ginger extract and bacterial inoculants for the suppression of Alternaria solani causing early blight disease in Tomato.

Early blight (EB), caused by Alternaria solani, is a serious problem in tomato production. Plant growth-promoting rhizobacteria promote plant growth and inhibit plant disease. The present study explored the bio-efficacy of synergistic effect of rhizobacterial isolates and ginger powder extract (GPE) against tomato EB disease, singly and in combination. Six fungal isolates from symptomatic tomato plants were identified as A. solani on the basis of morphological features i.e., horizontal septation (6.96 to 7.93 µm), vertical septation (1.50 to 2.22 µm), conidia length (174.2 to 187.6 µm), conidial width (14.09 to 16.52 µm), beak length (93.06 to 102.26 µm), and sporulation. Five of the twenty-three bacterial isolates recovered from tomato rhizosphere soil were nonpathogenic to tomato seedlings and were compatible with each other and with GPE. Out of five isolates tested individually, three isolates (St-149D, Hyd-13Z, and Gb-T23) showed maximum inhibition (56.3%, 48.3%, and 42.0% respectively) against mycelial growth of A. solani. Among combinations, St-149D + GPE had the highest mycelial growth inhibition (76.9%) over the untreated control. Bacterial strains molecularly characterized as Pseudomonas putida, Bacillus subtilis, and Bacillus cereus and were further tested in pot trials through seed bacterization for disease control. Seeds treated with bacterial consortia + GPE had the highest disease suppression percentage (78.1%), followed by St-149D + GPE (72.2%) and Hyd-13Z + GPE (67.5%). Maximum seed germination was obtained in the bacterial consortia + GPE (95.0 ± 2.04) followed by St-149D + GPE (92.5 ± 1.44) and Hyd-13Z + GPE (90.0 ± 2.04) over control (73.8 ± 2.39) and chemical control as standard treatment (90.0 ± 2). Ginger powder extracts also induce the activation of defence-related enzymes (TPC, PO, PPO, PAL, and CAT) activity in tomato plants. These were highly significant in the testing bacterial inoculants against A. solani infection in tomato crops.

Competitive fitness and stability of ammonium-excreting Azotobacter vinelandii strains in the soil.

Non-symbiotic N2-fixation would greatly increase the versatility of N-biofertilizers for sustainable agriculture. Genetic modification of diazotrophic bacteria has successfully enhanced NH4+ release. In this study, we compared the competitive fitness of A. vinelandii mutant strains, which allowed us to analyze the burden of NH4+ release under a broad dynamic range. Long-term competition assays under regular culture conditions confirmed a large burden for NH4+ release, exclusion by the wt strain, phenotypic instability, and loss of the ability to release NH4+. In contrast, co-inoculation in mild autoclaved soil showed a much longer co-existence with the wt strain and a stable NH4+ release phenotype. All genetically modified strains increased the N content and changed its chemical speciation in the soil. This study contributes one step forward towards bridging a knowledge gap between molecular biology laboratory research and the incorporation of N from the air into the soil in a molecular species suitable for plant nutrition, a crucial requirement for developing improved bacterial inoculants for economic and environmentally sustainable agriculture. KEY POINTS: • Genetic engineering for NH4+ excretion imposes a fitness burden on the culture medium • Large phenotypic instability for NH4+-excreting bacteria in culture medium • Lower fitness burden and phenotypic instability for NH4+-excreting bacteria in soil.

Microbeads as carriers for Bacillus pumilus: a biofertilizer focus on auxin production.

The study aimed to develop a solid biofertilizer using Bacillus pumilus, focusing on auxin production to enhance plant drought tolerance. Methods involved immobilising B. pumilus in alginate-starch beads, focusing on microbial concentration, biopolymer types, and environmental conditions. The optimal formulation showed a diameter of 3.58 mm ± 0.18, a uniform size distribution after 15 h of drying at 30 °C, a stable bacterial concentration (1.99 × 109 CFU g-1 ± 1.03 × 109 over 180 days at room temperature), a high auxin production (748.8 µg g-1 ± 10.3 of IAA in 7 days), and a water retention capacity of 37% ± 4.07. In conclusion, this new formulation of alginate + starch + L-tryptophan + B. pumilus has the potential for use in crops due to its compelling water retention, high viability in storage at room temperature, and high auxin production, which provides commercial advantages.

Improvement of straw decomposition and rice growth through co-application of straw-decomposing inoculants and ammonium nitrogen fertilizer.

BACKGROUND: The growth of rice is reduced by the slow decomposition of accumulated straw, which competes with rice for soil nitrogen nutrient. In recent year, straw-decomposing inoculants (SDIs) that can accelerate straw decomposition and ammonium nitrogen (N) fertilizer that can quickly generate available N is increasingly adopted in China. However, it is still unknown whether the N demand of straw decomposition and crop growth can be simultaneously met through the co-application of SDIs and ammonium N fertilizer. RESULTS: In this study, we investigated the effect of the co-application of SDIs and ammonium bicarbonate on decomposition rate of wheat straw, rice growth and rice yield over two consecutive years in rice-wheat rotation system. Compound fertilizer (A0) was used as control. The ratios of ammonium bicarbonate addition were 20% (A2), 30% (A3) and 40% (A4), respectively, without SDIs or with SDIs (IA2, IA3, IA4). Our results revealed that without SDIs, compared with A0, straw decomposition rate, rice growth and yield were improved under A2; However, under A3, rice yield was decreased due to the slow decomposition rate of straw and limited growth of rice during late growth stage. Combining SDIs and N fertilizer increased straw decomposition rate, rice growth rate and yield more than that of N fertilizer alone, especially under IA3. Compared with A0, straw decomposition rate, tiller number, aboveground biomass, leaf area index, root length, and nitrogen use efficiency were significantly increased by 16%, 8%, 27%, 12%, 17%, and 15% under IA3. Consequently, the average rice yield of IA3 was increased to 10,856 kg/ha, which was 13% and 9% higher, respectively, than of A0 and A2. CONCLUSION: Our results indicated that ammonium bicarbonate application alone carried a risk of nutrient deficiency during late growth stage and yield decline. Therefore, the co-application of SDIs and 30% ammonium N fertilizer substitution can be a favorable practice to simultaneously accelerate straw decomposition and increase rice crop growth.

Harnessing Native Bacillus spp. for Sustainable Wheat Production.

The genus Bacillus has been widely applied in contemporary agriculture as an environmentally-friendly biological agent. However, the real effect of commercial Bacillus-based fertilizers and pesticides varies immensely in the field. To harness Bacillus for efficient wheat production, we reviewed the diversity, functionality, and applicability of wheat-associated native Bacillus for the first time. Our main findings are: (i) Bacillus spp. inhabit the rhizosphere, root, stem, leaf, and kernel of wheat; (ii) B. subtilis and B. velezensis are the most widely endophytic species that can be isolated from both below and aboveground tissues; (iii) major functions of these representative strains are promotion of plant growth and alleviation of both abiotic and biotic stresses in wheat; (iv) stability and effectiveness are 2 major challenges during field application; (v) a STVAE pipeline that includes 5 processes, namely, Screen, Test, Validation, Application, and Evaluation, has been proposed for the capture and refinement of wheat-associated Bacillus spp. In particular, this review comprehensively addresses possible solutions, concerns, and criteria during the development of native Bacillus-based inoculants for sustainable wheat production.

Effects of sublethal stress application on the survival of bacterial inoculants: a systematic review.

The use of commercial bacterial inoculants formulated with plant-growth promoting bacteria (PGPB) in agriculture has shown significant prominence in recent years due to growth-promotion benefits provided to plants through different mechanisms. However, the survival and viability of bacterial cells in inoculants are affected during use and may decrease their effectiveness. Physiological adaptation strategies have attracted attention to solve the viability problem. This review aims to provide an overview of research on selecting sublethal stress strategies to increase the effectiveness of bacterial inoculants. The searches were performed in November 2021 using Web of Science, Scopus, PubMed, and Proquest databases. The keywords "nitrogen-fixing bacteria", "plant growth-promoting rhizobacteria", "azospirillum", "pseudomonas", "rhizobium", "stress pre-conditioning", "adaptation", "metabolic physiological adaptation", "cellular adaptation", "increasing survival", "protective agent" and "protective strategy" were used in the searches. A total of 2573 publications were found, and 34 studies were selected for a deeper study of the subject. Based on the studies analysis, gaps and potential applications related to sublethal stress were identified. The most used strategies included osmotic, thermal, oxidative, and nutritional stress, and the primary cell response mechanism to stress was the accumulation of osmolytes, phytohormones, and exopolysaccharides (EPS). Under sublethal stress, the inoculant survival showed positive increments after lyophilization, desiccation, and long-term storage processes. The effectiveness of inoculant-plants interaction also had positive increments after sublethal stress, improving plant development, disease control, and tolerance to environmental stresses compared to unappealed inoculants.

Dry matter content and inoculant alter the metabolome and bacterial community of alfalfa ensiled at high temperature.

Alfalfa silage fermentation quality, metabolome, bacterial interactions, and successions as well as their predicted metabolic pathways were explored under different dry matter contents (DM) and lactic acid bacteria (LAB) inoculations. Silages were prepared from alfalfa with DM contents of 304 (LDM) and 433 (HDM) g/kg fresh weight and inoculated with Lactiplantibacillus plantarum (L. plantarum, LP), Pediococcus pentosaceus (P. pentosaceus, PP), or sterile water (control). The silages were stored at a simulated hot climate condition (35°C) and sampled at 0, 7, 14, 30, and 60 days of fermentation. The results revealed that HDM significantly improved the alfalfa silage quality and altered microbial community composition. The GC-TOF-MS analysis discovered 200 metabolites in both LDM and HDM alfalfa silage, mainly consisting of amino acids, carbohydrates, fatty acids, and alcohols. Compared with LP and control, PP-inoculated silages had increased concentrations of lactic acid (P < 0.05) and essential amino acids (threonine and tryptophan) as well as decreased pH, putrescine content, and amino acid metabolism. However, alfalfa silage inoculated with LP had higher proteolytic activities than control and PP-inoculated silage, as revealed by a higher concentration of ammonia nitrogen (NH3-N), and also upregulated amino acid and energy metabolism. HDM content and P. pentosaceus inoculation significantly altered the composition of alfalfa silage microbiota from 7 to 60 days of ensiling. Conclusively, these results indicated that inoculation with PP exhibited great potential in enhancing the fermentation of silage with LDM and HDM via altering the microbiome and metabolome of the ensiled alfalfa, which could help in understanding and improving the ensiling practices under hot climate conditions. KEY POINTS: • HDM improved fermentation quality and declined putrescine content of alfalfa silage • P. pentosaceus inoculation enhanced the fermentation quality of alfalfa silage • P. pentosaceus is an ideal inoculant for alfalfa silage under high temperature.

Rice Big Grain1 enhances biomass and plant growth-promoting traits in rhizospheric yeast Candida tropicalis.

The Big Grain1 (BG1) gene of rice (Oryza sativa L.) is reported to increase the yield of rice crops; however, its molecular mechanism is largely concealed. To explore its functional prospects, we have taken a structure-function-based approach. In silico analyses suggest OsBG1 is a DNA- and phytohormone-binding protein. Heterologous expression of OsBG1 with galactose-inducible promoter GAL1p in the rhizospheric yeast Candida tropicalis SY005 revealed 7.9- and 1.5-fold higher expression of the gene at 12 and 24 h, respectively, compared to the expression at 36 h post-galactose induction. Functional activity of the induced OsBG1 in engineered yeast increased cell density, specific growth rate, and biomass by 28.5%, 29.8%, and 14.1%, respectively, and decreased the generation time by 21.25%. Flow cytometry-based cell cycle analysis of OsBG1-expressing yeast cells exhibited an increase in the cells of the G2/M population by 15.8% after 12 h of post-galactose induction. The gene expression study of yeast transformants disclosed that OsBG1 regulates cell division by upregulating the expression of the endogenous gene cyclin B1 (CtCYB1) by 1.3- and 1.9-folds at 10 and 12 h, respectively, compared to the control, and is positively influenced by the phytohormone indole acetic acid (IAA). Further, the study revealed that OsBG1 significantly increases biofilm formation, stress tolerance, and IAA production in C. tropicalis SY005, implying its prospective role in enhancing plant growth-promoting traits in microbes. OsBG1-expressing rhizospheric yeast cells significantly improved the germination and growth parameters of the bio-inoculated rice seeds. Altogether, this study suggests OsBG1 can be employed to genetically improve suitable bio-inoculants for their plant growth-promoting traits to augment crop productivity. KEY POINTS: • In silico analyses suggested OsBG1 is a phytohormone-binding transcription factor. • OsBG1 enhanced growth in rhizospheric Candida tropicalis by upregulating CtCYB1. • OsBG1 improved plant growth-promoting traits of the rhizospheric yeast C. tropicalis.

The Use of Synthetic Microbial Communities to Improve Plant Health.

Despite the numerous benefits plants receive from probiotics, maintaining consistent results across applications is still a challenge. Cultivation-independent methods associated with reduced sequencing costs have considerably improved the overall understanding of microbial ecology in the plant environment. As a result, now, it is possible to engineer a consortium of microbes aiming for improved plant health. Such synthetic microbial communities (SynComs) contain carefully chosen microbial species to produce the desired microbiome function. Microbial biofilm formation, production of secondary metabolites, and ability to induce plant resistance are some of the microbial traits to consider when designing SynComs. Plant-associated microbial communities are not assembled randomly. Ecological theories suggest that these communities have a defined phylogenetic organization structured by general community assembly rules. Using machine learning, we can study these rules and target microbial functions that generate desired plant phenotypes. Well-structured assemblages are more likely to lead to a stable SynCom that thrives under environmental stressors as compared with the classical selection of single microbial activities or taxonomy. However, ensuring microbial colonization and long-term plant phenotype stability is still one of the challenges to overcome with SynComs, as the synthetic community may change over time with microbial horizontal gene transfer and retained mutations. Here, we explored the advances made in SynCom research regarding plant health, focusing on bacteria, as they are the most dominant microbial form compared with other members of the microbiome and the most commonly found in SynCom studies.

Characterization of rhizobia for beneficial traits that promote nodulation in legumes under abiotically stressed conditions.

The growing interest in using rhizobia as inoculants in sustainable agricultural systems has prompted the screening of rhizobia species for beneficial traits that enhance nodulation and nitrogen fixation under abiotic stressed conditions. This study reports phenotypic and phylogenetic characterization of rhizobia strains previously isolated from the root nodules of several indigenous and exotic legumes growing in South Africa and other countries. The Rhizobia strains were screened for their ability to tolerate various abiotic stresses (temperature 16, 28, and 36 °C; acidity/alkalinity pH 5, 7, and 9; heavy metals 50, 100, and 150 mM AlCl3.6H2O; and salinity 50, 100, and 150 mM NaCl). Phylogenetic characterization of the isolates was determined using multilocus sequence analysis of the 16S rRNA, recA, acdS, exoR, nodA, and nodC genes. The analysis indicated that the isolates are phylogenetically related to Sinorhizobium, Bradyrhizobium, Rhizobium, Mesorhizobium, and Aminobacter genera and exhibited significant variations in their tolerance to abiotic stresses. Amid the increasing threats of the global stresses, these current results provide baseline information in the selection of rhizobia for use as inoculants under extreme temperatures, acidity/alkalinity, and salinity stress conditions in South Africa.

Do commercial arbuscular mycorrhizal inoculants contain the species that they claim?

The use of arbuscular mycorrhizal (AM) fungal inoculants as a means to promote plant growth is gaining momentum worldwide. Although there is an increasing number of commercial products available for various applications, the quality of these remains uncertain. We determined the AM fungal species composition in eleven inoculants from four producers by using DNA metabarcoding and compared them to the AM fungal species declared on the product labels. Our DNA metabarcoding of the inoculants revealed a concerning discrepancy between the declared and detected AM fungal species compositions of the products. While nine products contained at least one declared species, two did not contain any matching species and all inoculants but one contained additional species not declared on the product label. These findings highlight the need for better guidelines and industry standards to ensure consumer protection in the AM fungal inoculum market. Additionally, we call for caution when using commercial AM fungal inoculants in scientific experiments without confirmatory information about their species composition.

SCAR marker: A potential tool for authentication of agriculturally important microorganisms.

Microbial inoculants are globally recommended for plant growth promotion and control of plant pathogens. These inoculants require stringent quality checks for sustainable field efficacy. Questionable regulatory frameworks constantly deteriorate the reliability of bio-inoculant technology. Existing global regulations do not involve any rapid molecular technique for the routine inspection of microbial preparations. Sequence characterized amplified region (SCAR) marker offers rapid and precise strain-level authentication of target microbes. Such advanced molecular techniques must be exploited to accurately validate the microbial formulations. Besides, the global dissemination of plant pathogenic microbes has always been an alarming threat to food security. SCAR markers could be used at the plant quarantine centers to rapidly detect catastrophic pathogens, thereby circumventing the import and export of contagious plant materials. The current review is focused on promoting the SCAR marker technology to validate commercial bio-inoculants and predict plant pandemics.

Mechanism of sulfur-oxidizing inoculants and nitrate on regulating sulfur functional genes and bacterial community at the thermophilic compost stage.

The emission of H2S odors predominantly occurred at the thermophilic phase of composting, which could cause odorous gas pollution and reduce the fertilizer value of composting products. And sulfur-oxidizing bacteria (SOB) possess oxidative capacities for inorganic sulfur compounds with nitrate applied as electron acceptors. Therefore, this study aimed to assess the effectiveness of combined additives (SOB inoculants and nitrate) on the bacterial community diversity, sulfur-oxidizing gene abundances, and metabolic function prediction at the thermophilic stage of sewage sludge composting. The highest sulfate contents were increased by 1.02-1.34 folds, and the abundances of the sulfur-oxidizing genes (sqr, pdo, sox, and sor) were also enhanced by adding the combined additives. Network patterns revealed a strengthened interaction of inoculants and sulfur functional genes. Microbial functional pathways predicted higher metabolic levels of carbohydrate and amino acid metabolisms with the addition of combined additives, and the predicted relative abundances of sulfur metabolism and nitrogen metabolism were increased by 19.3 ± 2.5% and 24.7 ± 4.1%, respectively. Heatmap analysis showed that the SOB might have a competitive advantage over the indigenous denitrifying bacteria in using nitrate for biochemical reactions. Correlation analyses suggested that sulfur-oxidizing efficacy could be indirectly affected by the environmental parameters through changing the structure of bacterial community. These findings provide new insights toward an optimized inoculation strategy of using SOB and nitrate to enhance sulfur preservation and modulate the bacterial communities at the thermophilic phase of sewage sludge composting.

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.

Co-inoculation with tropical strains of Azospirillum and Bacillus is more efficient than single inoculation for improving plant growth and nutrient uptake in maize.

Usage of Bacillus and Azospirillum as new eco-friendly microbial consortium inoculants is a promising strategy to increase plant growth and crop yield by improving nutrient availability in agricultural sustainable systems. In this study, we designed a multispecies inoculum containing B. thuringiensis (strain B116), B. subtillis (strain B2084) and Azospirillum sp. (strains A1626 and A2142) to investigate their individual or co-inoculated ability to solubilize and mineralize phosphate, produce indole acetic acid (IAA) and their effect on maize growth promotion in hydroponics and in a non-sterile soil. All strains showed significant IAA production, P mineralization (sodium phytate) and Ca-P, Fe-P (tricalcium phosphate and iron phosphate, respectively) solubilization. In hydroponics, co-inoculation with A1626 x A2142, B2084 x A2142, B2084 x A1626 resulted in higher root total length, total surface area, and surface area of roots with diameter between 0 and 1 mm than other treatments with single inoculant, except B2084. In a greenhouse experiment, maize inoculated with the two Azospirillum strains exhibited enhanced shoot dry weight, shoot P and K content, root dry weight, root N and K content and acid and alkaline phosphatase activities than the other treatments. There was a significant correlation between soil P and P shoot, alkaline phosphatase and P shoot and between acid phosphatase and root dry weight. It may be concluded that co-inoculations are most effective than single inoculants strains, mainly between two selected Azospirillum strains. Thus, they could have synergistic interactions during maize growth, and be useful in the formulation of new inoculants to improve the tropical cropping systems sustainability.

Whole-Genome Resequencing of Spontaneous Oxidative Stress-Resistant Mutants Reveals an Antioxidant System of Bradyrhizobium japonicum Involved in Soybean Colonization.

Soybean is the most inoculant-consuming crop in the world, carrying strains belonging to the extremely related species Bradyrhizobium japonicum and Bradyrhizobium diazoefficiens. Currently, it is well known that B. japonicum has higher efficiency of soybean colonization than B. diazoefficiens, but the molecular mechanism underlying this differential symbiotic performance remains unclear. In the present study, genome resequencing of four spontaneous oxidative stress-resistant mutants derived from the commercial strain B. japonicum E109 combined with molecular and physiological studies allowed identifying an antioxidant cluster (BjAC) containing a transcriptional regulator (glxA) that controls the expression of a catalase (catA) and a phosphohydrolase (yfbR) related to the hydrolysis of hydrogen peroxide and oxidized nucleotides, respectively. Integrated synteny and phylogenetic analyses supported the fact that BjAC emergence in the B. japonicum lineage occurred after its divergence from the B. diazoefficiens lineage. The transformation of the model bacterium B. diazoefficiens USDA110 with BjAC from E109 significantly increased its ability to colonize soybean roots, experimentally recapitulating the beneficial effects of the occurrence of BjAC in B. japonicum. In addition, the glxA mutation significantly increased the nodulation competitiveness and plant growth-promoting efficiency of E109. Finally, the potential applications of these types of non-genetically modified mutant microbes in soybean production worldwide are discussed.

Does Organomineral Fertilizer Combined with Phosphate-Solubilizing Bacteria in Sugarcane Modulate Soil Microbial Community and Functions?

Soil bacterial and fungal communities are suitable soil ecosystem health indicators due to their sensitivity to management practices and their role in soil ecosystem processes. Here, information on composition and functions of bacterial and fungal communities were evaluated at two phenological stages of sugarcane (six and twelve months, equivalent to the most intensive vegetative stage and to final maturation, respectively) when organomineral fertilizer, combined with phosphate-solubilizing bacteria (PSB), was added into the soil. Organic compost enriched with apatite (C + A) or phosphorite (C + P) and compost without phosphate enrichment (C) were used in the presence or absence of PSB. In addition, we used a control fertilized with soluble triple superphosphate. The differences were more related to the sampling period than to the type of organomineral fertilizer, being observed higher available phosphorus at six months than at twelve months. Only in the C treatment we observed the presence of Bacillaceae and Planococcaceae, while Pseudomonadaceae were only prevalent in inoculated C + A. As for fungi, the genera Chaetomium and Achroiostachys were only present in inoculated C + P, while the genus Naganishia was most evident in inoculated C + A and in uninoculated C + P. Soliccocozyma represented 75% of the total fungal abundance in uninoculated C while in inoculated C, it represented 45%. The bacterial community was more related to the degradation of easily decomposable organic compounds, while the fungal community was more related to degradation of complex organic compounds. Although the microbial community showed a resilient trait, subtle changes were detected in microbial community composition and function, and this may be related to the increase in yield observed.

Metaprofiling of the bacterial community in sorghum silages inoculated with lactic acid bacteria.

AIMS: To characterize the fermentation process and bacterial diversity of sorghum silage inoculated with Lactiplantibacillus plantarum LpAv, Pediococcus pentosaceus PpM and Lacticaseibacillus paracasei LcAv. METHODS AND RESULTS: Chopped sorghum was ensiled using the selected strains. Physicochemical parameters (Ammonia Nitrogen/Total Nitrogen, Dry Matter, Crude Protein, Acid Detergent Fibre, Neutral Detergent Fibre, Acid Detergent Lignin, Ether Extract and Ashes), bacterial counts, cell cytometry and 16sRNA sequencing were performed to characterize the ensiling process and an animal trial (BALB/c mice) was conducted in order to preliminary explore the potential of sorghum silage to promote animal gut health. After 30 days of ensiling, the genus Lactobacillus comprised 68.4 ± 2.3% and 73.5 ± 1.8% of relative abundance, in control and inoculated silages respectively. Richness (Chao1 index) in inoculated samples, but not in control silages, diminished along ensiling, suggesting the domination of fermentation by the inoculated LAB. A trend in conferring enhanced protection against Salmonella infection was observed in the mouse model used to explore the potential to promote gut health of sorghum silage. CONCLUSIONS: The LAB strains used in this study were able to dominate sorghum fermentation. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report using metaprofiling of 16sRNA to characterize sorghum silage, showing a microbiological insight where resident and inoculated LAB strains overwhelmed the epiphytic microbiota, inhibiting potential pathogens of the genus Klebsiella.

The introduced strain Mesorhizobium ciceri USDA 3378 is more competitive than an indigenous strain in nodulation of chickpea in newly introduced areas of China.

The present study aimed to compare the competitive advantage of two chickpea nodulating rhizobia strains (an indigenous strain Mesorhizobium muleiense CCBAU 83963T and an introduced strain Mesorhizobium ciceri USDA 3378) in different soils originated from new chickpea cultivation areas of China. The results showed that USDA 3378 had a significant competitive advantage in nodulation, with nodulation occupation rates ranging from 84·6% to 100% in all the sampled soils. According to the efficiency of symbiosis under single inoculation, chickpea plants inoculated with USDA 3378 showed better symbiotic performance based on the plant dry weight, leaf chlorophyll content and nodule numbers. The chickpea plants inoculated with USDA 3378 formed nodules about 2 days earlier than those inoculated with CCBAU 83963T . The higher growth in media and the stronger adsorption on chickpea roots of USDA 3378 when mixed with CCBAU 83963T may explain why USDA 3378 shows a competitive advantage. The results from this study will contribute towards the development of effective chickpea rhizobial inoculants for soil conditioning and more environmentally friendly production of chickpeas in China.

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.