Genomic blueprints of soybean (Glycine max) pathogen resistance: revealing the key genes for sustainable agriculture.

Soybean (Glycine max ) is an important oilseed, protein and biodiesel crop. It faces significant threats from bacterial, fungal and viral pathogens, which cause economic losses and jeopardises global food security. In this article, we explore the relationship between soybeans and these pathogens, focusing on the molecular responses that are crucial for soybeans defence mechanisms. Molecular responses involve small RNAs and specific genes, including resistance (R) genes that are pivotal in triggering immune responses. Functional genomics, which makes use of cutting-edge technologies, such as CRISPR Cas9 gene editing, allows us to identify genes that provide insights into the defence mechanisms of soybeans with the focus on using genomics to understand the mechanisms involved in host pathogen interactions and ultimately improve the resilience of soybeans. Genes like GmKR3 and GmVQ58 have demonstrated resistance against soybean mosaic virus and common cutworm, respectively. Genetic studies have identified quantitative trait loci (QTLs) including those linked with soybean cyst nematode, root-knot nematode and Phytophthora root and stem rot resistance. Additionally, resistance against Asian soybean rust and soybean cyst nematode involves specific genes and their variations in terms of different copy numbers. To address the challenges posed by evolving pathogens and meet the demands of a growing population, accelerated soybean breeding efforts leveraging functional genomics are imperative. Targeted breeding strategies based on a deeper understanding of soybean gene function and regulation will enhance disease resistance, ensuring sustainable agriculture and global food security. Collaborative research and continued technological advancements are crucial for securing a resilient and productive agricultural future.

Graphitic carbon nitride alleviates cadmium toxicity to microbial communities in soybean rhizosphere.

Cadmium (Cd) contamination has led to various harmful impacts on soil microbial ecosystem, agricultural crops, and thus human health. Nanomaterials are promising candidates for reducing the accumulation of heavy metals in plants. In this study, graphitic carbon nitride (g-C3N4), a two-dimensional polymeric nanomaterial, was applied for ameliorating Cd phytotoxicity to soybean (Glycine max (L.) Merr.). Its impacts on rhizosphere variables, microorganisms, and metabolism were examined. It was found that g-C3N4 increased carbon/nitrogen/phosphorus (C/N/P) content, especially when N contents were averagely 4.2 times higher in the g-C3N4-treated groups. g-C3N4 significantly induced alterations in microbial community structures (P < 0.05). The abundance of the probiotics class Nitrososphaeria was enriched (on average 70% higher in the g-C3N4-treated groups) as was Actinobacteria (226% higher in the g-C3N4 group than in the CK group). At the genus level, g-C3N4 recruited more Bradyrhizobium (122% higher) in the Cd + g-C3N4 group than in the Cd group and more Sphingomonas (on average 24% higher) in the g-C3N4-treated groups. The changes of microbial clusters demonstrated the potential of g-C3N4 to shape microbial functions, promote plant growth, and enhance Cd resistance, despite observing less pronounced modifications in microbial communities in Cd-contaminated soil compared to Cd-free soil. Moreover, abundance of functional genes related to C/N/P transformation was more significantly promoted by g-C3N4 in Cd-contaminated soil (increased by 146%) than in Cd-free one (increased by 32.8%). Therefore, g-C3N4 facilitated enhanced microbial survival and adaptation through the amplification of functional genes. These results validated the alleviation of g-C3N4 on the microbial communities in the soybean rhizosphere and shed a new light on the application of environmental-friendly nanomaterials for secure production of the crop under soil Cd exposure.

Transgenic soybean of GsMYB10 shapes rhizosphere microbes to promote resistance to aluminum (Al) toxicity.

Plant resistance genes could affect rhizosphere microbiota, which in turn enhanced plant resistance to stresses. Our previous study found that overexpression of the GsMYB10 gene led to enhanced tolerance of soybean plants to aluminum (Al) toxicity. However, whether GsMYB10 gene could regulate rhizosphere microbiota to mitigate Al toxicity remains unclear. Here, we analyzed the rhizosphere microbiomes of HC6 soybean (WT) and transgenic soybean (trans-GsMYB10) at three Al concentrations, and constructed three different synthetic microbial communities (SynComs), including bacterial, fungal and cross-kingdom (bacteria and fungi) SynComs to verify their role in improving Al tolerance of soybean. Trans-GsMYB10 shaped the rhizosphere microbial communities and harbored some beneficial microbes, such as Bacillus, Aspergillus and Talaromyces under Al toxicity. Fungal and cross-kingdom SynComs showed a more effective role than the bacterial one in resistance to Al stress, and these SynComs helped soybean resist Al toxicity via affecting some functional genes that involved cell wall biosynthesis and organic acid transport etc. Overall, this study reveals the mechanism of soybean functional genes regulating the synergistic resistance of rhizosphere microbiota and plants to Al toxicity, and also highlights the possibility of focusing on the rhizobial microbial community as a potential molecular breeding target to produce crops.

Means, motive and opportunity for biological invasions: Genetic introgression in a fungal pathogen.

Invasions by fungal plant pathogens pose a significant threat to the health of agricultural ecosystems. Despite limited standing genetic variation, many invasive fungal species can adapt and spread rapidly, resulting in significant losses to crop yields. Here, we report on the population genomics of Colletotrichum truncatum, a polyphagous pathogen that can infect more than 460 plant species, and an invasive pathogen of soybean in Brazil. We study the whole-genome sequences of 18 isolates representing 10 fields from two major regions of soybean production. We show that Brazilian C. truncatum is subdivided into three phylogenetically distinct lineages that exchange genetic variation through hybridization. Introgression affects 2%-30% of the nucleotides of genomes and varies widely between the lineages. We find that introgressed regions comprise secreted protein-encoding genes, suggesting possible co-evolutionary targets for selection in those regions. We highlight the inherent vulnerability of genetically uniform crops in the agro-ecological environment, particularly when faced with pathogens that can take full advantage of the opportunities offered by an increasingly globalized world. Finally, we discuss "the means, motive and opportunity" of fungal pathogens and how they can become invasive species of crops. We call for more population genomic studies because such analyses can help identify geographical areas and pathogens that pose a risk, thereby helping to inform control strategies to better protect crops in the future.

Dynamic changes of rhizosphere soil bacterial community and nutrients in cadmium polluted soils with soybean-corn intercropping.

BACKGROUND: Soybean-corn intercropping is widely practised by farmers in Southwest China. Although rhizosphere microorganisms are important in nutrient cycling processes, the differences in rhizosphere microbial communities between intercropped soybean and corn and their monoculture are poorly known. Additionally, the effects of cadmium (Cd) pollution on these differences have not been examined. Therefore, a field experiment was conducted in Cd-polluted soil to determine the effects of monocultures and soybean-corn intercropping systems on Cd concentrations in plants, on rhizosphere bacterial communities, soil nutrients and Cd availability. Plants and soils were examined five times in the growing season, and Illumina sequencing of 16S rRNA genes was used to analyze the rhizosphere bacterial communities. RESULTS: Intercropping did not alter Cd concentrations in corn and soybean, but changed soil available Cd (ACd) concentrations and caused different effects in the rhizosphere soils of the two crop species. However, there was little difference in bacterial community diversity for the same crop species under the two planting modes. Proteobacteria, Chloroflexi, Acidobacteria, Actinobacteria and Firmicutes were the dominant phyla in the soybean and corn rhizospheres. In ecological networks of bacterial communities, intercropping soybean (IS) had more module hubs and connectors, whereas intercropped corn (IC) had fewer module hubs and connectors than those of corresponding monoculture crops. Soil organic matter (SOM) was the key factor affecting soybean rhizosphere bacterial communities, whereas available nutrients (N, P, K) were the key factors affecting those in corn rhizosphere. During the cropping season, the concentration of soil available phosphorus (AP) in the intercropped soybean-corn was significantly higher than that in corresponding monocultures. In addition, the soil available potassium (AK) concentration was higher in intercropped soybean than that in monocropped soybean. CONCLUSIONS: Intercropped soybean-corn lead to an increase in the AP concentration during the growing season, and although crop absorption of Cd was not affected in the Cd-contaminated soil, soil ACd concentration was affected. Intercropped soybean-corn also affected the soil physicochemical properties and rhizosphere bacterial community structure. Thus, intercropped soybean-corn was a key factor in determining changes in microbial community composition and networks. These results provide a basic ecological framework for soil microbial function in Cd-contaminated soil.

Regulation of the Emissions of the Greenhouse Gas Nitrous Oxide by the Soybean Endosymbiont Bradyrhizobium diazoefficiens.

The greenhouse gas nitrous oxide (N2O) has strong potential to drive climate change. Soils are a major source of N2O, with microbial nitrification and denitrification being the primary processes involved in such emissions. The soybean endosymbiont Bradyrhizobium diazoefficiens is a model microorganism to study denitrification, a process that depends on a set of reductases, encoded by the napEDABC, nirK, norCBQD, and nosRZDYFLX genes, which sequentially reduce nitrate (NO3-) to nitrite (NO2-), nitric oxide (NO), N2O, and dinitrogen (N2). In this bacterium, the regulatory network and environmental cues governing the expression of denitrification genes rely on the FixK2 and NnrR transcriptional regulators. To understand the role of FixK2 and NnrR proteins in N2O turnover, we monitored real-time kinetics of NO3-, NO2-, NO, N2O, N2, and oxygen (O2) in a fixK2 and nnrR mutant using a robotized incubation system. We confirmed that FixK2 and NnrR are regulatory determinants essential for NO3- respiration and N2O reduction. Furthermore, we demonstrated that N2O reduction by B. diazoefficiens is independent of canonical inducers of denitrification, such as the nitrogen oxide NO3-, and it is negatively affected by acidic and alkaline conditions. These findings advance the understanding of how specific environmental conditions and two single regulators modulate N2O turnover in B. diazoefficiens.

Alternative products to control late season diseases in soybeans / Produtos alternativos para o controle de doenças de final de ciclo na soja

In the last crop seasons, the complex of late season diseases (CLSD) of soybean (Glycine max L. (Merrill)), has been causing considerable reductions in the crop yield. Currently, there are no cultivars resistant to all pathogens that causes CLSD. The present study evaluated the effect of applying the acibenzolar-S-methyl resistance inducer, alternative products and fungicide on the severity of CLSD in the soybean cultivar BMX Potência RR during the 2013/2014 and 2014/2015 crops, in the field. The treatments for the experiments were: 1 - control (water); 2 - acibenzolar-S-methyl; 3 - calcium; 4 - micronutrients: copper, manganese and zinc; 5 - micronutrients: manganese, zinc and molybdenum; 6 - nitrogen-potassium fertilizer; 7 - Ascophyllum nodosum and 8 - azoxystrobin + cyproconazole with the addition of the adjuvant. Four applications of alternative products and two of fungicide were carried out in both harvests. A diagrammatic scale assessed the severity of CLSD at the phenological stage R7.1. The acibenzolar-S-methyl resistance inducer, alternative products (macro and micronutrients) and A. nodosum had no effect on the severity of CLSD in the two harvests. The fungicide (azoxystrobin + cyproconazole) reduced the severity of CLSD and prevented damage to productivity in both experiments.

Nas últimas safras, o complexo de doenças de final de ciclo (DFC) da soja (Glycine max L. (Merrill)), vem provocando reduções consideráveis no rendimento da cultura. Atualmente, não há variedades resistentes a todos os patógenos causadores das DFC. O objetivo do presente trabalho foi avaliar o efeito da aplicação do indutor de resistência acibenzolar-S-methyl, produtos alternativos e fungicida na severidade de DFC na cultivar de soja BMX Potência RR durante as safras 2013/2014 e 2014/2015, em campo. Os tratamentos para os experimentos foram: 1 - testemunha (água); 2 - acibenzolar-S-methyl; 3 - cálcio; 4 - micronutrientes: cobre, manganês e zinco; 5 - micronutrientes: manganês, zinco e molibdênio; 6 - adubo NK; 7 - Ascophyllum nodosum e 8 - azoxistrobina + ciproconazol com adição do adjuvante. Foram realizadas quatro aplicações dos produtos alternativos e duas do fungicida, nas duas safras. A severidade de DFC foi avaliada por escala diagramática no estádio fenológico R7.1. O indutor de resistência acibenzolar-S-methyl, os produtos alternativos (macro e micronutrientes) e A. nodosum não apresentaram efeito sobre a severidade das DFC nas duas safras. O fungicida (azoxistrobina + ciproconazol) reduziu a severidade das DFC e evitou danos a produtividade nos dois experimentos.

Five Surfactin Isomers Produced during Cheonggukjang Fermentation by Bacillus pumilus HY1 and Their Properties.

The cyclic lipopeptide produced from Bacillus pumilus strain HY1 was isolated from Korean soybean sauce cheonggukjang. The chemical structures of the surfactin isomers were analyzed using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) and electrospray ionization tandem mass spectrometry (ESI-MS/MS). The five potential surfactin isoforms were detected with protonated masses of m/z 994.7, 1008.7, 1022.7, 1036.7, and 1050.7 and different structures in combination with Na+, K+, and Ca2+ ions. ESI-MS/MS analysis revealed that the isolated surfactin possessed the precise amino acid sequence LLVDLL and hydroxyl fatty acids with 12 to 16 carbons. The surfactin content during cheonggukjang fermentation increased from 0.3 to 51.2 mg/kg over 60 h of fermentation. The mixture of five surfactin isoforms of cheonggukjang inhibited the growth of two cancer cell lines. The growth of both MCF-7 and Caco-2 cells was strongly inhibited with 100 µg/µL of surfactin. This study is the first-time report of five surfactin isomers of Bacillus pumilus strain HY1 during Korean soybean sauce cheonggukjang fermentation, which has cytotoxic properties.

Characteristics of Bacterial Strains with Desirable Flavor Compounds from Korean Traditional Fermented Soybean Paste (Doenjang).

To identify and analyze the characteristics of the microorganisms involved in the formation of the desirable flavor of Doenjang, a total of 179 strains were isolated from ninety-four Doenjang collected from six regions in South Korea, and fourteen strains were selected through a sensory evaluation of the aroma of each culture. The enzyme activities of amylase, protease and lipase was shown in the various strains. Bacillus sp.-K3, Bacillus sp.-K4 and Bacillus amyloliquefaciens-J2 showed relatively high protease activity, at 317.1 U, 317.3 U and 319.5 U, respectively. The Bacillus sp.-K1 showed the highest lipase activity at 2453.6 U. In the case of amylase, Bacillus subtilis-H6 showed the highest activity at 4105.5 U. The results of the PCA showed that Bacillus subtilis-H2, Bacillus subtilis-H3, and Bacillus sp.-K2 were closely related to the production of 3-hydroxy-2-butanone (23.51%~43.37%), and that Bacillus subtilis-H5 and Bacillus amyloliquefaciens-J2 were significantly associated with the production of phenethyl alcohol (0.39% and 0.37%). The production of peptides was observed to vary among the Bacillus cultures such as Val-Val-Pro-Pro-Phe-Leu and Pro-Ala-Glu-Val-Leu-Asp-Ile. These peptides are precursors of related volatile flavor compounds created in Doenjang via the enzymatic or non-enzymatic route; it is expected that these strains could be used to enhance the flavor of Doenjang.

Current Perspectives on the Physiological Activities of Fermented Soybean-Derived Cheonggukjang.

Cheonggukjang (CGJ, fermented soybean paste), a traditional Korean fermented dish, has recently emerged as a functional food that improves blood circulation and intestinal regulation. Considering that excessive consumption of refined salt is associated with increased incidence of gastric cancer, high blood pressure, and stroke in Koreans, consuming CGJ may be desirable, as it can be made without salt, unlike other pastes. Soybeans in CGJ are fermented by Bacillus strains (B. subtilis or B. licheniformis), Lactobacillus spp., Leuconostoc spp., and Enterococcus faecium, which weaken the activity of putrefactive bacteria in the intestines, act as antibacterial agents against pathogens, and facilitate the excretion of harmful substances. Studies on CGJ have either focused on improving product quality or evaluating the bioactive substances contained in CGJ. The fermentation process of CGJ results in the production of enzymes and various physiologically active substances that are not found in raw soybeans, including dietary fiber, phospholipids, isoflavones (e.g., genistein and daidzein), phenolic acids, saponins, trypsin inhibitors, and phytic acids. These components prevent atherosclerosis, oxidative stress-mediated heart disease and inflammation, obesity, diabetes, senile dementia, cancer (e.g., breast and lung), and osteoporosis. They have also been shown to have thrombolytic, blood pressure-lowering, lipid-lowering, antimutagenic, immunostimulatory, anti-allergic, antibacterial, anti-atopic dermatitis, anti-androgenetic alopecia, and anti-asthmatic activities, as well as skin improvement properties. In this review, we examined the physiological activities of CGJ and confirmed its potential as a functional food.

Identification of the Viral Determinant of Hypovirulence and Host Range in Sclerotiniaceae of a Genomovirus Reconstructed from the Plant Metagenome.

Uncharacterized viral genomes that encode circular replication-associated proteins of single-stranded DNA viruses have been discovered by metagenomics/metatranscriptomics approaches. Some of these novel viruses are classified in the newly formed family Genomoviridae. Here, we determined the host range of a novel genomovirus, SlaGemV-1, through the transfection of Sclerotinia sclerotiorum with infectious clones. Inoculating with the rescued virions, we further transfected Botrytis cinerea and Monilinia fructicola, two economically important members of the family Sclerotiniaceae, and Fusarium oxysporum. SlaGemV-1 causes hypovirulence in S. sclerotiorum, B. cinerea, and M. fructicola. SlaGemV-1 also replicates in Spodoptera frugiperda insect cells but not in Caenorhabditis elegans or plants. By expressing viral genes separately through site-specific integration, the replication protein alone was sufficient to cause debilitation. Our study is the first to demonstrate the reconstruction of a metagenomically discovered genomovirus without known hosts with the potential of inducing hypovirulence, and the infectious clone allows for studying mechanisms of genomovirus-host interactions that are conserved across genera. IMPORTANCE Little is known about the exact host range of widespread genomoviruses. The genome of soybean leaf-associated gemygorvirus-1 (SlaGemV-1) was originally assembled from a metagenomic/metatranscriptomic study without known hosts. Here, we rescued SlaGemV-1 and found that it could infect three important plant-pathogenic fungi and fall armyworm (S. frugiperda Sf9) insect cells but not a model nematode, C. elegans, or model plant species. Most importantly, SlaGemV-1 shows promise for inducing hypovirulence of the tested fungal species in the family Sclerotiniaceae, including Sclerotinia sclerotiorum, Botrytis cinerea, and Monilinia fructicola. The viral determinant of hypovirulence was further identified as replication initiation protein. As a proof of concept, we demonstrate that viromes discovered in plant metagenomes can be a valuable genetic resource when novel viruses are rescued and characterized for their host range.

Arabidopsis non-host resistance PSS30 gene enhances broad-spectrum disease resistance in the soybean cultivar Williams 82.

Non-host resistance (NHR), which protects all members of a plant species from non-adapted or non-host plant pathogens, is the most common form of plant immunity. NHR provides the most durable and robust form of broad-spectrum immunity against non-adaptive pathogens pathogenic to other crop species. In a mutant screen for loss of Arabidopsis (Arabidopsis thaliana) NHR against the soybean (Glycine max (L.) Merr.) pathogen Phytophthora sojae, the Phytophthora sojae-susceptible 30 (pss30) mutant was identified. The pss30 mutant is also susceptible to the soybean pathogen Fusarium virguliforme. PSS30 encodes a folate transporter, AtFOLT1, which was previously localized to chloroplasts and implicated in the transport of folate from the cytosol to plastids. We show that two Arabidopsis folate biosynthesis mutants with reduced folate levels exhibit a loss of non-host immunity against P. sojae. As compared to the wild-type Col-0 ecotype, the steady-state folate levels are reduced in the pss1, atfolt1 and two folate biosynthesis mutants, suggesting that folate is required for non-host immunity. Overexpression of AtFOLT1 enhances immunity of transgenic soybean lines against two serious soybean pathogens, the fungal pathogen F. virguliforme and the soybean cyst nematode (SCN) Heterodera glycines. Transgenic lines showing enhanced SCN resistance also showed increased levels of folate accumulation. This study thus suggests that folate contributes to non-host plant immunity and that overexpression of a non-host resistance gene could be a suitable strategy for generating broad-spectrum disease resistance in crop plants.

Elizabethkingia argenteiflava sp. nov., isolated from the pod of soybean, Glycine max.

Soybean pods, separated and enclosed from the outside environment, are considered a suitable place to find new microbes. A Gram-stain-negative, aerobic bacterium, bacterial strain (YB22T) was isolated from the pod of Glycine max (soybean) collected from a rural area in Republic of Korea and characterized by using polyphasic taxonomy. Cells of the strain were rod-shaped (approximately 0.4-0.6 µm wide and 4.0-5.0 µm long), non-flagellated and formed silver-yellow colonies. Cells grew at 25-35 °C (optimum, 28-30 °C), at pH 5.0-9.0 (optimum, pH 7.0) and with 0-2.0% NaCl (optimum, 0 % NaCl). 16S rRNA gene sequencing showed that strain YB22T was phylogenetically closest to the genus Elizabethkingia, and showed highest similarities to Elizabethkingia occulta G4070T (96.7 %), Elizabethkingia meningoseptica ATCC 13253T (96.7 %), Elizabethkingia miricola DSM 14571T (96.6 %), Elizabethkingia bruuniana G0146T (96.5 %), Elizabethkingia ursingii G4122T (96.4 %) and Elizabethkingia anophelis R26T (96.2 %). Average amino acid identity values between strain YB22T and other taxa in the genus Elizabethkingia were all above the threshold range of genus determination. Average nucleotide identity and digital DNA-DNA hybridization values between strain YB22T and other phylogenetic relatives were all found to be below the threshold range for species determination. The respiratory quinone of strain YB22T was menaquinone 6 (MK-6) and the predominant cellular fatty acids were iso-C15 : 0 (47.8 %) and iso-C17 : 0 3-OH (18.5 %). The major polar lipids were phosphatidylethanolamine, four unidentified aminolipids and three unidentified polar lipids. The phylogenetic analysis and physiological and biochemical data showed that strain YB22T should represent a novel species in the genus Elizabethkingia, for which the name Elizabethkingia argenteiflava sp. nov. is proposed. The type strain for this novel species is YB22T (=KCCM 43263T=JCM 32097T).

Halotolerant bacteria mitigate the effects of salinity stress on soybean growth by regulating secondary metabolites and molecular responses.

BACKGROUND: Salinity is a major threat to the agriculture industry due to the negative impact of salinity stress on crop productivity. In the present study, we isolated rhizobacteria and evaluated their capacities to promote crop growth under salt stress conditions. RESULTS: We isolated rhizospheric bacteria from sand dune flora of Pohang beach, Korea, and screened them for plant growth-promoting (PGP) traits. Among 55 bacterial isolates, 14 produced indole-3-acetic acid (IAA), 10 produced siderophores, and 12 produced extracellular polymeric and phosphate solubilization. Based on these PGP traits, we selected 11 isolates to assess for salinity tolerance. Among them, ALT29 and ALT43 showed the highest tolerance to salinity stress. Next, we tested the culture filtrate of isolates ALT29 and ALT43 for IAA and organic acids to confirm the presence of these PGP products. To investigate the effects of ALT29 and ALT43 on salt tolerance in soybean, we grew seedlings in 0 mM, 80 mM, 160 mM, and 240 mM NaCl treatments, inoculating half with the bacterial isolates. Inoculation with ALT29 and ALT43 significantly increased shoot length (13%), root length (21%), shoot fresh and dry weight (44 and 35%), root fresh and dry weight (9%), chlorophyll content (16-24%), Chl a (8-43%), Chl b (13-46%), and carotenoid (14-39%) content of soybean grown under salt stress. Inoculation with ALT29 and ALT43 also significantly decreased endogenous ABA levels (0.77-fold) and increased endogenous SA contents (6-16%), increased total protein (10-20%) and glutathione contents, and reduced lipid peroxidation (0.8-5-fold), superoxide anion (21-68%), peroxidase (12.14-17.97%), and polyphenol oxidase (11.76-27.06%) contents in soybean under salinity stress. In addition, soybean treated with ALT29 and ALT43 exhibited higher K+ uptake (9.34-67.03%) and reduced Na+ content (2-4.5-fold). Genes involved in salt tolerance, GmFLD19 and GmNARK, were upregulated under NaCl stress; however, significant decreases in GmFLD19 (3-12-fold) and GmNARK (1.8-3.7-fold) expression were observed in bacterial inoculated plants. CONCLUSION: In conclusion, bacterial isolates ALT29 and ALT43 can mitigate salinity stress and increase plant growth, providing an eco-friendly approach for addressing saline conditions in agricultural production systems.

Application of ultrasound technology in the field of solid-state fermentation: increasing peptide yield through ultrasound-treated bacterial strain.

BACKGROUND: The increase of peptide yield contributed to reducing the usage of antibiotics in solid-state fermented feed. Ultrasound technology is used in the field of liquid-state fermentation to improve yield of fermented products but has not been utilized in the field of solid-state fermentation (SSF). The main objective of this study was to investigate the feasibility of improving peptide yield in SSF products through ultrasound-treated bacterial strain. RESULTS: The highest peptides content in soybean meal SSF products reached 153.28 mg g-1 , which increased by 15.05% compared with the control. This content value was acquired through treating the bacteria of Bacillus amyloliquefaciens by ultrasound before inoculating into soybean meal under the optimized mode and parameters (simultaneous dual-frequency ultrasound mode, frequency combination of 40/60 kHz, total power density of 40 W L-1 , time of 20 min, pulse-on and pulse-off times of 40 and 60 s, delayed inoculation time of 0 h). Fermenting with ultrasound-treated bacterial strain can effectively increase peptide yield, biomass and protease activity of soybean meal fermented products during the SSF prophase. After treating by ultrasound, the latent phase and logarithmic phase of the bacterial strain shortened by 1 and 3 h while the generation time reduced by 23.64%. In qualitative test of protease activity, diameter ratio (DR) value of ultrasound-treated bacterial cells enlarged by 12.0% compared with the control. CONCLUSION: Peptide yield of soybean meal SSF products can be improved through ultrasound-treated bacterial inoculum, which attributed to the promoting effect of ultrasound treatment on growth activity and protease production capability of bacterial cells. © 2021 Society of Chemical Industry.

Bradyrhizobium diazoefficiens Requires Chemical Chaperones To Cope with Osmotic Stress during Soybean Infection.

When engaging in symbiosis with legume hosts, rhizobia are confronted with environmental changes, including nutrient availability and stress exposure. Genetic circuits allow responding to these environmental stimuli to optimize physiological adaptations during the switch from the free-living to the symbiotic life style. A pivotal regulatory system of the nitrogen-fixing soybean endosymbiont Bradyrhizobium diazoefficiens for efficient symbiosis is the general stress response (GSR), which relies on the alternative sigma factor σEcfG However, the GSR-controlled process required for symbiosis has not been identified. Here, we demonstrate that biosynthesis of trehalose is under GSR control, and mutants lacking the respective biosynthetic genes otsA and/or otsB phenocopy GSR-deficient mutants under symbiotic and selected free-living stress conditions. The role of trehalose as a cytoplasmic chemical chaperone and stress protectant can be functionally replaced in an otsA or otsB mutant by introducing heterologous genetic pathways for biosynthesis of the chemically unrelated compatible solutes glycine betaine and (hydroxy)ectoine. Alternatively, uptake of exogenously provided trehalose also restores efficient symbiosis and tolerance to hyperosmotic and hyperionic stress of otsA mutants. Hence, elevated cytoplasmic trehalose levels resulting from GSR-controlled biosynthesis are crucial for B. diazoefficiens cells to overcome adverse conditions during early stages of host infection and ensure synchronization with root nodule development.IMPORTANCE The Bradyrhizobium-soybean symbiosis is of great agricultural significance and serves as a model system for fundamental research in bacterium-plant interactions. While detailed molecular insight is available about mutual recognition and early nodule organogenesis, our understanding of the host-imposed conditions and the physiology of infecting rhizobia during the transition from a free-living state in the rhizosphere to endosymbiotic bacteroids is currently limited. In this study, we show that the requirement of the rhizobial general stress response (GSR) during host infection is attributable to GSR-controlled biosynthesis of trehalose. Specifically, trehalose is crucial for an efficient symbiosis by acting as a chemical chaperone to protect rhizobia from osmostress during host infection.

Fg12 ribonuclease secretion contributes to Fusarium graminearum virulence and induces plant cell death.

Filamentous fungal pathogens secrete effectors that modulate host immunity and facilitate infection. Fusarium graminearum is an important plant pathogen responsible for various devastating diseases. However, little is known about the function of effector proteins secreted by F. graminearum. Herein, we identified several effector candidates in the F. graminearum secretome. Among them, the secreted ribonuclease Fg12 was highly upregulated during the early stages of F. graminearum infection in soybean; its deletion compromised the virulence of F. graminearum. Transient expression of Fg12 in Nicotiana benthamiana induced cell death in a light-dependent manner. Fg12 possessed ribonuclease (RNase) activity, degrading total RNA. The enzymatic activity of Fg12 was required for its cell death-promoting effects. Importantly, the ability of Fg12 to induce cell death was independent of BAK1/SOBIR1, and treatment of soybean with recombinant Fg12 protein induced resistance to various pathogens, including F. graminearum and Phytophthora sojae. Overall, our results provide evidence that RNase effectors not only contribute to pathogen virulence but also induce plant cell death.

Enhancement of isoflavone aglycone, amino acid, and CLA contents in fermented soybean yogurts using different strains: Screening of antioxidant and digestive enzyme inhibition properties.

This study was the first to evaluate changes in isoflavone, amino acid, conjugated linoleic acid (CLA), antioxidant effect, and digestive enzyme inhibition during fermentation of soy-milk to soy-yogurt with L. brevis and L. plantarum. Total average isoflavones were reduced (1318.2 â†’ 971.1 µg/g) with an increase of aglycones (60.2 â†’ 804.9 µg/g, genistein > daidzein > glycitein) in soy powder yogurts (SPYs). Amino acids increased considerably, as did ornithine (average 4.1 â†’ 551.0 mg/g), and CLA showed high variations from not-detected (ND) to 0.5, 0.9 mg/g (cis-9, trans-11) and ND to 0.3, 0.2 mg/g (trans-10, cis-12). Digestive enzyme inhibitions (α-glucosidase, α-amylase, and pancreatic lipase) displayed high activities (average 50.6 â†’ 67.2, 5.2 â†’ 46.4, 10.6 â†’ 51.4%). Moreover, the antioxidant abilities against radicals were elevated as follows: ABTS > DPPH > hydroxyl (average 63.5 â†’ 86.5, 50.2 â†’ 70.3, 39.3 â†’ 55.2%). Specifically, SPY using mixed strains exhibited the greatest enzymatic inhibition and antioxidant capacities.

Facets of rhizospheric microflora in biocontrol of phytopathogen Macrophomina phaseolina in oil crop soybean.

The use of microbial bioinoculants for managing plant diseases and promoting plant growth is an effective alternative approach to integrated farming. One of the devastating phytopathogens is Macrophomina phaseolina (Tassi) Goid. It is an omnipresent fungus infecting more than 500 plant species. It causes charcoal rot disease in soybean leading to 30-50% yield loss. Soybean Glycine max (L.) oil seed crop produced globally is highly susceptible to M. phaseolina. India is the fifth largest producer of soybean in the world. Madhya Pradesh is the largest soybean-producing state in India; Around 70% yield loss of soybean is accounted to M. phaseolina infection in India. Control of charcoal rot is the requisite of the current situation. Chemical control is not feasible due to saprophytic nature and prolonged survival of Macrophomina phaseolina. Chemical fungicides are expensive, toxic, hazardous, and cause pollution. Biological control is an effective approach to control this devastating fungus. The rhizosphere of soil is rich in beneficial microflora competent to suppress plant pathogens and also promote plant growth. PGPR have well-developed mechanisms that impart antagonistic traits to them. PGPR produces various antifungal metabolites siderophores and HCN which inhibit fungal growth, and can be used as potent BCA. Pseudomonas and Bacillus species have been reported effective against M. phaseolina. The mechanisms and antifungal compounds produced by these bacteria to control charcoal rot can be studied extensively. BCA or the metabolites secreted by them have the potential to develop effective bioformulations for soybean at the commercial level for sustainable agriculture.

Effects of rhizobia and arbuscular mycorrhizal fungi on yield, size distribution and fatty acid of soybean seeds grown under drought stress.

Soybean (Glycine max L.) is among the most economically important legumes that provide more than 1/4 of food (for man) and animal feed. However, its yield is comparatively low, most especially under drought stress. The aim of this study therefore was to assess the ability of Rhizobium spp. and mycorrhizal fungi to enhance the yield, seed size and fatty acid content of soybean grown under semi-arid environment. Rhizobium sp. strain R1 was found to possess nitrogen-fixing gene coniferyl aldehyde dehydrogenase function while Rhizobium cellulosilyticum strain R3 was found to have nitrogen-fixing genes cysteine desulfurase SufS and cysteine desulfurase IscS activity. Soybean (Glycine max L) seeds inoculated with Rhizobium spp. and mycorrhizal fungi were cultivated in soil exposed to drought stress. Rhizobium spp. inoculation and mycorrhization alleviate drought stress and increase yield, size and fat content of soybean seeds. This increase in the aboveground parameters was accompanied with an increase in belowground mycorrhizal spore number, percentage root mycorrhization and aboveground shoot relative water content (RWC) in the dually inoculated (R1 + R3MY) soybean plants. In particular, the dually inoculated (R1 + R3MY) soybean plants revealed 34.3 g fresh weight, 15.1 g dry weight and soybean plants singly inoculated with Rhizobium sp. strain R1 (R1) produced more large seeds with 12.03 g dry weight. The non-inoculated (control) seeds contained a higher percentage of moisture content compared to the microbially amended seeds while seeds co-inoculated with Rhizobium cellulosilyticum strain R3 and mycorrhizal consortium revealed the highest percent (8.4 %) of fat. Several fatty acids that are of significant health benefits to humans were observed in the soybean seeds. In order to gain insights into the bacterial communities of rhizospheric soil collected at different stages of soybean growth, class-based Heat-map analysis was performed on the Miseq sequenced data. The core bacteria that were found in the rhizospheric soil were Verrumicrobia, Proteobacteria, Gemmatimonadetes, Firmicutes, Cyanobacteria, Chloroflexi, Bacteroidetes, Actinobacteria, Acidobacteria, Planctomycetes, Deinococcus thermus and Nitrospira suggesting that the rhizobia and fungi used in this study can also improve soil microbial diversity.