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1.
Nucleic Acids Res ; 52(2): 844-855, 2024 Jan 25.
Article in English | MEDLINE | ID: mdl-38048327

ABSTRACT

Prokaryotic Argonautes (pAgos) play a vital role in host defense by utilizing short nucleic acid guides to recognize and target complementary nucleic acids. Despite being the majority of pAgos, short pAgos have only recently received attention. Short pAgos are often associated with proteins containing an APAZ domain and a nuclease domain including DUF4365, SMEK, or HNH domain. In contrast to long pAgos that specifically cleave the target DNA, our study demonstrates that the short pAgo from Thermocrispum municipal, along with its associated DUF4365-APAZ protein, forms a heterodimeric complex. Upon RNA-guided target DNA recognition, this complex is activated to nonspecifically cleave DNA. Additionally, we found that the TmuRE-Ago complex shows a preference for 5'-OH guide RNA, specifically requires a uridine nucleotide at the 5' end of the guide RNA, and is sensitive to single-nucleotide mismatches between the guide RNA and target DNA. Based on its catalytic properties, our study has established a novel nucleic acid detection method and demonstrated its feasibility. This study not only expands our understanding of the defense mechanism employed by short pAgo systems but also suggests their potential applications in nucleic acid detection.


Subject(s)
Actinobacteria , Argonaute Proteins , DNA , RNA, Bacterial , Argonaute Proteins/metabolism , DNA/metabolism , Endonucleases/metabolism , Nucleic Acids/metabolism , Prokaryotic Cells/metabolism , Actinobacteria/physiology , RNA, Bacterial/metabolism
2.
Cell ; 186(22): 4803-4817.e13, 2023 10 26.
Article in English | MEDLINE | ID: mdl-37683634

ABSTRACT

Patescibacteria, also known as the candidate phyla radiation (CPR), are a diverse group of bacteria that constitute a disproportionately large fraction of microbial dark matter. Its few cultivated members, belonging mostly to Saccharibacteria, grow as epibionts on host Actinobacteria. Due to a lack of suitable tools, the genetic basis of this lifestyle and other unique features of Patescibacteira remain unexplored. Here, we show that Saccharibacteria exhibit natural competence, and we exploit this property for their genetic manipulation. Imaging of fluorescent protein-labeled Saccharibacteria provides high spatiotemporal resolution of phenomena accompanying epibiotic growth, and a transposon-insertion sequencing (Tn-seq) genome-wide screen reveals the contribution of enigmatic Saccharibacterial genes to growth on their hosts. Finally, we leverage metagenomic data to provide cutting-edge protein structure-based bioinformatic resources that support the strain Southlakia epibionticum and its corresponding host, Actinomyces israelii, as a model system for unlocking the molecular underpinnings of the epibiotic lifestyle.


Subject(s)
Bacteria , Bacteria/classification , Bacteria/genetics , Bacteria/growth & development , Metagenome , Metagenomics , Phylogeny , Actinobacteria/physiology
3.
Plant Physiol Biochem ; 196: 472-483, 2023 Mar.
Article in English | MEDLINE | ID: mdl-36764263

ABSTRACT

Atrazine is used to control broad-leaved weeds in farmland and has negative impacts on soybean growth. Legume-rhizobium symbiosis plays an important role in regulating abiotic stress tolerance of plants, however, the mechanisms of rhizobia regulate the tolerance of soybean to atrazine based on the biochemical responses of the plant-soil system are limited. In this experiment, Glycine max (L.) Merr. Dongnong 252, planted in 20 mg kg-1 of atrazine-contaminated soil, was inoculated with Bradyrhizobium japonicum AC20, and the plant growth, rhizosphere soil microbial diversity and the expression of the genes related to soybean carbon and nitrogen metabolism were assessed. The results indicated that strain AC20 inoculation alleviated atrazine-induced growth inhibition via increasing the contents of leghemoglobin and total nitrogen in soybean seedlings. The psbA gene expression level of the soybean seedlings that inoculated strain AC20 was 1.4 times than that of no rhizobium inoculating treatments. Moreover, the inoculated AC20 increased the abundance of Acidobacteria and Actinobacteria in soybean rhizosphere. Transcriptome analysis demonstrated that strain AC20 regulated the genes expression of amino acid metabolism and carbohydrate metabolism of soybean seedlings. Correlation analysis between 16S rRNA and transcriptome showed that strain AC20 reduced Planctomycetes abundance so as to down-regulated the expression of genes Glyma. 13G087800, Glyma. 12G005100 and Glyma.12G098900 involved in starch synthesis pathway of soybean leaves. These results provide available information for the rhizobia application to enhance the atrazine tolerate in soybean seedlings.


Subject(s)
Atrazine , Bradyrhizobium , Drug Resistance , Glycine max , Herbicides , Rhizosphere , Bradyrhizobium/physiology , Glycine max/drug effects , Glycine max/genetics , Glycine max/growth & development , Glycine max/microbiology , Atrazine/toxicity , Herbicides/toxicity , Soil Microbiology , Microbiota/physiology , Carbohydrate Metabolism/genetics , Acidobacteria/physiology , Actinobacteria/physiology , Transcriptome , Weed Control , Gene Expression Regulation, Plant , Amino Acids/metabolism
4.
Invest Ophthalmol Vis Sci ; 63(2): 21, 2022 02 01.
Article in English | MEDLINE | ID: mdl-35142786

ABSTRACT

Purpose: To determine the possible microbiome related to Vogt-Koyanagi-Harada (VKH) disease in comparison to patients with noninfectious anterior scleritis and healthy people. Methods: Fecal samples were extracted from 42 individuals, including 11 patients with active VKH, 11 healthy people, and 20 patients with noninfectious anterior scleritis. We amplified the V3 to V4 16S ribosomal DNA (rDNA) region to obtain the target sequence. Then, the target sequence was amplified by polymerase chain reaction. The obtained target sequences were sequenced by high-throughput 16S rDNA analysis. Results: At the genus level, there were three enriched (Stomatobaculum, Pseudomonas, Lachnoanaerobaculum) and two depleted (Gordonibacter, Slackia) microbes that were detected only in patients with VKH. There were 10 enriched and 12 depleted microbes that were observed in both patients with VKH disease and noninfectious anterior scleritis (P < 0.05). The interactions of these microbes were graphed. Tyzzerella and Eggerthella were the nodes of interaction between these microorganisms, which were regulated by both positive and negative aspects, but the expression level in patients with active VKH was upregulated. Conclusions: Special or nonspecial enrichment and decreased intestinal microbes were observed in patients with active VKH. The action mechanism of these microbes needs further study.


Subject(s)
Actinobacteria/physiology , Clostridiales/physiology , Gastrointestinal Microbiome/physiology , Pseudomonas/physiology , Uveomeningoencephalitic Syndrome/microbiology , Adult , Case-Control Studies , DNA, Bacterial/genetics , Dysbiosis/microbiology , Feces/microbiology , Female , Genotyping Techniques , Healthy Volunteers , Humans , Male , Middle Aged , RNA, Ribosomal, 16S/genetics , Scleritis/microbiology
5.
Int J Mol Sci ; 23(3)2022 Jan 19.
Article in English | MEDLINE | ID: mdl-35163015

ABSTRACT

Nowadays, the worldwide agriculture is experiencing a transition process toward more sustainable production, which requires the reduction of chemical inputs and the preservation of microbiomes' richness and biodiversity. Plants are no longer considered as standalone entities, and the future of agriculture should be grounded on the study of plant-associated microorganisms and all their potentiality. Moreover, due to the climate change scenario and the resulting rising incidence of abiotic stresses, an innovative and environmentally friendly technique in agroecosystem management is required to support plants in facing hostile environments. Plant-associated microorganisms have shown a great attitude as a promising tool to improve agriculture sustainability and to deal with harsh environments. Several studies were carried out in recent years looking for some beneficial plant-associated microbes and, on the basis of them, it is evident that Actinomycetes and arbuscular mycorrhizal fungi (AMF) have shown a considerable number of positive effects on plants' fitness and health. Given the potential of these microorganisms and the effects of climate change, this review will be focused on their ability to support the plant during the interaction with abiotic stresses and on multi-omics techniques which can support researchers in unearthing the hidden world of plant-microbiome interactions. These associated microorganisms can increase plants' endurance of abiotic stresses through several mechanisms, such as growth-promoting traits or priming-mediated stress tolerance. Using a multi-omics approach, it will be possible to deepen these mechanisms and the dynamic of belowground microbiomes, gaining fundamental information to exploit them as staunch allies and innovative weapons against crop abiotic enemies threatening crops in the ongoing global climate change context.


Subject(s)
Actinobacteria/physiology , Computational Biology/methods , Crops, Agricultural/growth & development , Mycorrhizae/physiology , Climate Change , Crops, Agricultural/microbiology , Genomics , Metabolomics , Plant Development , Soil Microbiology , Stress, Physiological , Systems Biology
6.
Mol Microbiol ; 117(1): 179-192, 2022 01.
Article in English | MEDLINE | ID: mdl-34687261

ABSTRACT

WhiB7/WblC is a transcriptional factor of actinomycetes conferring intrinsic resistance to multiple translation-inhibitory antibiotics. It positively autoregulates its own transcription in response to the same antibiotics. The presence of a uORF and a potential Rho-independent transcription terminator in the 5' leader region has suggested a possibility that the whiB7/wblC gene is regulated via a uORF-mediated transcription attenuation. However, experimental evidence for the molecular mechanism to explain how antibiotic stress suppresses the attenuator, if any, and induces transcription of the whiB7/wblC gene has been lacking. Here we report that the 5' leader sequences of the whiB7/wblC genes in sub-clades of actinomycetes include conserved antiterminator RNA structures. We confirmed that the putative antiterminator in the whiB7/wblC leader sequences of both Streptomyces and Mycobacterium indeed suppresses Rho-independent transcription terminator and facilitates transcription readthrough, which is required for WhiB7/WblC-mediated antibiotic resistance. The antibiotic-mediated suppression of the attenuator can be recapitulated by amino acid starvation, indicating that translational inhibition of uORF by multiple signals is a key to induce whiB7/wblC expression. Our findings of a mechanism leading to intrinsic antibiotic resistance could provide an alternative to treat drug-resistant mycobacteria.


Subject(s)
5' Untranslated Regions/genetics , Actinobacteria/genetics , Anti-Bacterial Agents/pharmacokinetics , Bacterial Proteins/metabolism , Gene Expression Regulation, Bacterial , Mycobacterium/genetics , Streptomyces coelicolor/genetics , Actinobacteria/physiology , Anti-Bacterial Agents/pharmacology , Bacterial Proteins/genetics , Drug Resistance, Bacterial/genetics , Mycobacterium/physiology , Ribosomes/metabolism , Streptomyces coelicolor/physiology , Transcription Factors/genetics , Transcription Factors/metabolism , Transcription, Genetic
7.
BMC Microbiol ; 21(1): 335, 2021 12 07.
Article in English | MEDLINE | ID: mdl-34876006

ABSTRACT

BACKGROUND: The native potatoes (Solanum tuberosum subsp. tuberosum L.) grown in Chile (Chiloé) represent a new, unexplored source of endophytes to find potential biological control agents for the prevention of bacterial diseases, like blackleg and soft rot, in potato crops. RESULT: The objective of this study was the selection of endophytic actinobacteria from native potatoes for antagonistic activity against Pectobacterium carotovorum subsp. carotovorum and Pectobacterium atrosepticum, and their potential to suppress tissue maceration symptoms in potato tubers. This potential was determined through the quorum quenching activity using a Chromobacterium violaceaum ATCC 12472 Wild type (WT) bioassay and its colonization behavior of the potato plant root system (S. tuberosum) by means of the Double labeling of oligonucleotide probes for fluorescence in situ hybridization (DOPE-FISH) targeting technique. The results showed that although Streptomyces sp. TP199 and Streptomyces sp. A2R31 were able to inhibit the growth of the pathogens, only the Streptomyces sp. TP199 isolate inhibited Pectobacterium sp. growth and diminished tissue maceration in tubers (p ≤ 0.05). Streptomyces sp. TP199 had metal-dependent acyl homoserine lactones (AHL) quorum quenching activity in vitro and was able to colonize the root endosphere 10 days after inoculation. CONCLUSIONS: We concluded that native potatoes from southern Chile possess endophyte actinobacteria that are potential agents for the disease management of soft rot and blackleg.


Subject(s)
Actinobacteria/physiology , Antibiosis/physiology , Endophytes/physiology , Solanum tuberosum/microbiology , Actinobacteria/classification , Actinobacteria/genetics , Actinobacteria/isolation & purification , Biological Control Agents/isolation & purification , Chile , Endophytes/classification , Endophytes/genetics , Endophytes/isolation & purification , Pectobacterium/physiology , Plant Diseases/microbiology , Plant Diseases/prevention & control , Plant Tubers/microbiology , Quorum Sensing , Streptomyces/classification , Streptomyces/genetics , Streptomyces/isolation & purification , Streptomyces/physiology
8.
PLoS One ; 16(11): e0260451, 2021.
Article in English | MEDLINE | ID: mdl-34813629

ABSTRACT

The mortality rates of COVID-19 vary widely across countries, but the underlying mechanisms remain unelucidated. We aimed at the elucidation of relationship between gut microbiota and the mortality rates of COVID-19 across countries. Raw sequencing data of 16S rRNA V3-V5 regions of gut microbiota in 953 healthy subjects in ten countries were obtained from the public database. We made a generalized linear model (GLM) to predict the COVID-19 mortality rates using gut microbiota. GLM revealed that low genus Collinsella predicted high COVID-19 mortality rates with a markedly low p-value. Unsupervised clustering of gut microbiota in 953 subjects yielded five enterotypes. The mortality rates were increased from enterotypes 1 to 5, whereas the abundances of Collinsella were decreased from enterotypes 1 to 5 except for enterotype 2. Collinsella produces ursodeoxycholate. Ursodeoxycholate was previously reported to inhibit binding of SARS-CoV-2 to angiotensin-converting enzyme 2; suppress pro-inflammatory cytokines like TNF-α, IL-1ß, IL-2, IL-4, and IL-6; have antioxidant and anti-apoptotic effects; and increase alveolar fluid clearance in acute respiratory distress syndrome. Ursodeoxycholate produced by Collinsella may prevent COVID-19 infection and ameliorate acute respiratory distress syndrome in COVID-19 by suppressing cytokine storm syndrome.


Subject(s)
Actinobacteria/physiology , COVID-19/prevention & control , Gastrointestinal Microbiome , Intestines/microbiology , SARS-CoV-2/physiology , Ursodeoxycholic Acid/metabolism , COVID-19/etiology , COVID-19/pathology , Humans
9.
PLoS One ; 16(11): e0259465, 2021.
Article in English | MEDLINE | ID: mdl-34731210

ABSTRACT

Curtobacterium sp. GD1 was isolated from leaves of conventionally grown soybean in Brazil. It was noteworthy that among all bacteria previously isolated from the same origin, only Curtobacterium sp. GD1 showed a strong chitinase activity. The enzyme was secreted and its production was induced by the presence of colloidal chitin in the medium. The chitinase was partially purified and characterized: molecular weight was approximately 37 kDa and specific activity 90.8 U/mg. Furthermore, Curtobacterium sp. GD1 genome was sequenced and analyzed. Our isolate formed a phylogenetic cluster with four other Curtobacterium spp. strains, with ANIb/ANIm ≥ 98%, representing a new, still non described Curtobacterium species. The circular genome visualization and comparison of genome sequences of strains forming new cluster indicated that most regions within their genomes were highly conserved. The gene associated with chitinase production was identified and the distribution pattern of glycosyl hydrolases genes was assessed. Also, genes associated with catabolism of structural carbohydrates such as oligosaccharides, mixed polysaccharides, plant and animal polysaccharides, as well as genes or gene clusters associated with resistance to antibiotics, toxic compounds and auxin biosynthesis subsystem products were identified. The abundance of putative glycosyl hydrolases in the genome of Curtobacterium sp. GD1 suggests that it has the tools for the hydrolysis of different polysaccharides. Therefore, Curtobacterium sp. GD1 isolated from soybean might be a bioremediator, biocontrol agent, an elicitor of the plant defense responses or simply degrader.


Subject(s)
Actinobacteria/physiology , Chitin/chemistry , Chitinases/genetics , Glycine max/microbiology , Whole Genome Sequencing/methods , Actinobacteria/classification , Actinobacteria/isolation & purification , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Chitinases/metabolism , Genome, Bacterial , High-Throughput Nucleotide Sequencing , Hydrolysis , N-Glycosyl Hydrolases/genetics , N-Glycosyl Hydrolases/metabolism , Phylogeny , Plant Leaves/chemistry , Plant Leaves/microbiology , Glycine max/metabolism
11.
Cell Host Microbe ; 29(11): 1649-1662.e7, 2021 11 10.
Article in English | MEDLINE | ID: mdl-34637779

ABSTRACT

Saccharibacteria (TM7) are obligate epibionts living on the surface of their host bacteria and are strongly correlated with dysbiotic microbiomes during periodontitis and other inflammatory diseases, suggesting they are putative pathogens. However, due to the recalcitrance of TM7 cultivation, causal research to investigate their role in inflammatory diseases is lacking. Here, we isolated multiple TM7 species on their host bacteria from periodontitis patients. These TM7 species reduce inflammation and consequential bone loss by modulating host bacterial pathogenicity in a mouse ligature-induced periodontitis model. Two host bacterial functions involved in collagen binding and utilization of eukaryotic sialic acid are required for inducing bone loss and are altered by TM7 association. This TM7-mediated downregulation of host bacterial pathogenicity is shown for multiple TM7/host bacteria pairs, suggesting that, in contrast to their suspected pathogenic role, TM7 could protect mammalian hosts from inflammatory damage induced by their host bacteria.


Subject(s)
Actinobacteria/pathogenicity , Alveolar Bone Loss/microbiology , Bacterial Physiological Phenomena , Gingivitis/microbiology , Periodontitis/microbiology , Symbiosis , Actinobacteria/genetics , Actinobacteria/isolation & purification , Actinobacteria/physiology , Actinomyces/genetics , Actinomyces/isolation & purification , Actinomyces/pathogenicity , Actinomyces/physiology , Alveolar Bone Loss/prevention & control , Animals , Bacteria/classification , Bacteria/isolation & purification , Bacteria/pathogenicity , Bacterial Infections/microbiology , Bacterial Infections/prevention & control , Collagen/metabolism , Dental Plaque/microbiology , Down-Regulation , Genes, Bacterial , Gingivitis/prevention & control , Humans , Male , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Microbiota , N-Acetylneuraminic Acid/metabolism , Periodontitis/prevention & control , Propionibacteriaceae/genetics , Propionibacteriaceae/isolation & purification , Propionibacteriaceae/pathogenicity , Propionibacteriaceae/physiology , Virulence
12.
J Basic Microbiol ; 61(11): 1002-1015, 2021 Nov.
Article in English | MEDLINE | ID: mdl-34528722

ABSTRACT

The enzyme dextranase is widely used in the sugar and food industries, as well as in the medical field. Most land-derived dextranases are produced by fungi and have the disadvantages of long production cycles, low tolerance to environmental conditions, and low safety. The use of marine bacteria to produce dextranases may overcome these problems. In this study, a dextranase-producing bacterium was isolated from the Rizhao seacoast of Shandong, China. The bacterium, denoted as PX02, was identified as Cellulosimicrobium sp. and its growing conditions and the production and properties of its dextranase were investigated. The dextranase had a molecular weight of approximately 40 kDa, maximum activity at 40°C and pH 7.5, with a stability range of up to 45°C and pH 7.0-9.0. High-performance liquid chromatography showed that the dextranase hydrolyzed dextranT20 to isomaltotriose, maltopentaose, and isomaltooligosaccharides. Hydrolysis by dextranase produced excellent antioxidant effects, suggesting its potential use in the health food industry. Investigation of the action of the dextranase on Streptococcus mutans biofilm and scanning electron microscopy showed that it to be effective both for removing and inhibiting the formation of biofilms, suggesting its potential application in the dental industry.


Subject(s)
Actinobacteria/enzymology , Bacterial Proteins/metabolism , Dextranase/metabolism , Actinobacteria/classification , Actinobacteria/isolation & purification , Actinobacteria/physiology , Antioxidants/chemistry , Antioxidants/metabolism , Antioxidants/pharmacology , Bacterial Proteins/chemistry , Bacterial Proteins/pharmacology , Biofilms/drug effects , Biofilms/growth & development , China , Dextranase/chemistry , Dextranase/pharmacology , Hydrogen-Ion Concentration , Hydrolysis , Metals/metabolism , Molecular Weight , Seawater/microbiology , Streptococcus mutans/drug effects , Substrate Specificity , Temperature
13.
World J Microbiol Biotechnol ; 37(7): 109, 2021 May 31.
Article in English | MEDLINE | ID: mdl-34057641

ABSTRACT

Phosphate solubilizing microorganisms widely exist in plant rhizosphere soil, but report about the P solubilization and multiple growth-promoting properties of rare actinomycetes are scarce. In this paper, a phosphate solubilizing Tsukamurella tyrosinosolvens P9 strain was isolated from the rhizosphere soil of tea plants. Phosphorus-dissolving abilities of this strain were different under different carbon and nitrogen sources, the soluble phosphorus content was 442.41 mg/L with glucose and potassium nitrate as nutrient sources. The secretion of various organic acids, such as lactic acid, maleic acid, oxalic acid, etc., was the main mechanism for P solubilization and pH value in culture was very significant negative correlation with soluble P content. In addition, this strain had multiple growth-promoting characteristics with 37.26 µg/mL of IAA and 72.01% of siderophore relative content. Under pot experiments, P9 strain improved obviously the growth of peanut seedlings. The bacterial communities of peanut rhizoshpere soil were assessed after inoculated with P9 strain. It showed that there was no significant difference in alpha-diversity indices between the inoculation and control groups, but the P9 treatment group changed the composition of bacterial communities, which increased the relative abundance of beneficial and functional microbes, which relative abundances of Chitinophagaceae at the family level, and of Flavihumibacter, Ramlibacter and Microvirga at the genus level, were all siginificant increased. Specially, Tsukamurella tyrosinosolvens were only detected in the rhizosphere of the inoculated group. This study not only founded growth-promoting properties of T. tyrosinosolvens P9 strain and its possible phosphate solublizing mechanism, but also expected to afford an excellent strain resource in biological fertilizers.


Subject(s)
Actinobacteria/classification , Arachis/growth & development , Calcium Phosphates/chemistry , Actinobacteria/isolation & purification , Actinobacteria/physiology , Arachis/microbiology , Carbon/metabolism , Gas Chromatography-Mass Spectrometry , Glucose/metabolism , Nitrates/metabolism , Nitrogen/metabolism , Phylogeny , Potassium Compounds/metabolism , Rhizosphere , Soil Microbiology
14.
Nat Microbiol ; 6(6): 703-711, 2021 06.
Article in English | MEDLINE | ID: mdl-33927381

ABSTRACT

Many wastewater treatment plants around the world suffer from the operational problem of foaming. This is characterized by a persistent stable foam that forms on the aeration basin, which reduces effluent quality. The foam is often stabilized by a highly hydrophobic group of Actinobacteria known as the Mycolata1. Gordonia amarae is one of the most frequently reported foaming members1. With no currently reliable method for treating foams, phage biocontrol has been suggested as an attractive treatment strategy2. Phages isolated from related foaming bacteria can destabilize foams at the laboratory scale3,4; however, no phage has been isolated that lyses G. amarae. Here, we assemble the complete genomes of G. amarae and a previously undescribed species, Gordonia pseudoamarae, to examine mechanisms that encourage stable foam production. We show that both of these species are recalcitrant to phage infection via a number of antiviral mechanisms including restriction, CRISPR-Cas and bacteriophage exclusion. Instead, we isolate and cocultivate an environmental ultrasmall epiparasitic bacterium from the phylum Saccharibacteria that lyses G. amarae and G. pseudoamarae and several other Mycolata commonly associated with wastewater foams. The application of this parasitic bacterium, 'Candidatus Mycosynbacter amalyticus', may represent a promising strategy for the biocontrol of bacteria responsible for stabilizing wastewater foams.


Subject(s)
Actinobacteria/physiology , Bacteria/growth & development , Bacteria/isolation & purification , Wastewater/microbiology , Actinobacteria/virology , Bacteria/classification , Bacteria/genetics , Bacterial Physiological Phenomena , Bacteriophages/physiology , Genome, Bacterial , Phylogeny , Wastewater/chemistry
15.
PLoS One ; 16(2): e0245613, 2021.
Article in English | MEDLINE | ID: mdl-33529190

ABSTRACT

Sugarcane ratoon stunting disease (RSD) caused by Leifsonia xyli subsp. xyli (Lxx) is a common destructive disease that occurs around the world. Lxx is an obligate pathogen of sugarcane, and previous studies have reported some physiological responses of RSD-affected sugarcane. However, the molecular understanding of sugarcane response to Lxx infection remains unclear. In the present study, transcriptomes of healthy and Lxx-infected sugarcane stalks and leaves were studied to gain more insights into the gene activity in sugarcane in response to Lxx infection. RNA-Seq analysis of healthy and diseased plants transcriptomes identified 107,750 unigenes. Analysis of these unigenes showed a large number of differentially expressed genes (DEGs) occurring mostly in leaves of infected plants. Sugarcane responds to Lxx infection mainly via alteration of metabolic pathways such as photosynthesis, phytohormone biosynthesis, phytohormone action-mediated regulation, and plant-pathogen interactions. It was also found that cell wall defense pathways and protein phosphorylation/dephosphorylation pathways may play important roles in Lxx pathogeneis. In Lxx-infected plants, significant inhibition in photosynthetic processes through large number of differentially expressed genes involved in energy capture, energy metabolism and chloroplast structure. Also, Lxx infection caused down-regulation of gibberellin response through an increased activity of DELLA and down-regulation of GID1 proteins. This alteration in gibberellic acid response combined with the inhibition of photosynthetic processes may account for the majority of growth retardation occurring in RSD-affected plants. A number of genes associated with plant-pathogen interactions were also differentially expressed in Lxx-infected plants. These include those involved in secondary metabolite biosynthesis, protein phosphorylation/dephosphorylation, cell wall biosynthesis, and phagosomes, implicating an active defense response to Lxx infection. Considering the fact that RSD occurs worldwide and a significant cause of sugarcane productivity, a better understanding of Lxx resistance-related processes may help develop tools and technologies for producing RSD-resistant sugarcane varieties through conventional and/or molecular breeding.


Subject(s)
Actinobacteria/physiology , Gram-Positive Bacterial Infections/genetics , Host-Pathogen Interactions/genetics , Plant Diseases/genetics , Saccharum/genetics , Saccharum/microbiology , Transcriptome , Gene Expression Regulation, Plant , Genes, Plant , Gram-Positive Bacterial Infections/microbiology , Photosynthesis/genetics , Plant Diseases/microbiology , Plant Leaves/genetics , Plant Leaves/microbiology , RNA, Plant/genetics , RNA, Plant/isolation & purification , RNA-Seq , Reverse Transcriptase Polymerase Chain Reaction , Saccharum/metabolism , Signal Transduction/genetics
16.
Arch Microbiol ; 203(4): 1367-1374, 2021 May.
Article in English | MEDLINE | ID: mdl-33386868

ABSTRACT

An alkaliphilic actinomycete, designated HAJB-30 T, was isolated from a soda alkali-saline soil in Heilongjiang, Northeast China. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain HAJB-30 T was most closely related to type strains of the genus Phytoactinopolyspora with sequence similarities ranging between 93.5 and 98.9%. Strain HAJB-30 T grew at pH 8.0-11.0 (optimum pH 9.5-10.0) and in the presence of 0-7.0% NaCl (optimum 1.0-3.0%). Whole-cell hydrolysates of the isolate contained LL-diaminopimelic acid as the diagnostic diamino acid and mannose and rhamnose as diagnostic sugars. The major fatty acids identified were iso-C14:0, iso-C15:0, anteiso-C15:0, iso-C16:0 and anteiso-C17:0, while the menaquinone was MK-9(H4). The genome (6,589,901 bp), composed of 50 contigs, had a G + C content of 66.8%. Out of the 6074 predicted genes, 6020 were protein-coding genes, and 54 were ncRNAs. Digital DNA-DNA hybridization (dDDH) estimation and average nucleotide identity (ANI) of strain HAJB-30 T against genomes of the type strains of related species in the same family ranged between 19.7 and 22.0% and between 71.5 and 76.8%, respectively. From these results, it was concluded that strain HAJB-30 T possesses sufficient characteristics differentiated from all recognized Phytoactinopolyspora species, it is considered to be a novel species for which the name Phytoactinopolyspora limicola sp. nov. is proposed. The type strain is HAJB-30 T (= CGMCC 4.7591 T, = JCM 33694 T).


Subject(s)
Actinobacteria/classification , Actinobacteria/physiology , Soil Microbiology , Actinobacteria/chemistry , Actinobacteria/genetics , Base Composition , DNA, Bacterial/genetics , Diaminopimelic Acid/analysis , Fatty Acids/analysis , Genome, Fungal/genetics , Nucleic Acid Hybridization , Phospholipids/chemistry , Phylogeny , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Soil/chemistry , Sugars/analysis , Vitamin K 2/analysis
17.
J Microbiol ; 59(4): 360-368, 2021 Apr.
Article in English | MEDLINE | ID: mdl-33496938

ABSTRACT

Three novel bacterial strains, HDW9AT, HDW9BT, and HDW9CT, isolated from the intestine of the diving beetles Cybister lewisianus and Cybister brevis, were characterized as three novel species using a polyphasic approach. The isolates were Gram-staining-positive, strictly aerobic, non-motile, and rod-shaped. They grew optimally at 30°C (pH 7) in the presence of 0.5% (wt/vol) NaCl. Phylogenetic analysis based on the 16S rRNA gene sequences revealed that they belong to the genus Leucobacter and are closely related to L. denitrificans M1T8B10T (98.4-98.7% sequence similarity). Average nucleotide identity (ANI) values among the isolates were 76.4-84.1%. ANI values for the isolates and the closest taxonomic species, L. denitrificans KACC 14055T, were 72.3-73.1%. The isolates showed ANI values of < 76.5% with all analyzable Leucobacter strains in the EzBioCloud database. The genomic DNA G + C content of the isolates was 60.3-62.5%. The polar lipid components were phosphatidylglycerol, diphosphatidylglycerol, and other unidentified glycolipids, phospholipids, and lipids. The major cellular fatty acids were anteiso-C15:0, iso-C16:0, and anteiso-C17:0. MK-10 was the major respiratory quinone, and MK-7 and MK-11 were the minor respiratory quinones. The whole-cell sugar components of the isolates were ribose, glucose, galactose, and mannose. The isolates harbored L-2,4-diaminobutyric acid, L-serine, L-lysine, L-aspartic acid, glycine, and D-glutamic acid within the cell wall peptidoglycan. Based on phylogenetic, phenotypic, chemotaxonomic, and genotypic analyses, strains HDW9AT, HDW9BT, and HDW9CT represent three novel species within the genus Leucobacter. We propose the name Leucobacter coleopterorum sp. nov. for strain HDW9AT (= KACC 21331T = KCTC 49317T = JCM 33667T), the name Leucobacter insecticola sp. nov. for strain HDW9BT (= KACC 21332T = KCTC 49318T = JCM 33668T), and the name Leucobacter viscericola sp. nov. for strain HDW9CT (= KACC 21333T = KCTC 49319T = JCM 33669T).


Subject(s)
Actinobacteria/classification , Coleoptera/microbiology , Phylogeny , Actinobacteria/isolation & purification , Actinobacteria/physiology , Animals , Bacterial Typing Techniques , Base Composition , Cell Wall/chemistry , DNA, Bacterial/genetics , Fatty Acids/chemistry , Intestines/microbiology , Peptidoglycan/chemistry , RNA, Ribosomal, 16S/genetics , Vitamin K 2/chemistry
18.
J Appl Microbiol ; 130(5): 1508-1522, 2021 May.
Article in English | MEDLINE | ID: mdl-33010096

ABSTRACT

AIMS: This study aimed to isolate actinomycetes from marine environments and examine their antifungal activity against Talaromyces marneffei both in vitro and in vivo. METHODS AND RESULTS: Nineteen out of 101 actinomycete extracts were active and further determined for their minimum inhibitory concentrations (MIC). Three extracts of AMA50 that isolated from sediment showed strong antifungal activity against T. marneffei yeast (MICs ≤0·03-0·25 µg ml-1 ) and mould (MICs 0·5-16 µg ml-1 ) forms. The hexane extract from the cells of AMA50 (AMA50CH) exhibited the best activity against both the forms (MIC ≤ 1 µg ml-1 ). Three extracts from AMA50 killed the melanized yeast cells at 0·5 µg ml-1 . The AMA50CH was further tested for protective effects in Caenorhabditis elegans model. At concentrations of 1-8 µg ml-1 , the AMA50CH prolonged survival of T. marneffei-infected C. elegans with a 60-70% survival rate. The composition of AMA50CH was determined by gas chromatography-mass spectrometry. The major components were n-hexadecanoic acid, tetradecanoic acid and pentadecanoic acid. Sequencing analysis revealed that isolate AMA50 belonged to the genus Streptomyces. CONCLUSIONS: The AMA50CH from Streptomyces sp. AMA50 was the most effective extract against T. marneffei. SIGNIFICANCE AND IMPACT OF THE STUDY: Talaromyces marneffei is one of the most important thermally dimorphic pathogenic fungi. These results indicated the potency of marine-derived actinomycete extracts against T. marneffei both in vitro and in vivo.


Subject(s)
Actinobacteria/physiology , Antibiosis , Antifungal Agents/pharmacology , Talaromyces/drug effects , Talaromyces/physiology , Actinobacteria/chemistry , Actinobacteria/isolation & purification , Animals , Antifungal Agents/isolation & purification , Antifungal Agents/toxicity , Aquatic Organisms/microbiology , Caenorhabditis elegans/drug effects , Caenorhabditis elegans/microbiology , Geologic Sediments/microbiology , Microbial Sensitivity Tests , Talaromyces/ultrastructure
19.
Biomolecules ; 10(12)2020 12 15.
Article in English | MEDLINE | ID: mdl-33333896

ABSTRACT

The use of actinomycetes for improving soil fertility and plant production is an attractive strategy for developing sustainable agricultural systems due to their effectiveness, eco-friendliness, and low production cost. Out of 17 species isolated from the soil rhizosphere of legume crops, 4 bioactive isolates were selected and their impact on 5 legumes: soybean, kidney bean, chickpea, lentil, and pea were evaluated. According to the morphological and molecular identification, these isolates belong to the genus Streptomyces. Here, we showed that these isolates increased soil nutrients and organic matter content and improved soil microbial populations. At the plant level, soil enrichment with actinomycetes increased photosynthetic reactions and eventually increased legume yield. Actinomycetes also increased nitrogen availability in soil and legume tissue and seeds, which induced the activity of key nitrogen metabolizing enzymes, e.g., glutamine synthetase, glutamate synthase, and nitrate reductase. In addition to increased nitrogen-containing amino acids levels, we also report high sugar, organic acids, and fatty acids as well as antioxidant phenolics, mineral, and vitamins levels in actinomycete treated legume seeds, which in turn improved their seed quality. Overall, this study shed the light on the impact of actinomycetes on enhancing the quality and productivity of legume crops by boosting the bioactive primary and secondary metabolites. Moreover, our findings emphasize the positive role of actinomycetes in improving the soil by enriching its microbial population. Therefore, our data reinforce the usage of actinomycetes as biofertilizers to provide sustainable food production and achieve biosafety.


Subject(s)
Actinobacteria/physiology , Fabaceae/growth & development , Nitrogen/metabolism , Seeds/physiology , Soil , Actinobacteria/isolation & purification , Actinobacteria/ultrastructure , Amino Acids/analysis , Fatty Acids/analysis , Photosynthesis , Phylogeny , RNA, Ribosomal, 16S/genetics , Rhizosphere
20.
PLoS One ; 15(12): e0234893, 2020.
Article in English | MEDLINE | ID: mdl-33382695

ABSTRACT

Breast cancer is the second leading cause of cancer-related mortality in women. Various nutritional compounds possess anti-carcinogenic properties which may be mediated through their effects on the gut microbiota and its production of short-chain fatty acids (SCFAs) for the prevention of breast cancer. We evaluated the impact of broccoli sprouts (BSp), green tea polyphenols (GTPs) and their combination on the gut microbiota and SCFAs metabolism from the microbiota in Her2/neu transgenic mice that spontaneously develop estrogen receptor-negative [ER(-)] mammary tumors. The mice were grouped based on the dietary treatment: control, BSp, GTPs or their combination from beginning in early life (BE) or life-long from conception (LC). We found that the combination group showed the strongest inhibiting effect on tumor growth volume and a significant increase in tumor latency. BSp treatment was integrally more efficacious than the GTPs group when compared to the control group. There was similar clustering of microbiota of BSp-fed mice with combination-fed mice, and GTPs-fed mice with control-fed mice at pre-tumor in the BE group and at pre-tumor and post-tumor in the LC group. The mice on all dietary treatment groups incurred a significant increase of Adlercreutzia, Lactobacillus genus and Lachnospiraceae, S24-7 family in the both BE and LC groups. We found no change in SCFAs levels in the plasma of BSp-fed, GTPs-fed and combination-fed mice of the BE group. Marked changes were observed in the mice of the LC group consisting of significant increases in propionate and isobutyrate in GTPs-fed and combination-fed mice. These studies indicate that nutrients such as BSp and GTPs differentially affect the gut microbial composition in both the BE and LC groups and the key metabolites (SCFAs) levels in the LC group. The findings also suggest that temporal factors related to different time windows of consumption during the life-span can have a promising influence on the gut microbial composition, SCFAs profiles and ER(-) breast cancer prevention.


Subject(s)
Diet/methods , Fatty Acids, Volatile/blood , Gastrointestinal Microbiome/drug effects , Mammary Neoplasms, Experimental/prevention & control , Polyphenols/pharmacology , Seedlings/chemistry , Actinobacteria/drug effects , Actinobacteria/isolation & purification , Actinobacteria/physiology , Animals , Brassica/chemistry , Clostridiales/drug effects , Clostridiales/isolation & purification , Clostridiales/physiology , Female , Gastrointestinal Microbiome/physiology , Gene Expression , Lactobacillus/drug effects , Lactobacillus/isolation & purification , Lactobacillus/physiology , Mammary Glands, Animal/drug effects , Mammary Glands, Animal/metabolism , Mammary Glands, Animal/pathology , Mammary Neoplasms, Experimental/blood , Mammary Neoplasms, Experimental/genetics , Mammary Neoplasms, Experimental/pathology , Mice , Mice, Knockout , Polyphenols/chemistry , Receptor, ErbB-2/deficiency , Receptor, ErbB-2/genetics , Receptors, Estrogen/deficiency , Receptors, Estrogen/genetics , Tea/chemistry
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