ABSTRACT
There is an enormous diversity of life forms present in the extremely intricate marine environment. The growth and development of seaweeds in this particular environment are controlled by the bacteria that settle on their surfaces and generate a diverse range of inorganic and organic chemicals. The purpose of this work was to identify epiphytic and endophytic bacterial populations associated with ten common marine macroalgae from various areas along the Mediterranean Sea coast in Alexandria. This was done to target their distribution and possible functional aspects. Examine the effects of the algal habitat on the counting and phenotypic characterization of bacteria, which involves grouping bacteria based on characteristics such as shape, colour, mucoid nature, type of Gram stain, and their ability to generate spores. Furthermore, studying the physiological traits of the isolates under exploration provides insight into the optimum environmental circumstances for bacteria associated with the formation of algae. The majority of the bacterial isolates exhibited a wide range of enzyme activities, with cellulase, alginase, and caseinase being the most prevalent, according to the data. Nevertheless, 26% of the isolates displayed amylolytic activity, while certain isolates from Miami, Eastern Harbor, and Montaza lacked catalase activity. Geographical variations with the addition of algal extract may impact on the enumeration of the bacterial population, and this might have a relationship with host phylogeny. The most significant observation was that endophytic bacteria associated with green algae increased in all sites, while those associated with red algae increased in Abu Qir and Miami sites and decreased in Eastern Harbor. At the species level, the addition of algal extract led to a ninefold increase in the estimated number of epiphytic bacteria for Cladophora pellucida in Montaza. Notably, after adding algal extract, the number of presented endophytic bacteria associated with Codium sp. increased in Abu Qir while decreasing with the same species in Montaza. In addition to having the most different varieties of algae, Abu Qir has the most different bacterial isolates.
Subject(s)
Bacteria , Endophytes , Phylogeny , RNA, Ribosomal, 16S , Seaweed , Bacteria/classification , Bacteria/isolation & purification , Bacteria/genetics , Egypt , Seaweed/microbiology , Endophytes/classification , Endophytes/isolation & purification , Endophytes/genetics , Endophytes/physiology , Mediterranean Sea , RNA, Ribosomal, 16S/genetics , Biodiversity , Seawater/microbiology , DNA, Bacterial/genetics , EcosystemABSTRACT
BACKGROUND: Taxol, derived from Taxus trees, is a valuable natural resource for the development of anticancer drugs. Endophytic fungi from Taxus trees are a promising alternative source of Taxol. However, the impact of plant-endophytic microbial interaction on the host's Taxol biosynthesis is largely unknown. RESULTS: In the current study, the diversity of endophytic fungi in three different Taxus species was analyzed using Internal Transcribed Spacer sequencing. A total of 271 Operational Taxonomic Units (OTUs) were identified, grouping into 2 phyla, 8 classes, 16 orders, 19 families, and 19 genera. Alpha and beta diversity analysis indicated significant differences in endophytic fungal communities among the various Taxus trees. At the genus level, Alternaria and Davidiella were predominantly found in T. mairei and T. media, respectively. By utilizing a previously published dataset, a Pearson correlation analysis was conducted to predict the taxol biosynthesis-related fungal genera. Following screening, two isolates of Alternaria (L7 and M14) were obtained. Effect of inoculation with Alternaria isolates on the gene expression and metabolite accumulation of T. mairei was determined by transcriptomic and untargeted metabolomic studies. The co-inoculation assay suggests that the two Alternaria isolates may have a negative regulatory effect on taxol biosynthesis by influencing hormone signaling pathways. CONCLUSION: Our findings will serve as a foundation for advancing the production and utilization of Taxus and will also aid in screening endophytic fungi related to taxol production.
Subject(s)
Alternaria , Endophytes , Paclitaxel , Taxus , Taxus/microbiology , Paclitaxel/biosynthesis , Endophytes/genetics , Endophytes/metabolism , Endophytes/isolation & purification , Endophytes/classification , Alternaria/genetics , Alternaria/metabolism , Alternaria/classification , Alternaria/isolation & purification , Phylogeny , Fungi/genetics , Fungi/metabolism , Fungi/classification , Fungi/isolation & purification , DNA, Fungal/genetics , DNA, Ribosomal Spacer/geneticsABSTRACT
Endophytic bacteria found in marine macroalgae have been studied for their potential antimicrobial activity, consequently, they could serve as a valuable source of bioactive compounds to control pathogenic bacteria, yeasts, and fungi. Algae endophytic bacteria were isolated from Caulerpa sp., Ulva sp., Ahnfeltiopsis sp., and Chondracantus chamissoi from Yacila and Cangrejo Beaches (Piura, Peru). Antimicrobial assays against pathogenic bacteria were evaluated using cross-culture, over-plate, and volatile organic compound tests. Afterward, the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of selected crude extracts were determined, also ITS molecular analysis, antifungal activity, and PCR of iturin, fengycin, and surfactin genes were performed for bacteria strains exhibiting better activity. Forty-six algae endophytic bacteria were isolated from algae. Ten strains inhibited gram-positive pathogenic bacteria (Enterococcus faecalis, Staphylococcus epidermidis, S. aureus, and Listeria monocytogenes), and 12 inhibited gram-negative bacteria (Escherichia coli and Salmonella enteric sv typhimurium). Bacteria with better activity belong to Bacillus sp., Kluyvera ascorbata, Pantoea agglomerans, Leclercia adecarboxylata, and Enterobacter sp., which only four showed antifungal activities against Candida albicans, C. tropicalis, Colletotrichium sp., Fusarium sp., Fusarium oxysporum, and Alternaria sp. Furthermore, K. ascorbata YAFE21 and Bacillus sp. YCFE4 exhibited iturin and fengycin genes. The results indicate that the algae endophytic bacteria found in this study, particularly K. ascorbata YAFE21, Bacillus sp. YCFR6, L. adecarboxylata CUFE2, Bacillus sp. YUFE8, Enterobacter sp. YAFL1, and P. agglomerans YAFL6, could be investigated as potential producers of antimicrobial compounds due to their broad activity against various microorganisms.
Subject(s)
Endophytes , Microbial Sensitivity Tests , Seaweed , Endophytes/isolation & purification , Endophytes/genetics , Endophytes/metabolism , Endophytes/chemistry , Endophytes/classification , Seaweed/microbiology , Bacteria/drug effects , Bacteria/isolation & purification , Bacteria/classification , Anti-Infective Agents/pharmacology , Anti-Infective Agents/isolation & purification , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/isolation & purification , Antifungal Agents/pharmacology , Antifungal Agents/isolation & purification , Fungi/drug effects , Fungi/isolation & purification , Fungi/classification , Gram-Negative Bacteria/drug effects , Ulva/microbiology , Caulerpa/microbiology , Gram-Positive Bacteria/drug effectsABSTRACT
Understanding the resident microbial communities and their above and below ground interactions with plants will provide necessary information for crop disease protection and stress management. In this study, we show how diversity of core microbiome varies with disease susceptibility of a crop. To test this hypothesis, we have focused on identifying the core microbial species of cotton leaf curl disease (CLCuD) susceptible Gossypium hirsutum and CLCuD resistant Gossypium arboreum under viral infestation. Derivation of core membership is challenging as it depends on an occupancy threshold of microbial species in a sampling pool, whilst accounting for different plant compartments. We have used an abundance-occupancy distribution approach where we dynamically assess the threshold for core membership, whilst marginalizing for occupancy in four compartments of the cotton plant, namely, leaf epiphyte, leaf endophyte, rhizosphere, and root endophyte. Additionally, we also fit a neutral model to the returned core species to split them into three groups, those that are neutral, those that are selected by the plant environment, and finally those that are dispersal limited. We have found strong inverse relationship between diversity of core microbiome and disease susceptibility with the resistant variety, G. arboreum, possessing higher diversity of microbiota. A deeper understanding of this association will aid in the development of biocontrol agents for improving plant immunity against biotrophic pathogens.
Subject(s)
Disease Resistance , Gossypium , Microbiota , Plant Diseases , Gossypium/microbiology , Plant Diseases/microbiology , Plant Leaves/microbiology , Endophytes/classification , Endophytes/genetics , Endophytes/isolation & purification , Bacteria/classification , Bacteria/genetics , Bacteria/isolation & purification , Plant Roots/microbiology , Biodiversity , Rhizosphere , Soil MicrobiologyABSTRACT
Plants intimately coexist with diverse taxonomically structured microbial communities that influence host health and productivity. The coexistence of plant microbes in the phyllosphere benefits biodiversity maintenance, ecosystem function, and community stability. However, differences in community composition and network structures of phyllosphere epiphytic and endophytic fungi are widely unknown. Using Illumina Miseq sequencing of internal transcribed spacer (ITS) and 28S rRNA gene amplicons, we characterised the epiphytic and endophytic fungal communities associated with cashew phyllosphere (leaf, flower and fruit) from Kwale, Kilifi and Lamu counties in Kenya. The ITS and 28S rRNA gene sequences were clustered into 267 and 108 operational taxonomic units (OTUs) at 97% sequence similarity for both the epiphytes and endophytes. Phylum Ascomycota was abundant followed by Basidiomycota, while class Saccharomycetes was most dominant followed by Dothideomycetes. The major non-ascomycete fungi were associated only with class Tremellales. The fungal communities detected had notable ecological functions as saprotrophs and pathotrophs in class Saccharomyectes and Dothideomycetes. The community composition of epiphytic and endophytic fungi significantly differed between the phyllosphere organs which was statistically confirmed by the Analysis of Similarity test (ANOSIM Statistic R: 0.3273, for 28S rRNA gene and ANOSIM Statistic R: 0.3034 for ITS). The network analysis revealed that epiphytic and endophytic structures were more specialized, modular and had less connectance. Our results comprehensively describe the phyllosphere cashew-associated fungal community and serve as a foundation for understanding the host-specific microbial community structures among cashew trees.
Subject(s)
Anacardium , Endophytes , Kenya , Anacardium/microbiology , Endophytes/genetics , Endophytes/classification , Fungi/genetics , Fungi/classification , Fungi/isolation & purification , DNA Barcoding, Taxonomic , RNA, Ribosomal, 28S/genetics , Mycobiome/genetics , Biodiversity , Phylogeny , Plant Leaves/microbiology , DNA, Fungal/geneticsABSTRACT
This work presents the isolation of endophytic fungi from the leaves of Campomanesia adamantium (Cambess.) O. Berg (Myrtaceae), a native species found in Brazil and popularly known as "guabiroba-do-campo", with abundant distribution in the Brazilian Cerrado. It has been popularly used for its anti-rheumatic, anti-inflammatory, antidiarrheal, blood cholesterol-reducing, urinary antiseptic, and depurative properties. Theese fungi are microorganisms that live inside higher plants, at least for a period of their life cycle, occupying the intercellular spaces of plant tissues such as leaves and stems. These fungi are harmless to the host plant, and their secondary metabolites promote protection, regulate growth, combat bacteria, viruses, and fungi, and promote resistance to abiotic stress, as well as insecticidal effects. Endophytic fungi associated with the leaves of C. adamantium were isolated using the culture medium isolation technique. After growth, the fungi were divided into groups based on morphotypes. Fungal genomic DNA was extracted, and a polymerase chain reaction (PCR) was conducted to amplify the ITS1-5.8S-ITS2 regions of rRNA, and the nucleotide sequences obtained were compared with those available in the GenBank database for molecular identification of the isolates. The phylogenetic tree was constructed using MEGA 11 software. The results showed representatives of the Ascomycota phylum, and it was possible to identify at the genus level 18 fungi of the genera Colletotrichum, Diaporthe, Epicoccum, and Neofusicoccum.
Subject(s)
Endophytes , Fungi , Myrtaceae , Plant Leaves , Endophytes/isolation & purification , Endophytes/classification , Endophytes/genetics , Brazil , Myrtaceae/microbiology , Fungi/classification , Fungi/isolation & purification , Fungi/genetics , Plant Leaves/microbiology , DNA, Fungal/genetics , Polymerase Chain Reaction , PhylogenyABSTRACT
Beauveria bassiana (Bb) is a widespread entomopathogenic fungus widely used in agriculture for crop protection. Other than pest control, fungi belonging to the B. bassiana complex represent an important microbial resource in agroecosystems, considering their multiple interactions with other microorganisms as antagonists of phytopathogens, or with plants as endophytic colonizers and growth promoters. Here, we characterised field collected or commercial isolates of B. bassiana relative to the environmental factors that affect their growth. We further compared the metabolome, the entomopathogenic potential and biocontrol activity of the tested isolates respectively on the insect pest Spodoptera littoralis or against the fungal plant pathogen Fusarium oxysporum. Our analysis revealed that the B. bassiana complex is characterised by a high level of inter-isolate heterogeneity in terms of nutritional requirements, establishment of intra- or inter-kingdom interactions, and the nature of metabolites produced. Interestingly, certain B. bassiana isolates demonstrated a preference for low nutrient plant-derived media, which hints at their adaptation towards an endophytic lifestyle over a saprophytic one. In addition, there was a noticeable variation among different B. bassiana isolates in their capacity to kill S. littoralis larvae in a contact infection test, but not in an intrahaemocoelic injection experiment, suggesting a unique level of adaptability specific to the host. On the other hand, most B. bassiana isolates exhibited similar biocontrol efficacy against the soil-dwelling ascomycete F. oxysporum f. sp. lycopersici, a pathogen responsible for vascular wilt disease in tomato plants, effectively averting wilting. Overall, we show that the effectiveness of B. bassiana isolates can greatly vary, emphasising the importance of isolate selection and nutritional adaptability consideration for their use in sustainable agriculture.
Subject(s)
Beauveria , Fusarium , Larva , Pest Control, Biological , Spodoptera , Beauveria/physiology , Beauveria/isolation & purification , Beauveria/metabolism , Animals , Spodoptera/microbiology , Larva/microbiology , Plant Diseases/microbiology , Plant Diseases/prevention & control , Agriculture , Metabolome , Endophytes/isolation & purification , Endophytes/metabolism , Endophytes/physiology , Endophytes/classificationABSTRACT
Here, we report on a Cordyceps species entering into a multi-trophic, multi-kingdom association. Cordyceps cateniannulata, isolated from the stem of wild Coffea arabica in Ethiopia, is shown to function as an endophyte, a mycoparasite and an entomopathogen. A detailed polyphasic taxonomic study, including a multilocus phylogenetic analysis, confirmed its identity. An emended description of C. cateniannulata is provided herein. Previously, this species was known as a pathogen of various insect hosts in both the Old and New World. The endophytic status of C. cateniannulata was confirmed by re-isolating it from inoculated coffee plants. Inoculation studies have further shown that C. cateniannulata is a mycoparasite of Hemileia vastatrix, as well as an entomopathogen of major coffee pests; infecting and killing Hypothenemus hampei and Leucoptera coffeella. This is the first record of C. cateniannulata from Africa, as well as an endophyte and a mycoparasite. The implications for its use as a biocontrol agent are discussed.
Subject(s)
Coffea , Cordyceps , Endophytes , Phylogeny , Endophytes/classification , Endophytes/isolation & purification , Endophytes/genetics , Endophytes/physiology , Cordyceps/genetics , Cordyceps/classification , Coffea/microbiology , Coffea/parasitology , Animals , Plant Diseases/microbiology , Plant Diseases/parasitology , Ethiopia , DNA, Fungal/genetics , DNA, Fungal/chemistry , DNA, Ribosomal Spacer/genetics , DNA, Ribosomal Spacer/chemistry , Plant Stems/microbiology , Plant Stems/parasitology , Sequence Analysis, DNA , Cluster AnalysisABSTRACT
In the present study, eleven endophytic bacterial strains, Herbaspirillum sp. (GP-SGM1, GP-SGM2, GP-SGM3, and GP-SGM11), Pseudomonas sp. (GP-SGM4, GP-SGM5), Novosphingobium sp. GP-SGM6, Chryseobacterium sp. GP-SGM7, Labedella sp. GP-SGM8, Brevibacterium sp. GP-SGM9, and Pseudomonas sp. GP-SGM10, were isolated from the rhizomes of Gunnera perpensa L. The growth kinetics, assessed through maximum growth rates (µmax) and optical density (OD) values, revealed that GP-SGM7 exhibited highest µmax values of 0.33 ± 0.01 hours (h)-1 with an OD of 4.20 ± 0.04. In contrast, GP-SGM11 exhibited the lowest µmax of 0.12 ± 0.05 h-1 and the smallest OD of 1.50 ± 0.00. In addition, the endophyte crude extracts were tested for antibacterial activity against five pathogenic strains using the disk diffusion method, with GP-SGM7 crude extracts exhibiting promising antibacterial activity against Klebsiella pneumoniae and Staphylococcus aureus. Antioxidant activity was determined by DPPH (2, 2-diphenyl-1-picrylhydrazyl) and FRAP (ferric reducing antioxidant power) assays. The crude extracts of GP-SGM1, GP-SGM7, GP-SGM9, and GP-SGM10 were the most effective at scavenging DPPH radicals, with GP-SGM7 also exhibiting a high FRAP value of 0.54 ± 0.01. These findings emphasize the therapeutic potential of endophytic bacteria from G. perpensa L. in addressing skin-related issues, including bacterial infections and free radicals.
Subject(s)
Bacteria , Endophytes , Endophytes/isolation & purification , Endophytes/classification , Endophytes/genetics , Endophytes/metabolism , Endophytes/chemistry , Bacteria/isolation & purification , Bacteria/classification , Bacteria/drug effects , Bacteria/genetics , Antioxidants/pharmacology , Antioxidants/metabolism , Antioxidants/chemistry , Anti-Bacterial Agents/pharmacology , Phylogeny , RNA, Ribosomal, 16S/genetics , Microbial Sensitivity TestsABSTRACT
Apple ring rot, one of the most common apple postharvest diseases during storage, is caused by Botryosphaeria dothidea. Presently, the disease management is primarily dependent on chemical fungicide application. Here we demonstrated an endophyte bacterium Bacillus tequilensis QNF2, isolated from Chinese leek (Allium tuberosum) roots considerably suppressed B. dothidea mycelial growth, with the highest suppression of 73.56 % and 99.5 % in the PDA and PDB medium, respectively in vitro confront experiments. In in vivo experiments, B. tequilensis QNF2 exhibited a control efficacy of 88.52 % and 100 % on ring rot disease on postharvest apple fruits inoculated with B. dothidea disc and dipped into B. dothidea culture, respectively. In addition, B. tequilensis QNF2 volatile organic compounds (VOCs) also manifested markedly inhibition against B. dothidea mycelial growth and the ring rot on postharvest apple fruits. Moreover, B. tequilensis QNF2 severely damaged the mycelial morphology of B. dothidea. Finally, B. tequilensis QNF2 significantly repressed the expression of six pathogenicity-related genes, such as adh, aldh, aldh3, galm, pdc1, pdc2, involved in glycolysis/gluconeogenesis of B. dothidea. The findings of the study proved that B. tequilensis QNF2 was a promising alternative for controlling apple ring rot of postharvest apple fruit.
Subject(s)
Ascomycota , Bacillus , Endophytes , Fruit , Malus , Plant Diseases , Malus/microbiology , Plant Diseases/microbiology , Ascomycota/growth & development , Ascomycota/drug effects , Ascomycota/genetics , Ascomycota/physiology , Bacillus/genetics , Bacillus/physiology , Bacillus/isolation & purification , Endophytes/genetics , Endophytes/metabolism , Endophytes/isolation & purification , Endophytes/classification , Endophytes/physiology , Fruit/microbiology , Volatile Organic Compounds/pharmacology , Volatile Organic Compounds/metabolism , Volatile Organic Compounds/analysis , Antibiosis , Mycelium/growth & development , Mycelium/drug effectsABSTRACT
Endophytes play an important role in plant development, survival, and establishment, but their temporal dynamics in young conifer plants are still largely unknown. In this study, the bacterial community was determined by metabarcoding of the 16S rRNA gene in the rhizoplane, roots, and aerial parts of 1- and 5-month-old seedlings of natural populations of Abies religiosa (Kunth) Schltdl. & Cham. In 1-month-old seedlings, Pseudomonas dominated aerial parts (relative abundance 71.6%) and roots (37.9%). However, the roots exhibited significantly higher bacterial species richness than the aerial parts, with the dissimilarity between these plant sections mostly explained by the loss of bacterial amplification sequence variants. After 5 months, Mucilaginibacter dominated in the rhizoplane (9.0%), Streptomyces in the roots (12.2%), and Pseudomonas in the aerial parts (18.1%). The bacterial richness and community structure differed significantly between the plant sections, and these variations were explained mostly by 1-for-1 substitution. The relative abundance of putative metabolic pathways significantly differed between the plant sections at both 1 and 5 months. All the dominant bacterial genera (e.g., Pseudomonas and Burkholderia-Caballeronia-Paraburkholderia) have been reported to have plant growth-promoting capacities and/or antagonism against pathogens, but what defines their role for plant development has still to be determined. This investigation improves our understanding of the early plant-bacteria interactions essential for natural regeneration of A. religiosa forest.
Subject(s)
Abies , Bacteria , Endophytes , Plant Roots , RNA, Ribosomal, 16S , Seedlings , Seedlings/microbiology , Seedlings/growth & development , Bacteria/classification , Bacteria/genetics , Bacteria/isolation & purification , Endophytes/classification , Endophytes/isolation & purification , Endophytes/physiology , Endophytes/genetics , RNA, Ribosomal, 16S/genetics , Abies/microbiology , Plant Roots/microbiology , Soil Microbiology , Biodiversity , Microbiota , DNA, Bacterial/geneticsABSTRACT
This study investigated the potential of endophytic fungi to produce paclitaxel (Taxol®), a potent anticancer compound widely employed in chemotherapy. This research aimed to identify, confirm, and characterize endophytic fungi capable of paclitaxel (PTX) production and assess their paclitaxel yield. Additionally, it aimed to investigate factors influencing paclitaxel production. A total of 100 endophytic fungal isolates were collected and identified from the roots of Artemisia judaica. Aspergillus fumigatiaffinis exhibited the highest PTX production (26.373 µg L-1) among the isolated endophytic fungi. The strain was identified as A. fumigatiaffinis (Accession No. PP235788.1). Molecular identification confirmed its novelty, representing the first report of PTX production by A. fumigatiaffinis, an endophyte of Artemisia judaica. Optimization through full factorial design of experiments (DOE) and response surface methodology (RSM) significantly enhanced PTX production to 110.23 µg L-1 from 1 g of dry weight of the fungal culture under optimal conditions of pH 8.0, 150 µg L-1 becozyme supplementation, and 18 days of fermentation in potato dextrose broth. The presence of paclitaxel was confirmed using thin layer chromatography, high performance liquid chromatography, and gas chromatography-mass spectrometry. These findings maximize the role of endophytic fungus to produce a secondary metabolite that might be able to replace the chemically produced PTX and gives an opportunity to provide a sustainable source of PTX eco-friendly at high concentrations. KEY POINTS: ⢠Endophytic fungi, like A. fumigatiaffinis, show promise for eco-friendly paclitaxel production ⢠Optimization strategies boost paclitaxel yield significantly, reaching 110.23 µg L -1 ⢠Molecular identification confirms novelty, offering a sustainable PTX source.
Subject(s)
Aspergillus , Endophytes , Fermentation , Paclitaxel , Paclitaxel/biosynthesis , Aspergillus/metabolism , Aspergillus/genetics , Endophytes/metabolism , Endophytes/genetics , Endophytes/isolation & purification , Endophytes/classification , Plant Roots/microbiology , Culture Media/chemistry , Gas Chromatography-Mass Spectrometry , Chromatography, High Pressure LiquidABSTRACT
Two endophytic actinobacteria, strains MK5T and MK7, were isolated from the surface-sterilized root of Jasmine rice (Oryza sativa KDML 105). These strains were aerobic actinobacteria with a well-developed substrate and aerial mycelia that formed spiral spore chains. The type strains that shared the high 16S rRNA gene sequence similarity with both strains were Streptomyces naganishii NBRC 12892T (99.4%), "Streptomyces griseicoloratus" TRM S81-3T (99.2%), and Streptomyces spiralis NBRC 14215T (98.9%). Strains MK5T and MK7 are the same species sharing a digital DNA-DNA hybridization (dDDH) value of 95.3% and a 16S rRNA gene sequence similarity of 100%. Chemotaxonomic data confirmed the affiliation of strains MK5T and MK7 to the genus Streptomyces. Strains MK5T and MK7 contained MK-9(H4) as a major menaquinone; the whole-cell sugar of both strains was galactose and glucose. The strain MK5T shared 93.4% average nucleotide identity (ANI)-Blast, 95.5% ANI-MUMmer, 93% average amino acid identity, and 61.3% dDDH with S. spiralis NBRC 14215T. The polyphasic approach confirmed that strain MK5T represents a novel species, and the name Streptomyces mahasarakhamensis sp. nov. is proposed. The type strain is MK5T (= TBRC 17754 = NRRL B-65683). Genome mining, using an in silico approach and searching biosynthesis gene clusters of strains MK5T and MK7, revealed that the genomes contained genes encoding proteins relating to plant growth promotion, bioactive compounds, and beneficial enzymes. Strains MK5T and MK7 could produce indole acetic acid and solubilize phosphate in vitro.
Subject(s)
DNA, Bacterial , Endophytes , Oryza , Phylogeny , RNA, Ribosomal, 16S , Streptomyces , Oryza/microbiology , Streptomyces/genetics , Streptomyces/isolation & purification , Streptomyces/classification , Streptomyces/metabolism , RNA, Ribosomal, 16S/genetics , Endophytes/genetics , Endophytes/classification , Endophytes/isolation & purification , Endophytes/metabolism , DNA, Bacterial/genetics , Plant Roots/microbiology , Plant Growth Regulators/metabolism , Vitamin K 2/analogs & derivatives , Bacterial Typing Techniques , Sequence Analysis, DNA , Nucleic Acid Hybridization , Fatty Acids/metabolism , Base CompositionABSTRACT
Lycoris radiata is the main source of galanthamine, a clinical drug used in Alzheimer's disease; however, the galanthamine content in L. radiata is low. Lycoris aurea is another Lycoris species with high galanthamine content. Fungal endophytes can enhance plant secondary metabolite accumulation; thus, we compared the fungal communities in these two Lycoris species to identify certain fungal taxa in L. aurea capable of enhancing galanthamine accumulation. Several fungal endophytes, which were enriched in, exclusively isolated from L. aurea, or showed significant correlations with galanthamine, were demonstrated to enhance the accumulation of only galanthamine but no other Amaryllidaceae alkaloids (AAs) in L. radiata. These fungal endophytes mainly upregulated the downstream genes in the biosynthesis pathways of AAs in L. radiata, suggesting that they may allocate more precursors for galanthamine biosynthesis. This study demonstrated that fungal endophytes from L. aurea with higher galanthamine content can specifically enhance the accumulation of this medicinal alkaloid in other Lycoris species, thereby increasing the galanthamine source and reducing galanthamine separation and purification costs. This study broadens our understanding of the complex interactions between plant secondary metabolites and fungal endophytes.
Subject(s)
Endophytes , Fungi , Galantamine , Lycoris , Galantamine/metabolism , Lycoris/metabolism , Lycoris/microbiology , Endophytes/metabolism , Endophytes/isolation & purification , Endophytes/classification , Endophytes/genetics , Fungi/classification , Fungi/metabolism , Fungi/genetics , Fungi/isolation & purification , Secondary Metabolism , Amaryllidaceae Alkaloids/metabolism , Biosynthetic Pathways/genetics , MycobiomeABSTRACT
BACKGROUND: Rhizosphere bacterial community and endophytes are now known to influence plant health and response to environmental stress. Very few studies have reported the diversity of endophytic bacterial communities of Vanilla planifolia and their potential roles in promoting plant growth or contributing to aromatic quality. RESULTS: In this study, the composition and diversity of the Vanilla rhizosphere bacterial community were explored by analyzing rhizosphere soil and root tissue samples as well as green pods of three accessions of Vanilla planifolia grown on different types of substrates (compost and leaf litter). In addition, the endophytic bacterial diversity of roots and green pods as well as the evolution of endophytic bacteria after the curing process of vanilla green pods were analyzed based on a metabarcoding approach. The results showed that bacterial species richness and diversity were higher in the compost. The analysis of the soil bacterial composition displayed that Halomonas, Pseudoalteromonas, Enterobacter and Bradyrhizobium were the most abundant genera. Moreover, the results indicated that the soil bacterial community structure was linked to the host plant genotype. Regarding the roots endophytic bacteria composition, the genera Halomonas, Pseudoalteromonas, Bacillus and Carboxydocella genera were present in all samples, independently from the substrate nature. Several genera including Bacillus, Bradyrhizobium, Burkholderia and Halomonas were transmitted internally from the roots to the green pods. The curing process reduced the bacterial richness and bacterial diversity associated with the green pods. Halomonas, Pseudoalteromonas, Bacillus, and Carboxydocella are the dominant genera in the pods after the curing process. CONCLUSIONS: This study provides an overview of changes of the bacterial communities dynamics especially endophytic in the roots and the green pods. It highlighted bacterial genera (Halomonas, Pseudoalteromonas, Bacillus, and Carboxydocella) potentially implicated in the formation of aroma compounds of vanilla beans.
Subject(s)
Bacteria , Biodiversity , Endophytes , Plant Roots , Rhizosphere , Soil Microbiology , Vanilla , Endophytes/classification , Endophytes/genetics , Endophytes/isolation & purification , Bacteria/classification , Bacteria/genetics , Bacteria/isolation & purification , Plant Roots/microbiology , Vanilla/microbiology , RNA, Ribosomal, 16S/genetics , Phylogeny , Microbiota/genetics , DNA, Bacterial/geneticsABSTRACT
In order to provide a highly feasible research pathway for the control of larch shoot blight, healthy larch branches and leaves were collected from 13 sampling sites in 8 provinces in China. The antagonistic endophytic bacteria obtained from the screening were used to carry out disease control experiments in potted seedlings. The safety evaluation test was conducted on the antagonistic bacteria. Subsequently, the strains with better preventive effect and high safety were identified by morphological and molecular methods. A total of 391 strains of endophytic bacteria were isolated from healthy larch branches and leaves. Seventy-eight strains of larch endophytic bacteria with antagonistic effect were obtained by primary sieving. Ten strains of endophytic bacteria with obvious antagonism were further obtained by secondary sieving, and all of them had an inhibition rate of more than 57%. Among them, strains YN 2, JL 6, NMG 23, and JL 54 showed the highest inhibition rate of 63.16%-65.08%, which was significantly different from the other treatments. The results of the pot test showed that 14 days after inoculation with the pathogen, strains YN 2 and JL 54 were more effective in the control of larch shoot blight, with the preventive effects reaching 57.7% and 50.0%, respectively. Strains JL 6 and JL 54 were biologically safe in the safety evaluation test. Therefore, strain JL 54 was selected for identification. It was identified as Bacillus amyloliquefaciens through morphological observation, 16S rDNA sequence, gyrB gene sequence and 16S rDNA-gyrB tandem feature sequence analysis. IMPORTANCE: Larch shoot blight is a widely distributed, damaging, and rapidly spreading fungal disease of forest trees that poses a serious threat to larch plantations. Endophytic bacteria have biological effects on host plants against pests and diseases, and they have a growth-promoting effect on plants. In this paper, we investigated for the first time the biocontrol effect of endophytic bacteria on larch shoot blight by screening endophytic bacteria with the function of antagonizing dieback fungi. Bacillus amyloliquefaciens JL 54 has a better prospect of biocontrol against larch shoot blight, which lays the foundation for the application of this bacterium in the future.
Subject(s)
Antibiosis , Endophytes , Larix , Plant Diseases , Endophytes/isolation & purification , Endophytes/classification , Endophytes/physiology , Endophytes/genetics , Plant Diseases/microbiology , Plant Diseases/prevention & control , Larix/microbiology , Bacteria/isolation & purification , Bacteria/classification , Bacteria/genetics , Ascomycota/genetics , Ascomycota/growth & development , Ascomycota/isolation & purification , Ascomycota/physiology , China , Plant Leaves/microbiology , RNA, Ribosomal, 16S/genetics , PhylogenyABSTRACT
Endophytic fungi, pivotal in facilitating plant co-evolution, significantly enhance plant growth, stress resistance, and environmental adaptability. Despite their importance, the spatial distribution of stem endophytic fungi (SEF) within host plants remains poorly characterized. Here, we employed high-throughput sequencing to conduct a comparative analysis of SEF communities in Mussaenda pubescens on a regional scale. Our findings reveal that whole-SEF communities were overwhelmingly dominated by members of the phylum Ascomycota, accounting for 85.9 %, followed by Basidiomycota at 13.9 %, and that alpha diversity within the whole-SEF community of M. pubescens remains relatively consistent across sampling sites. However, significant variation was observed within conditionally abundant taxa (CAT), conditionally rare or abundant taxa (CRAT), and conditionally rare taxa (CRT). Climatic factors emerged as the primary influence on SEF community distribution, followed by spatial distance and stem chemical properties. Neutral community modeling results suggested that both stochastic and deterministic processes play a role in shaping whole-SEF communities, with deterministic processes having a stronger influence on CRT subcommunities. Furthermore, the CRT co-occurrence network exhibited a more complex structure, characterized by higher values of network betweenness and degree relative to CAT and CRAT subcommunities. These findings enhance our understanding of community assembly and ecological interactions between stem fungal endophytes, presenting opportunities for harnessing fungal resources for the benefit of humanity.
Subject(s)
Endophytes , Plant Stems , Endophytes/classification , Endophytes/isolation & purification , Endophytes/genetics , Plant Stems/microbiology , Ascomycota/classification , Ascomycota/genetics , Ascomycota/isolation & purification , Fungi/classification , Fungi/isolation & purification , Fungi/genetics , High-Throughput Nucleotide Sequencing , Basidiomycota/classification , Basidiomycota/genetics , Basidiomycota/isolation & purification , BiodiversityABSTRACT
Post-harvest decay of fresh agricultural produce is a major threat to food security globally. Synthetic fungicides, commonly used in practice for managing the post-harvest losses, have negative impacts on consumers' health. Studies have reported the effectiveness of fungal isolates from plants as biocontrol agents of post-harvest diseases, although this is still poorly established in tomatoes (Solanum lycopersicum L. cv. Jasmine). In this study, 800 endophytic fungi were isolated from mature green and ripe untreated and fungicide-treated tomato fruits grown in open soil and hydroponics systems. Of these, five isolates (Aureobasidium pullulans SUG4.1, Coprinellus micaceus SUG4.3, Epicoccum nigrum SGT8.6, Fusarium oxysporum HTR8.4, Preussia africana SUG3.1) showed antagonistic properties against selected post-harvest pathogens of tomatoes (Alternaria alternata, Fusarium solani, Fusarium oxysporum, Geotrichum candidum, Rhizopus stolonifera, Rhizoctonia solani), with Lactiplantibacillus plantarum as a positive control. P. africana SUG3.1 and C. micaceus SUG4.3 significantly inhibited growth of all the pathogens, with antagonistic capabilities comparable to that exhibited by L. plantarum. Furthermore, the isolates produced an array of enzymes, including among others, amylase, cellulose and protease; and were able to utilize several carbohydrates (glucose, lactose, maltose, mannitol, sucrose). In conclusion, P. africana SUG3.1 and C. micaceus SUG4.3 may complement L. plantarum as biocontrol agents against post-harvest pathogens of tomatoes.
Subject(s)
Endophytes , Fruit , Fungi , Plant Diseases , Solanum lycopersicum , Solanum lycopersicum/microbiology , Plant Diseases/microbiology , Plant Diseases/prevention & control , Fruit/microbiology , Endophytes/isolation & purification , Endophytes/physiology , Endophytes/classification , Fungi/isolation & purification , Fungi/physiology , Fungi/classification , Fungi/drug effects , Antibiosis , Biological Control Agents , Fungicides, Industrial/pharmacologyABSTRACT
Fungal endophytes inhabit a similar ecological niche to that occupied by many phytopathogens, with several pathogens isolated from healthy tissues in their latent phase. This study aimed to evaluate the pathogenicity, the colonisation ability, and the enzyme activity of 37 endophytic fungal isolates recovered from apparently healthy apple shoot and leaf tissues. The pathogenicity of the isolates was assessed on 'Royal Gala' and 'Braeburn' fruit and detached 'Royal Gala' shoots. For the non-pathogenic isolates, their ability to endophytically colonise detached 'Royal Gala' shoots was evaluated. Enzyme activity assays were undertaken to determine whether the pathogenicity of the endophytes was related to the production of the extracellular enzymes, amylase, cellulase, pectinase, protease, and xylanase. Of the 37 isolates studied, eight isolates, representing the genera Colletotrichum, Diaporthe, Fusarium, and Penicillium, were shown to be pathogenic on both apple shoots and fruit. Two isolates identified as Trichoderma atroviride, were pathogenic only on shoots, and three isolates, representing the genus Diaporthe, were pathogenic only on fruit. Of the remaining 24 isolates, 22 (Biscogniauxia (n = 8), Chaetomium (n = 4), Trichoderma (n = 3), Epicoccum (n = 2), Neosetophoma (n = 2), Xylaria (n = 1), Daldinia (n = 1), and Paraphaeosphaeria (n = 1)) were recovered from the inoculated apple shoots but two failed to colonise the shoot tissues. Of the isolates tested, 20 produced amylase, 15 cellulase, 25 pectinase, 26 protease, and 13 xylanase. There was no correlation between the range and type of enzymes produced by the isolates and their pathogenicity or ability to endophytically colonise the shoot tissue. The study showed that approximately one-third (13/37) of the isolates recovered from the apparently healthy apple shoot tissues were observed as latent pathogens. The isolates that did not cause disease symptoms may have the ability to reduce colonisation of apple tissues by pathogens including Neonectria ditissima associated with European canker of apple.
Subject(s)
Endophytes , Fungi , Malus , Plant Leaves , Malus/microbiology , Endophytes/isolation & purification , Endophytes/classification , Endophytes/genetics , Plant Leaves/microbiology , Fungi/isolation & purification , Fungi/classification , Fungi/genetics , Fungi/pathogenicity , Plant Diseases/microbiology , Plant Shoots/microbiology , Fruit/microbiologyABSTRACT
BACKGROUND: Rhizosphere and endophytic fungi play important roles in plant health and crop productivity. However, their community dynamics during the continuous cropping of Knoxia valerianoides have rarely been reported. K. valerianoides is a perennial herb of the family Rubiaceae and has been used in herbal medicines for ages. Here, we used high-throughput sequencing technology Illumina MiSeq to study the structural and functional dynamics of the rhizosphere and endophytic fungi of K. valerianoides. RESULTS: The findings indicate that continuous planting has led to an increase in the richness and diversity of rhizosphere fungi, while concomitantly resulting in a decrease in the richness and diversity of root fungi. The diversity of endophytic fungal communities in roots was lower than that of the rhizosphere fungi. Ascomycota and Basidiomycota were the dominant phyla detected during the continuous cropping of K. valerianoides. In addition, we found that root rot directly affected the structure and diversity of fungal communities in the rhizosphere and the roots of K. valerianoides. Consequently, both the rhizosphere and endophyte fungal communities of root rot-infected plants showed higher richness than the healthy plants. The relative abundance of Fusarium in two and three years old root rot-infected plants was significantly higher than the control, indicating that continuous planting negatively affected the health of K. valerianoides plants. Decision Curve Analysis showed that soil pH, organic matter (OM), available K, total K, soil sucrase (S_SC), soil catalase (S_CAT), and soil cellulase (S_CL) were significantly related (p < 0.05) to the fungal community dynamics. CONCLUSIONS: The diversity of fungal species in the rhizosphere and root of K. valerianoides was reported for the first time. The fungal diversity of rhizosphere soil was higher than that of root endophytic fungi. The fungal diversity of root rot plants was higher than that of healthy plants. Soil pH, OM, available K, total K, S_CAT, S_SC, and S_CL were significantly related to the fungal diversity. The occurrence of root rot had an effect on the community structure and diversity of rhizosphere and root endophytic fungi.