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1.
Curr Microbiol ; 81(11): 393, 2024 Oct 06.
Article in English | MEDLINE | ID: mdl-39369364

ABSTRACT

Endophytes have drawn attentions due to their effectiveness in providing benefits to host and non-host plants. In this study endophytic bacteria were isolated from stem and leaf samples of medicinally important plants Rhododendron griffithianum Wight and Rhododendron arboreum Smith subsp. cinnamomeum (Wall. ex G. Don) grown at higher altitudes of Darjeeling, India. Two endophytic bacteria, Pseudomonas lurida RGDS03 and Bacillus velezensis RCDL12 were identified based on 16S rRNA gene sequencing analysis. The endophytes exhibited indole acetic acid (IAA), gibberellic acid (GA), siderophore production, phosphate solubilization, nitrogen-fixing abilities, though B. velezensis RCDL12 showed superior production of IAA (126.04 ± 0.40 µg/mL), GA (241.00 ± 0.44 µg/mL), and phosphate (74.4 ± 0.41 µg/mL) solubilization as compared to P. lurida RGDS03. Purity of extracted IAA from these two endophytes was confirmed by HPLC and LC-MS analysis. In this study, P. lurida RGDS03 inhibited mycelial growth of two tested phytopathogens Phytophthora sp. and Pestalotiopsis sp. of broad host range. However, only against Pestalotiopsis sp. did B. velezensis RCDL12 exhibit antifungal activity. Study was conducted on growth promotion capabilities of isolates on rice and mung bean seedlings. P. lurida RGDS03, B. velezensis RCDL12 and consortium of both the strains reported with promising growth promotion on both rice (85-97%) and mung bean (86-99%) in terms of their seed germination, vegetative growth (root and shoot length, fresh and dry weight), and chlorophyll content as compared to the control plants (untreated). This study has emphasized growth-promoting and biocontrol activities of endophytic bacteria from rhododendrons, and application to enhance crop development for sustainable agriculture.


Subject(s)
Antifungal Agents , Bacillus , Endophytes , Indoleacetic Acids , Plant Growth Regulators , Pseudomonas , Rhododendron , Bacillus/isolation & purification , Bacillus/metabolism , Bacillus/genetics , Bacillus/classification , Endophytes/isolation & purification , Endophytes/classification , Endophytes/metabolism , Endophytes/genetics , Pseudomonas/isolation & purification , Pseudomonas/genetics , Pseudomonas/classification , India , Indoleacetic Acids/metabolism , Antifungal Agents/pharmacology , Antifungal Agents/metabolism , Plant Growth Regulators/metabolism , Rhododendron/microbiology , RNA, Ribosomal, 16S/genetics , Gibberellins/metabolism , Phytophthora/drug effects , Phytophthora/growth & development , Plant Leaves/microbiology , Phylogeny , Plant Diseases/microbiology , Plant Diseases/prevention & control , Nitrogen Fixation , Siderophores/metabolism , Phosphates/metabolism , Plant Stems/microbiology
2.
Curr Microbiol ; 81(12): 402, 2024 Oct 11.
Article in English | MEDLINE | ID: mdl-39392504

ABSTRACT

The isolation of endophytic actinobacteria from the roots of wild populations of Artemisia herba-alba Asso, a medicinal plant collected from the arid lands of Algeria, is reported for the first time. Forty-five actinobacterial isolates were identified by molecular analysis and in vitro evaluated for antimicrobial activity and plant growth-promoting (PGP) abilities (1-Aminocyclopropane-1-carboxylic acid (ACC) deaminase activity, nitrogen fixation, phosphate and potassium solubilization, ammonia, and siderophores production). The phylogenetic relationships based on 16S rRNA gene sequences show that the genus Nocardioides (n = 23) was dominant in the sampled localities. The remaining actinobacterial isolates were identified as Promicromonospora (n = 11), Streptomyces (n = 6), Micromonopora (n = 3), and Saccharothrix (n = 2). Only six (13.33%) strains (five Streptomyces and one Saccharothrix species) were antagonistic in vitro against at least one or more indicator microorganisms. The antimicrobial activity of actinobacterial strains targeted mainly Gram-positive bacteria. The results demonstrate that more than 73% of the isolated strains had ACC deaminase activity, could fix atmospheric nitrogen and were producers of ammonia and siderophores. However, only one (2.22%) strain named Saccharothrix sp. BT79 could solubilize phosphorus and potassium. Overall, many strains exhibited a broad spectrum of PGP abilities. Thus, A. herba-alba provides a source of endophytic actinobacteria that should be explored for their potential biological activities.


Subject(s)
Actinobacteria , Artemisia , Endophytes , Phylogeny , Plant Roots , RNA, Ribosomal, 16S , Algeria , Actinobacteria/genetics , Actinobacteria/classification , Actinobacteria/isolation & purification , Actinobacteria/metabolism , Plant Roots/microbiology , Endophytes/classification , Endophytes/isolation & purification , Endophytes/genetics , Endophytes/metabolism , Artemisia/microbiology , RNA, Ribosomal, 16S/genetics , Biodiversity , Nitrogen Fixation , Siderophores/metabolism
3.
Microb Cell Fact ; 23(1): 266, 2024 Oct 05.
Article in English | MEDLINE | ID: mdl-39369235

ABSTRACT

BACKGROUND: Hazardous synthetic dye wastes have become a growing threat to the environment and public health. Fungal enzymes are eco-friendly, compatible and cost-effective approach for diversity of applications. Therefore, this study aimed to screen, optimize fermentation conditions, and characterize laccase from fungal endophyte with elucidating its ability to decolorize several wastewater dyes. RESULTS: A new fungal endophyte capable of laccase-producing was firstly isolated from cladodes of Opuntia ficus-indica and identified as T. harzianum AUMC14897 using ITS-rRNA sequencing analysis. Furthermore, the response surface methodology (RSM) was utilized to optimize several fermentation parameters that increase laccase production. The isolated laccase was purified to 13.79-fold. GFC, SDS-PAGE revealed laccase molecular weight at 72 kDa and zymogram analysis elucidated a single band without any isozymes. The peak activity of the pure laccase was detected at 50 °C, pH 4.5, with thermal stability up to 50 °C and half life span for 4 h even after 24 h retained 30% of its activity. The Km and Vmax values were 0.1 mM, 22.22 µmol/min and activation energy (Ea) equal to 5.71 kcal/mol. Furthermore, the purified laccase effectively decolorized various synthetic and real wastewater dyes. CONCLUSION: Subsequently, the new endophytic strain produces high laccase activity that possesses a unique characteristic, it could be an appealing candidate for both environmental and industrial applications.


Subject(s)
Coloring Agents , Endophytes , Laccase , Opuntia , Wastewater , Laccase/metabolism , Laccase/genetics , Laccase/isolation & purification , Laccase/chemistry , Opuntia/microbiology , Wastewater/microbiology , Coloring Agents/metabolism , Endophytes/enzymology , Endophytes/genetics , Endophytes/isolation & purification , Hypocreales/enzymology , Hypocreales/genetics , Fermentation , Hydrogen-Ion Concentration , Enzyme Stability , Fungal Proteins/genetics , Fungal Proteins/metabolism , Fungal Proteins/chemistry , Fungal Proteins/isolation & purification
4.
Adv Appl Microbiol ; 129: 171-187, 2024.
Article in English | MEDLINE | ID: mdl-39389705

ABSTRACT

Anthropogenic activities have dramatically accelerated the release of toxic metal(loid)s into soil and water, which can be subsequently accumulated in plants and animals, threatening biodiversity, human health, and food security. Compared to physical and chemical remediation, bioremediation of metal(loid)-polluted soil using plants and/or plant symbiotic fungi is usually low-cost and environmentally friendly. Mycorrhizal fungi and endophytic fungi are two major plant fungal symbionts. Mycorrhizal fungi can immobilize metal(loid)s via constitutive mechanisms, including intracellular sequestration with vacuoles and vesicles and extracellular immobilization by cell wall components and extracellular polymeric substances such as glomalin. Mycorrhizal fungi can improve the efficacy of phytoremediation by promoting plant symplast and apoplast pathways. Endophytic fungi also use constitutive cellular components to immobilize metal(loid)s and to reduce the accumulation of metal(loid)s in plants by modifying plant physiological status. However, a specific mechanism for the removal of methylmercury pollution was recently discovered in the endophytic fungi Metarhizium, which could be acquired from bacteria via horizontal gene transfer. In contrast to mycorrhizal fungi that are obligate biotrophs, some endophytic fungi, such as Metarhizium and Trichoderma, can be massively and cost-effectively produced, so they seem to be well-placed for remediation of metal(loid)-polluted soil on a large scale.


Subject(s)
Biodegradation, Environmental , Fungi , Metalloids , Mycorrhizae , Plants , Soil Pollutants , Symbiosis , Plants/microbiology , Metalloids/metabolism , Metalloids/toxicity , Soil Pollutants/metabolism , Soil Pollutants/toxicity , Fungi/metabolism , Fungi/genetics , Mycorrhizae/metabolism , Mycorrhizae/physiology , Endophytes/metabolism , Endophytes/physiology , Endophytes/isolation & purification , Endophytes/genetics , Metals/metabolism , Metals/toxicity , Soil Microbiology
5.
Microb Ecol ; 87(1): 119, 2024 Sep 28.
Article in English | MEDLINE | ID: mdl-39340548

ABSTRACT

Microbiota associated with host-parasite relationships offer an opportunity to explore interactions among plants, parasites, and microbes, thereby contributing to the overall complexity of community structures. The dynamics of ecological interactions between parasitic plants and their hosts in arid environments remain largely understudied, especially in Africa. This study aimed to examine the bacterial communities of Cuscuta epithymum L. (clover dodder), an epiphytic parasitic plant, and its host, Ziziphus lotus L. (jujuba), in an arid environment. Our goal was to uncover the ecological complexities of microbial communities within the framework of plant-plant interactions. We conducted a comprehensive analysis of the bacterial composition and diversity within populations of the C. epithymum parasite, the infected- and non-infected jujuba host, and their interface at the shoots of the host. This involved amplicon sequencing, targeting the V5-V6 regions of the 16S rRNA gene. A total of 5680 amplicon sequence variants (ASVs) were identified, with Pseudomonadota, Bacillota, and Actinobacteriota being prevalent phyla. Among the bacterial communities, three genera were dominant: Cutibacterium, Staphylococcus, and Acinetobacter. Interestingly, analyses of alpha-diversity (p = 0.3 for Shannon index and p = 0.5 for Simplon index) and beta-diversity (PERMANOVA, with p-values of 0.6 and 0.3) revealed no significant differences between Cuscuta-infected and non-infected jujube shrubs, suggesting a shared shoot endophytic bacteriome. This finding advances our comprehension of microbial communities linked to plant-parasite interactions in the arid environments of Africa. Further research on various hosts is required to confirm plant-to-plant bacterial transmission through Cuscuta infection. Additionally, studies on functional diversity, cytology, ecophysiology and the mechanisms by which bacterial communities transferred between host and parasite are necessary.


Subject(s)
Bacteria , Cuscuta , Endophytes , Microbiota , RNA, Ribosomal, 16S , Ziziphus , Cuscuta/physiology , Cuscuta/microbiology , Cuscuta/genetics , Ziziphus/microbiology , RNA, Ribosomal, 16S/genetics , Endophytes/genetics , Endophytes/physiology , Endophytes/classification , Endophytes/isolation & purification , Bacteria/genetics , Bacteria/classification , Bacteria/isolation & purification , Host-Parasite Interactions , DNA, Bacterial/genetics
6.
BMC Microbiol ; 24(1): 367, 2024 Sep 28.
Article in English | MEDLINE | ID: mdl-39342140

ABSTRACT

BACKGROUND: The plant microbiome is one of the key determinants of healthy plant growth. However, the complexity of microbial diversity in plant microenvironments in different regions, especially the relationship between subsurface and aboveground microorganisms, is not fully understood. The present study investigated the diversity of soil microorganisms in different regions and the diversity of microorganisms within different ecological niches, and compared soil microorganisms and endophytic microorganisms. METHODS: 16 S and ITS sequencing was used to sequence the soil and endophytes microbiome of honeysuckle. Alpha diversity analysis and principal component analysis (PCoA) were used to study the soil and endophyte microbial communities, and the function of endophyte bacteria and fungi was predicted based on the PICRUST2 process and FUNGuild. RESULTS: In total, there were 382 common bacterial genera and 139 common fungal genera in the soil of different producing areas of honeysuckle. There were 398 common bacterial genera and 157 common fungal genera in rhizosphere soil. More beneficial bacteria were enriched in rhizosphere soil. Endophytic bacteria were classified into 34 phyla and 770 genera. Endophytic fungi were classified into 11 phyla and 581 genera, among which there were significant differences in the dominant genera of roots, stems, leaves, and flowers, as well as in community diversity and richness. Endophytic fungal functions were mainly dominated by genes related to saprophytes, functional genes that could fight microorganisms were also found in KEGG secondary functional genes. CONCLUSION: More beneficial bacteria were enriched in rhizosphere soil of honeysuckle, and the microbial network of the rhizosphere is more complex than that of the soil. Among the tissues of honeysuckle, the flowers have the richest diversity of endophytes. The endogenous dominant core bacteria in each part of honeysuckle plant have a high degree of overlap with the dominant bacteria in soil. Functional prediction suggested that some dominant core bacteria have antibacterial effects, providing a reference for further exploring the strains with antibacterial function of honeysuckle. Understanding the interaction between honeysuckle and microorganisms lays a foundation for the study of growth promotion, quality improvement, and disease and pests control of honeysuckle from the perspective of microorganisms.


Subject(s)
Bacteria , Endophytes , Fungi , Lonicera , Microbiota , Rhizosphere , Soil Microbiology , Endophytes/classification , Endophytes/genetics , Endophytes/isolation & purification , Bacteria/classification , Bacteria/genetics , Bacteria/isolation & purification , Fungi/classification , Fungi/genetics , Fungi/isolation & purification , Lonicera/microbiology , Biodiversity , Plant Roots/microbiology , Phylogeny , RNA, Ribosomal, 16S/genetics , Soil/chemistry
7.
An Acad Bras Cienc ; 96(suppl 1): e20230251, 2024.
Article in English | MEDLINE | ID: mdl-39292101

ABSTRACT

The diversity of endophytes and their ecological relationships with the endangered conifer Araucaria angustifolia (a critically endangered species) are unrevealed. This study aimed to characterize the diversity of endophytic fungi associated with A. angustifolia. To this end, we analyzed 90 fragments from five individuals collected from a mixed localized fragment in Guarapuava-PR, Brazil. The total DNA of 61 morphotypes was extracted and the Internal Transcribed Spacer (ITS) region was amplified and sequenced. The sequence analysis allowed the identification of 37 genera belonging to the phylum Ascomycota and the classes Eurotiomycetes, Dothideomycetes, and Sordariomycetes, divided into 11 orders and 13 families. Most of the isolated fungi belonged to the Sordariomycetes class (40%) and to the Xylaria genus (14%), while Eurotiomycetes was the minority class within the community. Our results reveal the high endophytic richness supporting the life cycle of A. angustifolia and reinforce the necessity for the conservation of this conifer, as many genetic resources can be lost owing to its irrational exploration.


Subject(s)
Araucaria , Endophytes , Endophytes/classification , Endophytes/genetics , Endophytes/isolation & purification , Brazil , Araucaria/microbiology , DNA, Fungal/genetics , Biodiversity , Ascomycota/classification , Ascomycota/isolation & purification , Ascomycota/genetics , Phylogeny , Fungi/classification , Fungi/isolation & purification , Fungi/genetics , Sequence Analysis, DNA
8.
World J Microbiol Biotechnol ; 40(10): 317, 2024 Sep 12.
Article in English | MEDLINE | ID: mdl-39261398

ABSTRACT

In this study, nine endophytic fungi capable of producing multiple phenolic compounds were screened and identified from 152 fungi isolated from pigeon pea in a natural habitat (Honghe, Yunnan Province, China). Talaromyces neorugulosus R-209 exhibited the highest potential for phenolic compound production. L-phenylalanine feeding was used to enhance phenolic compound production in T. neorugulosus R-209 cultures. Under the optimal feeding conditions (l-phenylalanine dose of 0.16 g/L and feeding phase of 6 days), the yields of genistein, apigenin, biochanin A, and cajaninstilbene acid increased by 15.59-fold, 7.20-fold, 25.93-fold, and 10.30-fold over control, respectively. T. neorugulosus R-209 fed with l-phenylalanine was found to be stable in the production of phenolic compounds during ten successive subcultures. Moreover, bioactivities of extracts of T. neorugulosus R-209 cultures were significantly increased by l-phenylalanine feeding. Overall, l-phenylalanine feeding strategy made T. neorugulosus R-209 more attractive as a promising alternative source for the production of health-beneficial phenolic compounds in the nutraceutical/medicinal industries.


Subject(s)
Cajanus , Endophytes , Phenols , Phenylalanine , Talaromyces , Talaromyces/metabolism , Phenylalanine/metabolism , Endophytes/metabolism , Endophytes/isolation & purification , Phenols/metabolism , Cajanus/microbiology , China , Ecosystem
9.
Ying Yong Sheng Tai Xue Bao ; 35(6): 1689-1694, 2024 Jun.
Article in Chinese | MEDLINE | ID: mdl-39235028

ABSTRACT

We isolated the dark septate endophytic (DSE) fungi from roots of typical plant species in the tundra of Changbai Mountains Nature Reserve, including Rhododendron aureum, R. conferentiatum, Vaccinium uliginosum, and Dryas octopetala, and studied their colonization. We further investigated the DSE community composition and species diversity of the four tundra plant species by using morphological characteristics combined with rDNA ITS sequence analysis. The results showed that DSE formed a typical structure of "microsclerotia" in roots of the four plant species. A total of 69 strains of DSE fungi were isolated from the root samples, belonging to 10 genera, and 12 species. They were Phialocephala fortinii, Alternaria alternata, A. tenuissima, Epicocum nigrum, Canariomyces microsporus, Colletotrichum spaethianum, C. camelliae, Leptophoria sp., Cladosporium cladosporioides, Phoma sp., Cadophora sp., and Discosia italica, respectively. The DSE fungal species diversity was rich, and all these fungal species were firstly reported as DSE fungi in the alpine tundra belt of China. Among them, Phialocephala fortinii was the common and dominant species of all tundra plants. The Simpson, Pielou, and Shannon diversity indices of DSE fungi of the four plant species of tundra differed significantly. Our results showed that tundra plants have rich diversity of DSE fungi, and they can form a good symbiotic relationship, which enhance the adaptability of tundra plants to the harsh environment.


Subject(s)
Biodiversity , Endophytes , Plant Roots , Endophytes/isolation & purification , Endophytes/classification , Endophytes/genetics , China , Plant Roots/microbiology , Rhododendron/microbiology , Fungi/classification , Fungi/isolation & purification , Fungi/genetics , Vaccinium/microbiology , Ascomycota/isolation & purification , Ascomycota/classification , Ascomycota/genetics , Ecosystem , Alternaria/isolation & purification , Alternaria/classification , Alternaria/genetics
10.
Curr Microbiol ; 81(11): 367, 2024 Sep 19.
Article in English | MEDLINE | ID: mdl-39299961

ABSTRACT

Alternaria species (Deuteromycetes, Ascomycota) as ubiquitous fungi and prolific producers of a variety of toxic compounds are a part of microbiomes of plants, humans, and animals, mainly causing disease, allergic reactions, and toxicosis. However, some species have also been reported as endophytic microorganisms with highly bioactive metabolites. Our previous results indicate that potentially endophytic Alternaria species from Cupressaceae produce bioactive metabolites that possibly contribute to plant holobiont's health. Here, a possible mechanism behind this bioactivity is elucidated. As some endophytic fungi are reported to produce cytotoxic taxane diterpenoids, eight potentially endophytic Alternaria isolates from our collection were analyzed for the presence of the key genes of the paclitaxel (Taxol) biosynthetic pathway, i.e., taxadin synthase (ts), 10-deacetylbaccatin III-10-O-acetyltransferase (dbat), and C-13-phenylpropanoid side-chain CoA acyltransferase (bapt). The presence of all genes, i.e., ts, dbat, and bapt, was detected by PCR in six isolates and dbat and bapt in two isolates. Chemical analyses of the fermentation broths by TLC and HPLC chromatography and IR spectroscopy indicated the synthesis of the final product, i.e., paclitaxel. So, we introduce the synthesis of taxane diterpenoids as a possible mechanism by which Alternaria occupies the plant niches and protects the plant holobiont in the presence of competing microorganisms.


Subject(s)
Alternaria , Biosynthetic Pathways , Taxoids , Alternaria/genetics , Alternaria/metabolism , Taxoids/metabolism , Biosynthetic Pathways/genetics , Endophytes/metabolism , Endophytes/genetics , Endophytes/isolation & purification , Endophytes/classification , Bridged-Ring Compounds/metabolism , Diterpenes/metabolism , Paclitaxel/biosynthesis , Fungal Proteins/genetics , Fungal Proteins/metabolism , Genomics , Phylogeny
11.
Microbiol Spectr ; 12(10): e0080724, 2024 Oct 03.
Article in English | MEDLINE | ID: mdl-39248476

ABSTRACT

Below-ground microorganisms, particularly endophytes, are pivotal for plant establishment and functioning through nutrient acquisition and enhancing resistance to abiotic and biotic stresses. The impact of host plant origin within a species on the composition and interaction networks of root endophytic fungi and bacteria has been less explored compared with plant phylogeny and biological distance. This study investigates the effect of geographic origin on the fungal and bacterial microbiomes of Pinus sylvestris L. root endophytes. Roots from plants grown in a common garden, originating from six locations, were harvested in two distinct seasons. Fungal and bacterial microbiomes were analyzed using Illumina MiSeq sequencing. The operational taxonomic unit (OTU) richness of endophytic fungi and bacteria showed no significant variation due to tree origin or season. However, the Shannon diversity index for endophytic fungi was seasonally influenced. The composition of endophytic fungal and bacterial communities was affected by both tree origin and season, correlating with host root biochemical parameters, such as starch, total non-structural carbohydrates, carbon, nitrogen, and climatic factors, such as mean annual precipitation and temperature. Moreover, the abundance of specific endophytic fungi and bacteria varied across different P. sylvestris origins, depending on the season. The complexity of the co-occurrence networks of fungal and bacterial endophytes within P. sylvestris also differed by geographical origin and season. This study highlights the significant role of biochemical and climatic factors associated with tree origin in shaping interactions with endophytic communities, potentially affecting plant health and adaptability across diverse environments. IMPORTANCE: This study advances our understanding of how plant ecotype and seasonal changes influence root endophytic communities in Scots pine (Pinus sylvestris). By examining trees from various origins grown in a common garden, it highlights the role of tree origin and season in shaping fungal and bacterial community and co-occurrence networks. Importantly, this research demonstrates that tree origin impacts the composition and interaction networks of root endophytes and depends on the season. The study's findings suggest that root biochemical traits and climatic conditions (e.g., temperature, precipitation) associated with tree origin are crucial in determining the assembly of endophytic communities. This understanding could lead to innovative strategies for enhancing plant health and adaptability across different environments, contributing to forestry and conservation efforts. The research underscores the complexity of plant-microbe interactions and the need for a comprehensive approach to studying them, highlighting the interplay between tree origin and microbial ecology in forest ecosystems.


Subject(s)
Bacteria , Endophytes , Fungi , Microbiota , Pinus sylvestris , Plant Roots , Pinus sylvestris/microbiology , Endophytes/classification , Endophytes/isolation & purification , Endophytes/genetics , Endophytes/physiology , Bacteria/classification , Bacteria/genetics , Bacteria/isolation & purification , Fungi/classification , Fungi/isolation & purification , Fungi/genetics , Fungi/physiology , Plant Roots/microbiology , Seasons , Phylogeny , Biodiversity
12.
Article in English | MEDLINE | ID: mdl-39212636

ABSTRACT

Three novel mycelium-forming actinobacteria, designated OC33-EN06T, OC33-EN07T, and OC33-EN08T, were isolated from wild orchid (Aerides multiflora Roxb), collected from a hill evergreen forest in Northern Thailand. Strains OC33-EN06T and OC33-EN07T showed the highest 16S rRNA gene similarity with Actinomycetospora lutea TT00-04T, 99.17 and 99.45%, respectively. Strain OC33-EN08T showed high similarity with four species, namely 'Actinomycetospora termitidis Odt1-22T' (99.37%), Actinomycetospora chiangmaiensis DSM 45062T (99.02%), Actinomycetospora corticicola 014-5T (99.02%), and Actinomycetospora soli SF1T (98.81%). Comparative genome analysis of OC33-EN06T, OC33-EN07T, and OC33-EN08T with the closely related type strains showed that average nucleotide identity (ANI) based on blast, ANI based on MUMmer, and average amino acid identity values were less than 95% and the digital DNA-DNA hybridization values were less than 70%, all below the thresholds for species demarcation. The digital G+C content of OC33-EN06T, OC33-EN07T, and OC33-EN08T were 74.5, 74, and 74 mol%, respectively. These three strains developed bud-like chains of non-motile cylindrical spores with a smooth surface. The cell-wall peptidoglycan contained meso-diaminopimelic acid. The whole-cell sugars contained ribose, arabinose, and galactose. The predominant menaquinone was MK-8(H4). The phospholipid profile included phosphatidylcholine, phosphatidylethanolamine, phosphatidylmethylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, and phosphatidylinositol. Based on comparative analysis of genotypic, phenotypic and chemotaxonomic data, strains OC33-EN06T (=TBRC 18349T=NBRC 116543T), OC33-EN07T (=TBRC 18350T=NBRC 116544T), and OC33-EN08T (=TBRC 18318T=NBRC 116542T) represent the type strains of three novel species of the genus Actinomycetospora for which the names Actinomycetospora aeridis sp. nov., Actinomycetospora flava sp. nov., and Actinomycetospora aurantiaca sp. nov., are proposed.


Subject(s)
Bacterial Typing Techniques , Base Composition , DNA, Bacterial , Fatty Acids , Orchidaceae , Phylogeny , RNA, Ribosomal, 16S , Sequence Analysis, DNA , RNA, Ribosomal, 16S/genetics , Thailand , Fatty Acids/analysis , Fatty Acids/chemistry , DNA, Bacterial/genetics , Orchidaceae/microbiology , Nucleic Acid Hybridization , Endophytes/classification , Endophytes/isolation & purification , Endophytes/genetics , Actinomycetales/isolation & purification , Actinomycetales/classification , Actinomycetales/genetics , Peptidoglycan , Forests , Vitamin K 2/analogs & derivatives , Vitamin K 2/analysis , Mycelium
13.
Curr Microbiol ; 81(10): 318, 2024 Aug 21.
Article in English | MEDLINE | ID: mdl-39164555

ABSTRACT

Two endophytic bacterial strains, designated S1-1-2 T and S1-1-8, were isolated from the leaves of a mangrove plant, Avicennia marina. The isolates were Gram-stain-negative, motile, rod-shaped bacteria with lateral flagella. Growth occurred at 4-41 °C, pH 4.0-11.0, and 0.5-25.0% NaCl. The predominant fatty acids of the novel strains were C18:1 ω6c/ω7c, C19:0 cyclo ω8c, and C16:0. The predominant respiratory quinone was Q-9. The DNA G + C contents of strains S1-1-2 T and S1-1-8 analyzed by genome sequences were 63.8%. Phylogenetic analysis based on 16S rRNA gene sequences obtained using sanger sequencing and whole-genome phylogenetic analysis revealed an affiliation between the two strains and the genus Salinicola in the class Gammaproteobacteria. Detailed genotypic, chemotaxonomic, and phenotypic data support the conclusion that these two strains should be described as a novel species in the genus Salinicola. Here, Salinicola avicenniae sp. nov. (type strain S1-1-2 T = LMG 32655 T = MCCC 1A19027T) is proposed.


Subject(s)
Avicennia , Base Composition , DNA, Bacterial , Gammaproteobacteria , Phylogeny , RNA, Ribosomal, 16S , Avicennia/microbiology , RNA, Ribosomal, 16S/genetics , China , DNA, Bacterial/genetics , Gammaproteobacteria/classification , Gammaproteobacteria/genetics , Gammaproteobacteria/isolation & purification , Fatty Acids/analysis , Bacterial Typing Techniques , Sequence Analysis, DNA , Plant Leaves/microbiology , Endophytes/genetics , Endophytes/classification , Endophytes/isolation & purification
14.
Arch Microbiol ; 206(9): 372, 2024 Aug 10.
Article in English | MEDLINE | ID: mdl-39126528

ABSTRACT

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 effects
15.
Microbiol Spectr ; 12(10): e0366223, 2024 Oct 03.
Article in English | MEDLINE | ID: mdl-39136439

ABSTRACT

The seed serves as the primary source for establishing microbial populations in plants across subsequent generations, influencing plant growth and overall health. Cropping conditions, especially farming practices, can influence the composition and functionality of the seed microbiome. Very little is known about the differences in seed microbiome between organic and conventional production systems. In this study, we characterized the endophytic microbial populations in seeds of rice grown under organic and conventional management practices through culture-dependent and -independent analyses. The V4 region of 16S rRNA was used for bacterial taxa identification, and the ITS1 region was used for the identification of fungal taxa. Our results revealed significantly higher Shannon and Simpson indices for bacterial diversity in the conventional farming system, whereas the fungal diversity was higher for observed, Shannon, and Simpson indices in the organic farming system. The cultivable endophytic bacteria were isolated and identified using the full-length 16S rRNA gene. There was no difference in culturable endophytic bacterial isolates in rice seeds grown under both conventional and organic farming systems. Among 33 unique isolates tested in vitro, three bacteria-Bacillus sp. ST24, Burkholderia sp. OR5, and Pantoea sp. ST25-showed antagonistic activities against Marasmius graminum, Rhizoctonia solani AG4, and R. solani AG11, the fungal pathogens causing seedling blight in rice. IMPORTANCE: In this paper, we studied the differences in the endophytic microbial composition of rice seeds grown in conventional and organic farming systems. Our results demonstrate a greater bacterial diversity in conventional farming, while organic farming showcases a higher fungal diversity. Additionally, our research reveals the ability of seed bacterial endophytes to inhibit the growth of three fungal pathogens responsible for causing seedling blight in rice. This study provides valuable insights into the potential use of beneficial seed microbial endophytes for developing a novel microbiome-based strategy in the management of rice diseases. Such an approach has the potential to enhance overall plant health and improve crop productivity.


Subject(s)
Bacteria , Endophytes , Fungi , Microbiota , Organic Agriculture , Oryza , RNA, Ribosomal, 16S , Seeds , Oryza/microbiology , Endophytes/isolation & purification , Endophytes/classification , Endophytes/genetics , Seeds/microbiology , Microbiota/genetics , RNA, Ribosomal, 16S/genetics , Bacteria/classification , Bacteria/isolation & purification , Bacteria/genetics , Fungi/isolation & purification , Fungi/classification , Fungi/genetics , Burkholderia/genetics , Burkholderia/isolation & purification , Burkholderia/classification , Rhizoctonia/isolation & purification , Rhizoctonia/genetics , Rhizoctonia/growth & development , Bacillus/isolation & purification , Bacillus/genetics , Bacillus/classification , Pantoea/isolation & purification , Pantoea/genetics , Pantoea/classification , Plant Diseases/microbiology , Agriculture/methods
16.
Microb Ecol ; 87(1): 108, 2024 Aug 28.
Article in English | MEDLINE | ID: mdl-39196422

ABSTRACT

The study of plant-microbe interactions is a rapidly growing research field, with increasing attention to the role of seed-borne microbial endophytes in protecting the plant during its development from abiotic and biotic stresses. Recent evidence suggests that seed microbiota is crucial in establishing the plant microbial community, affecting its composition and structure, and influencing plant physiology and ecology. For Theobroma cacao L., the diversity and composition of vertically transmitted microbes have yet to be addressed in detail. We explored the composition and diversity of seed-borne endophytes in cacao pods of commercial genotypes (ICS95, IMC67), recently liberated genotypes from AGROSAVIA (TCS01, TCS19), and landraces from Tumaco (Colombia) (AC9, ROS1, ROS2), to evaluate microbial vertical transmission and establishment in various tissues during plant development. We observed a higher abundance of Pseudomonas and Pantoea genera in the landraces and AGROSAVIA genotypes, while the commercial genotypes presented a higher number of bacteria species but in low abundance. In addition, all the genotypes and plant tissues showed a high percentage of fungi of the genus Penicillium. These results indicate that domestication in cacao has increased bacterial endophyte diversity but has reduced their abundance. We isolated some of these seed-borne endophytes to evaluate their potential as growth promoters and found that Bacillus, Pantoea, and Pseudomonas strains presented high production of indole acetic acid and ACC deaminase activity. Our results suggest that cacao domestication could lead to the loss of essential bacteria for seedling establishment and development. This study improves our understanding of the relationship and interaction between perennial plants and seed-borne microbiota.


Subject(s)
Bacteria , Cacao , Domestication , Endophytes , Seeds , Cacao/microbiology , Endophytes/genetics , Endophytes/classification , Endophytes/isolation & purification , Endophytes/physiology , Seeds/microbiology , Seeds/growth & development , Bacteria/classification , Bacteria/genetics , Bacteria/isolation & purification , Microbiota , Fungi/genetics , Fungi/classification , Fungi/isolation & purification , Genotype , Biodiversity
17.
J Microbiol Biotechnol ; 34(9): 1857-1866, 2024 Sep 28.
Article in English | MEDLINE | ID: mdl-39187457

ABSTRACT

Endophytic fungi have been shown to synthesize bioactive secondary metabolites, some of which promote plant growth through various mechanisms. In our previous study, endophytic fungi were isolated from mango trees (Mangifera indica L.). The present study examined fifty endophytic fungal isolates for mineral solubilization activity, ammonia production, and siderophore production. It was shown that these isolates could produce phytohormones indole-3-acetic acid and gibberellic acid, as well as inhibit plant pathogens, specifically Colletotrichum gloeosporioides and Lasiodiplodia theobromae. The results showed that all the isolated fungal endophytes exhibited various activities. Based on the findings, two fungal endophytes-Aureobasidium pullulans CY.OS 13 and Aspergillus tamarii CY.OS 144-were selected for dual inoculation in chili plants under pot-scale conditions to investigate their potential to improve growth-related traits such as seed germination, shoot and root length, biomass, and chlorophyll content. Seed treated with A. pullulans CY.OS 13 and/or A. tamarii CY.OS 144 showed a significant (p < 0.05) increase in seed germination and growth parameters of chili plants grown under pot-scale conditions. Particularly, chili plants whose seeds were injected with a combination of the two selected endophytic fungi showed the highest plant development traits. Therefore, the selected endophytic fungi have the potential to be used as biofertilizers, especially when combined. They could eventually replace chemical fertilizers because they are environmentally friendly, beneficial to humans, and can even promote sustainable agriculture.


Subject(s)
Colletotrichum , Endophytes , Indoleacetic Acids , Mangifera , Plant Growth Regulators , Endophytes/isolation & purification , Endophytes/metabolism , Mangifera/microbiology , Plant Growth Regulators/metabolism , Indoleacetic Acids/metabolism , Colletotrichum/growth & development , Gibberellins/metabolism , Plant Roots/microbiology , Plant Roots/growth & development , Siderophores/metabolism , Plant Development , Germination , Ascomycota/growth & development , Ascomycota/metabolism , Ascomycota/isolation & purification , Fungi/isolation & purification , Fungi/classification , Fungi/metabolism , Fungi/growth & development , Seeds/microbiology , Seeds/growth & development , Aspergillus/growth & development , Aspergillus/metabolism , Aspergillus/isolation & purification , Chlorophyll/metabolism , Ammonia/metabolism , Capsicum/microbiology , Capsicum/growth & development , Biomass
18.
BMC Microbiol ; 24(1): 291, 2024 Aug 03.
Article in English | MEDLINE | ID: mdl-39097685

ABSTRACT

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/genetics
19.
Microbiol Res ; 287: 127862, 2024 Oct.
Article in English | MEDLINE | ID: mdl-39121704

ABSTRACT

Endophytes, microorganisms inhabiting internal plant tissues, play a pivotal role in plant growth and disease resistance. Moreover, previous studies have established that Musa plants derive disease protective functions from their microbiome. Notably, one of the crop wild relatives of banana, the Calcutta 4 variety, exhibits resistance to various phytopathogens such as Pseudocercospora fijiensis (P. fijiensis), while the Williams commercial cultivar (cv.) is highly susceptible. Therefore, this study aims primarily to characterize and compare the endophytic microbiota composition of Calcutta 4 and Williams banana plants when grown sympatrically. Alongside, differences in endophytic microbiome between plant sections (shoot or roots), growth phases (in vitro or greenhouse) and fitness factors such as the addition of plant growth-promoting bacteria Bacillus subtilis EA-CB0575 (T2 treatment) or infection by P. fijiensis (T3 treatment) were examined. Both culture-dependent and -independent techniques were used to evaluate these differences and assess the culturability of banana endophytes under varying conditions. Microbial cultures resulted in 331 isolates distributed across 54 genera when all treatments were evaluated, whereas 16 S sequencing produced 9510 ASVs assigned in 1456 genera. Alpha and beta diversity exhibited significant differences based on plant section, with an increase in phylogenetic diversity observed in plants with pathogen infection (T3) compared to control plants (T1). Additionally, four differentially abundant genera associated with nitrogen metabolism were identified in T3 plants and seven genera showed differential abundance when comparing varieties. When culture-dependent and -independent methods were compared, it was found that isolates represented 3.7 % of the genera detected by culture-independent methods, accounting for 12-41 % of the total data depending on the treatment. These results are crucial for proposing management strategies derived from crop wild relatives to enhance the resilience of susceptible commercial varieties against fitness factors affecting crop development. Additionally, they help to decipher the pathogenic effects of P. fijiensis in banana plants and advance the understanding of how plant domestication influences the endosphere.


Subject(s)
Bacteria , Biodiversity , Endophytes , Microbiota , Musa , Plant Roots , Musa/microbiology , Endophytes/classification , Endophytes/isolation & purification , Endophytes/genetics , Plant Roots/microbiology , Bacteria/classification , Bacteria/genetics , Bacteria/isolation & purification , Plant Diseases/microbiology , RNA, Ribosomal, 16S/genetics , Phylogeny , Crops, Agricultural/microbiology , Plant Shoots/microbiology , Plant Shoots/growth & development , Disease Resistance
20.
BMC Microbiol ; 24(1): 293, 2024 Aug 06.
Article in English | MEDLINE | ID: mdl-39107684

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 , Ecosystem
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