Endophytic Strains Research Articles

Endophytic Bacillus velezensis GsB01 controls Gleditsia sinensis wilt by secreting antifungal metabolites and modulates symbiotic microbiota within trees.

Identifying effective biological control agents against fungal pathogens and determining their mechanisms of action are important in the control of plant diseases. In this study, we isolated an endophytic bacterial strain, GsB01, from the branches of asymptomatic Gleditsia sinensis. Multi-locus sequence analysis identified the strain as Bacillus velezensis. GsB01 exhibited significant antifungal activity against Thyronectria austroamericana, the causative agent of G. sinensis wilt. Liquid chromatography-mass spectrometry identified four consistently present antimicrobial compounds in GsB01 metabolite fractions with high antifungal activity: macrolactin A, bacillaene A, surfactin, and iturin. GsB01's active metabolite fractions altered the metabolic profiles of T. austroamericana, disrupting seven pathways, including arginine biosynthesis, nucleotide metabolism, purine metabolism, and the pentose phosphate pathway. Furthermore, absolute quantitative polymerase chain reaction analysis suggested that GsB01 may increase the abundance of endophytic bacteria in G. sinensis. The 16S rRNA amplicon sequencing revealed changes in the endophytic landscape in stems and roots following GsB01 introduction, particularly with significant variation in the dominant bacterial genera within the stems. The study highlights GsB01's potential against plant wilt and suggests that its antifungal activity is achieved by secreting antifungal metabolites. The study also recorded changes in the symbiotic microbiota within trees that had been infected with a pathogenic fungus and subsequently treated with an endophytic antagonistic bacterial strain. © 2024 Society of Chemical Industry.

Growth regulation in bread wheat via novel bioinoculant formulation

Wheat (Triticum aestivum L.) is one of the most significant crops and the backbone of food security worldwide. However, low wheat production remains a substantial concern in global agricultural systems. It can be attributed to several factors, including adverse climatic conditions, plant disease and poor soil quality. Recent efforts have explored bioinoculant applications as a promising approach to enhance wheat yield, trying to mitigate constraints essential for future wheat production and global food security. This study tested talc powder, wheat biochar, sugarcane bagasse biochar, and farmyard manure as carriers with two endophytic bacterial strains, Burkholderia phytofirmans PsJN and Bacillus spp. MN54 was applied to three wheat varieties (Ujala-16, Zincol-16, and Fathejang-16). The data was recorded at the seedling and maturity growth stages of plants. A pot experiment revealed significant improvements in plant growth following bioinoculant application compared to controls. Notably, the combination of sugarcane bagasse biochar with Bacillus sp. MN54 exhibited the most pronounced effects, promoting internodal length, spike length, tiller number per plant, grain yield per plant, and spikelets per spike. Additionally, talc powder with Bacillus sp. MN54 increased peduncle length, tiller number per plant, and spike length in Fathejang-16. These findings offer valuable insights into optimizing bioinoculant formulations for improved agricultural practices, adapting to climate change, and contributing to ensuring global food security.

ЭНДОФИТНЫЕ ШТАММЫ БАКТЕРИЙ В ТЕХНОЛОГИИ ВОЗДЕЛЫВАНИЯ КУКУРУЗЫ НА ЗЕРНО

The study of the effect of endophytic bacterial strains on the formation of the general habitus of the corn plant during the growing season, as well as the effect on grain yield, was carried out in the conditions of the Ancestral region of the Republic of Tatarstan in 2022-2023. The soil of the experimental site was represented by a subtype of gray forest, the variety was heavy loamy. Endophytic bacterial strains were used in corn crops in the form of leaf dressing with a rate of 1.0 l/ha during the growing season in the 7-8 leaf phase. The technology of cultivation of corn for grain was generally accepted for the Republic of Tatarstan. A two-factor experiment was laid: hybrids (factor A) - early maturing Nur and ROSS-195, mid-early ‒ Krasnodarskiy-230; preparations of endophytic bacterial strains (factor B) - without application (control), KS-25, KS-31, KS-38, KS-54, Consortium and PS-17 (standard). The years of research were characterized as moderately arid in terms of climatic conditions. The preparations had a positive effect on the height of plants, above-ground mass and leaf area, which were determined in the phases of paniculation, cob formation and milk ripeness. The Nur hybrid responded positively to the treatment of crops with preparations KS-25, KS-38 and KS-54, the ROSS-195 hybrid - Consortium, KS-31 and KS-38, Krasnodarskiy-230 - KS-31 and KS-54. The highest aboveground mass in the experiment in the milk ripeness phase of the Nur hybrid was noted in the variant with KS-38 (48.25 t/ha), in the ROSS-195 and Krasnodarskiy-230 hybrids ‒ with KS-54 (67.38 and 58.25 t/ha, respectively). The largest leaf area was observed in the cob formation phase of the Nur hybrid when used in the KS-31 variants, in the ROSS-195 and Krasnodarskiy-230 hybrids when treated with the Consortium preparation. The grain yield of the Nur hybrid in variants with the PS-17, KS-38 and KS-25 preparations increased by 9.6…27.6%, the ROSS-195 hybrid when sprayed with KS-38 and KS ‒ by 15.3…19.7%, the Krasnodarskiy-230 hybrid when using KS-25, KS-54 and PS-17 ‒ by 9.0…28.1%. Preparations with endophytic bacterial strains increased the protein content in the hybrid grain by 0.15…0.73%.

Whole-genome sequence of Bacillus paralicheniformis strain R243 isolated from rice (Oryza sativa L.) roots.

We isolated an endophytic strain R243 and sequenced its genome using the NextSeq 1000 (Illumina) sequencing platform. The sequenced genome contains 4,478 protein-coding genes with a mean GC content of 45.56%. The strain R243 was identified as Bacillus paralicheniformis. Gene clusters involved in the synthesis of biocontrol substances were identified.

New Isocoumarins from An Endophytic Fungal Strain Diaporthe arengae M2 and Their Antibacterial Activities.

Four new isocoumarin derivatives 12-O-acetyl-isocitreoisocoumarinol (1), (+)-(10R)-O-acetyl-diaportinol (2-a), (-)-(10S)-O-acetyl-diaportinol (2-b), peyroisocoumarin E (3) and new stereoconfigurations of three isocoumarin derivatives desmethyldichlorodiaportinol A (4), threo-monochlorodiaportinol A (5-a), erytheo-monochlorodiaportinol A (5-b), together with nine known ones (6-14), were separated from the rice fermentation of endophytic fungus Diaporthe arengae M2 isolated from Camellia oleifera. The structures of new compounds were determined by extensive spectroscopic analyses including nuclear magnetic resonance (NMR) and high resolution electrospray ionization mass spectroscopy (HR-ESI-MS). Compounds 4, 7, 8, 12, 13 exhibited definite inhibition against five strains of bacteria with the MIC values range from 16 μg/mL to 64 μg/mL.

Biocontrol potential of Burkholderia gladioli STPT16 on postharvest decay of litchi fruit caused by Peronophythora litchii and Colletotrichum fructicola

Litchi downy blight (LDB) caused by Peronophythora litchii and anthracnose (AD) caused by Colletotrichum species are major pre- and postharvest disease of litchi fruit. Biocontrol agents such as Burkholderia has been applied for controlling postharvest fruit decay. In this work, an endophytic strain SZPT16 isolated from health litchi branch was identified as Burkholderia gladioli and exhibited strong antagonistic activity against mycelial growth and conidia/sporangia germination of two postharvest pathogens. Dual incubation of pathogen and biocontrol agents caused alterations in hyphal and conidial/sporangial structures and severe cell morphology changes such as mycelia deformation, cell wall collapse and cellular disorganization. Cell-free culture filtrates (CF, diluted at 25-fold) of B. gladioli SZPT16 showed an efficient inhibition on postharvest litchi decay by preharvest application (74.77 % protection) and postharvest application (64.68 % and 71.14 % protection for LDB and AD, respectively). Furthermore, UPLC-MS/MS metabolomic analysis identified 41 up-regulated differential metabolites, which mainly included nucleosides, nucleotides, and analogues, organic acids and derivatives, organic oxygen compounds, and organoheterocyclic compounds. This study firstly reported the efficacy of CF of B. gladioli SZPT16 in protecting postharvest litchi fruit against natural and inoculated pathogen infections. The overall results provide a promising alternative to chemical fungicides to control litchi postharvest decay.

Diversity and Functional Roles of Root-Associated Endophytic Fungi in Two Dominant Pioneer Trees Reclaimed from a Metal Mine Slag Heap in Southwest China

The utilization of fast-growing, economically valuable woody plants with strong stress resistance, such as poplar and willow, to revegetate severely metal-contaminated mine tailings not only offers a productive and profitable use of abandoned polluted soil resources but also facilitates the phytoremediation of these polluted soils. This study examines the diversity and functional roles of endophytic fungi naturally colonizing the roots of an artificially established Populus yunnanensis forest and the naturally reclaimed pioneer species Coriaria sinica on an abandoned tailing dam in southwest China. Culture-independent analyses revealed that the root systems of both plant species were abundantly colonized by arbuscular mycorrhizal fungi and endophytic fungi, forming rich and diverse endophytic fungal communities predominantly represented by the genera Ilyonectria, Tetracladium, Auricularia, and unclassified members of Helotiales. However, the composition of root endophytic fungal communities differed significantly between the two plant species. Using a culture-dependent approach, a total of 192 culturable endophytic fungal strains were isolated from the roots. The dominant genera included Cadophora, Cladosporium, Cyphellophora, and Paraphoma, most of which were previously identified as dark septate endophytes (DSE). Six representative DSE strains were selected for further study, and significant cadmium tolerance and various plant growth-promoting traits were observed, including the solubilization of insoluble inorganic and organic phosphorus, indole-3-acetic acid (IAA) production, and siderophore synthesis. In greenhouse experiments, inoculating two DSE strains mitigated the inhibitory effects of metal-polluted tailing soil on the growth of P. yunnanensis. This was achieved by reducing heavy metal uptake in roots and limiting metal translocation to the above-ground tissues, thereby promoting plant growth and adaptability. Our findings suggest that as plants reclaim metal-polluted tailings, root-associated endophytic fungal communities also undergo natural succession, playing a critical role in enhancing the host plant’s tolerance to stress. Therefore, these restored root-associated fungi, particularly DSE, are essential functional components of the root systems in plants used for tailing reclamation.

Molecular Characterization of Endophytic Fungal Strains for Green Synthesis of ZnO and Au Nanoparticles

Abstract Introduction/Objective Introduction: Traditional methods for nanoparticle synthesis often involve hazardous compounds, posing environmental risks and limiting their biomedical applications due to instability. The exploration of green synthesis approaches using natural products and eco-friendly solvents presents a challenging yet imperative pursuit in nanoparticle research. Methods/Case Report Methods: Endophytic fungi were isolated from healthy Eucalyptus sideroxylon plants collected from the Desert Research Center. Cultivation of fungi in malt extract broth was conducted at 28°C with agitation for 96 hours. Subsequently, fungal DNA was extracted, and ITS-rDNA regions were PCR-amplified for identification. Sequencing and phylogenetic analysis were performed to characterize the fungal isolates. Fungal filtrates were utilized for the biosynthesis of ZnO and Au nanoparticles (NPs) from ZnSO4 and HAuCl4 solutions, respectively. Results (if a Case Study enter NA) Results: Among the 13 fungal isolates, Fusarium chlamydosporum (FI-03) exhibited the most efficient synthesis of ZnO and Au NPs. Sequencing confirmed the identity of the fungal strain. The synthesized NPs displayed uniform spherical shapes, with ZnO NPs averaging 19.3 nm and Au NPs averaging 22.1 nm in size. Transmission electron microscopy (TEM) analysis further elucidated the morphology of the NPs. Conclusion Conclusion: Fusarium chlamydosporum emerged as a promising candidate for the green synthesis of ZnO and Au NPs. This environmentally friendly approach offers a cost-effective and biocompatible alternative to conventional nanoparticle synthesis methods.

Comparison between bacterial bio-formulations and gibberellic acid effects on Stevia rebaudiana growth and production of steviol glycosides through regulating their encoding genes

Stevia rebaudiana is associated with the production of calorie-free steviol glycosides (SGs) sweetener, receiving worldwide interest as a sugar substitute for people with metabolic disorders. The aim of this investigation is to show the promising role of endophytic bacterial strains isolated from Stevia rebaudiana Egy1 leaves as a biofertilizer integrated with Azospirillum brasilense ATCC 29,145 and gibberellic acid (GA3) to improve another variety of stevia (S. rebaudiana Shou-2) growth, bioactive compound production, expression of SGs involved genes, and stevioside content. Endophytic bacteria isolated from S. rebaudiana Egy1 leaves were molecularly identified and assessed in vitro for plant growth promoting (PGP) traits. Isolated strains Bacillus licheniformis SrAM2, Bacillus paralicheniformis SrAM3 and Bacillus paramycoides SrAM4 with accession numbers MT066091, MW042693 and MT066092, respectively, induced notable variations in the majority of PGP traits production. B. licheniformis SrAM2 revealed the most phytohormones and hydrogen cyanide (HCN) production, while B. paralicheniformis SrAM3 was the most in exopolysaccharides (EPS) and ammonia production 290.96 ± 10.08 mg/l and 88.92 ± 2.96 mg/ml, respectively. Treated plants significantly increased in performance, and the dual treatment T7 (B. paramycoides SrAM4 + A. brasilense) exhibited the highest improvement in shoot and root length by 200% and 146.7%, respectively. On the other hand, T11 (Bacillus cereus SrAM1 + B. licheniformis SrAM2 + B. paralicheniformis SrAM3 + B. paramycoides SrAM4 + A. brasilense + GA3) showed the most elevation in number of leaves, total soluble sugars (TSS), and up-regulation in the expression of the four genes ent-KO, UGT85C2, UGT74G1 and UGT76G1 at 2.7, 3.3, 3.4 and 3.7, respectively. In High-Performance Liquid Chromatography (HPLC) analysis, stevioside content showed a progressive increase in all tested samples but the maximum was exhibited by dual and co-inoculations at 264.37% and 289.05%, respectively. It has been concluded that the PGP endophytes associated with S. rebaudiana leaves improved growth and SGs production, implying the usability of these strains as prospective tools to improve important crop production individually or in consortium.

Endophyte Bacillus vallismortis BL01 to Control Fungal and Bacterial Phytopathogens of Tomato (Solanum lycopersicum L.) Plants

Some strains of Bacillus vallismortis have been reported to be efficient biocontrol agents against tomato pathogens. The aim of our study was to assess the biocontrol ability of the endophytic strain BL01 Bacillus vallismortis through in vitro and field trials, as well as to verify its plant colonization ability and analyze the bacterial genome in order to find genes responsible for the biocontrol activity. We demonstrated in a gnotobiotic system and by confocal laser microscopy that the endophytic strain BL01 was able to colonize the endosphere and rhizosphere of tomato, winter wheat and oilseed rape. In vitro experiments demonstrated the inhibition activity of BL01 against a wide range of phytopathogenic fungi and bacteria. BL01 showed biological efficacy in two-year field experiments with tomato plants against black bacterial spotting by 40–70.8% and against late blight by 47.1% and increased tomato harvest by 24.9% or 10.9 tons per hectare compared to the control. Genome analysis revealed the presence of genes that are responsible for the synthesis of biologically active secondary metabolites, which could be responsible for the biocontrol action. Strain BL01 B. vallismortis can be considered an effective biocontrol agent to control both fungal and bacterial diseases in tomato plants.

A study on the potential of endophytic bacteria to promote plant growth: uses in agriculture and future directions

The escalating demand for chemical-free fertilizers in agriculture stems from the adverse impacts associated with chemical fertilizers on both human and animal health, as well as environmental pollution. To address this concern, plant-based microorganisms emerge as a promising solution for the development of environmentally sustainable biofertilizers. Among these microorganisms, endophytic bacteria, residing within plant tissues without causing harm to the host plant, exhibit exceptional attributes conducive to plant growth promotion. Notably, these bacteria demonstrate the production of phytohormones, ammonia, phosphate solubilization, and nitrogen fixation capabilities. Additionally, endophytic bacteria showcase the synthesis of hydrolytic enzymes, the production of siderophores, and antimicrobial activity against pathogens. Such characteristics contribute significantly to the robust growth and development of host plants, fostering tolerance to environmental stresses. This manuscript aims to comprehensively review the plant growth promotion activities of endophytic bacteria, elucidating their diversity and isolation from various plants. Furthermore, it explores the potential future directions in this burgeoning field of research, envisioning the development of endophytic bacterial strains capable of replacing traditional chemical fertilizers. Future research endeavors hold the promise of uncovering novel and effective endophytic bacterial strains, heralding a sustainable paradigm shift in the realm of agricultural fertilization.

Optimization, purification and characterization of laccase from a new endophytic Trichoderma harzianum AUMC14897 isolated from Opuntia ficus-indica and its applications in dye decolorization and wastewater treatment.

Hazardous synthetic dye wastes have become a growing threat to the environment and public health. Fungal enzymes are eco-friendly, compatible and cost-effective approachfor 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. 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 72kDa 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 4h even after 24h retained 30% of its activity. The Km and Vmax values were 0.1mM, 22.22µmol/min and activation energy (Ea) equal to 5.71kcal/mol. Furthermore, the purified laccase effectively decolorized various synthetic and real wastewater dyes. 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.

Endophytic bacilli from Cyamopsis tetragonoloba (L.) Taub. induces plant growth and drought tolerance.

Cyamopsis tetragonoloba (L.) Taub. (guar) is a commercially important crop known for its galactomannan content in seeds. Drought stress is a significant global concern that compromises the productivity of major legumes including guar. The endophytic microbes associated with plants play a significant role in enhancing plant growth and modulating the impact of abiotic stress(s). The present study involved the isolation of 73 endophytic bacteria from the guar seeds of drought-tolerant (RGC-1002 and RGC-1066) and sensitive (Sarada and Varsha) varieties. Based on multiple PGP attributes and drought tolerance, at 50% PEG6000 w/v, 11 efficient isolates were selected and identified through 16S rRNA gene sequencing. Isolates belonging to ten different species of bacilli including Cytobacillus oceanisediminis, Mesobacillus fermenti, Peribacillus simplex from sensitive and Bacillus zanthoxyli, B. safensis, B. velezensis, B. altitudinis, B. licheniformis, B. tequilensis, and B. paralicheniformis isolated from tolerant varieties. A greenhouse experiment with a drought-sensitive guar variety demonstrated that inoculation of selected isolates showed comparatively better plant growth, higher relative water content (RWC), decreased carbon isotope discrimination ratio (Δ13C), increased chlorophyll, carotenoids, anthocyanin, and proline content, decreased malondialdehyde (MDA) and modulated defense enzymes as compared to their uninoculated controls. Tolerant variety isolates B. tequilensis NBRI14G and B. safensis NBRI10R showed the most promising results in improving plant growth and also drought stress tolerance in guar plants. This study represents for the first time that seed endophytic bacterial strains from guar can be utilized to develop the formulation for improving the productivity of guar under drought-stress conditions.

Isolation and identification of endophytic fungi from Alhagi sparsifolia Shap. and their antibacterial activity

In order to explore the endophytic resources of Alhagi sparsifolia Shap. and identified novel antibacterial substances. Thirty endophytic fungal strains were isolated from the stems and roots of A. sparsifolia Shap. Morphological and molecular biology methods were used to identify ten strains of fungi, including four strains of Aspergillus niger, three strains of Alternaria alternata, two strains of Aspergillus flavus, and one strain of Fusarium incarnatum. All these strains were isolated from A. sparsifolia Shap. for the first time, and of these Aspergillus was the dominant genus. Antibacterial activity of the ten strains against Escherichia coli, Staphylococcus aureus, Candida albicans, and Pseudomonas aeruginosa were evaluated using the disc diffusion method. The results demonstrated that the metabolites from all the strains had inhibitory effects on at least one indicator bacterium. Notably, the endophytic fungi AFJ3 and AFG6 demonstrated strong broad-spectrum antibacterial activity, particularly against E. coli, with inhibition zones measuring 32.0 ± 0.3 and 31.3 ± 0.3 mm, respectively. The three endophytic fungi (AFG1, AFG2, and AFG3) isolated from the roots demonstrated significant antibacterial activity against P. aeruginosa forming an inhibition zone of diameter 31.3 ± 0.1, 25.6 ± 0.2, and 25.6 ± 0.2 mm, respectively. However, the strains of endophytic fungi demonstrated no significant inhibitory effects on C. albicans. Ultra-high performance liquid chromatography with quadrupole time-of-flight mass spectrometry/mass spectrometry (UPLC-QTOF-MS/MS) analysis depicted that the ethyl acetate phase of AFJ3 and AFG6 fermentation broth predominantly contained organic acids, phenolic acids, flavonoids, and fatty acids. These secondary metabolites often exhibited good antibacterial activity. This study broadens our understanding of endophytic fungi in A. sparsifolia Shap. The antibacterial activity of some strains of endophytic fungi was significant, making it worthy of further research on their active material.

Comparative genome analysis of endophytic Bacillus amyloliquefaciens MR4: a potential biocontrol agent isolated from wild medicinal plant root tissue.

In this investigation, 396 endophytic bacterial strains from six indigenous medicinal plant species within the Xinjiang Tumor Peak National Nature Reserve were subjected to screening. The strain MR4 emerged as a noteworthy contender, demonstrating pronounced biocontrol capabilities coupled with exceptional cold tolerance. Through morphological scrutiny and comprehensive genomic sequencing, MR4 was identified as Bacillus amyloliquefaciens. Antagonistic assays revealed MR4's efficacy in suppressing the causative agents of cotton wilt and verticillium wilt, achieving inhibition rates surpassing 50%. Analyses, underpinned by PCR methodologies, indicated MR4's capacity to biosynthesize a minimum of eight distinct antimicrobial agents. The whole-genome sequencing data indicated that B. amyloliquefaciens MR4 had the genome size and GC content of 4,017,872bp and 47.14%, respectively, and 4191 coding genes were identified. The genome consists of a single chromosome and one plasmid. Moreover, it was augmented by annotations from various databases, including GO, KEGG, and COG. The pathogenicity of MR4 undergoes evaluation, while predictions concerning MR4's secondary metabolites have disclosed gene clusters for 13 varieties of these compounds, with particular emphasis on surfactins and fengycin. Comparative analyses with four paradigmatic strains shed light on MR4's genomic composition and its phylogenetic lineage within the Bacillus genus. The genomic data pertaining to MR4 have been duly submitted to the NCBI GenBank, bearing the accession numbers CP146236 (Chr1) and CP146237 (plas1). This study endeavors to furnish potent microbial resources for the biocontrol and enhancement of plant growth, thereby providing a theoretical groundwork for MR4's agronomic utilization.

Response of Plant Endophyte Communities to Heavy Metal Stress and Plant Growth Promotion by the Endophyte Serratia marcescens (Strain JG1)

Effects of heavy metals on soil microbial communities have been extensively studied due to their persistence in the environment and imposed threats to living organisms; however, there is a lack of in-depth studies of the impacts of heavy metals on plant endophyte communities. Therefore, the responses of plant endophyte communities to different concentrations of heavy metals were investigated in this study. The endophyte communities of plants existing in severely (W1, Pb, 110.49 mg/kg, Cd, 1.11 mg/kg), moderately (W2, Pb, 55.06 mg/kg, Cd, 0.48 mg/kg), and mildly (W3, Pb, 39.06 mg/kg, Cd, 0.20 mg/kg) contaminated soils were analyzed by 16s rRNA high-throughput Illumina sequencing. Furthermore, networks were constructed to illustrate the relationships between microorganisms and environmental factors. High-quality sequences were clustered at a 97% similarity level. Results revealed that the diversity of the community and relative abundance of Cyanobacteria phylum increased with decreasing levels of pollution. Cyanobacteria and Proteobacteria were found to be the dominant phylum, while Methylobacterium and Sphingomonas were observed as the dominant genus. Tukey’s HSD test showed that the relative abundances of Cyanobacteria and Proteobacteria phyla and Methylobacterium and Sphingomonas genera differed significantly (p < 0.01) among the plants of the three sample sites. Environmental factor analysis revealed a significant negative correlation (p < 0.01) of Cyanobacteria and a significant positive correlation (p < 0.01) of Methylobacterium with the heavy metal content in the environment. These findings suggest that Cyanobacteria and Methylobacterium may be phylum and genus indicators, respectively, of heavy metal toxicity. Tax4Fun analysis showed the effect of heavy metal toxicity on the abundance of genes involved in plant metabolism. In addition, culturable endophytic strains were isolated to study their resistance to heavy metal stress and their ability to promote plant growth. The potting tests showed that the JG1 strain was tolerant to heavy metals, and it could significantly promote the growth of the host plant under stress caused by multiple heavy metals. Compared to the control, the JG1-treated plants showed a 23.14% increase in height and a 12.84% increase in biomass. Moreover, AP, AK, and HN contents in JG1-treated plants were 20.87%, 12.55%, and 9.03% higher, respectively, under heavy metal stress. The results of this study provide a scientific basis for the construction of an efficient plant endophyte restoration system.

Antibacterial, Antifungal, and Cytotoxic Effects of Endophytic Streptomyces Species Isolated from the Himalayan Regions of Nepal and Their Metabolite Study.

Background/Objectives: Recently, antimicrobial-resistant pathogens and cancers have emerged as serious global health problems, highlighting the immediate need for novel therapeutics. Consequently, we aimed to isolate and characterize endophytic Streptomyces strains from the rhizospheres of the Himalayan region of Nepal and identify specialized metabolites with antibacterial, antifungal, and cytotoxic potential. Methods: To isolate Streptomyces sp., we collected two soil samples and cultured them on an ISP4 medium after pretreatment. We isolated and identified the strains PY108 and PY109 using a combination of morphological observations and 16S rRNA gene sequencing. Results: The BLAST results showed that PY108 and PY109 resembled Streptomyces hundungensis PSB170 and Streptomyces sp. Ed-065 with 99.28% and 99.36% nucleotide similarity, respectively. Antibacterial assays of ethyl acetate (EA) extracts from both isolates PY108 and PY109 in a tryptic soy broth (TSB) medium were conducted against four pathogenic bacteria. They showed significant antibacterial potential against Staphylococcus aureus and Klebsiella pneumoniae. Similarly, these extracts exhibited moderate antifungal activities against Saccharomyces cerevisiae and Aspergillus niger. Cytotoxicity assays on cervical cancer cells (HeLa) and breast cancer cells (MCF-7) revealed significant potential for both extracts. LC-MS/MS profiling of the EA extracts identified 27 specialized metabolites, including diketopiperazine derivatives, aureolic acid derivatives such as chromomycin A, and lipopeptide derivatives. In comparison, GC-MS analysis detected 34 metabolites, including actinomycin D and γ-sitosterol. Furthermore, a global natural product social molecular networking (GNPS)-based molecular networking analysis dereplicated 24 metabolites in both extracts. Conclusions: These findings underscore the potential of endophytic Streptomyces sp. PY108 and PY109 to develop new therapeutics in the future.

Endophytic Bacterial Biofilm-Formers Associated with Antarctic Vascular Plants.

Deschampsia antarctica and Colobantus quitensis are the only two vascular plants colonized on the Antarctic continent, which is usually exposed to extreme environments. Endophytic bacteria residing within plant tissues can exhibit diverse adaptations that contribute to their ecological success and potential benefits for their plant hosts. This study aimed to characterize 12 endophytic bacterial strains isolated from these plants, focusing on their ecological adaptations and functional roles like plant growth promotion, antifungal activities, tolerance to salt and low-carbon environments, wide temperature range, and biofilm formation. Using 16S rRNA sequencing, we identified several strains, including novel species like Hafnia and Agreia. Many strains exhibited nitrogen-fixing ability, phosphate solubilization, ammonia, and IAA production, potentially benefiting their hosts. Additionally, halotolerance and carbon oligotrophy were also shown by studied bacteria. While some Antarctic bacteria remain strictly psychrophilic, others demonstrate a remarkable ability to tolerate a wider range of temperatures, suggesting that they have acquired mechanisms to cope with fluctuations in environmental temperature and developed adaptations to survive in intermediate hosts like mammals and/or birds. Such adaptations and high plasticity of metabolism of Antarctic endophytic bacteria provide a foundation for research and development of new promising products or mechanisms for use in agriculture and technology.

Sodium alginate improves phytase stability and enhances soil phosphorous utilization

A plant growth-promoting endophytic strain Bacillus sp. WR11 produces high-level phytase. Phytase degrades phytate, which is the major form of organic phosphate in soils that cannot be readily utilized by plants. To strengthen the strain or its phytase for cost-efficient biodegradation of phytate, we compared the impacts of different chemical additives on phytase production and preservation. Phytate degradation ability in soils and growth promotion effect in wheat were evaluated. To ensure safe application, potential pathogenicity of the strain was also predicted. Phytase activity assay suggests that sodium alginate (SA) significantly improves phytase production to as higher as 2.21-fold, but has little influence on the growth of WR11 during liquid fermentation. During liquid storage of cell-free supernatant (CFS) at room temperature, SA maintains 68% phytase activity indicating significantly improved stability within four weeks. Determination of water-soluble P and phytate-P indicates Bacillus sp. WR11 and CFS effectively degrade 20.17–38.55% phytate in soils. Quantification of biomass, phosphorus and iron contents shows positive effect of WR11 and its CFS on wheat growth. Particularly, the degradation ability (63.2%) and growth promoting effect of CFS is much stronger in the presence of 0.8% SA. Genomic prediction demonstrates strain WR11 has a large number of genes in the databases of Pathogen-Host Interactions, Virulence Factors of pathogenic bacteria, Antibiotic Resistance Genes, and the Comprehensive Antibiotic Research Database, respectively. In conclusion, SA significantly improves phytase production and phytate degradation. Phytase accompanied with SA may be an alternative strategy for phytate degradation and plant growth promotion, when microorganisms have safety issues.

Profiling Metabolites with Antifungal Activities from Endophytic Plant-Beneficial Strains of Pseudomonas chlororaphis Isolated from Chamaecytisus albus (Hack.) Rothm.

Fungal phytopathogens represent a large and economically significant challenge to food production worldwide. Thus, the application of biocontrol agents can be an alternative. In the present study, we carried out biological, metabolomic, and genetic analyses of three endophytic isolates from nodules of Chamaecytisus albus, classified as Pseudomonas chlororaphis acting as antifungal agents. The efficiency of production of their diffusible and volatile antifungal compounds (VOCs) was verified in antagonistic assays with the use of soil-borne phytopathogens: B. cinerea, F. oxysporum, and S. sclerotiorum. Diffusible metabolites were identified using chromatographic and spectrometric analyses (HPTLC, GC-MS, and LC-MS/MS). The phzF, phzO, and prnC genes in the genomes of bacterial strains were confirmed by PCR. In turn, the plant growth promotion (PGP) properties (production of HCN, auxins, siderophores, and hydrolytic enzymes, phosphate solubilization) of pseudomonads were bioassayed. The data analysis showed that all tested strains have broad-range antifungal activity with varying degrees of antagonism. The most abundant bioactive compounds were phenazine derivatives: phenazine-1-carboxylic acid (PCA), 2-hydroxy-phenazine, and diketopiperazine derivatives as well as ortho-dialkyl-aromatic acids, pyrrolnitrin, siderophores, and HCN. The results indicate that the tested P. chlororaphis isolates exhibit characteristics of biocontrol organisms; therefore, they have potential to be used in sustainable agriculture and as commercial postharvest fungicides to be used in fruits and vegetables.