Enrichment Culture Research Articles

Draft genome of an anaerobic nitrate-reducing, benzene-degrading member of the order Thermincolales.

We present a metagenome assembled genome (MAG) of an anaerobic bacterium from a nitrate-reducing, benzene-degrading enrichment culture (NRBC). The draft Thermincolales genome consists of 20 contigs with a total length of 4.09 Mbp and includes putative carboxylase genes likely involved in benzene activation.

Metagenomic analyses of aerobic bacterial enrichment cultures that degraded Tris(2-chloroethyl) phosphate (TCEP) and its transformation products

Tris(2-chloroethyl) phosphate (TCEP) is of growing public concern worldwide because of its ubiquitous contamination, toxicity, and persistence. In this study, we investigated bacterial communities in aerobic enrichment cultures with TCEP and its two major transformation products bis(2-chloroethyl) phosphate (BCEP) and 2-chloroethanol (2-CE) as the sole carbon source. Burkholderiales and Rhizobiales were likely two main bacterial guilds involved in the hydrolysis of TCEP, while Rhizobiales and Sphingomonadales may play an important role in the hydrolysis of BCEP, given the increase of Rhizobiales and Sphingomonadales-related phosphoesterase genes when the carbon source was switched from TCEP to BCEP. All Burkholderiales, Rhizobiales, Sphingomonadales were probably efficient in 2-CE metabolism, because their dehydrogenase genes and dehalogenase genes increased after 2-CE enrichment. The different substrate preference of different bacterial guilds highlighted the importance to understand the diversity and collaboration among functional bacteria. Meanwhile, two TCEP-degrading strains affiliated with Xanthobacter and Ancylobacter were isolated. Xanthobacter sp. strain T2-1 was able to degrade both TCEP and BCEP following the pseudo-first-order kinetics with reaction rates of 1.66 h−1 for TCEP and 1.02 h−1 for BCEP, respectively. Ancylobacter sp. strain T3-4 could degrade TCEP following the pseudo-first-order kinetics with a reaction rate of 2.54 h−1, but is unable to degrade BCEP. Additionally, strains that were phylogenetically closely related were found to have different degradation capabilities toward TCEP and/or BCEP, indicating the importance to investigate functional genes such as phosphoesterase genes.

Description and genome analysis of a novel archaeon isolated from a syntrophic pyrite-forming enrichment culture and reclassification of Methanospirillum hungatei strains GP1 and SK as Methanospirillum purgamenti sp. nov.

The archaeal isolate J.3.6.1-F.2.7.3T was obtained from an anaerobic enrichment culture, where it may play an important role in methane production during pyrite formation. The new isolate formed a species-level clade with Methanospirillum hungatei strains GP1 and SK, which is separate from the type strain JF-1T. Cultivation-independent surveys indicate the occurrence of this phylogenetic group in sediments and anaerobic digesters. The abundance of this clade appears to be negatively affected by high nitrogen loads, indicating a sensitivity to certain nitrogen compounds that is not known in M. hungatei JF-1T. The relatively large core genome of this Methanospirillum clade is indicative of niche specialization and efficient control of horizontal gene transfer. Genes for nitrogenase and F420-dependent secondary alcohol dehydrogenase contribute to the metabolic versatility of this lineage. Characteristics of the new isolate such as the ability to utilize 2-propanol as an electron donor or the requirement for acetate as a carbon source are found also in the strains GP1 and SK, but not in the type strain M. hungatei JF-1T. Based on the genomic differences to related species, a new species within the genus Methanospirillum is proposed with the name M. purgamenti sp. nov. The determined phenotypic characteristics support this proposal and indicate a metabolic adaptation to a separate ecological niche.

Continuous exposure to doxorubicin induces stem cell-like characteristics and plasticity in MDA-MB-231 breast cancer cells identified with the SORE6 reporter

PurposeCancer stem cells (CSCs) account for recurrence and resistance to breast cancer drugs, rendering them a cause of mortality and therapeutic failure. In this study, we examined the effects of exposure to low concentrations of doxorubicin (Dox) on CSCs and non-CSCs from TNBC.MethodsThe effects of Dox were studied using the SORE6 reporter system. We examined the enrichment of the CSCs population, as well as the proliferation, and death of the reporter-positive fraction (GFP + cells) by flow cytometry. The resistant and stemness phenotypes were analyzed by viability and mammosphere formation assay, respectively. We identified differentially expressed and coregulated genes by RNA-seq analysis, and the correlation between gene expression and clinical outcome was evaluated by Kaplan-Mayer analysis using public databases.ResultsIn MDAMB231 and Hs578t cells, we identified enriched subsets in the CSCs population after continuous exposure to low concentrations of Dox. Cells from these enriched cultures showed resistance to toxic concentrations of Dox and increased efficiency of mammosphere formation. In purified GFP + or GFP- cells, Dox increased the mammosphere-forming efficiency, promoted phenotypic switches in non-CSCs populations to a CSC-like state, reduced proliferation, and induced differential gene expression. We identified several biological processes and molecular functions that partially explain the development of doxorubicin-resistant cells and cellular plasticity. Among the genes that were regulated by Dox exposure, the expression of ITGB1, SNAI1, NOTCH4, STAT5B, RAPGEF3, LAMA2, and GNAI1 was significantly associated with poor survival, the stemness phenotype, and chemoresistance.ConclusionThe generation of chemoresistant cells that have characteristics of CSCs, after exposure to low concentrations of Dox, involves the differential expression of genes that have a clinical impact.

Complete genomic sequence of a Marinobacter species, a potential polyethylene degrader isolated from surface seawater.

The bacterium Marinobacter sp. RI1 was isolated from surface seawater through an enrichment culture using low-density polyethylene as the sole carbon source. Herein, we report its complete genomic sequence. Genomic annotation revealed that the strain harbors the genes encoding enzymes involved in alkane degradation, thus supporting polyethylene degradation.

Batch studies on the biodegradation of paracetamol and 1,4-hydroquinone by novel bacterial strains isolated from extreme environmental samples and the identification of candidate catabolic genes

The emerging pollutant paracetamol (APAP) is one of the most prescribed drugs worldwide. In addition, APAP and its main metabolites, namely, 4-aminophenol (4-AP), hydroquinone (H2Q), benzoquinone (BQ), and 2,5-dihydroxy-1,4-benzoquinone (2,5-OH-BQ), among others, are frequently detected in wastewater treatment plants (WWTPs) influents, effluents, and the environment. Thus, continuous release into the environment, especially aquatic environments, is a source of general concern. Six APAP-degrading bacterial strains were isolated from two mine samples from the Iberian Pyrite Belt (Lousal and Poderosa mines). Mycolicibacterium aubagnense HPB1.1, which was isolated using enrichment cultures from the Poderosa mine sample in the presence of H2Q as the sole carbon source, also showed APAP biodegrading capabilities. Pure cultures of this strain degraded 34.3 mg L−1 of APAP in 5 days and 9.4 mg L−1 of H2Q in 4 days. Interestingly, BQ and 2,5-OH-BQ were detected as metabolites resulting from H2Q abiotic degradation, but these compounds were removed in the strain’s cultures. Furthermore, M. aubagnense HPB1.1 whole-genome was sequenced, and its encoded proteins were aligned with enzymes of APAP-degrading bacteria recovered from databases and literature aiming to identify candidate catabolic genes. Putative amidases, deaminases, hydroxylases, and dioxygenases, responsible for the degradation of APAP by the HPB1.1 strain, were identified by similarity, corroborating its ability to transform APAP and its intermediate metabolite H2Q into less toxic metabolic compounds due to their capacity to break the aromatic ring of these molecules.

New insights on the degradation of polystyrene and polypropylene by larvae of the superworm Zophobas atratus and gut bacterial consortium enrichments obtained under different culture conditions

This study aims to deepen knowledge of the biodegradation of plastics, focusing on polypropylene (PP) fabric from surgical masks and polystyrene (PS) by larvae of Zophobas atratus as well as of specialized bacterial consortia from their gut, which were obtained in different enrichment conditions (aerobic, anaerobic, presence or absence of combined nitrogen). Plastics ingested by larvae obtained in Spain did not show any signs of oxidation but only limited depolymerization, preferably from the lowest molecular weight chains. Gut microbiota composition changed as an effect of plastic feeding. Such differences were more evident in bacterial enrichment cultures, where the polymer type influenced the composition more than by culture conditions, with an increase in the presence of nitrogen-fixers in anaerobic conditions. PS and PP degradation by different enrichment cultures was confirmed under aerobic and anaerobic conditions by respirometry tests, with anaerobic conditions favouring a more active plastic degradation. In addition, exposure to selected bacterial consortia in aerobiosis induced limited surface oxidation of PS. This possibly indicates that different biochemical routes are being utilized in the anaerobic gut and in aerobic conditions to degrade the polymer.

The Dissemination, Impact and Effectiveness of Jameson’s Cultural Theory: A Survey-Based Study in the Chinese Context

This study explores the dissemination, impact, and effectiveness of Fredric Jameson’s cultural theory within Chinese mass culture by conducting a comprehensive survey across various social groups and regions. The paper examines the acceptance and communication of Jameson’s theories, focusing on how his insights into cultural expansion and ideology resonate with different demographics in China. The results indicate that the primary audience for Jameson’s works consists of individuals aged 25-35 with high educational backgrounds, with significant interest in regions like Zhejiang, Hubei, Guangzhou, Beijing, Shanghai, and Shenzhen. The survey also reveals varying degrees of familiarity with Jameson’s theories among respondents, highlighting a growing influence yet uneven understanding. These findings suggest that a systematic interpretation of Jameson’s cultural theory can significantly contribute to the development and enrichment of Chinese mass culture.

Isolation and Molecular Identification of Novel Chicken Feather Degrading Bacteria Bacillus tropicus KRKJVR 5

The isolation and molecular identification of a novel chicken feather-degrading bacterium, Bacillus tropicus KRKJVR 5, were conducted to explore its potential in biotechnological applications. Soil samples collected from various sites in Suryapet district, Telangana, were screened using the enrichment culture method to identify potential keratinolytic producers. A total of ten bacterial isolates were initially identified, out of which five isolates demonstrated significant feather degradation on feather-containing agar media. Among these, Isolate 5 exhibited the highest keratinase activity, completely degrading feathers within 48 hours and producing 8.66 U/ml of keratinase, as determined by the Azocasein assay. Morphological and biochemical tests characterized Isolate 5 as a rod-shaped, Gram-positive Bacillus species with positive results for spore staining, Voges-Proskauer, catalase, starch hydrolysis, and carbohydrate fermentation tests, and negative results for indole, methyl red, citrate, H2S, and urease tests. Definitive genetic identification through 16S rRNA sequencing and comparison with the NCBI database confirmed Isolate 5 as Bacillus tropicus KRKJVR 5, with a 92% similarity to other Bacillus tropicus strains. The sequence data was submitted to NCBI and assigned the accession number PP859287. These findings highlight the potential of Bacillus tropicus KRKJVR 5 for efficient keratin degradation, making it a promising candidate for various industrial and environmental applications.

Impact of additives on syntrophic propionate and acetate enrichments under high-ammonia conditions

High ammonia concentrations in anaerobic degradation systems cause volatile fatty acid accumulation and reduced methane yield, which often derive from restricted activity of syntrophic acid-oxidising bacteria and hydrogenotrophic methanogens. Inclusion of additives that facilitate the electron transfer or increase cell proximity of syntrophic species by flocculation can be a suitable strategy to counteract these problems, but its actual impact on syntrophic interactions has yet to be determined. In this study, microbial cultivation and molecular and microscopic analysis were performed to evaluate the impact of conductive (graphene, iron oxide) and non-conductive (zeolite) additives on the degradation rate of acetate and propionate to methane by highly enriched ammonia-tolerant syntrophic cultures derived from a biogas process. All additives had a low impact on the lag phase but resulted in a higher rate of acetate (except graphene) and propionate degradation. The syntrophic bacteria ‘Candidatus Syntrophopropionicum ammoniitolerans’, Syntrophaceticus schinkii and a novel hydrogenotrophic methanogen were found in higher relative abundance and higher gene copy numbers in flocculating communities than in planktonic communities in the cultures, indicating benefits to syntrophs of living in close proximity to their cooperating partner. Microscopy and element analysis showed precipitation of phosphates and biofilm formation in all batches except on the graphene batches, possibly enhancing the rate of acetate and propionate degradation. Overall, the concordance of responses observed in both acetate- and propionate-fed cultures highlight the suitability of the addition of iron oxide or zeolites to enhance acid conversion to methane in high-ammonia biogas processes.Key points• All additives promoted acetate (except graphene) and propionate degradation.• A preference for floc formation by ammonia-tolerant syntrophs was revealed.• Microbes colonised the surfaces of iron oxide and zeolite, but not graphene.Graphical

Cellulolytic characterization of the rumen-isolated Acinetobacter pittii ROBY and design of a potential controlled-release drug delivery system

A novel cellulolytic bacterial strain, ROBY, was isolated from a bovine rumen sample using the enrichment culture method. This isolate was found to be Acinetobacter pittii, with >99 % similarity according to 16S rRNA gene sequence analysis. The potential use of this strain in combination with doxorubicin (Dox)-integrated cellulose nanoparticles (Dox-CNPs) was evaluated as a proof-of-concept study for the further development of this approach as a novel controlled-release drug delivery strategy. The isolate can utilize CNPs as the sole carbon source for growth and degrade both Dox-CNPs and empty CNPs with high efficiency. Extracellular cellulases isolated from bacteria may also be used to trigger Dox release. The results also demonstrated that the release of Dox into the environment due to nanoparticle degradation in the samples incubated with Dox-CNPs significantly affected bacterial cell viability (∼75 % decrease), proving the release of Dox due to bacterial cellulase activity and suggesting the great potential of this approach for further development.

Reversed oxidative TCA (roTCA) for carbon fixation by an Acidimicrobiia strain from a saline lake.

Acidimicrobiia are widely distributed in nature and suggested to be autotrophic via the Calvin-Benson-Bassham (CBB) cycle. However, direct evidence of chemolithoautotrophy in Acidimicrobiia is lacking. Here, we report a chemolithoautotrophic enrichment from a saline lake, and the subsequent isolation and characterization of a chemolithoautotroph, Salinilacustristhrix flava EGI L10123T, which belongs to a new Acidimicrobiia family. Although strain EGI L10123T is autotrophic, neither its genome nor Acidimicrobiia metagenome-assembled genomes (MAGs) from the enrichment culture encode genes necessary for the CBB cycle. Instead, genomic, transcriptomic, enzymatic, and stable-isotope probing data hinted at the activity of the reversed oxidative TCA (roTCA) coupled with the oxidation of sulfide as the electron donor. Phylogenetic analysis and ancestral character reconstructions of Acidimicrobiia suggested that the essential CBB gene rbcL was acquired through multiple horizontal gene transfer events from diverse microbial taxa. In contrast, genes responsible for sulfide- or hydrogen-dependent roTCA carbon fixation were already present in the last common ancestor of extant Acidimicrobiia. These findings imply the possibility of roTCA carbon fixation in Acidimicrobiia and the ecological importance of Acidimicrobiia. Further research in the future is necessary to confirm whether these characteristics are truly widespread across the clade.

Biodegradation of Crude Oil by Nitrate-Reducing, Sulfate-Reducing, and Methanogenic Microbial Communities under High-Pressure Conditions.

Carbon capture, utilization, and storage (CCUS) is an important component in many national net-zero strategies, and ensuring that CO2 can be safely and economically stored in geological systems is critical. Recent discoveries have shown that microbial processes (e.g., methanogenesis) can modify fluid composition and fluid dynamics within the storage reservoir. Oil reservoirs are under high pressure, but the influence of pressure on the petroleum microbial community has been previously overlooked. To better understand microbial community dynamics in deep oil reservoirs, we designed an experiment to examine the effect of high pressure (12 megapascals [MPa], 60 °C) on nitrate-reducing, sulfate-reducing, and methanogenic enrichment cultures. Cultures were exposed to these conditions for 90 d and compared with a control exposed to atmospheric pressure (0.1 MPa, 60 °C). The degradation characteristic oil compounds were confirmed by thin-layer analysis of oil SARA (saturates, aromatics, resins, and asphaltenes) family component rods. We found that the asphaltene component in crude oil was biodegraded under high pressure, but the concentration of asphaltenes increased under atmospheric pressure. Gas chromatography analyses of saturates showed that short-chain saturates (C8-C12) were biodegraded under high and atmospheric pressure, especially in the methanogenic enrichment culture under high pressure (the ratio of change was -81%), resulting in an increased relative abundance of medium- and long-chain saturates. In the nitrate-reducing and sulfate-reducing enrichment cultures, long-chain saturates (C22-C32) were biodegraded in cultures exposed to high-pressure and anaerobic conditions, with a ratio of change of -8.0% and -2.3%, respectively. However, the relative proportion of long-chain saturates (C22-C32) increased under atmospheric pressure. Gas Chromatography Mass Spectrometry analyses of aromatics showed that several naphthalene series compounds (naphthalene, C1-naphthalene, and C2-naphthalene) were biodegraded in the sulfate-reducing enrichment under both atmospheric pressure and high pressure. Our study has discerned the linkages between the biodegradation characteristics of crude oil and pressures, which is important for the future application of bioenergy with CCUS (bio-CCUS).

Microbial Allies in Plastic Degradation: Specific bacterial genera as universal plastic-degraders in various environments

Microbiological degradation of polymers offers a promising approach for mitigating environmental plastic pollution. This study (i) elucidated the diversity and structure of bacterial microbiomes from distinct environments (landfill soil, sewage sludge, and river water) characterized by specific physicochemical parameters, and (ii) utilized environment-derived microbial cultures enriched with microplastics (MPs) to investigate the degradation of polymers and identify culturable bacterial strains contributing to the plastisphere. We found that alpha diversity was notably higher in river water (∼20%) compared to landfill soil and sewage sludge. Dominant phyla included Pseudomonadota in sewage sludge (39.1%) and water (23.7%), while Actinomycetota prevailed in soil (38.5%). A multistage experiment, involving successive subcultures of environmental microbiomes exposed to polypropylene (PP), polyvinyl chloride (PVC), polycarbonate (PC), and polylactic acid (PLA), facilitated the assessment of MPs degradation processes. Analysis of carbonyl indices CIs and FTIR spectra revealed substantial structural changes in the treatment PVC-landfill soil, as well as in PLA- and PC-sludge enriched cultures. Further, using enriched cultures as a source of microorganisms, the study obtained 17 strains of plastic degraders from landfill soil, 14 from sewage sludge, and 6 from river water. Remarkably, similar bacterial genera were isolated across environmental microbiomes regardless of the MPs substrate used in enriched cultures. Among the 37 identified strains, Pseudomonadota predominated (64.86%) and were accompanied by Bacteroidota (16.22%), Actinomycetota (13.51%), and Bacillota (5.41%). This study highlights the complex relationship between microbiome diversity and the biodegradation efficiency of plastics, showing the potential for using microbial communities in the plastic pollution management.

Exploring Listeria monocytogenes in Ewe Milk: ssrA Gene-based Real-time PCR Identification, Phylogenetic Analysis, and Antibacterial Assessment of Magnesium Oxide Nanoparticles Synthesized with Myrtus communis Leaf Extract

Background: Listeria monocytogenes, a zoonotic pathogen affecting humans and animals, exhibits a global distribution, including Iraq. This study focused on the rapid identification and phylogenetic analysis of L. monocytogenes in freshly collected ewe milk samples from various farms in Al-Qadisiyah province, Iraq.Methods: The study was conducted with care and precision, involving 150 milk samples. These samples were subjected to traditional bacterial isolation and identification using the enrichment culture method and biochemical tests, with the PCR technique confirming the results. The antibacterial activity of MgONPs was then assessed using the disc diffusion method, ensuring a comprehensive and reliable approach to the study.Result: The results show that 150 ewe milk samples underwent real-time PCR (RT-PCR) targeting the ssrA gene, followed by partial 16S rRNA gene sequencing (PSGS) of purified conventional PCR products. Furthermore, the study entails the biosynthesis of magnesium oxide nanoparticles using Myrtus communis leaf extract, followed by a comprehensive characterization utilizing UV-spectra, FTIR, SEM, and TEM techniques. The Agar well diffusion method assessed the antibacterial efficacy of these Biosynthesized nanoparticles against L. monocytogenes. The RT-PCR results revealed the presence of L. monocytogenes in 36 out of 150 samples (24%). Subsequent PCR analysis confirmed the presence of the pathogen in 30 out of these 36 positive samples (83.33%). Sequencing of two purified PCR products demonstrated 100% nucleotide identity with global isolates from Iraq and Turkey. Furthermore, the study demonstrated that L. monocytogenes exhibited substantial sensitivity (24.66 ± 0.3) to the biosynthesized magnesium oxide nanoparticles. These findings underscore the speed and precision of the RT-PCR method for detecting L. monocytogenes in fresh ewe milk samples.Conclusion: This comprehensive investigation enhances our understanding of L. monocytogenes prevalence in ewe milk and highlights the potential of Myrtus communis -derived nanoparticles for combating this pathogen.Keywords: Antibacterial nanoparticles; Listeria monocytogenes; Magnesium oxide; Myrtus communis sheep milk; ssrA gene

Methyl-reducing methanogenesis by a thermophilic culture of Korarchaeia.

Methanogenesis mediated by archaea is the main source of methane, a strong greenhouse gas, and thus is critical for understanding Earth's climate dynamics. Recently, genes encoding diverse methanogenesis pathways have been discovered in metagenome-assembled genomes affiliated with several archaeal phyla1-7. However, all experimental studies on methanogens are at present restricted to cultured representatives of the Euryarchaeota. Here we show methanogenic growth by a member of the lineage Korarchaeia within the phylum Thermoproteota (TACK superphylum)5-7. Following enrichment cultivation of 'Candidatus Methanodesulfokora washburnenis' strain LCB3, we used measurements of metabolic activity and isotope tracer conversion to demonstrate methanol reduction to methane using hydrogen as an electron donor. Analysis of the archaeon's circular genome and transcriptome revealed unique modifications in the energy conservation pathways linked to methanogenesis, including enzyme complexes involved in hydrogen and sulfur metabolism. The cultivation and characterization of this new group of archaea is critical for a deeper evaluation of the diversity, physiology and biochemistry of methanogens.

Synergistic material–microbe interface toward deeper anaerobic defluorination

Per- and polyfluoroalkyl substances (PFAS), particularly the perfluorinated ones, are recalcitrant to biodegradation. By integrating an enrichment culture of reductive defluorination with biocompatible electrodes for the electrochemical process, a deeper defluorination of a C6-perfluorinated unsaturated PFAS was achieved compared to the biological or electrochemical system alone. Two synergies in the bioelectrochemical system were identified: i) The in-series microbial-electrochemical defluorination and ii) the electrochemically enabled microbial defluorination of intermediates. These synergies at the material-microbe interfaces surpassed the limitation of microbial defluorination and further turned the biotransformation end products into less fluorinated products, which could be less toxic and more biodegradable in the environment. This material-microbe hybrid system brings opportunities in the bioremediation of PFAS driven by renewable electricity and warrants future research on mechanistic understanding of defluorinating and electroactive microorganisms at the material-microbe interface for system optimizations.

Polystyrene Microplastics Degradation by Microbial Consortium From Jakarta Bay

ABSTRACTThe continuously increasing accumulation of microplastics in the environment, such as polystyrene (PS), has posed an ecological and health threat. However, the information and biodegradation capability of plastics, including polystyrene, was still in the infancy stage in Indonesia. This study explored the potential of microbial consortia from Jakarta Bay to degrade polystyrene microplastics. The consortia were obtained from plastic waste with biofilms on their surfaces, collected from three stations in Jakarta Bay. The research enriched the biofilm's consortia using 1/10 Zobell marine broth media supplemented with 0.1% PS microplastics. Using enriched cultures, biodegradation tests were performed in a mineral salt medium with 0.2% PS microplastics as a sole carbon source for 60 days at 27°C, 120 rpm. Microbial consortia from Jakarta Bay showed an ability to perform polystyrene biodegradation. The calculated weight loss of the microplastics after 60 days of incubation was between 4% and 6.4%. The biodegradation of PS microplastics by the microbial consortium was also indicated by FTIR spectra, which showed changes in functional group transmittances and were confirmed by SEM data showing holes formation on PS surfaces. The results provide a baseline study for further research development in Indonesia's polystyrene biodegradation technology and management.

Impact of Paenibacillus elgii supernatant on screening bacterial strains with potential for biotechnological applications

The biotechnological industry faces a crucial demand for novel bioactive compounds, particularly antimicrobial agents, to address the rising challenge of bacterial resistance to current available antibiotics. Traditional strategies for cultivating naturally occurring microorganisms often limit the discovery of novel antimicrobial producers. This study presents a protocol for targeted selection of bacterial strains using the supernatant of Paenibacillus elgii, which produces abundant signal molecules and antimicrobial peptides. Soil samples were inoculated in these enriched culture media to selectively cultivate bacteria resistant to the supernatant, indicating their potential to produce similar compounds. The bacterial strains isolated through this method were assessed for their antibacterial activity. In addition, the functional annotation of the genome of one of these strains revealed several gene clusters of biotechnological interest. This study highlights the effectiveness of using this approach for selective cultivation of microorganisms with potential for biotechnological applications.

Alternaria alternata Pathogen from Cuscuta japonica Could Serve as a Potential Bioherbicide.

Dodder (Cuscuta spp.) is a dangerous parasitic plant that causes serious damage to crop production and is challenging to eliminate. Herbicide application is a common strategy to control dodder in the field, but it is costly, ineffective, and further results in hazardous outcomes. Therefore, our study aims to identify the potential pathogens in naturally occurring dodder infections which may provide efficient biocontrol options. In this regard, the pathogens were isolated from the infected plants, their pathogenicity was validated through inoculation, and the optimal culture conditions for their growth were identified by determining the pathogenicity difference. The pathogenicity range was determined in vitro using the leaves of common horticultural plants and crops. Furthermore, a small range of horticultural plants parasitized by Cuscuta reflexa in the field were inoculated with the pathogen to determine their biosafety and biocontrol potential, and the pathogens were identified by morphological and molecular characterization. We found 7 strains that were isolated after pathogen enrichment culture. Among them, Cbp6 and Cbp7 showed the highest pathogenicity against C. reflexa. After testing the inoculation of more than 50 species of plants, only 9 species showed varying degrees of lesions on leaves, which proved the high biosafety for common plants. Field spraying of these pathogens showed a good control effect on C. reflexa after 21 days; the disease severityreached 66.0%, while its host plant did not display obvious symptoms. In conclusion, the pathogens Cbp6 and Cbp7 were identified as Alternaria alternata, and the results of this study provide a theoretical basis for the biological control of dodder.