RESUMEN
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.
Asunto(s)
Avicennia , Composición de Base , ADN Bacteriano , Gammaproteobacteria , Filogenia , ARN Ribosómico 16S , Avicennia/microbiología , ARN Ribosómico 16S/genética , China , ADN Bacteriano/genética , Gammaproteobacteria/clasificación , Gammaproteobacteria/genética , Gammaproteobacteria/aislamiento & purificación , Ácidos Grasos/análisis , Técnicas de Tipificación Bacteriana , Análisis de Secuencia de ADN , Hojas de la Planta/microbiología , Endófitos/genética , Endófitos/clasificación , Endófitos/aislamiento & purificaciónRESUMEN
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 16â¯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 3.7â¯% of the genera detected by culture-independent methods, accounting for 12-41â¯% 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.
Asunto(s)
Bacterias , Biodiversidad , Endófitos , Microbiota , Musa , Raíces de Plantas , Musa/microbiología , Endófitos/clasificación , Endófitos/aislamiento & purificación , Endófitos/genética , Raíces de Plantas/microbiología , Bacterias/clasificación , Bacterias/genética , Bacterias/aislamiento & purificación , Enfermedades de las Plantas/microbiología , ARN Ribosómico 16S/genética , Filogenia , Productos Agrícolas/microbiología , Brotes de la Planta/microbiología , Brotes de la Planta/crecimiento & desarrollo , Resistencia a la EnfermedadRESUMEN
Understanding the resident microbial communities and their above and below ground interactions with plants will provide necessary information for crop disease protection and stress management. In this study, we show how diversity of core microbiome varies with disease susceptibility of a crop. To test this hypothesis, we have focused on identifying the core microbial species of cotton leaf curl disease (CLCuD) susceptible Gossypium hirsutum and CLCuD resistant Gossypium arboreum under viral infestation. Derivation of core membership is challenging as it depends on an occupancy threshold of microbial species in a sampling pool, whilst accounting for different plant compartments. We have used an abundance-occupancy distribution approach where we dynamically assess the threshold for core membership, whilst marginalizing for occupancy in four compartments of the cotton plant, namely, leaf epiphyte, leaf endophyte, rhizosphere, and root endophyte. Additionally, we also fit a neutral model to the returned core species to split them into three groups, those that are neutral, those that are selected by the plant environment, and finally those that are dispersal limited. We have found strong inverse relationship between diversity of core microbiome and disease susceptibility with the resistant variety, G. arboreum, possessing higher diversity of microbiota. A deeper understanding of this association will aid in the development of biocontrol agents for improving plant immunity against biotrophic pathogens.
Asunto(s)
Resistencia a la Enfermedad , Gossypium , Microbiota , Enfermedades de las Plantas , Gossypium/microbiología , Enfermedades de las Plantas/microbiología , Hojas de la Planta/microbiología , Endófitos/clasificación , Endófitos/genética , Endófitos/aislamiento & purificación , Bacterias/clasificación , Bacterias/genética , Bacterias/aislamiento & purificación , Raíces de Plantas/microbiología , Biodiversidad , Rizosfera , Microbiología del SueloRESUMEN
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.
Asunto(s)
Endófitos , Pruebas de Sensibilidad Microbiana , Algas Marinas , Endófitos/aislamiento & purificación , Endófitos/genética , Endófitos/metabolismo , Endófitos/química , Endófitos/clasificación , Algas Marinas/microbiología , Bacterias/efectos de los fármacos , Bacterias/aislamiento & purificación , Bacterias/clasificación , Antiinfecciosos/farmacología , Antiinfecciosos/aislamiento & purificación , Antibacterianos/farmacología , Antibacterianos/aislamiento & purificación , Antifúngicos/farmacología , Antifúngicos/aislamiento & purificación , Hongos/efectos de los fármacos , Hongos/aislamiento & purificación , Hongos/clasificación , Bacterias Gramnegativas/efectos de los fármacos , Ulva/microbiología , Caulerpa/microbiología , Bacterias Grampositivas/efectos de los fármacosRESUMEN
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.
Asunto(s)
Bacterias , Endófitos , Filogenia , ARN Ribosómico 16S , Algas Marinas , Bacterias/clasificación , Bacterias/aislamiento & purificación , Bacterias/genética , Egipto , Algas Marinas/microbiología , Endófitos/clasificación , Endófitos/aislamiento & purificación , Endófitos/genética , Endófitos/fisiología , Mar Mediterráneo , ARN Ribosómico 16S/genética , Biodiversidad , Agua de Mar/microbiología , ADN Bacteriano/genética , EcosistemaRESUMEN
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.
Asunto(s)
Alternaria , Endófitos , Paclitaxel , Taxus , Taxus/microbiología , Paclitaxel/biosíntesis , Endófitos/genética , Endófitos/metabolismo , Endófitos/aislamiento & purificación , Endófitos/clasificación , Alternaria/genética , Alternaria/metabolismo , Alternaria/clasificación , Alternaria/aislamiento & purificación , Filogenia , Hongos/genética , Hongos/metabolismo , Hongos/clasificación , Hongos/aislamiento & purificación , ADN de Hongos/genética , ADN Espaciador Ribosómico/genéticaRESUMEN
BACKGROUND: Microbial growth during plant tissue culture is a common problem that causes significant losses in the plant micro-propagation system. Most of these endophytic microbes have the ability to propagate through horizontal and vertical transmission. On the one hand, these microbes provide a rich source of several beneficial metabolites. RESULTS: The present study reports on the isolation of fungal species from different in vitro medicinal plants (i.e., Breynia disticha major, Breynia disticha, Duranta plumieri, Thymus vulgaris, Salvia officinalis, Rosmarinus officinalis, and Ocimum basilicum l) cultures. These species were tested for their indole acetic acid (IAA) production capability. The most effective species for IAA production was that isolated from Thymus vulgaris plant (11.16 µg/mL) followed by that isolated from sweet basil plant (8.78 µg/mL). On screening for maximum IAA productivity, medium, "MOS + tryptophan" was chosen that gave 18.02 µg/mL. The macroscopic, microscopic examination and the 18S rRNA sequence analysis indicated that the isolate that given code T4 was identified as Neopestalotiopsis aotearoa (T4). The production of IAA by N. aotearoa was statistically modeled using the Box-Behnken design and optimized for maximum level, reaching 63.13 µg/mL. Also, IAA extract was administered to sweet basil seeds in vitro to determine its effect on plant growth traits. All concentrations of IAA extract boosted germination parameters as compared to controls, and 100 ppm of IAA extract exhibited a significant growth promotion effect for all seed germination measurements. CONCLUSIONS: The IAA produced from N. aotearoa (T4) demonstrated an essential role in the enhancement of sweet basil (Ocimum basilicum) growth, suggesting that it can be employed to promote the plant development while lowering the deleterious effect of using synthetic compounds in the environment.
Asunto(s)
Endófitos , Germinación , Ácidos Indolacéticos , Ocimum basilicum , Semillas , Thymus (Planta) , Ocimum basilicum/microbiología , Thymus (Planta)/química , Ácidos Indolacéticos/metabolismo , Endófitos/fisiología , Endófitos/metabolismo , Endófitos/aislamiento & purificación , Endófitos/genética , Germinación/efectos de los fármacos , Semillas/microbiología , Semillas/crecimiento & desarrollo , Semillas/efectos de los fármacosRESUMEN
Medicinal plant microbiomes undergo selection due to secondary metabolite presence. Resident endophytic/epiphytic microorganisms directly influence plant's bioactive compound synthesis. Hypothesizing low microbial diversity in Serjania erecta leaves, we assessed leaf colonization by epiphytic and endophytic fungi. Given its traditional medicinal importance, we estimated diversity in the endophytic fungal microbiome. Analyses included scanning electron microscopy (SEM), isolation of cultivable species, and metagenomics. Epiphytic fungi interacted with S. erecta leaf tissues, horizontally transmitted via stomata/trichome bases, expressing traits for nematode trapping. Cultivable endophytic fungi, known for phytopathogenic habits, didn't induce dysbiosis symptoms. This study confirms low leaf microbiome diversity in S. erecta, with a tendency towards more fungal species, likely due to antibacterial secondary metabolite selection. The classification of Halicephalobus sp. sequence corroborated the presence of nematode eggs on the epidermal surface of S. erecta by SEM. In addition, we confirmed the presence of methanogenic archaea and a considerable number of methanotrophs of the genus Methylobacterium. The metagenomic study of endophytic fungi highlighted plant growth-promoting yeasts, mainly Malassezia, Leucosporidium, Meyerozyma, and Hannaella. Studying endophytic fungi and S. erecta microbiomes can elucidate their impact on beneficial bioactive compound production, on the other hand, it is possible that the bioactive compounds produced by this plant can recruit specific microorganisms, impacting the biological system.
Asunto(s)
Hongos , Microbiota , Nematodos , Hojas de la Planta , Hojas de la Planta/microbiología , Hojas de la Planta/parasitología , Animales , Nematodos/microbiología , Hongos/clasificación , Hongos/genética , Hongos/aislamiento & purificación , Endófitos/genética , Endófitos/aislamiento & purificación , Levaduras/clasificación , Levaduras/aislamiento & purificación , Levaduras/genética , Metagenómica/métodos , BiodiversidadRESUMEN
Endophytes play an important role in plant development, survival, and establishment, but their temporal dynamics in young conifer plants are still largely unknown. In this study, the bacterial community was determined by metabarcoding of the 16S rRNA gene in the rhizoplane, roots, and aerial parts of 1- and 5-month-old seedlings of natural populations of Abies religiosa (Kunth) Schltdl. & Cham. In 1-month-old seedlings, Pseudomonas dominated aerial parts (relative abundance 71.6%) and roots (37.9%). However, the roots exhibited significantly higher bacterial species richness than the aerial parts, with the dissimilarity between these plant sections mostly explained by the loss of bacterial amplification sequence variants. After 5 months, Mucilaginibacter dominated in the rhizoplane (9.0%), Streptomyces in the roots (12.2%), and Pseudomonas in the aerial parts (18.1%). The bacterial richness and community structure differed significantly between the plant sections, and these variations were explained mostly by 1-for-1 substitution. The relative abundance of putative metabolic pathways significantly differed between the plant sections at both 1 and 5 months. All the dominant bacterial genera (e.g., Pseudomonas and Burkholderia-Caballeronia-Paraburkholderia) have been reported to have plant growth-promoting capacities and/or antagonism against pathogens, but what defines their role for plant development has still to be determined. This investigation improves our understanding of the early plant-bacteria interactions essential for natural regeneration of A. religiosa forest.
Asunto(s)
Abies , Bacterias , Endófitos , Raíces de Plantas , ARN Ribosómico 16S , Plantones , Plantones/microbiología , Plantones/crecimiento & desarrollo , Bacterias/clasificación , Bacterias/genética , Bacterias/aislamiento & purificación , Endófitos/clasificación , Endófitos/aislamiento & purificación , Endófitos/fisiología , Endófitos/genética , ARN Ribosómico 16S/genética , Abies/microbiología , Raíces de Plantas/microbiología , Microbiología del Suelo , Biodiversidad , Microbiota , ADN Bacteriano/genéticaRESUMEN
This study investigated the potential of endophytic fungi to produce paclitaxel (Taxol®), a potent anticancer compound widely employed in chemotherapy. This research aimed to identify, confirm, and characterize endophytic fungi capable of paclitaxel (PTX) production and assess their paclitaxel yield. Additionally, it aimed to investigate factors influencing paclitaxel production. A total of 100 endophytic fungal isolates were collected and identified from the roots of Artemisia judaica. Aspergillus fumigatiaffinis exhibited the highest PTX production (26.373 µg L-1) among the isolated endophytic fungi. The strain was identified as A. fumigatiaffinis (Accession No. PP235788.1). Molecular identification confirmed its novelty, representing the first report of PTX production by A. fumigatiaffinis, an endophyte of Artemisia judaica. Optimization through full factorial design of experiments (DOE) and response surface methodology (RSM) significantly enhanced PTX production to 110.23 µg L-1 from 1 g of dry weight of the fungal culture under optimal conditions of pH 8.0, 150 µg L-1 becozyme supplementation, and 18 days of fermentation in potato dextrose broth. The presence of paclitaxel was confirmed using thin layer chromatography, high performance liquid chromatography, and gas chromatography-mass spectrometry. These findings maximize the role of endophytic fungus to produce a secondary metabolite that might be able to replace the chemically produced PTX and gives an opportunity to provide a sustainable source of PTX eco-friendly at high concentrations. KEY POINTS: ⢠Endophytic fungi, like A. fumigatiaffinis, show promise for eco-friendly paclitaxel production ⢠Optimization strategies boost paclitaxel yield significantly, reaching 110.23 µg L -1 ⢠Molecular identification confirms novelty, offering a sustainable PTX source.
Asunto(s)
Aspergillus , Endófitos , Fermentación , Paclitaxel , Paclitaxel/biosíntesis , Aspergillus/metabolismo , Aspergillus/genética , Endófitos/metabolismo , Endófitos/genética , Endófitos/aislamiento & purificación , Endófitos/clasificación , Raíces de Plantas/microbiología , Medios de Cultivo/química , Cromatografía de Gases y Espectrometría de Masas , Cromatografía Líquida de Alta PresiónRESUMEN
BACKGROUND: Alternaria alternata is the primary pathogen of potato leaf spot disease, resulting in significant potato yield losses globally. Endophytic microorganism-based biological control, especially using microorganisms from host plants, has emerged as a promising and eco-friendly approach for managing plant diseases. Therefore, this study aimed to isolate, identify and characterize the endophytic fungi from healthy potato leaves which had great antifungal activity to the potato leaf spot pathogen of A. alternata in vitro and in vivo. RESULTS: An endophytic fungal strain SD1-4 was isolated from healthy potato leaves and was identified as Talaromyces muroii through morphological and sequencing analysis. The strain SD1-4 exhibited potent antifungal activity against the potato leaf spot pathogen A. alternata Lill, with a hyphal inhibition rate of 69.19%. Microscopic and scanning electron microscope observations revealed that the strain SD1-4 grew parallel to, coiled around, shrunk and deformed the mycelia of A. alternata Lill. Additionally, the enzyme activities of chitinase and ß-1, 3-glucanase significantly increased in the hyphae of A. alternata Lill when co-cultured with the strain SD1-4, indicating severe impairment of the cell wall function of A. alternata Lill. Furthermore, the mycelial growth and conidial germination of A. alternata Lill were significantly suppressed by the aseptic filtrate of the strain SD1-4, with inhibition rates of 79.00% and 80.67%, respectively. Decrease of leaf spot disease index from 78.36 to 37.03 was also observed in potato plants treated with the strain SD1-4, along with the significantly increased plant growth characters including plant height, root length, fresh weight, dry weight, chlorophyll content and photosynthetic rate of potato seedlings. CONCLUSION: The endophyte fungus of T. muroii SD1-4 isolated from healthy potato leaves in the present study showed high biocontrol potential against potato leaf spot disease caused by A. alternata via direct parasitism or antifungal metabolites, and had positive roles in promoting potato plant growth.
Asunto(s)
Alternaria , Endófitos , Enfermedades de las Plantas , Hojas de la Planta , Solanum tuberosum , Talaromyces , Alternaria/crecimiento & desarrollo , Alternaria/fisiología , Enfermedades de las Plantas/microbiología , Enfermedades de las Plantas/prevención & control , Solanum tuberosum/microbiología , Talaromyces/genética , Talaromyces/crecimiento & desarrollo , Endófitos/fisiología , Endófitos/aislamiento & purificación , Endófitos/genética , Hojas de la Planta/microbiología , Hifa/crecimiento & desarrollo , Antibiosis , Quitinasas/metabolismo , Agentes de Control Biológico , Control Biológico de Vectores/métodosRESUMEN
Here, we report on a Cordyceps species entering into a multi-trophic, multi-kingdom association. Cordyceps cateniannulata, isolated from the stem of wild Coffea arabica in Ethiopia, is shown to function as an endophyte, a mycoparasite and an entomopathogen. A detailed polyphasic taxonomic study, including a multilocus phylogenetic analysis, confirmed its identity. An emended description of C. cateniannulata is provided herein. Previously, this species was known as a pathogen of various insect hosts in both the Old and New World. The endophytic status of C. cateniannulata was confirmed by re-isolating it from inoculated coffee plants. Inoculation studies have further shown that C. cateniannulata is a mycoparasite of Hemileia vastatrix, as well as an entomopathogen of major coffee pests; infecting and killing Hypothenemus hampei and Leucoptera coffeella. This is the first record of C. cateniannulata from Africa, as well as an endophyte and a mycoparasite. The implications for its use as a biocontrol agent are discussed.
Asunto(s)
Coffea , Cordyceps , Endófitos , Filogenia , Endófitos/clasificación , Endófitos/aislamiento & purificación , Endófitos/genética , Endófitos/fisiología , Cordyceps/genética , Cordyceps/clasificación , Coffea/microbiología , Coffea/parasitología , Animales , Enfermedades de las Plantas/microbiología , Enfermedades de las Plantas/parasitología , Etiopía , ADN de Hongos/genética , ADN de Hongos/química , ADN Espaciador Ribosómico/genética , ADN Espaciador Ribosómico/química , Tallos de la Planta/microbiología , Tallos de la Planta/parasitología , Análisis de Secuencia de ADN , Análisis por ConglomeradosRESUMEN
Beauveria bassiana (Bb) is a widespread entomopathogenic fungus widely used in agriculture for crop protection. Other than pest control, fungi belonging to the B. bassiana complex represent an important microbial resource in agroecosystems, considering their multiple interactions with other microorganisms as antagonists of phytopathogens, or with plants as endophytic colonizers and growth promoters. Here, we characterised field collected or commercial isolates of B. bassiana relative to the environmental factors that affect their growth. We further compared the metabolome, the entomopathogenic potential and biocontrol activity of the tested isolates respectively on the insect pest Spodoptera littoralis or against the fungal plant pathogen Fusarium oxysporum. Our analysis revealed that the B. bassiana complex is characterised by a high level of inter-isolate heterogeneity in terms of nutritional requirements, establishment of intra- or inter-kingdom interactions, and the nature of metabolites produced. Interestingly, certain B. bassiana isolates demonstrated a preference for low nutrient plant-derived media, which hints at their adaptation towards an endophytic lifestyle over a saprophytic one. In addition, there was a noticeable variation among different B. bassiana isolates in their capacity to kill S. littoralis larvae in a contact infection test, but not in an intrahaemocoelic injection experiment, suggesting a unique level of adaptability specific to the host. On the other hand, most B. bassiana isolates exhibited similar biocontrol efficacy against the soil-dwelling ascomycete F. oxysporum f. sp. lycopersici, a pathogen responsible for vascular wilt disease in tomato plants, effectively averting wilting. Overall, we show that the effectiveness of B. bassiana isolates can greatly vary, emphasising the importance of isolate selection and nutritional adaptability consideration for their use in sustainable agriculture.
Asunto(s)
Beauveria , Fusarium , Larva , Control Biológico de Vectores , Spodoptera , Beauveria/fisiología , Beauveria/aislamiento & purificación , Beauveria/metabolismo , Animales , Spodoptera/microbiología , Larva/microbiología , Enfermedades de las Plantas/microbiología , Enfermedades de las Plantas/prevención & control , Agricultura , Metaboloma , Endófitos/aislamiento & purificación , Endófitos/metabolismo , Endófitos/fisiología , Endófitos/clasificaciónRESUMEN
Gray mold, caused by Botrytis cinerea, is a prevalent postharvest disease of apple that limits their shelf life, resulting in significant economic losses. The use of antagonistic microorganisms has been shown to be an effective approach for managing postharvest diseases of fruit. In the present study, an endophytic yeast strain PGY-2 was isolated from apples and evaluated for its biocontrol efficacy against gray mold and its mechanisms of action. Results indicated that strain PGY-2, identified as Bullera alba, reduced the occurrence of gray mold on apples and significantly inhibited lesion development in pathogen-inoculated wounds. Gray mold control increased with the use of increasing concentrations of PGY-2, with the best disease control observed at 108 cells/mL. Notably, Bullera alba PGY-2 did not inhibit the growth of Botrytis cinerea in vitro indicating that the yeast antagonist did not produce antimicrobial compounds. The rapid colonization and stable population of PGY-2 in apple wounds at 4 °C and 25 °C confirmed its ability to compete with pathogens for nutrients and space. PGY-2 also had a strong ability to form a biofilm and enhanced the activity of multiple defense-related enzymes (POD, PPO, APX, SOD, PAL) in host tissues. Our study is the first time to report the use of Bullera alba PGY-2 as a biocontrol agent for postharvest diseases of apple and provide evidence that Bullera alba PGY-2 represents an endophytic antagonistic yeast with promising biocontrol potential and alternative to the use of synthetic, chemical fungicides for the control of postharvest gray mold in apples.
Asunto(s)
Antibiosis , Botrytis , Endófitos , Frutas , Malus , Enfermedades de las Plantas , Malus/microbiología , Botrytis/crecimiento & desarrollo , Botrytis/fisiología , Botrytis/efectos de los fármacos , Enfermedades de las Plantas/microbiología , Enfermedades de las Plantas/prevención & control , Endófitos/fisiología , Endófitos/aislamiento & purificación , Frutas/microbiología , Levaduras/fisiología , Levaduras/aislamiento & purificación , Biopelículas/crecimiento & desarrolloRESUMEN
Apple ring rot, one of the most common apple postharvest diseases during storage, is caused by Botryosphaeria dothidea. Presently, the disease management is primarily dependent on chemical fungicide application. Here we demonstrated an endophyte bacterium Bacillus tequilensis QNF2, isolated from Chinese leek (Allium tuberosum) roots considerably suppressed B. dothidea mycelial growth, with the highest suppression of 73.56 % and 99.5 % in the PDA and PDB medium, respectively in vitro confront experiments. In in vivo experiments, B. tequilensis QNF2 exhibited a control efficacy of 88.52 % and 100 % on ring rot disease on postharvest apple fruits inoculated with B. dothidea disc and dipped into B. dothidea culture, respectively. In addition, B. tequilensis QNF2 volatile organic compounds (VOCs) also manifested markedly inhibition against B. dothidea mycelial growth and the ring rot on postharvest apple fruits. Moreover, B. tequilensis QNF2 severely damaged the mycelial morphology of B. dothidea. Finally, B. tequilensis QNF2 significantly repressed the expression of six pathogenicity-related genes, such as adh, aldh, aldh3, galm, pdc1, pdc2, involved in glycolysis/gluconeogenesis of B. dothidea. The findings of the study proved that B. tequilensis QNF2 was a promising alternative for controlling apple ring rot of postharvest apple fruit.
Asunto(s)
Ascomicetos , Bacillus , Endófitos , Frutas , Malus , Enfermedades de las Plantas , Malus/microbiología , Enfermedades de las Plantas/microbiología , Ascomicetos/crecimiento & desarrollo , Ascomicetos/efectos de los fármacos , Ascomicetos/genética , Ascomicetos/fisiología , Bacillus/genética , Bacillus/fisiología , Bacillus/aislamiento & purificación , Endófitos/genética , Endófitos/metabolismo , Endófitos/aislamiento & purificación , Endófitos/clasificación , Endófitos/fisiología , Frutas/microbiología , Compuestos Orgánicos Volátiles/farmacología , Compuestos Orgánicos Volátiles/metabolismo , Compuestos Orgánicos Volátiles/análisis , Antibiosis , Micelio/crecimiento & desarrollo , Micelio/efectos de los fármacosRESUMEN
In the present study, eleven endophytic bacterial strains, Herbaspirillum sp. (GP-SGM1, GP-SGM2, GP-SGM3, and GP-SGM11), Pseudomonas sp. (GP-SGM4, GP-SGM5), Novosphingobium sp. GP-SGM6, Chryseobacterium sp. GP-SGM7, Labedella sp. GP-SGM8, Brevibacterium sp. GP-SGM9, and Pseudomonas sp. GP-SGM10, were isolated from the rhizomes of Gunnera perpensa L. The growth kinetics, assessed through maximum growth rates (µmax) and optical density (OD) values, revealed that GP-SGM7 exhibited highest µmax values of 0.33 ± 0.01 hours (h)-1 with an OD of 4.20 ± 0.04. In contrast, GP-SGM11 exhibited the lowest µmax of 0.12 ± 0.05 h-1 and the smallest OD of 1.50 ± 0.00. In addition, the endophyte crude extracts were tested for antibacterial activity against five pathogenic strains using the disk diffusion method, with GP-SGM7 crude extracts exhibiting promising antibacterial activity against Klebsiella pneumoniae and Staphylococcus aureus. Antioxidant activity was determined by DPPH (2, 2-diphenyl-1-picrylhydrazyl) and FRAP (ferric reducing antioxidant power) assays. The crude extracts of GP-SGM1, GP-SGM7, GP-SGM9, and GP-SGM10 were the most effective at scavenging DPPH radicals, with GP-SGM7 also exhibiting a high FRAP value of 0.54 ± 0.01. These findings emphasize the therapeutic potential of endophytic bacteria from G. perpensa L. in addressing skin-related issues, including bacterial infections and free radicals.
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Bacterias , Endófitos , Endófitos/aislamiento & purificación , Endófitos/clasificación , Endófitos/genética , Endófitos/metabolismo , Endófitos/química , Bacterias/aislamiento & purificación , Bacterias/clasificación , Bacterias/efectos de los fármacos , Bacterias/genética , Antioxidantes/farmacología , Antioxidantes/metabolismo , Antioxidantes/química , Antibacterianos/farmacología , Filogenia , ARN Ribosómico 16S/genética , Pruebas de Sensibilidad MicrobianaRESUMEN
Argemone mexicana belonging to family Papaveraceae is a traditional medicinal plant widely utilized by tribal people in India for treating various ailments like skin infections, wounds and inflammation. This plant is very rich in alkaloidal content, which has a great potential in the treatment of anti-inflammatory disorders. Therapeutically promising bioactive molecules are often produced by endophytic fungi associated with medicinal plants. In this investigation, endophytic fungi were isolated from various parts of A. mexicana and screened for alkaloidal content. Among these, one of the fungal isolate, Acremonium alternatum AMEF-5 producing maximum alkaloids showed significant anti-inflammatory activity. Fractionation of this crude fungal extract through column chromatography yielded eight fractions, which were further screened for anti-inflammatory activities. Fraction 3 exhibited significant anti-inflammatory activity by the inhibition of lipoxygenase enzyme (IC50 15.2 ± 0.09 µg/ml), scavenging of the nitric oxide radicals (IC50 11.38 ± 0.35 µg/ml), protein denaturation (IC50 14.93 ± 0.4 µg/ml), trypsin inhibition (IC50 12.06 ± 0.64 µg/ml) and HRBC stabilization (IC50 11.9 ± 0.22 µg/ml). The bioactive alkaloid in fraction 3 was identified as aconitine which was confirmed by UV, FTIR, HPLC, HRMS, 1H NMR, and 13C NMR analysis. This study demonstrates that endophytic fungi serve a potential source for sustainable production of therapeutically important alkaloids.
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Aconitina , Acremonium , Antiinflamatorios , Endófitos , Acremonium/metabolismo , Acremonium/química , Antiinflamatorios/farmacología , Aconitina/farmacología , Aconitina/química , Endófitos/metabolismo , Endófitos/química , Endófitos/aislamiento & purificación , Animales , Óxido Nítrico/metabolismo , Ratones , Alcaloides/farmacología , Lipooxigenasa/metabolismo , Células RAW 264.7 , IndiaRESUMEN
This study investigates the synthesis of vinblastine by endophytic fungi isolated from leaf of C. roseus. A total of 10 endophytic fungi were selected for secretion of vinca alkaloids based on the initial screening by biochemical tests and thin-layer chromatography (TLC). Out of these ten, only four fungal extracts showed positive results for presence of vinblastine at same retention time (10 min.) compared to reference compound on HPLC analysis. The detected concentration of vinblastine was maximum (17 µg/ml) in isolate no. CRL 22 followed by CRL 52, CRL 17 and CRL 28. To validate the presence of vinblastine, ultra-high-performance liquid chromatography coupled with high-resolution accurate mass spectrometry (HRMS) was employed. This analysis confirmed the presence of anhydrovinblastine, a precursor of vinblastine through the detection of molecular ions at m/z 793.4185 in extract of CRL 17. In addition to anhydrovinblastine, the intermediate compounds essential to the biosynthetic pathway of vinblastine were also detected in the extract of CRL 17. These host-origin compounds strongly suggest the presence of a biosynthetic pathway within the endophytic fungus. Based on morphological observation and sequence analysis of the ITS region of rDNA, endophytic fungi were identified as Alternaria alternata (CRL 17), Curvularia lunata (CRL 28), Aspergillus terrus (CRL 52), and Aspergillus clavatonanicus (CRL 22).
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Catharanthus , Endófitos , Hongos , Hojas de la Planta , Vinblastina , Catharanthus/microbiología , Vinblastina/metabolismo , Endófitos/metabolismo , Endófitos/aislamiento & purificación , Cromatografía Líquida de Alta Presión , Hongos/metabolismo , Hongos/aislamiento & purificación , Hongos/clasificación , Hongos/genética , Hojas de la Planta/microbiología , Cromatografía en Capa Delgada , Vías Biosintéticas , Espectrometría de MasasRESUMEN
Endophytic fungi are widely known as fungi that infect internal tissues of host plants for all or part of their life cycles, without causing visible symptoms of disease. The present study was carried out to identify and investigate the pathogenicity of endophytic fungi residing in husks, silks, and kernels of corn. Endophytic fungi were isolated from surface-sterilised silks, kernels, and husks of healthy corn plants and identified using sequencing of multiple markers comprising TEF-1α, ß-tubulin, calmodulin, ITS, LSU, and ACT. A total of 56 isolates of endophytic fungi belonging to 17 species, namely Fusarium pseudocircinatum (n = 8), F. verticillioides (n = 2), F. andiyazi (n = 4), F. sacchari (n = 1), F. mangiferae (n = 1), F. fujikuroi (n = 1), F. proliferatum (n = 3), F. incarnatum (n = 2), Penicillium oxalicum (n = 2), P. polonicum (n = 2), P. citrinum (n = 11), Aspergillus flavus (n = 10), A. tubingensis (n = 1), Cladosporium tenuissimum (n = 3), Aureobasidium pullulans (n = 3), Curvularia lunata (n = 1), and Epicoccum sorghinum (n = 1) were identified. Pathogenicity test showed that all endophytic fungi induced varying severities of disease symptoms on corn plants such as leaf chlorosis and necrosis, stem malformation, wilt, and stunted growth with F. verticillioides being the most virulent. The study revealed that corn tissues harbour diverse genera of endophytic fungi that can infect corn plants and may cause harmful effects to the host plants.
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Endófitos , Enfermedades de las Plantas , Zea mays , Zea mays/microbiología , Endófitos/genética , Endófitos/aislamiento & purificación , Endófitos/patogenicidad , Enfermedades de las Plantas/microbiología , Hongos/patogenicidad , Hongos/aislamiento & purificación , Hongos/genética , Hongos/clasificación , Filogenia , Fusarium/patogenicidad , Fusarium/genética , Fusarium/aislamiento & purificación , VirulenciaRESUMEN
Phellinus caribaeo-quercicola is a basidiomycetous fungus, isolated as an endophyte in this study from the healthy and symptomless leaves of Inula racemosa Hook. f., an important medicinal herb growing in Kashmir Himalaya. This study combines morphological, molecular and phylogenetic techniques to identify the fungal endophyte, using the ITS sequence of nrDNA. A detached leaf assay was conducted to assess the pathogenicity of the fungal endophyte suggesting its mutually symbiotic relationship with the host. The authors also investigated the antifungal potential of the isolated endophytic strain to ascertain its use as a biocontrol agent. The study shows that P. caribaeo-quercicola INL3-2 strain exhibits biocontrol activity against four key fungal phytopathogens that cause significant agronomic and economic losses: Aspergillus flavus, Aspergillus niger, Fusarium solani, and Fusarium oxysporum. Notably, P. caribaeo-quercicola INL3-2 strain is highly effective against A. flavus, with an inhibition percentage of 57.63%. In addition, this study investigates the antioxidant activity of P. caribaeo-quercicola INL3-2 strain crude extracts using ethyl acetate and methanol as solvents. The results showed that the methanolic fraction of P. caribaeo-quercicola exhibits potential as an antioxidant agent, with an IC50 value of 171.90 ± 1.15 µg/mL. This investigation is first of its kind and marks the initial report of this fungal basidiomycete, P. caribaeo-quercicola, as an endophyte associated with a medicinal plant. The findings of this study highlight the potential of P. caribaeo-quercicola INL3-2 strain as a dual-action agent with both biocontrol and antioxidant properties consistent with the medicinal properties of Inula racemosa. This endophytic fungus could be a promising source of natural compounds for use in agriculture, medicine, and beyond.