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1.
Front Plant Sci ; 13: 874819, 2022.
Article in English | MEDLINE | ID: mdl-35646017

ABSTRACT

Banana (Musa spp.) is an important fruit crop cultivated in most tropical countries. Banana Fusarium wilt caused by Fusarium oxysporum f. sp. cubense tropical race 4 (Foc TR4) is the most destructive fungal disease. Biocontrol using endophytic microorganisms is considered as a safety and sustainable strategy. Actinomycetes have a potential for the production of diverse metabolites. Isolation of endophytic actinomycetes with high efficiency and broad-spectrum antagonism is key for exploring biocontrol agents. Our previous study showed that a total of 144 endophytic actinomycetes were isolated from different tissues of medicinal plants in Hainan, China. Especially, strain 8ZJF-21 exhibited a broad-spectrum antifungal activity. Its morphological, physiological, and biochemical characteristics were consistent with the genus Streptomyces. The phylogenetic tree demonstrated that strain 8ZJF-21 formed a distinct clade with Streptomyces malaysiensis. Average nucleotide identity (ANI) was 98.49% above the threshold of novel species. The pot experiment revealed that endophytic Streptomyces malaysiensis 8ZJF-21 could improve the plant resistance to Foc TR4 by enhancing the expression levels of defense-related and antioxidant enzyme genes. It also promoted the plant growth by producing several extracellular enzymes and metabolites. Antifungal mechanism assays showed that S. malaysiensis 8ZJF-21 extract inhibited mycelial growth and spore germination of Foc TR4 in vitro. Pathogenic cells occurred cytoplasmic heterogeneity, disappeared organelles, and ruptured ultrastructure. Sequencing and annotation of genome suggested that S. malaysiensis 8ZJF-21 had a potential of producing novel metabolites. Nineteen volatile organic compounds were obtained from the extract by Gas Chromatography-Mass Spectrometry (GC-MS). Hence, endophytic Streptomyces strains will become essential biocontrol agents of modern agricultural practice.

2.
Phytopathology ; 112(9): 1877-1885, 2022 Sep.
Article in English | MEDLINE | ID: mdl-35471064

ABSTRACT

Fusarium wilt of banana caused by Fusarium oxysporum f. sp. cubense tropical race 4 (Foc TR4) is one of the most disastrous fungal diseases. Biological control is a promising strategy for controlling Fusarium wilt of banana. To explore endophytic actinomycetes as biocontrol resources against Foc TR4, antagonistic strains were isolated from different tissues of medicinal plants. Here, a total of 144 actinomycetes were isolated and belonged to Nonomuraea, Kitasatospora, and Streptomyces. Forty-three isolates exhibited antifungal activities against Foc TR4. The strain labeled with 5-4 isolated from roots of Piper austrosinense had a broad-spectrum antifungal activity by the production of chitinase and ß-1,3-glucanase and was identified as Streptomyces hygroscopicus subsp. hygroscopicus 5-4. Furthermore, disease index of banana wilt was significantly reduced by application of strain 5-4 in comparison with application of Foc TR4 alone. Exogenous application of strain 5-4 increased the expression levels of defense genes such as (PAL), peroxidase (POD), pathogenesis-related protein 1 (PR-1), hydrolytic enzymes (ß-1,3-glucanase), lysin motif receptor kinase 1 (LYK-1), and mitogen-activated protein kinase 1 (MPK-1). The antifungal mechanism assay demonstrated that extracts of strain 5-4 inhibited spore gemination and hyphal growth of Foc TR4, and caused abnormally swollen, deformity, and rupture of Foc TR4 hypha. Thus, S. hygroscopicus subsp. hygroscopicus 5-4 could be used as a potential biological agent for controlling Fusarium wilt of banana.


Subject(s)
Fusarium , Musa , Streptomyces , Antifungal Agents/pharmacology , Fusarium/genetics , Gene Expression Profiling , Musa/microbiology , Plant Diseases/microbiology , Streptomyces/genetics
3.
Front Microbiol ; 11: 610698, 2020.
Article in English | MEDLINE | ID: mdl-33552022

ABSTRACT

Fusarium wilt of banana caused by Fusarium oxysporum f. sp. cubense (Foc) is a disastrous soil-borne fungal disease. Foc tropical race 4 (Foc TR4) can infect almost all banana cultivars. Until now, there is a shortage of safety and effective control methods and commercial banana cultivars with a resistance against Foc TR4. Biocontrol using environmentally friendly microbes is a promising strategy for the management of Foc TR4. Here, a strain 5-10, newly isolated from a medicinal plant (Curculigo capitulata), exhibited a high antifungal activity against Foc TR4. Combing the morphological characteristics and molecular identification, strain 5-10 was classified as a Streptomyces genus. The sequenced genome revealed that more than 39 gene clusters were involved in the biosynthesis of secondary metabolites. Some multidrug resistance gene clusters were also identified such as mdtD, vatB, and vgaE. To improve the anti-Foc TR4 activity of the strain 5-10 extracts, an optimization method of fermentation broth was established. Antifungal activity increased by 72.13% under the fermentation system containing 2.86 g/L of NaCl and 11.57% of inoculation amount. After being treated with the strain 5-10 extracts, the Foc TR4 hyphae shrinked, deformed, and ruptured. The membrane integrity and cell ultrastructure incurred irreversible damage. Streptomyces sp. 5-10 extracts play a fungicidal role in Foc TR4. Hence, Streptomyces sp. 5-10 will be a potential biocontrol agent to manage fungal diseases by exploring the microbial fertilizer.

4.
Int J Biol Macromol ; 117: 1314-1325, 2018 Oct 01.
Article in English | MEDLINE | ID: mdl-29859842

ABSTRACT

Response surface methodology was used to optimize the extraction conditions for ultrasonic-assisted extraction of polysaccharides from mango pomace. The Optimum extraction conditions consisted of extraction temperature of 74 °C, ultrasonic power of 170 W, extraction time of 100 min, and raw material-to-water ratio of 1:40 g/mL. Under these conditions, the extraction yield was 3.71 ±â€¯0.07%. Three novel polysaccharide fractions, MG-1, MG-2 and MG-3 were purified from the crude polysaccharides by using DEAE-52 cellulose and Sephadex G-100 column chromatography. The molecular weight and monosaccharide composition of polysaccharide fractions (MPFs) were analyzed by high performance liquid gel permeation chromatography (HPGPC) and HPLC analysis, respectively. The characterizations of MPFs were conducted with FT-IR, 1H NMR and SEM. Furthermore, the anticancer activities of the polysaccharide fractions were also investigated in vitro. Results showed that MG-1, MG-2 and MG-3 exhibited significant anticancer activities against HepG2, MCF-7, A549, HeLa, A2780, HCT-116 and BGC-823 cells in a dose-dependent manner. MPFs were also showed to promote apoptosis as seen in the nuclear morphological examination study using calcein acetyl methoxy methyl easter (calcein-AM) and propidium iodide (PI) staining. This research could serve as a theoretical reference for the efficient utilization of MPFs in biomedical and functional food.


Subject(s)
Antineoplastic Agents, Phytogenic/chemistry , Antineoplastic Agents, Phytogenic/pharmacology , Mangifera/chemistry , Polysaccharides/chemistry , Polysaccharides/pharmacology , Antineoplastic Agents, Phytogenic/isolation & purification , Apoptosis/drug effects , Cell Line, Tumor , Cell Survival/drug effects , Chemical Fractionation/methods , Chromatography, High Pressure Liquid , Humans , Molecular Weight , Phytochemicals/chemistry , Plant Extracts/chemistry , Plant Extracts/isolation & purification , Plant Extracts/pharmacology , Polysaccharides/isolation & purification , Temperature
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