The Potential of a Fluorescent-Based Approach for Bioassay of Antifungal Agents against Chili Anthracnose Disease in Thailand

Chutrakul, Chanikul; Khaokhajorn, Pratoomporn; Auncharoen, Patchanee; Boonruengprapa, Tanapong; Mongkolporn, Orarat

doi:10.1271/bbb.120669

Cited by 16 publications

(5 citation statements)

References 8 publications

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“…The broad spectrum antifungal agent Amphotericin B (AmB) was tested as the positive control. This natural polyene inhibits human and plant pathogens by binding with ergosterol [ 37 , 38 , 39 , 40 , 41 ]. Different stock concentrations of AmB dissolved in 100% DMSO were tested in a DRC to determine the IC 50 (µg/mL) and the sensitivity of the assay.…”

Section: Resultsmentioning

confidence: 99%

“…Although no significant differences were observed between the assay window at both concentrations (S/B was 47.18 and 55.12 for absorbance and 15.68 and 18.76 for fluorescence, respectively), the RZ' factors were slightly higher with 1 × 10 6 The broad spectrum antifungal agent Amphotericin B (AmB) was tested as the positive control. This natural polyene inhibits human and plant pathogens by binding with ergosterol [37][38][39][40][41]. Different stock concentrations of AmB dissolved in 100% DMSO were tested in a DRC to determine the IC 50 (µg/mL) and the sensitivity of the assay.…”

Section: Colletotrichum Acutatum Microdilution Assaymentioning

confidence: 99%

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Development and Validation of a HTS Platform for the Discovery of New Antifungal Agents against Four Relevant Fungal Phytopathogens

Serrano,

González-Menéndez,

Tormo

et al. 2023

JoF

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Fungal phytopathogens are the major agents responsible for causing severe damage to and losses in agricultural crops worldwide. Botrytis cinerea, Colletotrichum acutatum, Fusarium proliferatum, and Magnaporthe grisea are included in the top ten fungal phytopathogens that impose important plant diseases on a broad range of crops. Microbial natural products can be an attractive alternative for the biological control of phytopathogens. The objective of this work was to develop and validate a High-throughput Screening (HTS) platform to evaluate the antifungal potential of chemicals and natural products against these four important plant pathogens. Several experiments were performed to establish the optimal assay conditions that provide the best reproducibility and robustness. For this purpose, we have evaluated two media formulations (SDB and RPMI-1640), several inoculum concentrations (1 × 106, 5 × 105 and 5 × 106 conidia/mL), the germination curves for each strain, each strain’s tolerance to dimethyl sulfoxide (DMSO), and the Dose Response Curves (DRC) of the antifungal control (Amphotericin B). The assays were performed in 96-well plate format, where absorbance at 620 nm was measured before and after incubation to evaluate growth inhibition, and fluorescence intensity at 570 nm excitation and 615 nm emission was monitored after resazurin addition for cell viability evaluation. Quality control parameters (RZ’ Factors and Signal to Background (S/B) ratios) were determined for each assay batch. The assay conditions were finally validated by titrating 40 known relevant antifungal agents and testing 2400 microbial natural product extracts from the MEDINA Library through both HTS agar-based and HTS microdilution-based set-ups on the four phytopathogens.

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Section: Resultsmentioning

confidence: 99%

Section: Colletotrichum Acutatum Microdilution Assaymentioning

confidence: 99%

Development and Validation of a HTS Platform for the Discovery of New Antifungal Agents against Four Relevant Fungal Phytopathogens

Serrano,

González-Menéndez,

Tormo

et al. 2023

JoF

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“…Ellipticine and doxorubicin were used as positive controls for anti-KB and anti-NCI-H187 cell lines, while doxorubicin and tamoxifen were used as positive controls for the anti-MCF-7 cell line. The anti-Mycobacterium tuberculosis H37Ra (ATCC 25177), cytotoxic against Vero cells, anti-herpes simplex type-1 (HSV-1), and anti-phytopathogenic fungi, including Curvularia lunata and Magnaporthe grisea BCC 10261, were determined using the green fluorescent protein microplate assay (GFPMA) [25,26]. Amphotericin B was used as the positive control for antifungal activity.…”

Section: Biological Activity Of the Pure Compoundmentioning

confidence: 99%

The lichen-derived Streptomyces isolated from Pyxine cocoes produces the antibiotic with potent antimicrobial and antitumor activities

Somphong¹,

Poengsungnoen²,

Buaruang³

et al. 2023

ScienceAsia

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The actinobacteria strain Lp03 was isolated from the lichen, Pyxine cocoes, collected from Lumpini Park, Bangkok, Thailand. Based on the polyphasic approach, strain Lp03 was identified as Streptomyces parvulus.Actinomycin D produced by this strain exhibited antibacterial activity against Bacillus cereus (MIC 1.56 µg/ml), Bacillus subtilis (MIC 5 µg/ml), Escherichia coli (MIC 12.5 µg/ml), Kocuria rhizophila (MIC 0.076 ng/ml), Klebsiella pneumoniae (MIC 0.78 µg/ml), Staphylococcus aureus (MIC 9.76 ng/ml), Staphylococcus epidermidis (MIC 9.76 ng/ml), and Mycobacterium tuberculosis (MIC 12.5 µg/ml). Actinomycin D was inactive against Acinetobacter baumannii, but the new finding revealed that it showed activity against A. baumannii when combined with β-naphthylamide (PNβN), the efflux inhibitor. Antiparasitic activity was observed against Plasmodium falciparum with an IC 50 value of 14.8 ng/ml. It showed cytotoxicity against Vero cells, NCI-H187, KB, and MCF-7 with IC 50 values of 125, 0.694, 18.4 ng/ml, and 21.94 µg/ml, respectively. The genome of S. parvulus harbors a high diversity of biosynthetic gene clusters for secondary metabolites (BGCs). The actinomycin BGCs could be detected in all publicly available genomes of S. parvulus but could be varied among the strains. This study is the first record for the complete taxonomic study and bioactive compound produced by S. parvulus isolated from the lichen.

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“…The antimicrobial activities were tested against Bacillus cereus ATCC 11778, Acinetobacter baumanii ATCC 19606 T , Enterococcus faecium ATCC 51559, Escherichia coli ATCC 25922, Klebsiella pneumoniae ATCC 700603, Pseudomonas aeruginosa K2733 and Candida albicans BCC 58146 using the broth microdilution method [45]. The green fluorescent protein microplate assay was used for the evaluation of cytotoxicity against Vero (African green monkey kidney fibroblasts, ATCC CCL-81), antituberculosis against M. tuberculosis strain H37Ra [46], and antiphytopathogenic fungi including Alternaria brassicicola BCC 42724 , Colletotrichum capsici BMGC 106, Colletotrichum gloeosporioides BMGC107, Curvularia lunata BCC 15558 and Magnaporthe grisea BCC10261 [47]. The cytotoxicity against KB (oral human epidermoid carcinoma, ATCC CCL-17), MCF-7 (human breast cancer, ATCC HTC-22) and NCIH187 (human small-cell lung cancer, ATCC CRL-5804) cell lines was evaluated by using the resazurin microplate assay [48, 49].…”

Section: Biological Activitiesmentioning

confidence: 99%

Streptomyces macrolidinus sp. nov., a novel soil actinobacterium with potential anticancer and antimalarial activity

Kanchanasin

Sripreechasak

Suriyachadkun

et al. 2023

International Journal of Systematic and Evolutionary Microbiology

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A novel actinomycete, strain RY43-2T, belonging to the genus Streptomyces , was isolated from a peat swamp forest soil collected from Rayong Province, Thailand. The strain was characterized by using a polyphasic approach. The cell-wall peptidoglycan contained ll-diaminopimelic. Ribose and glucose were detected in its whole-cell hydrolysates. The strain contained anteiso-C15:0, iso-C14:0 and iso-C16:0 as the predominant fatty acids, and MK-9(H4), MK-9(H6) and MK-9(H8) as the major menaquinones. The phospholipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylinositol, phosphatidylinositol mannoside, three unidentified ninhydrin-positive phospholipids and two unidentified phospholipids. Strain RY43-2T showed the highest 16S rRNA gene similarity to Streptomyces misionensis JCM 4497T (98.9 %) and Streptomyces lichenis LCR6-01T (98.9 %). The draft genome of RY43-2T was 6.7 Mb with 6078 coding sequences with an average G+C content of 70.8 mol%. Genomic analysis revealed that the average nucleotide identity (ANI) values based on blast (ANIb) and MUMmer (ANIm) between strain RY43-2T and S. misionensis JCM 4497T were 80.1 and 86.1%, respectively. The ANIb and ANIm values between strain RY43-2T and S. lichenis LCR6-01T were 77.0 and 85.5%, respectively. The digital DNA–DNA hybridization values were 25.2 and 23.0% in comparison with the draft genomes of S. misionensis JCM 4497T and S. lichenis LCR6-01T, respectively. The results of taxonomic analysis suggested that strain RY43-2T represented a novel species of the genus Streptomyces for which the name Streptomyces macrolidinus sp. nov. is proposed. The type strain is RY43-2T (=TBRC 7286T=NBRC 115640T). Strain RY43-2T exhibited antimicrobial activity against Enterococcus faecium ATCC 51559, Colletotrichum capsici BMGC 106 and Colletotrichum gloeosporioides BMGC 107 with the minimum inhibitory concentration values of 25.0, 12.5, and 6.25 µg ml−1. It also exhibited potent antimalarial activity against Plasmodium falciparum K1 with IC50 of 0.0031 µg ml−1. In addition, it showed cytotoxicity against Vero, KB, MCF-7 and NCI-H187 with IC50 values of 0.0347, 6.15, 3.36 and 0.0352 µg ml−1, respectively.

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The Potential of a Fluorescent-Based Approach for Bioassay of Antifungal Agents against Chili Anthracnose Disease in Thailand

Cited by 16 publications

References 8 publications

Development and Validation of a HTS Platform for the Discovery of New Antifungal Agents against Four Relevant Fungal Phytopathogens

Development and Validation of a HTS Platform for the Discovery of New Antifungal Agents against Four Relevant Fungal Phytopathogens

The lichen-derived Streptomyces isolated from Pyxine cocoes produces the antibiotic with potent antimicrobial and antitumor activities

Streptomyces macrolidinus sp. nov., a novel soil actinobacterium with potential anticancer and antimalarial activity

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