title: The potential of a fluorescent-based approach for bioassay of antifungal agents against chili anthracnose disease in Thailand.
abstract: Severe chili anthracnose disease in Thailand is caused by Colletotrichum gloeosporioides and C. capsici. To discover anti-anthracnose substances we developed an efficient dual-fluorescent labeling bioassay based on a microdilution approach. Indicator strains used in the assay were constructed by integrating synthetic green fluorescent protein (sGFP) and Discosoma sp. red fluorescent protein (DsRedExp) genes into the genomes of C. gloeosporioides or C. capsici respectively. Survival of co-spore cultures in the presence of inhibitors was determined by the expression levels of these fluorescent proteins. This developed assay has high potential for utilization in the investigation of selective inhibition activity to either one of the pathogens as well as the broad-range inhibitory effect against both pathogens. The value of using the dual-fluorescent assay is rapid, reliable, and consistent identification of anti-anthracnose agents. Most of all, the assay enables the identification of specific inhibitors under the co-cultivation condition.
authors: National Center for Genetic Engineering and Biotechnology (BIOTEC), Thailand Science Park, Phahon Yothin Read, Klong Nueng, Klong Luang, Pathum Thani, Tahiland. chanikul@biotec.or.th
authors: Chanikul
authors: C
authors: Chutrakul
authors: Pratoomporn
authors: P
authors: Khaokhajorn
authors: Patchanee
authors: P
authors: Auncharoen
authors: Tanapong
authors: T
authors: Boonruengprapa
authors: Orarat
authors: O
authors: Mongkolporn
mesh_terms: D000935:Antifungal Agents / Q000737:chemistry / Q000494:pharmacology; D001681:Biological Assay; D002212:Capsicum / Q000382:microbiology*; D020231:Colletotrichum / Q000145:classification / Q000235:genetics / Q000302:isolation & purification* / Q000472:pathogenicity; D017930:Genes, Reporter; D005818:Genetic Engineering; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D008856:Microscopy, Fluorescence; D010935:Plant Diseases / Q000382:microbiology / Q000517:prevention & control*; D013172:Spores, Fungal / Q000145:classification / Q000235:genetics / Q000302:isolation & purification* / Q000472:pathogenicity; D013785:Thailand; D000097573:Red Fluorescent Protein
chemical_list: D000935:Antifungal Agents; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D000097573:Red Fluorescent Protein