Isolation and Screening of Antibacterial Activity of Actinomycetota from the medicinal plant, Anthemis pseudocotula Boiss

Antibiotic resistance is rising dramatically worldwide, and thus the production of new antibiotics is indispensable. Recent scientific initiatives have focused on the bioprospecting of microorganisms' secondary metabolites, with a particular focus on the look for natural products with antimicrobial properties derived from endophytes. All plant species, regardless of their type, are thought to anchor endophytic bacteria (EB). There are many potential uses for the natural therapeutic compounds made by EB in medicine, agriculture, and the pharmaceutical industry. To investigate antibacterial properties in this study, Actinomycetota (formerly, Actinobacteria) were isolated from Anthemis pseudocotula Boiss., identified, and underwent bioprospecting by morphological and molecular methods. Samples were collected from Ilam, Iran, and then divided into roots, leaves, stems, and flowers. After disinfection, they were cut into ۲ mm pieces, cultured on casein agar culture medium, and incubated at ۲۸°C for up to four weeks. Actinomycetota was identified using the polymerase chain reaction method targeting the ۱۶S rRNA gene. To evaluate the antibacterial properties of the isolated Actinomycetota, the agar diffusion method was used. In parallel, the frequencies of biosynthetic gene clusters, including polyketide synthase (PKS-I and PKS-II) and nonribosomal peptide synthetase (NRPS) genes, were determined in the isolated Actinomycetota. Ninety bacteria were isolated from different parts of Anthemis flowers. Thirty-eight (۴۲.۲%) of these bacteria belonged to the phylum Actinomycetota, and out of these ۳۸, ۱۵ isolates (۳۹.۵%) had antibacterial properties. Of these, ۱۱ isolates (۷۳.۳%) exhibited antibacterial effects against Staphylococcus aureus, ۲ (۱۳.۳%) against Pseudomonas aeruginosa, ۳ (۲۰%) against Escherichia coli, and two isolates (۱۳.۳%) against Salmonella enterica sub-species of enterica serovar Typhimurium. The results of the molecular analysis of PKS-I, PKS-II, and NRPS genes showed that out of ۳۸ isolated Actinomycetota strains, ۲۳ isolates (۶۰.۵%) carried PKS-I gene, ۶ (۱۵.۸%) harbored PKS-II gene, and ۲۰ isolates (۵۲.۶%) had NRPS gene. This study indicates that Anthemis pseudocotula Boiss. has a number of active Actinomycetota that produce secondary metabolites with antibacterial properties.

Keywords:

Actinomycetota , Anthemis pseudocotula Boiss , Antibacterial activity , Bioprospecting , Isolation