کنگره بین المللی زیست فناوری گیاهان دارویی و قارچ های کوهی

صفحه اصلی / چهارمین کنگره بین المللی و پنجمین همایش ملی زیست فناوری گیاهان دارویی و قارچ های کوهی

Isolation and molecular identification of endophytic bacteria from Nepeta crispa

نویسندگان :

Farnoosh Rasoul sharifi¹ Reza Habibipoor² Mohsen Rajabi³ Melika Esfandiari⁴

1- دانشگاه آزاد اسلامی واحد همدان 2- دانشگاه آزاد اسلامی واحد همدان 3- دانشگاه آزاد اسلامی واحد همدان 4- دانشگاه آزاد اسلامی واحد همدان

کلمات کلیدی :

Endophyte،Nepeta crispa،PCR،Molecular Identification

چکیده :

Introduction: Endophytic bacteria are non-pathogenic microorganisms living in plant tissues that increase metabolism, growth and plant resistance against pathogens and environmental stresses. endophyte microorganisms activate the components and biochemical pathways of plant cells by producing secondary metabolites, extracellular enzymes and other substances. These microorganisms are a new source of bio-chemical active compounds with high potential to be used in medical, agricultural and industrial fields. This study was conducted in order to molecular identification of endophytic bacteria from Nepeta crispa L. in Hamadan province. Materials and methods: 12 healthy plants were collected from three different areas of the N. crispa habitat in Hamadan province. Explants of roots, stems and leaves were surface disinfected and cultured on Nutrient Agar (NA) medium. After isolation of endophytic bacteria, purification was done by single clone method and for molecular identification, DNA extraction was done using Denazist extraction kit. PCR process was performed to amplify the gene region of 16SrRNA (using 27F and 1492R primers). Sequencing of PCR products was done by Microsynth company, Switzerland. The studied isolates were analyzed based on the results of BLAST search on the NCBI site and phylogeny based on the 116SrRNA region and were identified based on their matching with the reference species with the mentioned similarity percentage. Results: Finally, in this research, 11 isolates were isolated and molecularly identified, and the results showed that all isolated bacteria were belong to Bacillales order. The maximum number of endophyte bacteria isolated in this research, were Bacillus subtilis bacteria (%18.18) (%97.17 similarity with the access number MH040981.1) and Bacillus pumilus (%18.18) (%94.83 similarity with the access number OP223406.1) that isolated from the tissue of root and stem, respectively. The following bacteria were also isolated and identified from this plant: Bacillus cereus (%89.64 similarity with the accession number MN793064.1), Providencia rettgeri (%95.85 similarity with the accession number MN589673.1), Bacillus amyloliquefaciens (%96.93 similarity with the accession number MT397274.1), Bacillus velezensis (%97.74 similarity with accession number OP060623.1), Bacillus mobilis (%98.21 similarity with accession number CP031443.1), Bacillus licheniformis (%97.61 similarity with accession number MT679253.1), Bacillus thuringensis (%100.00 similarity with access number KF054891.1). Conclusion: Bacillus is the most abundant endophytic bacteria isolated from N. crispa in the mountainous ecosystem of Hamadan province. This study showed that endophytic actinobacteria are a rich source of biologically active secondary metabolites with diverse chemical structures that have significant potential in the production of biotechnology, pharmaceutical and medical products. This is the first study that reports the presence of the mentioned endophytic bacteria in N. crispa.