همسانه‌سازی و تجزیه و تحلیل تبارزایی پوتی‌ویروس جدید (SYMV) آلوده کننده زعفران براساس ژن CP

Saffron (Crocus sativus) is a strategic product for the country, highly valued in various industries, including pharmaceuticals and food, due to its medicinal properties and unique flavor. Viral diseases of saffron have significantly reduced its production and caused economic damage worldwide. Recently, a new potyvirus called saffron yellow mosaic virus (SYMV) was reported in the main saffron-producing areas of South Khorasan and Razavi Khorasan provinces. To investigate this virus, samples were collected from the leaves and flower petals of saffron plants showing symptoms of chlorosis, mosaic patterns, and flower color breaking. Total RNA was extracted from these samples, and the coat protein (CP) gene of the virus (from nucleotide 8572 to 9382) was amplified using specific CP-F/R SYMV primers in a RT-PCR test. The amplified fragment was ligated into the pUCM-T plasmid using T4 DNA ligase enzyme. The recombinant plasmids were transformed into the competent cells of Escherichia coli strain DH5α via heat shock method. After selecting white colonies on LB agar medium containing ampicillin, IPTG, and X-Gal; colony PCR was performed for initial confirmation using M13 primers. The selected colonies were grown in liquid LB medium overnight at 37°C. Following plasmid DNA extraction, the recombinant plasmids were verified for accuracy through DNA sequencing. BLAST results of the 811 bp from the CP gene of the new potyvirus indicated its phylogenetic relationship with viruses in the bean yellow mosaic virus (BYMV) subgroup. Sequenced isolates had the highest identity to BYMV isolates at the nucleotide (69-72%) and amino acid (79-81%) levels.