تشخیص کمی RNA-viruses گامی در تحلیل هولوبیونت و ارتباط متقابل ویروس، همزیست و میزبان

The use of biological control agents (BCAs) in different agricultural systems, including conventional, IPM, IP and organic agriculture, has requirements such as host rearing in appropriate quality and quantity, as well as most of genetically control methods and related research and development projects. To keep the host rearing quality, the interactions of pathogen, symbiont and host model (six species of Golosina genus in a FAO/IAEA program) were investigated in detailed genomics, transcriptomics and proteomics. For identify the RNA viruses as Iflavirus and Negevirus and their role in the holobiont and host microbiota, RNA extracting from the complete insect sample were conducted. In the next step after adjusting and stabilizing the method from only one (middle) leg of the insect, two-step RT-PCR (while preparing cDNA), designing specific primers (pairs), multiple gradient PCRs at a temperature gradient of 40-70°C and checking the quantity of the viruses in the samples with qPCR were conducted for creating a virus-free colony for next studies to check their effects in the holobiont. Considering the absence of these viruses in Golosina pallidipes and its sensitivity to salivary gland hypertrophy viruses (SGHVs), the possibility of the symbiotic role of the two studied viruses should investigated in further transmission studies in diferent tissues was also checked by qPCR method. These viruses are transferred to the new host through hemolymph and their quantitative level increased in the new host in four weeks. Such interaction studies can clarify nexus of BCAs-host rearing quality.