VIROME PROFILING IN GLADIOLUS AND TUBEROSE CROPS USING NEXT GENERATION SEQUENCING (NGS)
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Date
2023-03-02
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University of Agricultural Sciences, Bangalore
Abstract
Viral diseases becoming major constraints for production of gladiolus and tuberose resulting in huge losses to growers in our country. Diagnosis of viruses infecting gladiolus and tuberose is very crucial to design management strategies. Next-generation sequencing (NGS) based virome profiling, an advanced sequencing technology has become popular method for the accurate detection of viruses present in the samples. A total 20 different virus-infected leaf samples from gladiolus and tuberose showing mottling, mosaic and yellowing symptoms were collected during the year 2021–22. Total RNA was isolated from all collected samples and used as an input for mRNAome sequencing. Raw data obtained were subjected to De-novo assembly. Further,individual contigs were subjected to standalone MEGABLAST against viral reference genomes available in NCBI database. Mapping of contigs against reference viral genome through BioEdit, Clustal X was carried out to reconstruct the complete/near-complete genome viruses. However, bean yellow mosaic virus (BYMV) and ornithogalum mosaic virus (OrMV) in gladiolus whereas, tuberose mild mosaic virus (TuMMV) and tuberose mild mottle virus (TuMMoV) were identified in tuberose, however, CMV infection was found in both gladiolus and tuberose. Further, recombination analysis using RDP5 indicated presence of recombination only in BYMV. Molecular tools i.e., RT-PCR and RT-LAMP were optimized for rapid detection of identified viruses. Infection of CMV in tuberose is the first report from our study in India