Infectious clone construction and pathogenicity confirmation of Cotton leaf curl Multan virus (CLCuMuV), Ramie mosaic virus (RamV) and Corchorus yellow vein Vietnam virus (CoYVV) by southern blot analysis

Geminiviruses are insect-transmissible, economically vital group of plant viruses, which cause significant losses to crop production and ornamental plants across the world. During this study, infectious clones of three devastating begomoviruses, i.e., Cotton leaf curl Multan virus (CLCuMuV), Ramie mosaic virus (RamV) and Corchorus yellow vein Vietnam virus (CoYVV) were constructed by following novel protocol. All infectious clones were confirmed by cloning and sequencing. All of the infectious clones were agro-inoculated in Agrobacterium. After the agro-infiltrations, all clones were injected into Nicotiana benthamiana and jute plants under controlled condition. After 28 days of inoculation, plants exhibited typical symptoms of their corresponding viruses. All the symptomatic and asymptomatic leaves were collected from inoculated plants for further analysis. The southern blot analysis was used to confirm the infection of studied begomoviruses. At the end, all the products were sequenced and analyzed.