Construction of an Infectious Chimeric Geminivirus by Molecular Cloning Based on Coinfection and Recombination

摘要

In the evolution of geminiviruses, genetic recombination plays a key role in ecological, biochemical and evolutionary processes. They evolve as results of prehistoric inter-genus recombination whereas intra-genus recombination of geminiviruses leads to the emergence of agriculturally important plant pathogens. Geminiviruses are transmitted by leafhoppers, treehoppers and whiteflies. An isolate of Chickpea chlorotic dwarf virus (CpCDV), a monopartite dicot infecting mastrevirus is transmitted by leafhopper. In contrast, Cotton leaf curl burewala virus (CLCuBuV), a monopartite begomovirus, was acquired and transmissible by whitefly to various host plants. In this study, we remove the coat protein (CP) gene of CpCDV by PCR and replaced with that of PCR amplified CLCuBuV-CP. The resultant infectious clone of chimeric CpCDV is produced later named mastrebegomo chimera (pGII0000MBC). By using Agroinfiltration technique (Agrobacterium-mediated inoculation) the chimeric clone along with wild type Clone CpCDV and CLCuBuV are injected on tomato (Lycopersicon esculentum Mill.) and tobacco (Nicotiana benthamiana Domin.) seedlings and get symptoms after 21dpi (days post inoculation). Later, the symptoms appeared on plants were compared with healthy plants. This study elaborates our awareness about the genetic recombination and coexistence of mastreviruses with begomovirus globally including Pakistan and will provide the basic information regarding their management.