Detection and Cloning of Begomovirus(es) from Different Plant Samples

Abstract

Geminiviruses (family Geminiviridae) are small circular single stranded DNA viruses infecting plants. Geminiviruses are comprises of seven genera of which the members of genus Begomovirus are most studied and dangerous species. They are exclusively transmitted by a single species of whitefly in circulative, non-propagative manner and infect dicotyledonous plants only. Leaf sample from hollyhock (Alcea rosea; isolate 3AK), showing vein yellowing symptom, was collected from areas around Chakri during 2014. Full length amplification product (genomic component) of approx. 2.8kb was obtained from the isolate 3AK. The isolate 3AK showed highest nucleotide sequence identity (86.6%) with Pedilanthus leaf curl virus. Based on present species demarcation criteria (89% nucleotide sequence identity), the isolate 3AK represent a new species of begomovirus for which the name “Hollyhock yellow vein mosaic Islamabad virus” HYVMIV) is proposed. The HYVMIV develop as a result of recombination with a major parent, Pedilanthus leaf curl virus (fragment between 31st to 2386bp) and minor parent Cotton leaf curl Multan virus (CLCuMV) (fragment between 2387 to 30bp). The complete nucleotide sequence of betasatellite from isolate 3AK was determined to be 1352bp and showed maximum nucleotide sequence identity (90%) to Kenaf leaf curl betasatellite. Cotton (family Malvaceae) is one of the major crops which accounts for sixty percent of the foreign exchange and affected by cotton leaf curl disease with no solution to date. Hollyhock also belongs to family Malvaceae and the occurrence of recombinant virus with CLCuMV as one of the parents is a new challenge for scientist and to national economy