Molecular Characterization and Transmission Study of the Begomovirus on Hollyhock Plant Causing Leaf Curl Disease for the First Time in Province Punjab, Pakistan

Abstract I. Background: Hollyhock (Alcea rosea) is an ornamental plant belonging to the Malvaceae family and has a remarkable aesthetic and medicinal value. Previously in Pakistan, the hollyhock plant was not found to be infected by begomovirus and the plant first time showed the symptoms of typical leaf curling, puckering as well as thickened veins. II. Methods and Results: During the year 2018, symptomatic samples of the hollyhock plants were collected that exhibited characteristic typical leaf curling, puckering as well as thickened veins. DNA was extracted from the samples and the PCR technique was optimized for the detection of begomovirus followed by sequencing. The samples were detected to be infected with begomovirus by using Av/Ac core, Begomo 01/02, and CLCV 01/02 primer showed positive results with 579bp, 2.8kb, and 1.1kb nucleotide respectively. The betasatellite was amplified by using beta01/02 and CLCuMuBF11/R33 showed positive results with 1400bp and 481bp respectively. Sequencing results showed that diseased hollyhock plants were associated with Cotton leaf curl Multan virus-Rajasthan strain along with Cotton leaf curl Multan betasatellite. III. Conclusion: Hollyhock plants infected by begomovirus has been reported for the first time as a possible source of virus inoculum from Pakistan.