Abstract:Kenaf leaf curl disease is a new disease occurred in Haikou, Hainan Province of China. The symptoms of the infected plants exhibits leaf curling upwards, vein swelling and vein dark green. PCR analysis indicates that the symptomatic plants were infected by begomoviruses and sequence analysis indicates that the isolate HN08 only contains the DNA-A component.The full length of DNA-A was 2 738 nucleotides. Multiple sequence alignment showed that the DNA-A of HN08 had more than 89.0% sequence identity with that of all other isolates of Cotton leaf curl Multan virus (CLCuMuV) deposited in GenBank, and more than 99.0% sequence identity with that of all isolates from China. The virus was also accompanied by betasatellite molecular,which was determined to be 1 346 nucleotides. Pairwise comparison indicated that the betasatellite molecular of HN08 had more than 83.0% sequence identity with that of Cotton leaf curl Multan betasatellite (CLCuMuB), and shared the highest sequence identity (99.7%) with that of isolate Fz1 from Fujian. The infectious clones of HN08 DNA-A and its betasatellite molecular, pGreenⅡ049-1.6A and pGreenⅡ049-2.0β were constructed, and were agro-inoculated into Hibiscus cannabinus plants. At 30 dpi, the new leaves of the H. cannabinus plants inoculated with the recombination of pGreenⅡ049-1.6A and pGreenⅡ049-2.0β displayed leaf curl symptoms. At 60 dpi, most of the leaves of the inoculated H. cannabinus plants showed severe leaf curl symptoms, similar to that observed in the fields. However, neither the plants inoculated with pGreenⅡ049-1.6A or pGreenⅡ049-2.0β showed visible symptoms. The results of PCR and Southern blot detection confirmed these symptoms caused by co-infection of HN08 DNA-A and its betasatellite molecular. Taken together, Kenaf leaf curl disease in Hainan province was caused by co-infection of CLCuMuV and its betasatellite (CLCuMuB). This is the first report of CLCuMuV on H. cannabinus .