Cotton leaf curl disease in Sindh province of Pakistan is associated with recombinant begomovirus components

Cotton leaf curl disease (CLCuD) is a devastating disease of cotton causing severe losses to cotton across the Punjab province of Pakistan and northeastern India. Although the disease has been reported as occurring sporadically in Sindh province, Pakistan, this has not caused significant losses. However, in the last few years the disease has become more significant in Sindh province. CLCuD is caused by begomoviruses in association with a disease-specific symptom determining satellite (Cotton leaf curl Multan betasaellite [CLCuMB]) and, in some cases, a non-essential alphasatellite (the function of which remains unclear). These components were cloned from six samples collected in Sindh. Analysis of the full-length sequences of six begomovirus clones showed one to be an isolate of Cotton leaf curl Kokhran virus (CLCuKV), a virus previously shown to be associated with CLCuD in the Punjab, whereas the other five clones showed less than ∼90% nucleotide sequence identity to several known begomoviruses associated with CLCuD. We take this to indicate that these are isolates of a newly identified begomovirus, for which we propose the name Cotton leaf curl Shahdadpur virus (CLCuShV). Closer inspection of the sequence of CLCuShV showed it to have a recombinant origin. For only two of the cotton samples was the presence of an alphasatellite detected. The sequences of clones of these alphasatellites indicate them to be newly identified species. A betasatellite was shown to be present in all six plants examined and sequence analysis of seven full-length clones indicated that two types of CLCuMB are present in Sindh and both are recombinant. These results indicate that the virus complex causing CLCuD in Sindh is distinct from that in the adjacent Punjab province. Possible reasons for these differences are discussed.