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Vol 55(2021) N 1 p. 83-91; DOI 10.1134/S0026893321010076 Full Text

Z. Iqbal1* **, M.N. Sattar2, M. Khurshid3

Cotton Leaf Curl Multan Betasatellite as a Tool to Study the Localization of Geminiviruses in Plants

1Central Laboratories, King Faisal University, Al-Ahsa, 31982 Kingdom of Saudi Arabia
2Department of Agricultural Biotechnology, College of Agriculture and Food Sciences, King Faisal University, Al-Ahsa, 31982 Kingdom of Saudi Arabia
3Institute of Biochemistry and Biotechnology, University of the Punjab, Quaid-e-Azam Campus, Lahore, 54590 Pakistan

*zafar@kfu.edu.sa
**zafariqbal2009@gmail.com
Received - 2020-03-11; Revised - 2020-04-30; Accepted - 2020-05-27

Cotton leaf curl Multan betasatellite (CLCuMB) is a ubiquitous betasatellite commonly found along with cotton leaf curl disease (CLCuD) associated begomoviruses in the Old World. It has a promiscuous replicative nature and trans-replicated by a diverse range of geminiviruses. CLCuMB encodes a single ORF, βCl, in the complementary direction and has pathogenicity, symptoms determinant, suppressor of post-transcription and transcription gene silencing functions. After substituting the βC1 gene with the target gene, it has been used successfully as a gene delivery vector. In the present study, the βC1 gene of CLCuMB was substituted with the green fluorescent protein (GFP) gene, and the resulting construct utilized as a reporter vector to decipher in planta localization of geminiviruses. The recombinant CLCuMB expressing GFP (CLCuMB-GFP) was co-inoculated to Nicotiana benthamiana plants either with Cotton leaf curl Kokharan virus (CLCuKoV) alone or in a combination with the wild type CLCuMB to investigate the objectives of the study. Results showed that CLCuKoV successfully supported the replication and systemic movement of CLCuMB-GFP either alone or in the presence of wild type CLCuMB. The presence of CLCuMB-GFP was readily detected with PCR and Southern blot hybridization. The modified CLCuMB may serve as a tool useful for in planta localization of geminiviruses.

cellular localization, CLCuKoV, CLCuMB, expression vector, green fluorescent protein



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