Abstract

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, βC1, 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.

中文翻译：

---

棉卷毛Multan Betasatellite作为研究植物中双子病毒定位的工具

摘要

棉叶卷曲木尔坦β卫星（CLCuMB）是一种普遍存在的β卫星，在旧世界中通常与棉叶卷曲病（CLCuD）相关的begomoviruses一起发现。它具有混杂的复制性质，并通过多种双生病毒进行反式复制。CLCuMB在互补方向上编码单个ORFβC1，并具有致病性，症状决定因素，转录后抑制因子和转录基因沉默功能。的β代后C1基因与靶基因中，已经成功地用作基因递送载体。在本研究中，β C1 CLCuMB的基因与绿色荧光蛋白（GFP）基因取代，并且所得到的构建体用作报告载体破译在植物双子病毒的本地化。将重组表达CLCuMB的GFP（CLCuMB-GFP）单独或与野生型CLCuMB一起与棉叶卷曲可卡兰病毒（CLCuKoV）共接种到本氏烟草植物中，以研究目的。结果显示，CLCuKoV成功地支持了单独或在野生型CLCuMB存在下CLCuMB-GFP的复制和系统运动。通过PCR和Southern印迹杂交很容易检测到CLCuMB-GFP的存在。修饰的CLCuMB可以用作在双生病毒的植物定位中有用的工具。