National Institute for Biotechnology and Genetic Engineering (NIBGE), Jhang Road, Faisalabad, Pakistan; Pakistan Institute of Engineering and Applied Sciences (PIEAS), Nilore, Islamabad, Pakistan.
National Institute for Biotechnology and Genetic Engineering (NIBGE), Jhang Road, Faisalabad, Pakistan; Department of Plant Pathology, Bahauddin Zakariya University, Multan, Pakistan.
Virus Res. 2018 Sep 2;256:174-182. doi: 10.1016/j.virusres.2018.08.017. Epub 2018 Aug 24.
Spinach is a common vegetable crop and very little data is available about its virus infection. Symptomatic leaves of spinach were collected during field survey. Circular DNA molecules were amplified from symptomatic samples using rolling circle amplification (RCA). After restriction analysis, presumed bands of virus and satellites were cloned, sequenced and analyzed. Analysis of sequenced RCA product revealed the presence of chickpea chlorotic dwarf virus (CpCDV; Mastrevirus). Further analyses of the cloned virus showed that strain "C" of CpCDV was present in symptomatic samples of spinach collected from field associated with vein darkening, curling and enations on leaves. Amplification of alpha- and betasatellites with universal primers was performed. CpCDV showed association with cotton leaf curl Multan betasatellite (CLCuMB) and cotton leaf curl Multan alphasatellites (CLCuMA). Infectivity analysis of CpCDV and CpCDV/CLCuMB were done in N. benthamiana using particle bombardment method and the results showed that CpCDV was able to transreplicates CLCuMB in this host. To our knowledge, this is the first report of a dicot infecting mastrevirus (CpCDV) along with CLCuMB and CLCuMA associated with leaf curl disease of spinach in Pakistan. The significance of the results is discussed.
菠菜是一种常见的蔬菜作物，但关于其病毒感染的资料很少。在田间调查中收集了有症状的菠菜叶片。使用滚环扩增（RCA）从有症状的样本中扩增出环状 DNA 分子。经过限制性分析，克隆、测序和分析了假定的病毒和卫星带。对 RCA 产物测序分析表明，存在野豌豆黄花叶矮化病毒（CpCDV；Mastrevirus）。对克隆病毒的进一步分析表明，CpCDV 的“C”株存在于田间菠菜叶片上叶脉变暗、卷曲和起皱的有症状样本中。使用通用引物进行了α-和β-卫星的扩增。CpCDV 与棉花曲叶多卫星（CLCuMB）和棉花曲叶多卫星α卫星（CLCuMA）有关。使用粒子轰击法在 N. benthamiana 中进行了 CpCDV 和 CpCDV/CLCuMB 的感染性分析，结果表明 CpCDV 能够在该宿主中转录复制 CLCuMB。据我们所知，这是首次报道在巴基斯坦菠菜的卷叶病中，双子叶植物感染 Mastrevirus（CpCDV）以及 CLCuMB 和 CLCuMA。讨论了结果的意义。
