Fonseca M E, Marcellino L H, Kitajima E W, Boiteux L S
Laboratório de Biologia Molecular, Centro Nacional de Pesquisa de Recursos Genéticos e Biotecnologia (CENARGEN)/EMBRAPA, Brasília (DF), Brazil.
J Gen Virol. 1994 Jun;75 ( Pt 6):1447-9. doi: 10.1099/0022-1317-75-6-1447.
The complete nucleotide (nt) sequence of the original coleus yellow viroid (CYVd) from Solenostemon scutellarioides, 'Golden Bedder', has been determined. The covalently closed single-stranded CYVd RNA molecule consists of 248 nt residues which assumes a rod-like secondary structure when folded in the model of lowest free energy. The sequence was determined by direct sequencing of RNA and from three overlapping cDNA clones. Comparison of the CYVd sequence with that of Coleus blumei viroid 1 (CbVd 1) from Germany demonstrated that they are closely related. The differences observed in the genome organization of CYVd relative to CbVd 1 were at three sites: position 25 (one U deletion), position 26 (a U was replaced by an A) and position 241 (one A insertion). The first two mutations were detected in one A-rich segment of eight nt (between positions 25 and 34). Northern blot hybridization of partially purified nucleic acids from the leaf tissue of S. scutellarioides 'Frilled Fantasy', inoculated with double-stranded cDNA, demonstrated that this fragment was infectious. These data enable CYVd to be assigned to the viroid class of plant pathogens, based on its biological properties and molecular structure. This work also gives additional support to the present classification system, in which the viroids isolated from S. scutellarioides form a distinct subgroup.
已确定来自彩叶草‘金色锦缎’(Solenostemon scutellarioides)的原始彩叶草黄化类病毒(CYVd)的完整核苷酸(nt)序列。共价闭合的单链CYVd RNA分子由248个nt残基组成,当以最低自由能模型折叠时呈棒状二级结构。该序列通过RNA直接测序和三个重叠的cDNA克隆确定。将CYVd序列与来自德国的彩叶草类病毒1(CbVd 1)的序列进行比较表明,它们密切相关。CYVd相对于CbVd 1在基因组组织上观察到的差异位于三个位点:第25位(缺失一个U)、第26位(一个U被A取代)和第241位(插入一个A)。前两个突变在一个富含A的8 nt片段(第25位和第34位之间)中检测到。用双链cDNA接种的彩叶草‘皱边幻想’(S. scutellarioides‘Frilled Fantasy’)叶片组织中部分纯化核酸的Northern印迹杂交表明,该片段具有感染性。基于其生物学特性和分子结构,这些数据使CYVd能够被归为植物病原体类病毒类别。这项工作也为目前的分类系统提供了额外支持,其中从彩叶草分离的类病毒形成一个独特的亚组。