Skotnicki M L, Mackenzie A M, Gibbs A J
Research School of Biological Sciences, Australian National University, Canberra.
Arch Virol. 1992;127(1-4):25-35. doi: 10.1007/BF01309572.
Full-length dsDNA clones that encode the genomes of two Australian turnip yellow mosaic isolates, TYMV-BL and TYMV-CL have been constructed. These clones were transcribed to give 6.3 kb capped ssRNA which infects Chinese cabbages to give symptoms indistinguishable from those produced by the parental viruses. Extensions of up to 26 nucleotides at the 3' end of the TYMV-BL clone delay infections, but virus particles isolated from these plants 4 weeks after inoculation contain RNA with the original TYMV-BL 3' terminus. A 90 nucleotide-long portion of the virion protein gene of TYMV-BL was replaced by a synthetic 90-mer primer with 16 nucleotide changes to decrease the large cytosine content (34-42%) characteristic of tymovirus genomic RNA. No reversion of any of the mutated nucleotides to cytosine occurred during 7 passages in Chinese cabbage. Hybrids between the TYMV-BL and TYMV-CL clones were also constructed, by exchanging various portions of the genome. However, it was not possible to determine definitively which part of the viral genome is responsible for the more severe symptoms caused by TYMV-BL as the hybrids gave intermediate symptoms.
已构建出编码两种澳大利亚芜菁黄花叶病毒分离株TYMV - BL和TYMV - CL基因组的全长双链DNA克隆。这些克隆被转录产生6.3 kb的加帽单链RNA,其感染大白菜后产生的症状与亲本病毒产生的症状无法区分。TYMV - BL克隆3'端长达26个核苷酸的延伸会延迟感染,但接种4周后从这些植物中分离出的病毒粒子含有具有原始TYMV - BL 3'末端的RNA。用一个有16个核苷酸变化的合成90聚体引物替换了TYMV - BL病毒粒子蛋白基因90个核苷酸长的部分,以降低芜菁黄花叶病毒基因组RNA特有的高胞嘧啶含量(34 - 42%)。在大白菜中传代7次期间,没有任何一个突变核苷酸回复为胞嘧啶。通过交换基因组的不同部分,还构建了TYMV - BL和TYMV - CL克隆之间的杂种。然而,由于杂种产生的是中间症状,所以无法确切确定病毒基因组的哪一部分导致了TYMV - BL引起的更严重症状。