This work was supported by a grant from The National Natural Sciences Foundation of China (30270744).
以32P标记的黄瓜花叶病毒(CMV)RNA3 cDNA片段和卫星RNA全长cDNA作为探针,定量测定CMV基因组RNA和卫星RNA的含量变化,结果显示:二者均具有明显的寄主效应和时间效应.在16~20℃条件下,接种不携带卫星RNA的分离物CMV-R3,15天、30天和75天时,CMV基因组RNA负荷量呈显著下降的趋势.在第15天,RNA3的负荷量以烟草>心叶烟>克里夫兰烟>番茄的顺序表现为不同寄主的显著性差异.相同条件下接种携带高拷贝卫星RNA的分离物CMV-RS,在5天和15天之间基因组RNA和卫星RNA负荷量均呈现上升的趋势,同时测得其基因组RNA和卫星的负荷量具有相似的寄主效应和时间效应,但程度不同.第15天时,二者负荷量以烟草>心叶烟>番茄的顺序表现寄主效应的显著性差异.在18~21℃条件下,接种携带坏死卫星RNA的CMV强毒株HC4,第5天、第10天和第15天时,基因组RNA和卫星RNA的负荷量均以番茄>心叶烟>烟草的顺序表现出显著性差异,并表现出明显的时间效应.不同来源CMV分离物还存在寄主选择性差异.
Radioactive dsDNA probes were prepared by labeling cDNA fragments of CMV RNA3 partial sequence and the full-length satellite RNA (satRNA) with 32P, respectively. By using nucleic acid spot hybridization (NASH), relative RNA loading (RRL) of both gemonic and satellite RNAs were quantitatively determined from the systemically infected hosts tissue. At 16~20℃, the radish-derived CMV-R3 containing no satRNA was inoculated on four host plants and examined 15 days, 30 days, and 75 days post inoculation, respectively. The resulting RRL showed a declining trend during the test period. On day 15, the RRL for genomic RNA differed obviously among the hosts in the order of Nicotiana.tobacum>N.glutinosa>N.clevelandii>tomato. Meanwhile, the RRL for the CMV-RS, another radish isolate containing high copies of satRNA, was examined 5 days and 15 days after inoculation on the same hosts with CMV-R3 being included as a non-satRNA control. The RRLs for both genomic RNA and satellite RNA of the CMV-RS displayed a similar host- and time-effect trend. On all the inoculated hosts, the RRL increased from day 5 to day 15 and RRL of CMV-RS for both genomic RNA and satRNA was in the quantitative order of N.tobacum>N.glutinosa>Nicandra.physalodesand tomato. At 18~21℃, CMV-HC4, a severe tomato isolate containing a necrosis satRNA, was tested after 5 days, 10 days, and 15 days inoculation on 5 hosts. The RRLs of HC4 genomic and satellite RNAs were under the influence of host and inoculation time. The RRLs for satRNA and genomic RNA were similar but had some degree differences among the hosts. On day 10 post inoculation, the relative amount of both genomic RNA and satRNA was ordered as tomato>N.glutinosa>N.tobacum. The results also showed that different CMV isolates have obvious preference among hosts for replication and accumulation of viral RNAs.
陈集双,柴立红,吴平.系统侵染寄主中黄瓜花叶病毒及其卫星RNA的动态变化[J].生物化学与生物物理进展,2003,30(2):285-289
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