Evaluation of the genetic diversity of Plum pox virus in a single plum tree
Evaluation of the genetic diversity of Plum pox virus in a single plum tree
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DOI：
10.1016/j.virusres.2012.04.002
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发表时间：
2012-07-01
期刊：
影响因子：
5
通讯作者：
Glasa, Miroslav
中科院分区：
文献类型：
--
作者：
Predajna, Lukas;Subr, Zdeno;Glasa, Miroslav
研究了梅痘病毒（Plum pox virus， PPV）的遗传多样性及其在单一多年生木本寄主（李子，Prunus domestica）中的分布。2003年，用PPV-M、PPV-D和PPV-Rec三种分离株侵染了一棵李树，使其在露天条件下生长。2010年9月，在树冠的不同部位采集了叶片和果实样本。采用RT-PCR扩增PPV基因组745bp的NIb-CP片段，其中包含编码CP n末端的高变区，并直接测序以确定优势序列。同时对PCR产物进行克隆，共对105个单克隆进行测序。序列分析显示，在感染7年后，树中仅检测到PPV-M，其他两个分离株（PPV-Rec和PPV-D）已被取代。尽管该分析针对的是相对较短的基因组部分，但在PPV-M中观察到大量的分离内变异性。从105个个体序列中共鉴定出51个不同的单倍型，其中2个为显性。然而，尽管获得的结果表明病毒群体可能在叶内/果实内分化，但没有明确的树结构可以突出显示病毒群体的结构。一致序列与原始源分离物的比较结果没有差异，表明该原始分离物在野外条件下具有植物内稳定性。(C) 2012 Elsevier B.V.版权所有
Genetic diversity of Plum pox virus (PPV) and its distribution within a single perennial woody host (plum, Prunus domestica) has been evaluated. A plum tree was triply infected by chip-budding with PPV-M, PPV-D and PPV-Rec isolates in 2003 and left to develop untreated under open field conditions. In September 2010 leaf and fruit samples were collected from different parts of the tree canopy. A 745-bp NIb-CP fragment of PPV genome, containing the hypervariable region encoding the CP N-terminal end was amplified by RT-PCR from each sample and directly sequenced to determine the dominant sequence. In parallel, the PCR products were cloned and a total of 105 individual clones were sequenced. Sequence analysis revealed that after 7 years of infection, only PPV-M was still detectable in the tree and that the two other isolates (PPV-Rec and PPV-D) had been displaced. Despite the fact that the analysis targeted a relatively short portion of the genome, a substantial amount of intra-isolate variability was observed for PPV-M. A total of 51 different haplotypes could be identified from the 105 individual sequences, two of which were largely dominant. However, no clear-cut structuration of the viral population by the tree architecture could be highlighted although the results obtained suggest the possibility of intra-leaf/fruit differentiation of the viral population. Comparison of the consensus sequence with the original source isolate showed no difference, suggesting within-plant stability of this original isolate under open field conditions. (C) 2012 Elsevier B.V. All rights reserved.