Abstract

Molecular analysis of new isolates of the plum pox virus (PPV) found in the gene pool collections of peach, apricot, and plum from the Nikitsky Botanical Garden (National Science Center, Russian Academy of Sciences), as well as on wild cherry plum trees in the surrounding area, has been carried out. Using the enzyme-linked immunosorbent assay and reverse transcription polymerase chain reaction, 19 new PPV isolates have been detected. Most of them belonged to the strain D; single isolates of the strains Rec and M were also found. The 3'-terminal sequences of genome of the new isolates were sequenced, including the coat protein (CP) gene and adjacent sequences of the NIb gene and the 3'-nontranslated region. Phylogenetic analysis of these sequences revealed the distribution of PPV-D isolates into several clusters or clades. An average divergence rate between sequences of Crimean PPV-D isolates was 2.0%. A relatively high level of genetic variation indicates a long-standing introduction of the ancestors of the current PPV-D isolates to Nikitsky Botanical Garden (NBG). Isolate H23, identified in the apricot collection, belonged to strain M. This is the first detection of PPV-M in Crimea. Isolate AL-Ch (strain Rec) was found on wild cherry plum. In the CP of isolate K87 (strain D), the K59T mutation was detected, which prevents its recognition by PPV-D-specific 4DG5 monoclonal antibodies. In CP of AL-Ch and previously characterized isolates K28 and Kisl-1pl (strain Rec), amino acid substitutions were detected at position 68, which did not affect the recognition of these isolates by AL monoclonal antibodies. Apparently, position 68 is outside the PPV-M/Rec-specific epitope. The results of the work show that PPV most likely entered the NBG with contaminated drupaceous fruit material as a result of several independent introductions from different regions of the world and spread over plantations by aphids.

中文翻译：

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李子痘病毒新克里米亚分离株的分子分析

摘要

从Nikitsky植物园（俄罗斯科学院国家科学中心）以及野生樱桃李树上桃，杏和李的基因库中发现的李子痘病毒（PPV）新分离株的分子分析在周边地区，已经进行了。使用酶联免疫吸附测定和逆转录聚合酶链反应，已检测到19种新的PPV分离株。它们大多数属于菌株D。还发现了菌株Rec和M的单个分离物。对新分离株基因组的3'-末端序列进行了测序，包括外壳蛋白（CP）基因和NIb的相邻序列基因和3'非翻译区。这些序列的系统发生分析表明PPV-D分离株分布成几个簇或进化枝。克里米亚半岛PPV-D分离株序列之间的平均差异为2.0％。较高水平的遗传变异表明，将目前PPV-D分离株的祖先长期引入了Nikitsky植物园（NBG）。在杏子中鉴定出的分离株H23属于菌株M。这是克里米亚首次检测到PPV-M。在野樱桃李上发现了分离株AL-Ch（Rec菌株）。在分离株K87（菌株D）的CP中，检测到K59T突变，这阻止了其被PPV-D特异性4DG5单克隆抗体识别。在AL-Ch的CP和先前鉴定的分离株K28和Kisl-1pl（Rec菌株）中，在68位检测到氨基酸取代，这不影响AL单克隆抗体对这些分离物的识别。显然，位置68在PPV-M / Rec特异性表位之外。工作结果表明，由于来自世界不同地区的几次独立引种并导致蚜虫在人工林中传播，PPV最有可能进入到污染了含糖果果实材料的NBG中。