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Reverse transcriptase domain sequences from tree peony (Paeonia suffruticosa) long terminal repeat retrotransposons: sequence characterization and phylogenetic analysis

机译:牡丹(Paeonia suffruticosa)长末端重复反转录转座子的逆转录酶结构域序列:序列表征和系统发育分析

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摘要

Tree peony is an important horticultural plant worldwide of great ornamental and medicinal value. Long terminal repeat retrotransposons (LTR-retrotransposons) are the major components of most plant genomes and can substantially impact the genome in many ways. It is therefore crucial to understand their sequence characteristics, genetic distribution and transcriptional activity; however, no information about them is available in tree peony. Ty1-copia-like reverse transcriptase sequences were amplified from tree peony genomic DNA by polymerase chain reaction (PCR) with degenerate oligonucleotide primers corresponding to highly conserved domains of the Ty1-copia-like retrotransposons in this study. PCR fragments of roughly 270 bp were isolated and cloned, and 33 sequences were obtained. According to alignment and phylogenetic analysis, all sequences were divided into six families. The observed difference in the degree of nucleotide sequence similarity is an indication for high level of sequence heterogeneity among these clones. Most of these sequences have a frame shift, a stop codon, or both. Dot-blot analysis revealed distribution of these sequences in all the studied tree peony species. However, different hybridization signals were detected among them, which is in agreement with previous systematics studies. Reverse transcriptase PCR (RT-PCR) indicated that Ty1-copia retrotransposons in tree peony were transcriptionally inactive. The results provide basic genetic and evolutionary information of tree peony genome, and will provide valuable information for the further utilization of retrotransposons in tree peony.
机译:牡丹是世界范围内重要的园艺植物,具有重要的观赏和药用价值。长末端重复逆转录转座子(LTR-retrotransposons)是大多数植物基因组的主要组成部分,可以从许多方面显着影响基因组。因此,了解其序列特征,遗传分布和转录活性至关重要。但是,牡丹没有有关它们的信息。通过聚合酶链反应(PCR),用简并的寡核苷酸引物(对应于Ty1-copia样逆转座子的高度保守的结构域),从牡丹基因组DNA中扩增出Ty1-copia样的逆转录酶序列。分离并克隆了大约270 bp的PCR片段,并获得了33个序列。根据比对和系统发育分析,所有序列分为六个家族。所观察到的核苷酸序列相似性程度的差异表明这些克隆之间具有高水平的序列异质性。这些序列中的大多数都有移码,终止密码子或两者兼有。点印迹分析揭示了这些序列在所有研究的牡丹品种中的分布。然而,在其中检测到不同的杂交信号,这与先前的系统研究一致。逆转录聚合酶链反应(RT-PCR)表明,牡丹中的Ty1-copia逆转座子是转录失活的。研究结果为牡丹基因组提供了基础的遗传和进化信息,为牡丹逆转录转座子的进一步利用提供了有价值的信息。

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