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蒙古蒲公英完整线粒体基因组的特征分析揭示了五次重复介导的重组。

Characterisation of the complete mitochondrial genome of Taraxacum mongolicum revealed five repeat-mediated recombinations.

作者信息

Jiang Mei, Ni Yang, Li Jingling, Liu Chang

机构信息

School of Pharmaceutical Sciences, Qilu University of Technology (Shandong Academy of Sciences), Jinan, People's Republic of China.

Key Laboratory for Applied Technology of Sophisticated Analytical Instruments of Shandong Province, Shandong Analysis and Test Center, Qilu University of Technology (Shandong Academy of Sciences), Jinan, People's Republic of China.

出版信息

Plant Cell Rep. 2023 Apr;42(4):775-789. doi: 10.1007/s00299-023-02994-y. Epub 2023 Feb 12.

Abstract

We reported the mitochondrial genome of Taraxacum mongolicum for the first time. Five pairs of repeats that can mediate recombination were validated, leading to multiple conformations of genome. Taraxacum mongolicum belongs to the Asteraceae family and has important pharmaceutical value. To explore the possible interaction between the organelle genomes, we assembled the complete mitochondrial genome (mitogenome) of T. mongolicum using Illumina and Oxford Nanopore sequencing data. This genome corresponded to a circular molecule 304,467 bp long. It encodes 52 unique genes including 31 protein-coding, 3 ribosomal RNA (rRNA) and 18 transfer RNA (tRNA) genes. In addition to the single circular conformation, the existence of alternative conformations mediated by five repetitive sequences in the mitogenome was identified and validated. Recombination mediated by the inverted repeats resulted in two conformations. Conversely, recombination mediated by the two direct repeats broke one large circular molecule into two subgenomic circular molecules. Furthermore, we identified 12 homologous fragments by comparing the sequences of mitogenome and plastome, including eight complete tRNA genes. Lastly, we identified a total of 278 RNA-editing sites in protein-coding sequences based on RNA-seq data. Among them, cox1 and nad5 gene has the most sites (21), followed by the nad2 gene with 19 sites. We successfully validated 213 predicted RNA-editing sites using PCR amplification and Sanger sequencing. This project reported the first mitogenome of T. mongolicum and demonstrated its multiple conformations generated by repeat-mediated recombination. This genome could provide critical information for the molecular breeding of T. mongolicum, and also be used as a reference genome for other species of the genus Taraxacum.

摘要

我们首次报道了蒙古蒲公英的线粒体基因组。验证了五对可介导重组的重复序列,导致基因组出现多种构象。蒙古蒲公英属于菊科,具有重要的药用价值。为了探索细胞器基因组之间可能的相互作用,我们利用Illumina和牛津纳米孔测序数据组装了蒙古蒲公英的完整线粒体基因组(线粒体基因组)。该基因组对应一个长度为304,467 bp的环状分子。它编码52个独特基因,包括31个蛋白质编码基因、3个核糖体RNA(rRNA)基因和18个转移RNA(tRNA)基因。除了单一的环状构象外,还鉴定并验证了线粒体基因组中由五个重复序列介导的替代构象的存在。由反向重复序列介导的重组产生了两种构象。相反,由两个正向重复序列介导的重组将一个大的环状分子分解为两个亚基因组环状分子。此外,通过比较线粒体基因组和质体基因组的序列,我们鉴定出12个同源片段,包括8个完整的tRNA基因。最后,基于RNA-seq数据,我们在蛋白质编码序列中总共鉴定出278个RNA编辑位点。其中,cox1和nad5基因的位点最多(21个),其次是nad2基因,有19个位点。我们通过PCR扩增和Sanger测序成功验证了213个预测的RNA编辑位点。该项目报道了蒙古蒲公英的首个线粒体基因组,并展示了其由重复序列介导的重组产生的多种构象。该基因组可为蒙古蒲公英的分子育种提供关键信息,也可作为蒲公英属其他物种的参考基因组。

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