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Organellar genome assembly methods and comparative analysis of horticultural plants

机译:园艺植物基因组组装方法及比较分析

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摘要

Although organellar genomes (including chloroplast and mitochondrial genomes) are smaller than nuclear genomes in size and gene number, organellar genomes are very important for the investigation of plant evolution and molecular ecology mechanisms. Few studies have focused on the organellar genomes of horticultural plants. Approximately 1193 chloroplast genomes and 199 mitochondrial genomes of land plants are available in the National Center for Biotechnology Information (NCBI), of which only 39 are from horticultural plants. In this paper, we report an innovative and efficient method for high-quality horticultural organellar genome assembly from next-generation sequencing (NGS) data. Sequencing reads were first assembled by Newbler, Amos, and Minimus software with default parameters. The remaining gaps were then filled through BLASTN search and PCR. The complete DNA sequence was corrected based on Illumina sequencing data using BWA (Burrows–Wheeler Alignment tool) software. The advantage of this approach is that there is no need to isolate organellar DNA from total DNA during sample preparation. Using this procedure, the complete mitochondrial and chloroplast genomes of an ornamental plant, Salix suchowensis, and a fruit tree, Ziziphus jujuba, were identified. This study shows that horticultural plants have similar mitochondrial and chloroplast sequence organization to other seed plants. Most horticultural plants demonstrate a slight bias toward A+T rich features in the mitochondrial genome. In addition, a phylogenetic analysis of 39 horticultural plants based on 15 protein-coding genes showed that some mitochondrial genes are horizontally transferred from chloroplast DNA. Our study will provide an important reference for organellar genome assembly in other horticultural plants. Furthermore, phylogenetic analysis of the organellar genomes of horticultural plants could accurately clarify the unanticipated relationships among these plants.
机译:尽管细胞器基因组(包括叶绿体和线粒体基因组)的大小和基因数量都小于核基因组,但细胞器基因组对于研究植物进化和分子生态机制非常重要。很少有研究集中在园艺植物的细胞器基因组上。国家生物技术信息中心(NCBI)提供了大约1193个陆地植物的叶绿体基因组和199个线粒体基因组,其中只有39个来自园艺植物。在本文中,我们报告了一种基于下一代测序(NGS)数据的高质量园艺细胞器基因组组装的创新有效方法。测序读取首先由具有默认参数的Newbler,Amos和Minimus软件组装而成。然后通过BLASTN搜索和PCR填补其余的缺口。使用BWA(Burrows–Wheeler比对工具)软件根据Illumina测序数据校正了完整的DNA序列。这种方法的优点是在样品制备过程中无需从总DNA中分离细胞器DNA。使用此程序,可以确定观赏植物柳柳和果树枣的完整线粒体和叶绿体基因组。这项研究表明,园艺植物的线粒体和叶绿体序列组织与其他种子植物相似。大多数园艺植物对线粒体基因组中富含A + T的特征表现出轻微的偏向。此外,基于15种蛋白质编码基因的39种园艺植物的系统发育分析表明,某些线粒体基因从叶绿体DNA水平转移。我们的研究将为其他园艺植物的细胞器基因组组装提供重要参考。此外,对园艺植物的细胞器基因组进行的系统发育分析可以准确阐明这些植物之间意料之外的关系。

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