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园艺植物细胞器基因组组装方法及比较分析

Organellar genome assembly methods and comparative analysis of horticultural plants.

作者信息

Wang Xuelin, Cheng Feng, Rohlsen Dekai, Bi Changwei, Wang Chunyan, Xu Yiqing, Wei Suyun, Ye Qiaolin, Yin Tongming, Ye Ning

机构信息

1College of Information Science and Technology, Nanjing Forestry University, Nanjing, Jiangsu China.

2Department of Pharmaceutical Science, College of Pharmacy, University of South Florida, Tampa, FL 33612 USA.

出版信息

Hortic Res. 2018 Jan 10;5:3. doi: 10.1038/s41438-017-0002-1. eCollection 2018.

Abstract

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, , and a fruit tree, , 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.

摘要

尽管细胞器基因组(包括叶绿体和线粒体基因组)在大小和基因数量上比核基因组小,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。很少有研究关注园艺植物的细胞器基因组。美国国立生物技术信息中心(NCBI)中大约有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,我们报告了一种从下一代测序(NGS)数据中高质量组装园艺细胞器基因组的创新高效方法。测序读段首先使用Newbler、Amos和Minimus软件以默认参数进行组装。然后通过BLASTN搜索和PCR填补剩余的缺口。使用BWA(Burrows-Wheeler比对工具)软件基于Illumina测序数据对完整的DNA序列进行校正。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。使用该程序,鉴定出了一种观赏植物和一种果树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。大多数园艺植物的线粒体基因组对富含A+T的特征有轻微偏向。此外,基于15个蛋白质编码基因对39种园艺植物进行的系统发育分析表明,一些线粒体基因是从叶绿体DNA水平转移而来的。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,对园艺植物细胞器基因组的系统发育分析可以准确阐明这些植物之间意想不到的关系。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b9e1/5798811/286d6a16807e/41438_2017_2_Fig1_HTML.jpg

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