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Single-Molecule Long-Read Sequencing of Purslane (Portulaca oleracea) and Differential Gene Expression Related with Biosynthesis of Unsaturated Fatty Acids

机译:与不饱和脂肪酸的生物合成相关的马冠(Portulaca Oleracea)的单分子长读取测序和差异基因表达

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摘要

This study aimed to obtain the full-length transcriptome of purslane (Portulaca oleracea); assorted plant samples were used for single-molecule real-time (SMRT) sequencing. Based on SMRT, functional annotation of transcripts, transcript factors (TFs) analysis, simple sequence repeat analysis and long non-coding RNAs (LncRNAs) prediction were accomplished. Total 15.33-GB reads were produced; with 9,350,222 subreads and the average length of subreads, 1640 bp was counted. With 99.99% accuracy, after clustering, 132,536 transcripts and 78,559 genes were detected. All unique SMART transcripts were annotated in seven functional databases. 4180 TFs (including transcript regulators) and 7289 LncRNAs were predicted. The results of RNA-seq were confirmed with qRT–PCR analysis. Illumina sequencing of leaves and roots of two purslane genotypes was carried out. Amounts of differential expression genes and related KEGG pathways were found. The expression profiles of related genes in the biosynthesis of unsaturated fatty acids pathway in leaves and roots of two genotypes of purslane were analyzed. Differential expression of genes in this pathway built the foundation of ω-3 fatty acid accumulation in different organs and genotypes of purslane. The aforementioned results provide sequence information and may be a valuable resource for whole-genome sequencing of purslane in the future.
机译:本研究旨在获得追踪(Portulaca Oleracea)的全长转录组;什锦植物样品用于单分子实时(SMRT)测序。基于SMRT,作用注释,成绩称因子(TFS)分析,简单的序列重复分析和长期非编码RNA(LNCRNA)预测。产生了15.33-GB的读数;含有9,350,222个底次和底次的平均长度,计算了1640年的BP。精度为99.99%,在聚类后,检测到132,536份转录物和78,559个基因。所有独特的智能成绩单都在七个功能数据库中注释。预测了4180个TFS(包括转录式调节器)和7289 LNCRNA。用QRT-PCR分析证实RNA-SEQ的结果。进行了叶片和两种追踪基因型的叶片序列。发现了差异表达基因和相关的Kegg途径。分析了在叶片中不饱和脂肪酸途径的生物合成中的相关基因的表达谱,并进行了两种基因分型的叶片。该途径中基因的差异表达为不同器官和追踪基因型的ω-3脂肪酸积累的基础。上述结果提供了序列信息,并且可以是未来主基因组测序的有价值资源。

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