Genome-wide Identification and Analysis of Long non-coding RNAs Involved in Fatty acid Biosynthesis in Young Soybean pods

Here, the low-linolenic acid mutant “MT72” and the wild-type control “JN18” were used as materials. The lncRNAs in young pods at 30 and 40 days (d) after flowering were systematically identified and analyzed using transcriptome sequencing technology combined with bioinformatics tools. A total of 39,324 lncRNAs and 561 differentially expressed lncRNAs were identified. A lncRNAs-miRNAs- protein-coding genes (mRNAs) network was constructed, and 46 lncRNAs, 46 miRNAs and 137 mRNAs were found to be correlated. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis of 12 targeted mRNAs in the competing endogenous RNA network showed that these lncRNAs may be involved in the biological processes of fatty acid transport, lipid synthesis and cell division. Finally, the expression levels of differentially expressed lncRNAs, miRNAs and mRNAs were verified using qRT-PCR. The expression patterns of most genes were consistent with the sequencing results. In conclusion, new information was provided for the study of fatty acid synthesis by lncRNAs in young soybean pods. Overall design: Studies on low linolenic acid mutants “MT72” and wild-type “JN18” at 30d and 40d after flowering of soybean

本研究以低亚麻酸突变体"MT72"与野生型对照"JN18"为实验材料。利用转录组测序技术结合生物信息学工具，系统鉴定并分析了开花后30天与40天大豆幼荚中的长链非编码RNA（lncRNAs）。共鉴定得到39324条lncRNAs以及561个差异表达lncRNAs。构建了lncRNAs-微小RNA（miRNAs）-蛋白编码基因（信使RNA，mRNAs）调控网络，筛选得到46个lncRNAs、46个miRNAs及137个mRNAs存在调控关联。对该内源竞争RNA网络中的12个靶标mRNAs进行基因本体（Gene Ontology, GO）与京都基因与基因组百科全书（Kyoto Encyclopedia of Genes and Genomes, KEGG）通路富集分析，结果显示上述lncRNAs可能参与脂肪酸转运、脂质合成及细胞分裂等生物学过程。最后采用实时定量聚合酶链式反应（qRT-PCR）验证了差异表达lncRNAs、miRNAs及mRNAs的表达水平，多数基因的表达模式与测序结果一致。综上，本研究为大豆幼荚中lncRNAs参与脂肪酸合成的相关研究提供了新的研究线索。整体实验设计：针对大豆开花后30天与40天的低亚麻酸突变体"MT72"及野生型"JN18"开展相关研究。