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Analysis of miRNAs Targeted Storage Regulatory Genes during Soybean Seed Development Based on Transcriptome Sequencing

机译:基于转录组测序的大豆种子发育过程中靶向存储调控基因的miRNA分析

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摘要

Soybeans are an important cash crop and are widely used as a source of vegetable protein and edible oil. MicroRNAs (miRNA) are endogenous small RNA that play an important regulatory role in the evolutionarily conserved system of gene expression. In this study, we selected four lines with extreme phenotypes, as well as high or low protein and oil content, from the chromosome segment substitution line (CSSL) constructed from suinong (SN14) and ZYD00006, and planted and sampled at three stages of grain development for small RNA sequencing and expression analysis. The sequencing results revealed the expression pattern of miRNA in the materials, and predicted miRNA-targeted regulatory genes, including 1967 pairs of corresponding relationships between known-miRNA and their target genes, as well as 597 pairs of corresponding relationships between novel-miRNA and their target genes. After screening and annotating genes that were targeted for regulation, five specific genes were identified to be differentially expressed during seed development and subsequently analyzed for their regulatory relationship with miRNAs. The expression pattern of the targeted gene was verified by Real-time Quantitative PCR (RT-qPCR). Our research provides more information about the miRNA regulatory network in soybeans and further identifies useful genes that regulate storage during soy grain development, providing a theoretical basis for the regulation of soybean quality traits.
机译:大豆是重要的经济作物,被广泛用作植物蛋白和食用油的来源。微小RNA(miRNA)是内源性小RNA,在基因表达的进化保守系统中起重要的调节作用。在这项研究中,我们从遂农(SN14)和ZYD00006构建的染色体片段替代品系(CSSL)中选择了四个具有极端表型以及蛋白质和油脂含量高或低的品系,并在三个阶段的谷物中进行种植和采样小RNA测序和表达分析的开发。测序结果揭示了miRNA在材料中的表达模式,并预测了靶向miRNA的调控基因,包括1967对已知miRNA与其靶基因之间的对应关系,以及597对新颖miRNA与其靶基因之间的对应关系。靶基因。在筛选并注释了调控基因后,确定了五个特定基因在种子发育过程中差异表达,随后对其与miRNA的调控关系进行了分析。通过实时定量PCR(RT-qPCR)验证了靶基因的表达模式。我们的研究提供了有关大豆中miRNA调控网络的更多信息,并进一步鉴定了在大豆籽粒发育过程中调控贮藏的有用基因,为调控大豆品质性状提供了理论基础。

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