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基于转录组测序的大豆种子发育过程中 miRNA 靶向贮藏调控基因分析。

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

机构信息

College of Agriculture, Northeast Agricultural University, Harbin 150030, Heilongjiang, China.

Department of Horticulture, Michigan State University, East Lansing 48824, MI, USA.

出版信息

Genes (Basel). 2019 May 28;10(6):408. doi: 10.3390/genes10060408.

Abstract

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.

摘要

大豆是一种重要的经济作物,广泛用作植物蛋白和食用油的来源。MicroRNAs (miRNA) 是内源性的小 RNA,在基因表达的进化保守系统中发挥重要的调节作用。在这项研究中,我们从苏农(SN14)和中油杂 6 号构建的染色体片段代换系(CSSL)中选择了四个表型极端、高蛋白和高油含量的品系,以及低蛋白和低油含量的品系,在三个籽粒发育阶段进行种植和采样,进行小 RNA 测序和表达分析。测序结果揭示了材料中 miRNA 的表达模式,并预测了 miRNA 靶向的调节基因,包括已知-miRNA 和其靶基因之间的 1967 对对应关系,以及 novel-miRNA 和其靶基因之间的 597 对对应关系。在筛选和注释受调控的基因后,鉴定出了五个在种子发育过程中差异表达的特定基因,并进一步分析了它们与 miRNAs 的调控关系。通过 Real-time Quantitative PCR (RT-qPCR) 验证了靶基因的表达模式。我们的研究提供了更多关于大豆中 miRNA 调控网络的信息,并进一步鉴定了在大豆籽粒发育过程中调节储存的有用基因,为大豆品质性状的调控提供了理论基础。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d68a/6628032/63bccbc468ec/genes-10-00408-g001.jpg

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