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龙眼早期体胚发生过程中 miRNA 及其靶基因的全基因组鉴定

Genome-wide identification of miRNAs and their targets during early somatic embryogenesis in Dimocarpus longan Lour.

机构信息

Institute of Horticultural Biotechnology, Fujian Agriculture and Forestry University, Fuzhou, 350002, China.

出版信息

Sci Rep. 2020 Mar 13;10(1):4626. doi: 10.1038/s41598-020-60946-y.

Abstract

miRNAs are endogenous regulatory factors that play pivotal roles in post-transcriptional regulation. However, their specific roles in early somatic embryogenesis (SE) remain unclear. Study of the SE system is fundamental for clarifying the molecular mechanisms in Dimocarpus longan. We identified 289 known miRNAs from 106 different miRNA families and 1087 novel miRNAs during early longan SE, including embryogenic callus (EC), incomplete pro-embryogenic culture (ICpEC), globular embryo (GE), and non-embryogenic callus (NEC). The abundances of known miRNAs were concentrated in GE. The differentially expression (DE) miRNAs showed five expression patterns during early SE. Largely miRNAs were expressed highly and specially in EC, ICpEC, and GE, respectively. Some miRNAs and putative target genes were enriched in lignin metabolism. Most potential targets were related to the pathways of plant hormone signal transduction, alternative splicing, tyrosine metabolism and sulfur metabolism in early longan SE. The regulatory relationships between dlo-miR166a-3p and DlHD-zip8, dlo-miR397a and DlLAC7, dlo-miR408-3p and DlLAC12 were confirmed by RNA ligase-mediated rapid amplification of cDNA ends. The expression patterns of eight DE miRNAs detected by qRT-PCR were consistent with RNA-seq. Finally, the miRNA regulatory network in early SE was constructed, which provided new insight into molecular mechanism of early SE in longan.

摘要

miRNAs 是内源性调控因子,在转录后调控中发挥关键作用。然而,它们在早期体胚发生 (SE) 中的具体作用尚不清楚。研究 SE 系统对于阐明龙眼的分子机制是基础。我们在早期龙眼 SE 中从 106 个不同的 miRNA 家族和 1087 个新的 miRNA 中鉴定出 289 个已知的 miRNA,包括胚性愈伤组织 (EC)、不完全原胚培养物 (ICpEC)、球形胚 (GE) 和非胚性愈伤组织 (NEC)。已知 miRNA 的丰度集中在 GE 中。差异表达 (DE) 的 miRNA 在早期 SE 期间表现出五种表达模式。大体上,miRNA 在 EC、ICpEC 和 GE 中表达水平高且特异性表达。一些 miRNA 和推定的靶基因富集在木质素代谢中。大多数潜在的靶基因与早期龙眼 SE 中植物激素信号转导、可变剪接、酪氨酸代谢和硫代谢途径有关。dlo-miR166a-3p 和 DlHD-zip8、dlo-miR397a 和 DlLAC7、dlo-miR408-3p 和 DlLAC12 之间的调控关系通过 RNA 连接酶介导的 cDNA 末端快速扩增得到证实。qRT-PCR 检测的 8 个 DE miRNA 的表达模式与 RNA-seq 一致。最后,构建了早期 SE 中的 miRNA 调控网络,为龙眼早期 SE 的分子机制提供了新的见解。

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