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西伯利亚鲟的RNA测序为研究形态发生分化和发育调控基因在发育早期与晚期阶段的表达动态提供了见解。

RNA-sequencing of the sturgeon Acipenser baeri provides insights into expression dynamics of morphogenic differentiation and developmental regulatory genes in early versus late developmental stages.

作者信息

Song Wei, Jiang Keji, Zhang Fengying, Lin Yu, Ma Lingbo

机构信息

Key Laboratory of East China Sea and Oceanic Fishery Resources Exploitation and Utilization, Ministry of Agriculture, Shanghai, 200090, China.

East China Sea fisheries Research Institute, Chinese Academy of Fishery Sciences, Shanghai, 200090, China.

出版信息

BMC Genomics. 2016 Aug 8;17:564. doi: 10.1186/s12864-016-2839-3.

Abstract

BACKGROUND

Acipenser baeri, one of the critically endangered animals on the verge of extinction, is a key species for evolutionary, developmental, physiology and conservation studies and a standout amongst the most important food products worldwide. Though the transcriptome of the early development of A. baeri has been published recently, the transcriptome changes occurring in the transition from embryonic to late stages are still unknown. The aim of this work was to analyze the transcriptomes of embryonic and post-embryonic stages of A. baeri and identify differentially expressed genes (DEGs) and their expression patterns using mRNA collected from specimens at big yolk plug, wide neural plate and 64 day old sturgeon developmental stages for RNA-Seq.

RESULTS

The paired-end sequencing of the transcriptome of samples of A. baeri collected at two early (big yolk plug (T1, 32 h after fertilization) and wide neural plate formation (T2, 45 h after fertilization)) and one late (T22, 64 day old sturgeon) developmental stages using Illumina Hiseq2000 platform generated 64039846, 64635214 and 75293762 clean paired-end reads for T1, T2 and T22, respectively. After quality control, the sequencing reads were de novo assembled to generate a set of 149,265 unigenes with N50 value of 1277 bp. Functional annotation indicated that a substantial number of these unigenes had significant similarity with proteins in public databases. Differential expression profiling allowed the identification of 2789, 12,819 and 10,824 DEGs from the respective T1 vs. T2, T1 vs. T22 and T2 vs. T22 comparisons. High correlation of DEGs' features was recorded among early stages while significant divergences were observed when comparing the late stage with early stages. GO and KEGG enrichment analyses revealed the biological processes, cellular component, molecular functions and metabolic pathways associated with identified DEGs. The qRT-PCR performed for candidate genes in specimens confirmed the validity of the RNA-seq data.

CONCLUSIONS

This study presents, for the first time, an extensive overview of RNA-Seq based characterization of the early and post-embryonic developmental transcriptomes of A. baeri and provided 149,265 gene sequences that will be potentially valuable for future molecular and genetic studies in A. baeri.

摘要

背景

西伯利亚鲟是极度濒危且濒临灭绝的动物之一,是进化、发育、生理学及保护研究的关键物种,也是全球最重要的食品之一。尽管西伯利亚鲟早期发育的转录组最近已发表,但从胚胎期到后期发生的转录组变化仍不清楚。本研究的目的是分析西伯利亚鲟胚胎期和胚后期的转录组,并使用从大卵黄栓、宽神经板和64日龄鲟鱼发育阶段的标本中收集的mRNA进行RNA测序,以鉴定差异表达基因(DEGs)及其表达模式。

结果

使用Illumina Hiseq2000平台对在两个早期(大卵黄栓(T1,受精后32小时)和宽神经板形成期(T2,受精后45小时))和一个后期(T22,64日龄鲟鱼)发育阶段收集的西伯利亚鲟样本的转录组进行双末端测序,分别为T1、T2和T22生成了64039846、64635214和75293762条干净的双末端读段。经过质量控制后,对测序读段进行从头组装,生成了一组149,265个单基因,N50值为1277 bp。功能注释表明,这些单基因中有大量与公共数据库中的蛋白质具有显著相似性。差异表达谱分析允许从各自的T1与T2、T1与T22以及T2与T22比较中鉴定出2789、12819和10824个DEG。在早期阶段记录到DEG特征的高度相关性,而在将后期与早期阶段进行比较时观察到显著差异。GO和KEGG富集分析揭示了与已鉴定的DEG相关的生物学过程、细胞成分、分子功能和代谢途径。对标本中候选基因进行的qRT-PCR证实了RNA测序数据的有效性。

结论

本研究首次全面概述了基于RNA测序的西伯利亚鲟胚胎期和胚后期发育转录组特征,并提供了149,265个基因序列,这些序列对未来西伯利亚鲟的分子和遗传研究可能具有重要价值。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8970/4977659/30633ee0dbb8/12864_2016_2839_Fig1_HTML.jpg

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