山麻鸭开产期和产蛋高峰期卵巢组织转录组分析

doi:10.3864/j.issn.0578-1752.2016.05.020

摘要/Abstract

摘要： 【目的】探明山麻鸭开产期和产蛋高峰期卵巢组织的转录组差异，丰富蛋鸭转录组数据信息。【方法】选取6份山麻鸭卵巢组织样品（开产期和产蛋高峰期各3份），利用Illumina HiSeq^TM 2000进行高通量测序，构建山麻鸭开产期和产蛋高峰期卵巢组织转录组文库，并用测序评估、基因功能注释等生物信息学方法进行分析。【结果】经过测序获得质量不低于20的碱基比例（Q20）均高于93%；开产期获得了43 489 798条reads，4 380 847 061 bp数据量；高峰期获得了42 782 676条reads，4 307 499 083 bp数据量；分别有70.92%和72.70%的reads能比对到鸭参考基因组序列上。对开产期与产蛋高峰期转录组进行比较，发现开产期有26 808个表达基因，产蛋高峰期有27 013个表达基因，与开产期为参考，在高峰期卵巢组织中共获得1 929个差异表达基因，其中上调基因989个，下调基因940个；进一步将这些差异基因在Nr数据库进行注释，发现获得注释的基因有1 423个，其中上调基因695个，下调基因728个；COG功能注释分析发现这些差异表达基因共获得663个功能注释，涉及25个功能分类；GO 功能注释分析表明，有1 122个基因获得GO 功能注释，可以分为61个功能分类，这些分类主要涉及到分子绑定、催化活性、细胞过程、生物调节等诸多生理生化过程，其中涉及到发育繁殖生物学过程的相关注释基因有406个；KEGG分析发现共有425个基因注释到160个代谢通路中，其中GnRH信号通路、GPI-anchor生物合成、TGF-β信号通路、核糖体生物合成等通路显著富集。GnRH信号通路和TGF-β信号通路在卵巢的生理生殖活动发挥了重要作用，而GPI-anchor生物合成和核糖体生物合成，这两类代谢通路在卵巢生理生殖活动的作用尚不明确。【结论】利用高通量测序技术对山麻鸭开产期和产蛋高峰期卵巢组织的转录组进行测序和分析，揭示了山麻鸭不同生理状态下卵巢组织差异表达基因的数量，获得了差异表达基因的功能、分类和代谢通路。为丰富蛋鸭卵巢组织转录组信息，为开展蛋鸭产蛋性状相关基因的研究及分子调控机制研究奠定基础。

关键词: 山麻鸭, 卵巢, 转录组, 差异表达基因, 功能分类

Abstract: 【Objective】The objective of this study was to find the transcriptome differences between ovary tissue in early laying period and egg laying peak period of Shanma ducks, and enrich the laying duck transcriptome data.【Method】Selecting 6 Shanma duck ovarian tissues as a sample (early laying period and peak period 3 samples respectively), then the transcriptome libraries of ovary tissue in the early laying period and the egg laying peak period of Shanma ducks were constructed by using an Illumina HiSeq^TM 2000 sequencing technique, and the bioinformatics methods were analyzed subsequently, such as sequencing assess and gene function annotation. 【Result】 The results showed that the base ratios with quality value higher than 20 in reads (Q20) were more than 93%. 43 489 798 reads and 438 084 706 bp transcriptome data were obtained in the early laying period, and 42 782 676 reads, 4 307 499 083 bp transcriptome data were obtained in the egg laying peak period. Respectively, 70.92% and 72.70% reads could be compared to the duck reference genome sequence. Comparison of the transcriptome of ovary tissue in the early laying period and the egg laying peak period, 26 808 expressed genes were found in the early laying period, 27 013 expressed genes were found in the egg laying peak period. As the early laying period for reference, 1 929 differentially expressed genes were found in the egg laying peak period, including 989 up-regulated genes and 940 down-regulated genes. Annotation analysis indicated that 1 423 genes were annotated in Nr data, including 695 up-regulated genes and 728 down-regulated genes. Through COG analysis, there were 663 functional annotations of these differentially expressed genes, involving 25 functional classifications. With GO function annotation classifications, a total of 1 122 genes were divided into 61 function categories, in which many functional categories were mainly involved, such as molecular binding, catalytic activity, cell process, biological regulation, which including 406 related annotation genes involved development and reproduction biological process. KEGG analysis showed a total of 425 genes were annotated to 160 metabolic pathways, and GnRH signaling pathway, GPI-anchor biosynthesis, TGF-beta signaling pathway, ribosome biosynthesis pathway were significantly enriched. The GnRH and TGF-β signaling pathway played an important role in the ovarian physiology and reproduction activities, but the role of GPI-anchor and ribosome biosynthesis was not clear in ovarian physiology and reproductive activity.【Conclusion】The transcriptome study of the ovary tissue in the early laying period and the egg laying peak period of Shanma ducks by using high-throughput sequencing technology revealed the number of differentially expressed genes under different physiological status of the ovarian tissue in Shanma ducks, obtained the function, classification and metabolic pathway of differentially expressed genes, which enriched the ovary transcriptome information of laying ducks, and laid a foundation for the study on related genes with reproductive traits and molecular mechanisms in laying ducks.

Key words: Shanma duck, ovary, transcriptome, differentially expressed genes, functional classification

朱志明，陈红萍，林如龙，缪中纬，辛清武，李丽，张丹青，郑嫩珠. 山麻鸭开产期和产蛋高峰期卵巢组织转录组分析[J]. 中国农业科学, 2016, 49(5): 998-1007.

ZHU Zhi-ming, CHEN Hong-ping, LIN Ru-long, MIAO Zhong-wei, XIN Qing-wu, LI Li, ZHANG Dan-qing, ZHENG Nen-zhu. Transcriptome Analysis of Ovary Tissue in Early Laying Period and Egg Laying Peak Period of Shanma Ducks[J]. Scientia Agricultura Sinica, 2016, 49(5): 998-1007.

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