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利用Illumina测序技术对牵牛进行从头转录组组装以进行基因发现和SSR标记鉴定。

De novo transcriptome assembly of Ipomoea nil using Illumina sequencing for gene discovery and SSR marker identification.

作者信息

Wei Changhe, Tao Xiang, Li Ming, He Bin, Yan Lang, Tan Xuemei, Zhang Yizheng

机构信息

Key Laboratory of Bio-resources and Eco-environment, Ministry of Education, Sichuan Key Laboratory of Molecular Biology and Biotechnology, College of Life Sciences, Sichuan University, Chengdu, 610064, China.

Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu, 610041, China.

出版信息

Mol Genet Genomics. 2015 Oct;290(5):1873-84. doi: 10.1007/s00438-015-1034-6. Epub 2015 Apr 16.

Abstract

Ipomoea nil is widely used as an ornamental plant due to its abundance of flower color, but the limited transcriptome and genomic data hinder research on it. Using illumina platform, transcriptome profiling of I. nil was performed through high-throughput sequencing, which was proven to be a rapid and cost-effective means to characterize gene content. Our goal is to use the resulting information to facilitate the relevant research on flowering and flower color formation in I. nil. In total, 268 million unique illumina RNA-Seq reads were produced and used in the transcriptome assembly. These reads were assembled into 220,117 contigs, of which 137,307 contigs were annotated using the GO and KEGG database. Based on the result of functional annotations, a total of 89,781 contigs were assigned 455,335 GO term annotations. Meanwhile, 17,418 contigs were identified with pathway annotation and they were functionally assigned to 144 KEGG pathways. Our transcriptome revealed at least 55 contigs as probably flowering-related genes in I. nil, and we also identified 25 contigs that encode key enzymes in the phenylpropanoid biosynthesis pathway. Based on the analysis relating to gene expression profiles, in the phenylpropanoid biosynthesis pathway of I. nil, the repression of lignin biosynthesis might lead to the redirection of the metabolic flux into anthocyanin biosynthesis. This may be the most likely reason that I. nil has high anthocyanins content, especially in its flowers. Additionally, 15,537 simple sequence repeats (SSRs) were detected using the MISA software, and these SSRs will undoubtedly benefit future breeding work. Moreover, the information uncovered in this study will also serve as a valuable resource for understanding the flowering and flower color formation mechanisms in I. nil.

摘要

由于花色丰富,矮牵牛被广泛用作观赏植物,但转录组和基因组数据有限阻碍了对其的研究。利用Illumina平台,通过高通量测序对矮牵牛进行转录组分析,这被证明是一种快速且经济高效的基因特征鉴定方法。我们的目标是利用所得信息促进对矮牵牛开花和花色形成的相关研究。总共产生了2.68亿条独特的Illumina RNA-Seq读数并用于转录组组装。这些读数被组装成220,117个重叠群,其中137,307个重叠群使用GO和KEGG数据库进行注释。基于功能注释结果,共有89,781个重叠群被赋予455,335个GO术语注释。同时,17,418个重叠群通过通路注释被鉴定出来,并在功能上被分配到144个KEGG通路。我们的转录组显示矮牵牛中至少有55个重叠群可能是与开花相关的基因,我们还鉴定出25个重叠群,它们编码苯丙烷生物合成途径中的关键酶。基于与基因表达谱相关的分析,在矮牵牛的苯丙烷生物合成途径中,木质素生物合成的抑制可能导致代谢通量重新导向花青素生物合成。这可能是矮牵牛花青素含量高,尤其是在其花朵中的最可能原因。此外,使用MISA软件检测到15,537个简单序列重复(SSR),这些SSR无疑将有利于未来的育种工作。此外,本研究中发现的信息也将作为理解矮牵牛开花和花色形成机制的宝贵资源。

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