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澳洲山龙眼(Dodonaea viscosa)的转录组测序、注释及多态性检测

Transcriptome sequencing, annotation and polymorphism detection in the hop bush, Dodonaea viscosa.

作者信息

Christmas Matthew J, Biffin Ed, Lowe Andrew J

机构信息

Environment Institute and School of Biological Sciences, The University of Adelaide, North Terrace, Adelaide, 5005, SA, Australia.

出版信息

BMC Genomics. 2015 Oct 16;16:803. doi: 10.1186/s12864-015-1987-1.

Abstract

BACKGROUND

The hop bush, Dodonaea viscosa, is a trans-oceanic species distributed oversix continents. It evolved in Australia where it is found over a wide range of habitat types and is an ecologically important species. Limited genomic resources are currently available for this species, thus our understanding of its evolutionary history and ecological adaptation is restricted. Here, we present a comprehensive transcriptome dataset for future genomic studies into this species.

METHODS

We performed Illumina sequencing of cDNA prepared from leaf tissue collected from seven populations of D. viscosa ssp. angustissima and spatulata distributed along an environmental gradient in South Australia. Sequenced reads were assembled to provide a transcriptome resource. Contiguous sequences (contigs) were annotated using BLAST searches against the NCBI non-redundant database and gene ontology definitions were assigned. Single nucleotide polymorphisms were detected for the establishment of a genetic marker set. A comparison between the two subspecies was also carried out.

RESULTS

Illumina sequencing returned 268,672,818 sequence reads, which were de novoassembled into 105,125 contigs. Contigs with significant BLAST alignments (E value < 1e(-5))numbered at 44,191, with 38,311 of these having their most significant hits to sequences from land plant species. Gene Ontology terms were assigned to 28,440 contigs and KEGG analysis identified 146 pathways that the gene products from 5,070 contigs are potentially involved in. The subspecies comparison identified 8,494 fixed SNP differences across 3,979 contiguous sequences, indicating a level of genetic differentiation between them. Across all samples, 248,235 SNPs were detected.

CONCLUSIONS

We have established a significant genomic data resource for D. viscosa,providing a comprehensive transcriptomic reference. Genetic differences among morphologically distinct subspecies were found. A wide range of putative gene regions were identified along with a large set of variable SNP markers, providing a basis for studies into the evolution and ecological adaptation of D. viscosa.

摘要

背景

澳洲瓶刷树(Dodonaea viscosa)是一种分布于六大洲的跨洋物种。它在澳大利亚进化,在那里可发现于广泛的栖息地类型中,是一种具有重要生态意义的物种。目前该物种的基因组资源有限,因此我们对其进化历史和生态适应性的理解受到限制。在此,我们提供一个全面的转录组数据集,以供未来对该物种进行基因组研究。

方法

我们对从南澳大利亚沿环境梯度分布的七个澳洲瓶刷树窄叶亚种(D. viscosa ssp. angustissima)和匙叶亚种(spatulata)种群的叶片组织制备的cDNA进行了Illumina测序。对测序读数进行组装以提供转录组资源。使用针对NCBI非冗余数据库的BLAST搜索对连续序列(重叠群)进行注释,并指定基因本体定义。检测单核苷酸多态性以建立遗传标记集。还对两个亚种进行了比较。

结果

Illumina测序返回了268,672,818个序列读数,这些读数被从头组装成105,125个重叠群。与BLAST有显著比对(E值<1e(-5))的重叠群有44,191个,其中38,311个与陆地植物物种的序列有最显著的匹配。基因本体术语被指定给28,440个重叠群,KEGG分析确定了5,070个重叠群的基因产物可能参与的146条途径。亚种比较在3,979个连续序列中鉴定出8,494个固定的SNP差异,表明它们之间存在一定程度的遗传分化。在所有样本中,共检测到248,235个SNP。

结论

我们为澳洲瓶刷树建立了一个重要的基因组数据资源,提供了一个全面的转录组参考。发现了形态上不同的亚种之间的遗传差异。鉴定出了广泛的假定基因区域以及大量可变的SNP标记,为研究澳洲瓶刷树的进化和生态适应性提供了基础。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8e20/4609105/a54e36aff27e/12864_2015_1987_Fig1_HTML.jpg

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