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基于深度测序的建兰花卉转录组分析。

Deep sequencing-based analysis of the Cymbidium ensifolium floral transcriptome.

作者信息

Li Xiaobai, Luo Jie, Yan Tianlian, Xiang Lin, Jin Feng, Qin Dehui, Sun Chongbo, Xie Ming

机构信息

Institute of Horticulture, Zhejiang Academy of Agricultural Sciences, Hangzhou, People's Republic of China.

Department of Gastroenterology, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, People's Republic of China.

出版信息

PLoS One. 2013 Dec 31;8(12):e85480. doi: 10.1371/journal.pone.0085480. eCollection 2013.

Abstract

Cymbidium ensifolium is a Chinese Cymbidium with an elegant shape, beautiful appearance, and a fragrant aroma. C. ensifolium has a long history of cultivation in China and it has excellent commercial value as a potted plant and cut flower. The development of C. ensifolium genomic resources has been delayed because of its large genome size. Taking advantage of technical and cost improvement of RNA-Seq, we extracted total mRNA from flower buds and mature flowers and obtained a total of 9.52 Gb of filtered nucleotides comprising 98,819,349 filtered reads. The filtered reads were assembled into 101,423 isotigs, representing 51,696 genes. Of the 101,423 isotigs, 41,873 were putative homologs of annotated sequences in the public databases, of which 158 were associated with floral development and 119 were associated with flowering. The isotigs were categorized according to their putative functions. In total, 10,212 of the isotigs were assigned into 25 eukaryotic orthologous groups (KOGs), 41,690 into 58 gene ontology (GO) terms, and 9,830 into 126 Arabidopsis Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, and 9,539 isotigs into 123 rice pathways. Comparison of the isotigs with those of the two related orchid species P. equestris and C. sinense showed that 17,906 isotigs are unique to C. ensifolium. In addition, a total of 7,936 SSRs and 16,676 putative SNPs were identified. To our knowledge, this transcriptome database is the first major genomic resource for C. ensifolium and the most comprehensive transcriptomic resource for genus Cymbidium. These sequences provide valuable information for understanding the molecular mechanisms of floral development and flowering. Sequences predicted to be unique to C. ensifolium would provide more insights into C. ensifolium gene diversity. The numerous SNPs and SSRs identified in the present study will contribute to marker development for C. ensifolium.

摘要

建兰是一种中国兰花,形态优雅,外观美丽,香气芬芳。建兰在中国有着悠久的栽培历史,作为盆栽植物和切花具有很高的商业价值。由于建兰基因组规模较大,其基因组资源的开发一直滞后。利用RNA测序技术的技术改进和成本降低,我们从花芽和成熟花中提取了总mRNA,共获得9.52 Gb的过滤核苷酸,包含98,819,349条过滤读段。这些过滤读段被组装成101,423个重叠群,代表51,696个基因。在101,423个重叠群中,41,873个是公共数据库中注释序列的推定同源物,其中158个与花发育相关,119个与开花相关。这些重叠群根据其推定功能进行分类。总共10,212个重叠群被归入25个真核直系同源组(KOG),41,690个归入58个基因本体(GO)术语中,9,830个归入126条拟南芥京都基因与基因组百科全书(KEGG)途径,9,539个重叠群归入123条水稻途径。将这些重叠群与两种相关兰花物种蝴蝶兰和墨兰的重叠群进行比较,发现17,906个重叠群是建兰独有的。此外,共鉴定出7,936个简单序列重复(SSR)和16,676个推定单核苷酸多态性(SNP)。据我们所知,这个转录组数据库是建兰的首个主要基因组资源,也是兰花属最全面的转录组资源。这些序列为理解花发育和开花的分子机制提供了有价值的信息。预测为建兰独有的序列将为深入了解建兰基因多样性提供更多见解。本研究中鉴定出的大量SNP和SSR将有助于建兰的分子标记开发。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c813/3877369/2f30b64b5da1/pone.0085480.g001.jpg

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