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转录组测序和差异基因表达分析澳棉种子萌发过程中延迟的腺体形态发生。

Transcriptome sequencing and differential gene expression analysis of delayed gland morphogenesis in Gossypium australe during seed germination.

机构信息

State Key Laboratory of Crop Genetics & Germplasm Enhancement, MOE Hybrid Cotton R&D Engineering Research Center, Nanjing Agricultural University, Nanjing, Jiangsu, People's Republic of China.

出版信息

PLoS One. 2013 Sep 20;8(9):e75323. doi: 10.1371/journal.pone.0075323. eCollection 2013.

Abstract

The genus Gossypium is a globally important crop that is used to produce textiles, oil and protein. However, gossypol, which is found in cultivated cottonseed, is toxic to humans and non-ruminant animals. Efforts have been made to breed improved cultivated cotton with lower gossypol content. The delayed gland morphogenesis trait possessed by some Australian wild cotton species may enable the widespread, direct usage of cottonseed. However, the mechanisms about the delayed gland morphogenesis are still unknown. Here, we sequenced the first Australian wild cotton species (Gossypiumaustrale) and a diploid cotton species (Gossypiumarboreum) using the Illumina Hiseq 2000 RNA-seq platform to help elucidate the mechanisms underlying gossypol synthesis and gland development. Paired-end Illumina short reads were de novo assembled into 226,184, 213,257 and 275,434 transcripts, clustering into 61,048, 47,908 and 72,985 individual clusters with N50 lengths of 1,710 bp, 1544 BP and 1,743 bp, respectively. The clustered Unigenes were searched against three public protein databases (TrEMBL, SwissProt and RefSeq) and the nucleotide and protein sequences of Gossypiumraimondii using BLASTx and BLASTn. A total of 21,987, 17,209 and 25,325 Unigenes were annotated. Of these, 18,766 (85.4%), 14,552 (84.6%) and 21,374 (84.4%) Unigenes could be assigned to GO-term classifications. We identified and analyzed 13,884 differentially expressed Unigenes by clustering and functional enrichment. Terpenoid-related biosynthesis pathways showed differentially regulated expression patterns between the two cotton species. Phylogenetic analysis of the terpene synthases family was also carried out to clarify the classifications of TPSs. RNA-seq data from two distinct cotton species provide comprehensive transcriptome annotation resources and global gene expression profiles during seed germination and gland and gossypol formation. These data may be used to further elucidate various mechanisms and help promote the usage of cottonseed.

摘要

陆地棉属是一种全球重要的作物,用于生产纺织品、油和蛋白质。然而,在栽培棉籽中发现的棉酚对人类和非反刍动物有毒。人们一直致力于培育低棉酚含量的改良栽培棉。一些澳大利亚野生棉种所具有的延迟腺体形态发生特性可能使棉籽能够广泛、直接地使用。然而,关于延迟腺体形态发生的机制尚不清楚。在这里,我们使用 Illumina Hiseq 2000 RNA-seq 平台对第一个澳大利亚野生棉种(Gossypiumaustrale)和二倍体棉花种(Gossypiumarboreum)进行了测序,以帮助阐明棉酚合成和腺体发育的机制。Illumina 短读对被从头组装成 226184、213257 和 275434 个转录本,聚类成 61048、47908 和 72985 个个体簇,N50 长度分别为 1710bp、1544bp 和 1743bp。聚类后的 Unigenes 被搜索了三个公共蛋白质数据库(TrEMBL、SwissProt 和 RefSeq)和 Gossypiumraimondii 的核苷酸和蛋白质序列,使用 BLASTx 和 BLASTn。总共注释了 21987、17209 和 25325 个 Unigenes。其中,18766(85.4%)、14552(84.6%)和 21374(84.4%)个 Unigenes可以被分配到 GO-term 分类。我们通过聚类和功能富集鉴定和分析了 13884 个差异表达的 Unigenes。萜烯相关生物合成途径在两个棉种之间表现出不同的表达模式。萜烯合酶家族的系统发育分析也进行了,以阐明 TPSs 的分类。来自两个不同棉花种的 RNA-seq 数据提供了全面的转录组注释资源和种子萌发、腺体和棉酚形成过程中的全基因表达谱。这些数据可能用于进一步阐明各种机制,并有助于促进棉籽的使用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0659/3779162/261cf3d8495c/pone.0075323.g001.jpg

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