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Comparative transcriptome analysis of cotton fiber development of Upland cotton ( Gossypium hirsutum ) and Chromosome Segment Substitution Lines from G. hirsutum × G. barbadense

机译:陆地棉×陆地棉×陆地棉和染色体片段代换系棉纤维发育的转录组比较分析。

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Background How to develop new cotton varieties possessing high yield traits of Upland cotton and superior fiber quality traits of Sea Island cotton remains a key task for cotton breeders and researchers. While multiple attempts bring in little significant progresses, the development of Chromosome Segment Substitution Lines (CSSLs) from Gossypium barbadense in G. hirsutum background provided ideal materials for aforementioned breeding purposes in upland?cotton improvement. Based on the excellent fiber performance and relatively clear chromosome substitution segments information identified by Simple Sequence Repeat (SSR) markers, two CSSLs, MBI9915 and MBI9749, together with the recurrent parent CCRI36 were chosen to conduct transcriptome sequencing during the development stages of fiber elongation and Secondary Cell Wall (SCW) synthesis (from 10DPA and 28DPA), aiming at revealing the mechanism of fiber development and the potential contribution of chromosome substitution segments from Sea Island cotton to fiber development of Upland cotton. Results In total, 15 RNA-seq libraries were constructed and sequenced separately, generating 705.433 million clean reads with mean GC content of 45.13% and average Q30 of 90.26%. Through multiple comparisons between libraries, 1801 differentially expressed genes (DEGs) were identified, of which the 902 up-regulated DEGs were mainly involved in cell wall organization and response to oxidative stress and auxin, while the 898 down-regulated ones participated in translation, regulation of transcription, DNA-templated and cytoplasmic translation based on GO annotation and KEGG enrichment analysis. Subsequently, STEM software was performed to explicate the temporal expression pattern of DEGs. Two peroxidases and four flavonoid pathway-related genes were identified in the “oxidation-reduction process”, which could play a role in fiber development and quality formation. Finally, the reliability of RNA-seq data was validated by quantitative real-time PCR of randomly selected 20 genes. Conclusions The present report focuses on the similarities and differences of transcriptome profiles between the two CSSLs and the recurrent parent CCRI36 and provides novel insights into the molecular mechanism of fiber development, and into further exploration of the feasible contribution of G. barbadense substitution segments to fiber quality formation, which will lay solid foundation for simultaneously improving fiber yield and quality of upland cotton through CSSLs.
机译:背景技术如何开发具有陆地棉高产性状和海岛棉纤维品质优良性状的棉花新品种,仍然是棉花育种人员和研究人员的关键任务。尽管多次尝试几乎没有取得重大进展,但是从陆地棉背景下的巴巴棉(Gossypium barbadense)染色体片段替代系(CSSLs)的开发为上述育种目的提供了理想的材料,以改良陆地棉。基于出色的纤维性能和通过简单序列重复(SSR)标记鉴定的相对清晰的染色体取代片段信息,选择了两个CSSL,MBI9915和MBI9749以及循环亲本CCRI36在纤维伸长和发育的发育阶段进行转录组测序。二次细胞壁(SCW)合成(来自10DPA和28DPA),旨在揭示纤维发育的机理以及海岛棉染色体替代片段对陆地棉纤维发育的潜在贡献。结果总共构建了15个RNA-seq文库,并分别测序,产生了70543.3百万个干净读段,平均GC含量为45.13%,平均Q30为90.26%。通过文库之间的多次比较,鉴定出1801个差异表达基因(DEG),其中902个上调的DEG主要参与细胞壁的组织以及对氧化应激和生长素的反应,而898个下调的基因参与翻译, GO注释和KEGG富集分析对转录,DNA模板化和细胞质翻译的调控。随后,执行STEM软件以阐明DEG的时间表达模式。在“氧化还原过程”中鉴定出两个过氧化物酶和四个类黄酮途径相关基因,它们可能在纤维发育和质量形成中起作用。最后,通过随机选择的20个基因的实时定量PCR验证了RNA-seq数据的可靠性。结论本报告着眼于两个CSSLs与轮回亲本CCRI36之间的转录组谱的异同,并为纤维发育的分子机制提供了新见解,并进一步探索了巴氏假单胞菌替代片段对纤维的可行贡献。质量形成,这将为同时通过CSSL提高纤维产量和陆地棉质量奠定坚实的基础。

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