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利用甲基化DNA免疫沉淀测序技术对棉花(陆地棉)盐胁迫诱导的DNA甲基化变化进行全基因组分析。

Genome-wide analysis of salinity-stress induced DNA methylation alterations in cotton (Gossypium hirsutum L.) using the Me-DIP sequencing technology.

作者信息

Lu X K, Shu N, Wang J J, Chen X G, Wang D L, Wang S, Fan W L, Guo X N, Guo L X, Ye W W

机构信息

College of Agronomy, , , China.

State Key Laboratory of Cotton Biology/Key Laboratory for Cotton Genetic Improvement, Ministry/Institute of Cotton Research of Chinese Academy of Agricultural Sciences, Anyang, , China.

出版信息

Genet Mol Res. 2017 Jun 29;16(2):gmr-16-02-gmr.16029673. doi: 10.4238/gmr16029673.

DOI:10.4238/gmr16029673
PMID:28671256
Abstract

Cytosine DNA methylation is a significant form of DNA modification closely associated with gene expression in eukaryotes, fungi, animals, and plants. Although the reference genomes of cotton (Gossypium hirsutum L.) have been publically available, the salinity-stress-induced DNA methylome alterations in cotton are not well understood. Here, we constructed a map of genome-wide DNA methylation characteristics of cotton leaves under salt stress using the methylated DNA immunoprecipitation sequencing method. The results showed that the methylation reads on chromosome 9 were most comparable with those on the other chromosomes, but the greatest changes occurred on chromosome 8 under salt stress. The DNA methylation pattern analysis indicated that a relatively higher methylation density was found in the upstream2k and downstream2k elements of the CDS region and CG-islands. Almost 94% of the reads belonged to LTR-gspsy and LTR-copia, and the number of methylation reads in LTR-gypsy was four times greater than that in LTR-copia in both control and stressed samples. The analysis of differentially methylated regions (DMRs) showed that the gene elements upstream2k, intron, and downstream2k were hypomethylated, but the CDS regions were hypermethylated. The GO (Gene Ontology) analysis suggested that the methylated genes were most enriched in cellular processes, metabolic processes, cell parts and catalytic activities, which might be closely correlated with response to NaCl stress. In this study, we completed a genomic DNA methylation profile and conducted a DMR analysis under salt stress, which provided valuable information for the better understanding of epigenetics in response to salt stress in cotton.

摘要

胞嘧啶DNA甲基化是一种重要的DNA修饰形式,与真核生物、真菌、动物和植物中的基因表达密切相关。尽管棉花(陆地棉)的参考基因组已公开可用,但盐分胁迫诱导的棉花DNA甲基化组变化仍未得到充分了解。在此,我们使用甲基化DNA免疫沉淀测序方法构建了盐胁迫下棉花叶片全基因组DNA甲基化特征图谱。结果表明,9号染色体上的甲基化读数与其他染色体上的最为可比,但在盐胁迫下8号染色体上的变化最大。DNA甲基化模式分析表明,在CDS区域和CG岛的上游2k和下游2k元件中发现了相对较高的甲基化密度。几乎94%的读数属于LTR - gspsy和LTR - copia,在对照和胁迫样本中,LTR - gypsy中的甲基化读数数量是LTR - copia中的四倍。差异甲基化区域(DMR)分析表明,上游2k、内含子和下游2k基因元件发生低甲基化,但CDS区域发生高甲基化。基因本体(GO)分析表明,甲基化基因在细胞过程、代谢过程、细胞部分和催化活性中富集程度最高,这可能与对NaCl胁迫的响应密切相关。在本研究中,我们完成了基因组DNA甲基化图谱,并在盐胁迫下进行了DMR分析,为更好地理解棉花对盐胁迫的表观遗传学提供了有价值的信息。

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