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纳米孔直接RNA测序揭示了玉米根中的短期盐胁迫反应。

Nanopore Direct RNA Sequencing Reveals the Short-Term Salt Stress Response in Maize Roots.

作者信息

He Shidong, Wang Hui, Lv Minghao, Li Shun, Song Junhui, Wang Rongxin, Jiang Shaolong, Jiang Lijun, Zhang Shuxin, Li Xiang

机构信息

National Key Laboratory of Wheat Improvement, College of Life Sciences, Shandong Agricultural University, Tai'an 271018, China.

出版信息

Plants (Basel). 2024 Jan 30;13(3):405. doi: 10.3390/plants13030405.

Abstract

Transcriptome analysis, relying on the cutting-edge sequencing of cDNA libraries, has become increasingly prevalent within functional genome studies. However, the dependence on cDNA in most RNA sequencing technologies restricts their ability to detect RNA base modifications. To address this limitation, the latest Oxford Nanopore Direct RNA Sequencing (ONT DRS) technology was employed to investigate the transcriptome of maize seedling roots under salt stress. This approach aimed to unveil both the RNA transcriptional profiles and alterations in base modifications. The analysis of the differential expression revealed a total of 1398 genes and 2223 transcripts that exhibited significant variation within the maize root system following brief exposure to salt stress. Enrichment analyses, such as the Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway assessments, highlighted the predominant involvement of these differentially expressed genes (DEGs) in regulating ion homeostasis, nitrogen metabolism, amino acid metabolism, and the phytohormone signaling pathways. The protein-protein interaction (PPI) analysis showed the participation of various proteins related to glycolytic metabolism, nitrogen metabolism, amino acid metabolism, abscisic acid signaling, and the jasmonate signaling pathways. It was through this intricate molecular network that these proteins collaborated to safeguard root cells against salt-induced damage. Moreover, under salt stress conditions, the occurrence of variable shear events (AS) in RNA modifications diminished, the average length of poly(A) tails underwent a slight decrease, and the number of genes at the majority of the variable polyadenylation (APA) sites decreased. Additionally, the levels of N5-methylcytosine (m5C) and N6-methyladenosine (m6A) showed a reduction. These results provide insights into the mechanisms of early salt tolerance in maize.

摘要

转录组分析依赖于cDNA文库的前沿测序技术,在功能基因组研究中越来越普遍。然而,大多数RNA测序技术对cDNA的依赖限制了它们检测RNA碱基修饰的能力。为了解决这一限制,我们采用了最新的牛津纳米孔直接RNA测序(ONT DRS)技术来研究盐胁迫下玉米幼苗根的转录组。该方法旨在揭示RNA转录谱和碱基修饰的变化。差异表达分析显示,在短暂暴露于盐胁迫后,玉米根系中共有1398个基因和2223个转录本表现出显著变化。基因本体论(GO)和京都基因与基因组百科全书(KEGG)通路评估等富集分析强调,这些差异表达基因(DEG)主要参与调节离子稳态、氮代谢、氨基酸代谢和植物激素信号通路。蛋白质-蛋白质相互作用(PPI)分析表明,各种与糖酵解代谢、氮代谢、氨基酸代谢、脱落酸信号和茉莉酸信号通路相关的蛋白质参与其中。正是通过这个复杂的分子网络,这些蛋白质协同保护根细胞免受盐诱导的损伤。此外,在盐胁迫条件下,RNA修饰中的可变剪切事件(AS)发生率降低,多聚腺苷酸(poly(A))尾巴的平均长度略有下降,大多数可变聚腺苷酸化(APA)位点的基因数量减少。此外,N5-甲基胞嘧啶(m5C)和N6-甲基腺苷(m6A)的水平也有所降低。这些结果为玉米早期耐盐机制提供了见解。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0a69/10857558/ab325912b61e/plants-13-00405-g001.jpg

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