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利用全长异构体测序技术对茶树(Camellia sinensis)转录组复杂性进行调查。

A survey of transcriptome complexity using full-length isoform sequencing in the tea plant Camellia sinensis.

机构信息

Shenzhen Branch, Guangdong Laboratory of Lingnan Modern Agriculture, Genome Analysis Laboratory of the Ministry of Agriculture and Rural Affairs, Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences, Shenzhen, China.

Key Laboratory of the Ministry of Education for Coastal and Wetland Ecosystems, College of the Environment and Ecology, Xiamen University, Xiamen, China.

出版信息

Mol Genet Genomics. 2022 Sep;297(5):1243-1255. doi: 10.1007/s00438-022-01913-2. Epub 2022 Jun 28.

Abstract

Tea is one of the most popular beverages and its leaves are rich in catechins, contributing to the diverse flavor as well as beneficial for human health. However, the study of the post-transcriptional regulatory mechanism affecting the synthesis of catechins remains insufficient. Here, we sequenced the transcriptome using PacBio sequencing technology and obtained 63,111 full-length high-quality isoforms, including 1302 potential novel genes and 583 highly reliable fusion transcripts. We also identified 1204 lncRNAs with high quality, containing 188 known and 1016 novel lncRNAs. In addition, 311 mis-annotated genes were corrected based on the high-quality Isoseq reads. A large number of alternative splicing (AS) events (3784) and alternative polyadenylation (APA) genes (18,714) were analyzed, accounting for 8.84% and 43.7% of the total annotated genes, respectively. We also found that 2884 genes containing AS and APA features exhibited higher expression levels than other genes. These genes are mainly involved in amino acid biosynthesis, carbon fixation in photosynthetic organisms, phenylalanine, tyrosine, tryptophan biosynthesis, and pyruvate metabolism, suggesting that they play an essential role in the catechins content of tea polyphenols. Our results further improved the level of genome annotation and indicated that post-transcriptional regulation plays a crucial part in synthesizing catechins.

摘要

茶是最受欢迎的饮料之一,其叶子富含儿茶素,为其多样的风味做出贡献,对人体健康也有益。然而,影响儿茶素合成的转录后调控机制的研究还不够充分。在这里,我们使用 PacBio 测序技术对转录组进行了测序,获得了 63111 条全长高质量的异构体,包括 1302 个潜在的新基因和 583 个高度可靠的融合转录本。我们还鉴定了 1204 个高质量的 lncRNA,其中包含 188 个已知和 1016 个新的 lncRNA。此外,基于高质量的 Isoseq 读数,纠正了 311 个错误注释的基因。分析了大量的可变剪接(AS)事件(3784 个)和可变多聚腺苷酸化(APA)基因(18714 个),分别占总注释基因的 8.84%和 43.7%。我们还发现,2884 个含有 AS 和 APA 特征的基因表达水平高于其他基因。这些基因主要参与氨基酸生物合成、光合生物中的碳固定、苯丙氨酸、酪氨酸、色氨酸生物合成和丙酮酸代谢,表明它们在茶多酚儿茶素含量中起着重要作用。我们的研究结果进一步提高了基因组注释水平,并表明转录后调控在合成儿茶素中起着至关重要的作用。

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