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基于 SMRT 测序平台的Clematis calyx 转录组测序和 SSR 预测。

Transcriptome sequencing and SSR prediction of Clematis calyx based on SMRT sequencing platform.

机构信息

Jiangsu Vocational College of Agriculture and Forestry, Zhengjiang, 212400, China.

出版信息

Sci Rep. 2024 Nov 22;14(1):28949. doi: 10.1038/s41598-024-80504-0.

DOI:10.1038/s41598-024-80504-0
PMID:39578612
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11584640/
Abstract

Clematis is an excellent vertical greening plant for garden viewing vines, with great ornamental and high medicinal value. To obtain transcriptome information and functional gene data for the Clematis calyx, this study utilized three biological replicates of the calyx from each of the three Clematis varieties: 'Henryi', 'Polish spirit', and 'Mme Julia Correvon' Single-molecule real-time sequencing technology was employed for full-length transcriptome sequencing. The study revealed that 21,673,173 non-chimeric sequences were identified through full-length transcriptome sequencing. After clustering, correction, and redundancy removal, 40,465 high-quality, full-length transcripts were obtained. Among these transcripts, there were 15,488 long non-coding RNA (lncRNA), 9,212 simple sequence repeats (SSR) sites, 1,247 transcription factors (TFs), 1,228 alternative splicing events, and 7,189 selectively polyadenylation sites predicted. In addition, 7,442 primer pairs were designed based on the SSR sites, covering 80.76% of the total SSR. 15 primer pairs were randomly selected for amplification, and 73.3% of them were successfully amplified. Transcript annotation results showed that 38,439, 38,094, 26,815, and 33,407 transcripts were annotated to the Nr, KEGG, KOG, and SwissProt databases, respectively. Among these, the KEGG database identified 137 metabolic pathways, including biosynthesis of secondary metabolites, carbon metabolism, and amino acid biosynthesis. 104 and 7 transcripts are involved in the flavonoid and anthocyanin metabolism pathways, respectively. The above results provide initial insights into the transcriptome information and functional characteristics of Clematis calyx. This critical data supports future research on marker primers for the color mechanism of Clematis calyx, the pathways and regulatory mechanisms of flavonoids and other products, as well as specific trait genes.

摘要

铁线莲是一种优良的观花攀援绿化植物,具有很高的观赏价值和药用价值。为了获得铁线莲花萼的转录组信息和功能基因数据,本研究利用 3 个铁线莲品种(‘亨利’、‘波兰精神’和‘Mme Julia Correvon’)的 3 个生物学重复的花萼进行单分子实时测序技术进行全长转录组测序。研究表明,通过全长转录组测序共鉴定出 21,673,173 个非嵌合序列。经过聚类、校正和去冗余处理,获得了 40,465 个高质量的全长转录本。其中,有 15,488 个长非编码 RNA(lncRNA)、9,212 个简单序列重复(SSR)位点、1,247 个转录因子(TFs)、1,228 个可变剪接事件和 7,189 个选择性多聚腺苷酸化位点预测。此外,基于 SSR 位点设计了 7,442 对引物,覆盖了总 SSR 的 80.76%。随机选择了 15 对引物进行扩增,其中 73.3%的引物成功扩增。转录本注释结果表明,38,439、38,094、26,815 和 33,407 个转录本分别注释到 Nr、KEGG、KOG 和 SwissProt 数据库中。其中,KEGG 数据库鉴定出 137 条代谢途径,包括次生代谢物合成、碳代谢和氨基酸生物合成。104 和 7 个转录本分别参与黄酮类和花色苷代谢途径。以上结果初步揭示了铁线莲花萼的转录组信息和功能特征。这些关键数据为今后研究铁线莲花萼颜色机制的标记引物、黄酮类等产物的途径和调控机制以及特定性状基因提供了依据。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ee8d/11584640/9859c817dc99/41598_2024_80504_Fig11_HTML.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ee8d/11584640/485f89cc195f/41598_2024_80504_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ee8d/11584640/b6b8a7ae2cea/41598_2024_80504_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ee8d/11584640/2e12cd27c383/41598_2024_80504_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ee8d/11584640/ab234effcc23/41598_2024_80504_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ee8d/11584640/2c6f1166c987/41598_2024_80504_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ee8d/11584640/7aee8d1623c1/41598_2024_80504_Fig7_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ee8d/11584640/45d3f974c4ee/41598_2024_80504_Fig8_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ee8d/11584640/d17d6cc09a0e/41598_2024_80504_Fig9_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ee8d/11584640/015020909e9b/41598_2024_80504_Fig10_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ee8d/11584640/9859c817dc99/41598_2024_80504_Fig11_HTML.jpg

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本文引用的文献

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Widespread Polycistronic Transcripts in Fungi Revealed by Single-Molecule mRNA Sequencing.单细胞 mRNA 测序揭示真菌中广泛存在的多顺反子转录本。
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