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1.毕节医学高等专科学校(贵州 毕节 551700)
2.广州医科大学药学院(广东 广州 511495)
杨杰,男,硕士,教授,主要从事分子生物学在中医药中的应用研究
收稿:2024-10-29,
修回:2025-03-09,
录用:2025-04-08,
纸质出版:2025-05-25
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杨杰,郭步伐,彭启伦等.基于Illumina Novaseq 6000测序技术研究头花蓼不同入药部位转录组特征[J].上海中医药大学学报,2025,39(03):18-26.
YANG Jie,GUO Bufa,PENG Qilun,et al.Research on transcriptome characteristics of different medicinal parts of Polygonum Capitatum based on Illumina Novaseq 6000 sequencing technology[J].Academic Journal of Shanghai University of Traditional Chinese Medicine,2025,39(03):18-26.
杨杰,郭步伐,彭启伦等.基于Illumina Novaseq 6000测序技术研究头花蓼不同入药部位转录组特征[J].上海中医药大学学报,2025,39(03):18-26. DOI: 10.16306/j.1008-861x.2025.03.003.
YANG Jie,GUO Bufa,PENG Qilun,et al.Research on transcriptome characteristics of different medicinal parts of Polygonum Capitatum based on Illumina Novaseq 6000 sequencing technology[J].Academic Journal of Shanghai University of Traditional Chinese Medicine,2025,39(03):18-26. DOI: 10.16306/j.1008-861x.2025.03.003.
目的
2
获取头花蓼茎、花和叶的转录组特征。
方法
2
从头花蓼茎、花和叶中提取总RNA,质量检测合格后构建转录组文库。文库质检合格后,使用Illumina Novaseq 6000测序平台进行测序,生成150 bp的双端序列。随后,采用相关生物信息学方法分析其转录组特性。
结果
2
共获取122.11G clean data。经Trinity软件配对末端拼接与CD-HIT软件去冗余后得到66 021条unigene,平均长度为 1 358.84 bp。序列同源性比对分析表明,unigenes在7个数据库中注释的比对率为 90.82%~92.34%。已注释的unigene,可归入基因本体(GO)数据库分类的细胞组分、生物学过程、分子功能3大类64小类,涉及133条京都基因与基因组百科全书(KEGG)通路,含有15条次生代谢产物生物合成通路。KEGG通路富集分析结果显示,头花蓼茎、花、叶三者之间两两比较,差异表达unigenes 主要富集于苯丙素生物合成途径和黄酮类生物合成途径。蛋白编码框序列49 440条。利用MISA软件预测出16 643条简单重复序列(SSRs)。
结论
2
利用高通量测序技术挖掘出头花蓼不同入药部位的功能基因组信息,为后续头花蓼基因功能鉴定及次生代谢产物生物合成途径解析和调控机制研究奠定理论基础。
Objective: To obtain the transcriptome characteristics of the stem, flower and leaf of
Polygonum capitatum
(PC).
Methods
2
Total RNA was extracted from the stems, flowers and leaves of PC, and the transcriptome library was constructed after the quality inspection passed. After the library passed the quality inspection, it was sequenced on Illumina Novaseq 6000 sequencing platform, and 150 bp paired-end sequences were generated. Subsequently, relevant bioinformatics methods were used to analyze the transcriptome characteristics.
Results
2
A total of 122.11G clean data was obtained. 66 021 unigenes with the average length 1 358.84 bp were de novo assembled after paired end splicing by Trinity and redundancy elimination by CD-HIT. Basic local alignment search tool (Blast) analysis revealed that the alignment rate of unigenes annotated in seven databases was 90.82%~92.34%. The annotated unigenes can be categorized into 64 subcategories of 3 major categories, cellular component, biological process and molecular function, of Gene Ontology (GO) database, involving 133 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, including 15 secondary metabolite biosynthesis pathways. KEGG pathway enrichment analysis indicated that differentially expressed unigenes in the pairwise comparison among the stems, flowers and leaves of PC were primarily enriched in phenylpropanoid biosynthesis pathway and flavonoid biosynthesis pathway. 49 440 protein-coding frame sequences were identified, and 16 643 simple sequence repeats (SSRs) were predicted by MISA.
Conclusion
2
The functional genomic information of different medicinal parts of PC was explored by high-throughput sequencing technology, laying a theoretical basis for subsequent gene function identification and the analysis and regulatory mechanisms research of secondary metabolite biosynthetic pathways of PC.
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