[关键词]
[摘要]
目的 比较不同生长时期款冬叶中苯丙素类成分生物合成相关基因的表达,推测苯丙素类成分的最佳合成积累时期,为款冬叶资源的合理利用提供科学依据。方法 利用Illumina HiSeq2500测序技术对不同时期的款冬叶进行转录组测序,获得转录组数据后进行基因功能注释等生物信息学分析,对苯丙素类生物合成相关基因在不同时期的表达进行比较。结果 通过转录组测序技术获得46 793个Unigenes,平均长度为952.144 8 bp,其中有4 774个可以被NR、Swiss-Prot、eggNOG、GO、KEGG等公共数据库共同注释。对注释得到的Unigenes进行KEGG代谢通路分析,结果显示款冬叶转录组中有144条Unigenes参与萜类、黄酮类和苯丙素类生物合成,其中萜类65条,苯丙素类64条,黄酮类15条。进一步对参与苯丙素类生物合成相关基因进行动态比较,发现其关键酶PAL、4CL、HCT、CCoAOMT等在9月的表达量最高,即苯丙素类成分在9月的生物合成量可能最高。结论 为苯丙素类成分的生物合成途径及调控分析奠定了基础,也为款冬叶资源的开发利用奠定了科学依据。
[Key word]
[Abstract]
Objective To compare the expression of genes in the leaves of Tussilago farfara that involved in biosynthesis of phenylpropanoids in different developmental stages, and infer the accumulation period of biosynthesis of phenylpropanoids and provide a scientific basis for the resource utilization of leaves of T. farfara. Methods The Illumina HiSeq2500 highthroughput sequencing method was used to analyze the transcriptome of the leaves of T. farfara in different periods. After obtaining transcriptome data, bioinformatics analysis of gene function annotation was performed to compare the expression of genes related to phenylpropanoid biosynthesis in different periods. Results A total of 46 793 unigenes were obtained by transcriptome sequencing and the average length was 952.144 8 bp. Among them, 4 774 unigenes were annotated in the public databases NR, Swiss-Prot, eggNOG, GO, and KEGG. According to the assignment of KEGG pathway, 144 unigenes were involved in terpenoid biosynthesis, phenylpropanoid biosynthesis and flavonoids, 65 unigenes were involved in terpenoid biosynthesis, 64 unigenes were involved in phenylpropanoid and 15 unigenes were involved in flavonoids biosynthesis. The enzyme genes involved in the phenylpropanoid biosynthesis were also compared in different development stages, and the results indicated that the expression of PAL, 4CL, HCT, and CCoAOMT, which were closely related to biosynthesis of phenylpropanoids, were highest in September, which means that the contents of these compounds might be highest in September. Conclusion This study lays the foundation for the biosynthetic pathway and regulation analysis of phenylpropanoids, and provides a scientific basis for the development and the resource utilization of leaves of T. farfara.
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[基金项目]
国家自然科学基金资助项目(31270008);国家中药标准化项目(ZYBZH-Y-JIN-34);山西省科技创新重点团队(201605D131045-18);地产中药功效物质研究与利用山西省重点实验室(201605D111004)