款冬叶不同发育阶段的转录组学分析
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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 Hi Seq2500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。
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 Hi Seq2500 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, 4 CL, 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.
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 Hi Seq2500 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, 4 CL, 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.
引文
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[13]林懋怡,牛卉,刘晋杰,等.基于转录组分析华细辛甲基丁香酚生物合成途径的相关基因[J].中草药,2017,48(15):3160-3167.
[2]吴琪珍,张朝凤,许翔鸿,等.款冬花化学成分和药理活性研究进展[J].中国野生植物资源,2015(2):33-36.
[3]Xue S Y,Li Z Y,Zhi H J,et al.Metabolic fingerprinting investigation of Tussilago farfara L.by GC-MS and multivariate data analysis[J].Biochem System Ecol,2012,41(1):6-12.
[4]Zhi H J,Qin X M,Sun H F,et al.Metabolic fingerprinting of Tussilago farfara L.using1H-NMR spectroscopy and multivariate data analysis[J].Phytochem Anal,2012,23(5):492-501.
[5]Li Z Y,Zhi H J,Zhang F S,et al.Metabolomic profiling of the antitussive and expectorant plant Tussilago farfara L.by nuclear magnetic resonance spectroscopy and multivariate data analysis[J].J Pharm Biom Anal,2013,75(5):158-164.
[6]贾岩,张福生,肖淑贤,等.款冬花不同发育阶段的代谢组学和比较转录组学分析[J].中国生物化学与分子生物学报,2017,33(6):615-623.
[7]Xu Z,Peters R J,Weirather J,et al.Full-length transcriptome sequences and splice variants obtained by a combination of sequencing platforms applied to different root tissues of Salvia miltiorrhiza and tanshinone biosynthesis[J].Plant J,2015,82(6):951-961.
[8]Chen S,Luo H,Li Y,et al.454 EST analysis detects genes putatively involved in ginsenoside biosynthesis in Panax ginseng[J].Plant Cell Rep,2011,30(9):1593-1601.
[9]He L,Xu X L,Li Y,et al.Transcriptome analysis of buds and leaves using 454 pyrosequencing to discover genes associated with the biosynthesis of active ingredients in Lonicera japonica Thunb[J].PLo S One,2013,8(4):e62922.
[10]Liu X B,Ma L,Zhang A H,et al.High-throughput analysis and characterization of Astragalus membranaceus transcriptome using 454 GSFLX[J].PLo S One,2014,9(5):e95831.
[11]Grabherr M G,Haas B J,Yassour M,et al.Full-length transcriptome assembly from RNA-Seq data without a reference genome[J].Nat Biotechnol,2011,29:644-652.
[12]李静,李娟,贾金萍,等.基于UHPLC-QExtractive轨道阱高分辨质谱的款冬花、叶的化学比较[J].药学学报,2018,53(3):444-452.
[13]林懋怡,牛卉,刘晋杰,等.基于转录组分析华细辛甲基丁香酚生物合成途径的相关基因[J].中草药,2017,48(15):3160-3167.
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