[关键词]
[摘要]
目的 对不同干旱胁迫程度下党参Codonopsis pilosula叶、茎、根进行转录组测序,探究干旱胁迫对党参差异基因和多糖合成相关酶基因的调控。方法 利用Illumina HiSeq 2500高通量测序技术对正常水分(85%~90%)和轻度(65%~70%)、中度(50%~55%)、重度干旱胁迫(35%~40%)下生长旺盛期党参叶、茎、根组织进行测序并建立cDNA数据库,经de novo拼接后得到Unigene,并进一步开展生物信息学分析。结果 共获得Unigene 60 377条,分别有51 477、29 896、33 479、35 912、47 378、18 649、31 833条被非冗余数据库(non-redundant protein sequence database,NR)、真核生物蛋白相邻类的聚簇(clusters of orthologous group for eukaryotic complete,KOG)、基因本体(gene ontology,GO)、Swiss-Prot、eggNOG、京都基因与基因组数据库(Kyoto encyclopedia of genes and genomes,KEGG)、Pfam等数据库注释,筛选到9个差异组DEGs总和89 369条,GO富集分析发现,党参叶、茎、根中的DEGs在GO功能注释中分布基本一致,主要集中于细胞、细胞部分、细胞过程、代谢过程、结合蛋白、催化活性等功能。KEGG途径富集结果表明,干旱胁迫下叶中DEGs显著富集在不饱和脂肪酸、苯丙烷生物合成,茎中DEGs主要富集在苯丙烷、角质、琥珀和蜡、淀粉和蔗糖、单萜的生物合成,根中DEGs主要富集于黄酮、苯丙烷、二苯乙烯、二芳基庚烷和姜酚的生物合成及戊糖、葡萄糖醛酸转换。轻度干旱能引起叶中多糖合成途径关键酶基因的表达上调,是促进党参多糖成分积累的基础。结论 通过转录组测序,初步表明干旱胁迫影响党参不同组织差异基因的表达,并调控党参根中多糖合成的相关酶基因,以期为从分子基因培育优质党参提供参考和依据。
[Key word]
[Abstract]
Objective Transcriptome sequencing was performed on leaves, stems and roots of Codonopsis pilosula under different drought stress levels to explore the regulation of different genes and enzymes related to polysaccharide synthesis in C. pilosula under drought stress.Methods Illumina HiSeq 2500 high-throughput sequencing technology was used to sequence the leaves, stalks and roots of C. pilosula in the control (85%-90%), light (65%-70%), moderate (50%-55%), and severe (35%-40%) drought treatments and establish cDNA database. Unigene was obtained by de novo splicing, and further biochemistry analysis was carried out.Results A total of 51 477, 29 896, 33 479, 35 912, 47 378,18 649 and 31 833 were annotated by non-redundant protein sequence database (NR), clusters of orthologous group for eukaryotic complete (KOG), gene ontology (GO), Swiss-Prot, eggNOG, Kyoto Encyclopedia of genes and genomes (KEGG) and Pfam databases respectively. A total of 89 369 DEGs were screened from nine difference groups. GO enrichment analysis showed that DEGs of leaf, stem and root weight were distributed basically the same in GO functional annotations, mainly focusing on cells, cell parts, cell processes, metabolic processes, binding proteins, catalytic activity and other functions. KEGG pathway enrichment results showed that DEGs were significantly enriched in the biosynthesis of unsaturated fatty acids and phenylpropanol under drought stress. In the stem, DEGs were mainly enriched in the biosynthesis of phenylpropanoid, cutin, suberine and wax, starch and sucrose, monoterpene. Light drought could up-regulate the expression of key enzyme genes of polysaccharide synthesis pathway in leaves, which was the basis of promoting polysaccharide accumulation in C. pilosula. Conclusion Through transcriptome sequencing, the expression of differential genes in different tissues of C. pilosula under drought stress was reveal, and the regulation of drought on polysaccharide synthesis of C. pilosula was analyzed, providing irrigation reference and genetic basis for cultivating high-quality C. pilosula.
[中图分类号]
R286.12
[基金项目]
国家自然科学基金资助项目(81760683);国家自然科学基金资助项目(81960683);现代农业产业技术体系建设专项资金资助(CARS-21);甘肃省教育厅"双一流"科研重点项目(GSSYLXM-05);研究生创新基金资助项目(2021CX48)