摘要
目的:对乌头的6个器官组织,主根、侧根、茎、叶、花、果实进行转录组测序分析,以研究参与其萜类次级代谢生物合成相关基因的表达谱,挖掘其功能基因。方法:采用Illumina Hi Seq 2500平台测序、组装得unigenes,与公共数据库NR,Swiss-Prot,GO,COG,KOG,KEGG比对、注释以获得差异基因,使用实时荧光定量聚合酶链式反应(Real-time PCR)验证参与萜类次级代谢生物合成的关键基因。结果:本研究共获得156 967 635条Clean Reads(28 254 174 300 bp),经序列合并拼接后获得103 337个unigenes,通过核苷酸和蛋白序列等方面的同源性分析,表明其中37.31%(38 554个unigenes)与其他生物的已知基因具有不同程度的同源性,通过功能分类研究和代谢途径分析(KEGG)获得参与萜类合成158个unigenes(编码5个关键酶)。结论:这些发现的基因将为乌头萜类化合物的生物合成途径及分子机制提供基础数据。
Objective：To study on the expression of genes involved in the biosynthesis of terpenoid secondary metabolism,and explore the functional genes in Aconitum carmichaelii based on transcriptomics sequencing analysis of six A.carmichaelii organs（including main root,lateral root,stem,leaves,flower and fruit）.Method：Illumina Hiseq 2500 was used to detect the sequencing and assemble into unigennes.By comparing with public database,the genes were annotated and differential expressed genes were obtained.Subsequently,Real-time PCR method was used to verify the key genes involved in the terpenoid secondary metabolism biosynthesis.Result：Total 156 967 635 Clean Reads（28 254 174 300 bp） were obtained,and 103 337 unigenes were obtained after sequence merging.Homology analysis of nucleotide and protein sequences indicated that 37.31%（38 554 unigenes） had different degree of homology with other creatures.In analysis of the functional classification and metabolic pathway（KEGG）,158 unigenes（encoding 5 enzymes） were involved in terpenoid synthesis.Conclusion：These genes will provide the basic data for the terpenoids biosynthetic pathway and molecular mechanism of A.carmichaelii.
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2017年第16期45-50,共6页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(81630101)
国家发改委标准化项目(ZYBZH-C-SC-51)
四川省科技支撑计划项目(2015FY111500-140)
四川省科技厅应用基础计划项目(2016JY0089)
中药资源四川省青年科技创新研究团队项目(2015TD0028)
关键词
乌头
转录组
萜类化合物
次级代谢生物合成
Aconitum carmichaelii
transcriptome
terpenoid
secondary metabolism biosynthesis
