基于转录组挖掘球药隔重楼螺甾烷型重楼皂苷生物合成相关基因  

Identification of genes involved in biosynthesis of spirostane-type polyphyllin in Paris fargesii based on transcriptome analysis

作　　者：曲别阿香 曲别军长 俸明康 何斌 何晓勇 李娟 冯景秋 盛华春 黄艳菲 阎新佳 刘圆 张绍山 QUBIE A-xiang;QUBIE Jun-zhang;FENG Ming-kang;HE Bin;HE Xiao-yong;LI Juan;FENG Jing-qiu;SHENG Hua-chun;HUANG Yan-fei;YAN Xin-jia;LIU Yuan;ZHANG Shao-shan(College of Pharmacy,Southwest Minzu University,Chengdu 610041,China;Sichuan Technology and Engineering Laboratory of Qiang-Yi Medicinal Resources Protection and Utilization,Chengdu 610225,China;Key Laboratory of Tibetan Plateau Ethnic Medicinal Resources Protection and Utilization of National Ethnic Affairs Commission,Chengdu 610225,China)

机构地区：[1]西南民族大学药学院,四川成都610041 [2]四川省羌彝药用资源保护与利用技术工程实验室,四川成都610225 [3]青藏高原民族药用资源保护与利用国家民委重点实验室,四川成都610225

出　　处：《中草药》2023年第20期6798-6812,共15页Chinese Traditional and Herbal Drugs

基　　金：四川省自然科学基金资助项目(22NSFSC3136);西南民族大学2023年研究生创新型项目资助硕士重点项目(ZD2023459)。

摘　　要：目的以球药隔重楼Paris fargesii根茎、茎和叶为材料,探究螺甾烷型重楼皂苷的生物合成基因。方法采用HPLC分析6个螺甾烷型重楼皂苷含量,进一步利用Illumina HiSeq X Ten平台进行转录组测序,结合定量和转录组数据进行生信分析。结果6个螺甾烷型重楼皂苷在球药隔重楼根茎、茎和叶中的含量具有显著差异。Illumina转录组测序共获得61755条非冗余unigenes,其中30263条(49.0%)被成功注释。催化生成薯蓣皂苷元的31个关键基因均被成功鉴定,且基因表达模式与皂苷的积累水平总体一致,其中IDI、8,7SI-4、CYP90G4、SMO2-2、C5-SD1、CYP51G、C14-R-2、HMGCR、CPI-5、CYP94D108、CAS、HMGCS、SMO1-3、SSR1-3、SQS、ispH和DXR等基因与重楼皂苷的积累呈显著正相关。共获得了61条尿苷二磷酸糖基转移酶(UGT)基因,其中30条潜在参与重楼皂苷的糖基化修饰。结论球药隔重楼根茎、茎和叶中皂苷合成基因表达模式与皂苷积累规律存在一致性,鉴定的基因可指导该类活性产物的生物合成途径解析。Objective To investigate the biosynthetic genes of spirostane-type polyphyllins of Paris fargesii.Methods Based on the rhizome,stem and leaf materials of P.fargesii,the contents of six spirostane-type polyphyllins were analyzed by HPLC.The transcriptome sequencing was performed using Illumina HiSeq X Ten platform.The quantitative data and transcriptome data were combined for bioinformatics analysis.Results The content of six spirostane-type polyphyllins had significant differences in the rhizome,stem and leaf of P.fargesii.A total of 61755 non-redundant unigenes were obtained by Illumina sequencing,of which 30263(49.0%)were successfully annotated.Thirty-one key genes in the biosynthesis pathway of diosgenin had been successfully identified,and the gene expression pattern was generally consistent with the accumulation level of spirostane-type polyphyllins.The accumulation level of spirostane-type polyphyllins were positively correlated with IDI,8,7SI-4,CYP90G4,SMO2-2,C5-SD1,CYP51G,C14-R-2,HMGCR,CPI-5,CYP94D108,CAS,HMGCS,SMO1-3,SSR1-3,SQS and ispH.A total of 61 uridine diphosphate glycosyltransferase(UGT)genes were obtained,of which 30 were potentially involved in the glycosylation modification of spirostane-type polyphyllins.Conclusion There was a consistency between the expression pattern of genes involved in the biosynthesis pathway of polyphyllins and the accumulation level of polyphyllins in the rhizome,stems and leaves of P.fargesii.The genes identified in this study can help to elucidate the biosynthetic pathway of this kinds of active products.

关 键 词：球药隔重楼 转录组 螺甾烷型重楼皂苷 生物合成途径 尿苷二磷酸糖基转移酶 

分 类 号：R286.12[医药卫生—中药学]