中药材

目的：探究不同种质类型朝鲜淫羊藿的黄酮类成分含量差异，挖掘调控其黄酮类成分积累和植物器官颜色差异的关键基因与转录因子。方法：以朝鲜淫羊藿叶片为材料，采用HPLC法分析不同种质类型朝鲜淫羊藿黄酮类成分含量的差异，并利用Illumina HiSeq平台进行转录组测序，结合定量与转录组数据进行生物信息学分析。结果：紫杆褐边叶型(Z-H型)的黄酮类成分含量显著高于绿杆绿边叶型(L-L型);Z-H型和L-L型种质转录组测序共获得77 140条Unigene,鉴定出20 509个差异表达基因，其中12 777个基因在Z-H型中表达上调；差异表达基因的GO功能主要富集于有机氮化合物生物合成过程、细胞酰胺代谢过程、酰胺生物合成过程等；KEGG代谢主要富集于核糖体、碳代谢、光合作用、苯丙烷及类黄酮合成等通路；Z-H型相对L-L型而言，苯丙烷生物合成途径中的PAL、C4H和部分4CL基因均呈上调表达；类黄酮和花青素生物合成途径中的CHS、FG2、FG3、ANS基因均呈上调表达；鉴定出34个MYB-related、WRKY、bZIP、bHLH转录因子，多数表达上调。结论：挖掘出的PAL、C4H、4CL、CHS、FG2、FG3、ANS基因及MYB-related、bHLH等转录因子，对朝鲜淫羊藿的黄酮类成分积累及茎杆、叶缘颜色变化具有调控作用，该结果可为朝鲜淫羊藿的遗传改良和黄酮类成分合成机制研究提供参考依据。

Objective: To explore the differences of flavonoid content among different germplasm types of Epimedium koreanum,and to excavate the key genes and transcription factors that regulate the accumulation of flavonoids and the color differences of plant organs.Methods: The differences in flavonoid content of different germplasm types of Epimedium koreanum were analyzed by HPLC using Epimedium koreanum leaves as materials, and transcriptome sequencing was performed by using the Illumina HiSeq platform, combining quantitative and transcriptome data for bioinformatics analysis.Results: The flavonoid content of the purple-stemmed brown-edged leaf type(Z-H type) was significantly higher than that of the green-stemmed green-edged leaf type(L-L type);A total of 77 140 Unigenes were obtained from Z-H type and L-L type by transcriptome sequencing, and 20 509 differentially expressed genes were identified, of which 12 777 genes expressions were up-regulated in Z-H type; The GO functions of the differentially expressed genes were mainly enriched in organonitrogen compound biosynthetic process, cellular amide metabolic processes and amide biosynthetic processes; KEGG metabolism was mainly enriched in ribosomes, carbon metabolism, photosynthesis, phenylpropanoid and flavonoid biosynthesis pathways; Compared with L-L type, the expressions of PAL,C4H and part of 4CL genes in phenylpropanoid biosynthesis pathway of Z-H type were up-regulated; the expressions of CHS,FG2,FG3 and ANS genes in flavonoid and anthocyanin biosynthesis pathways were up-regulated; Thirty-four MYB-related, WRKY,bZIP and bHLH transcription factors were identified, and most were up-regulated in expression.Conclusion: The mined PAL,C4H,4CL,CHS,FG2,FG3,ANS genes and transcription factors, such as MYB-related and bHLH,have a regulatory effect on the accumulation of flavonoid components and the color change of stem and leaf margin in Epimedium koreanum,and the results can provide a reference basis for the study of genetic improvement and the mechanism of flavonoid component synthesis in Epimedium koreanum.