谢冬梅 俞年军 黄璐琦 彭代银 +刘丛彬 朱月健 黄浩

[摘要]牡丹皮為我国传统常用中药，安徽省铜陵地区栽培的“凤丹”根皮加工而成的药材牡丹皮被誉为道地药材，药用活性成分丰富多样，但目前尚不清楚 “凤丹”药用部位次生代谢过程中活性物质合成的遗传学基础。研究采用Illumina HiSeq 4000高通量测序平台对五年生“凤丹”根皮转录组进行测序，对测序结果进行de novo 拼接和功能注释，测序后获得72 997条unigene。进一步利用公共数据库进行同源比对，其中41 139条unigene被Nr数据库成功注释，34 952条unigene能被GO数据库成功注释，20 016条unigene被KEGG数据库成功舒注释，共涉及到5个大类、34个种类、352条代谢通路；在次生物质合成与代谢途径中，其中苯丙素类化合物、萜类化合物骨架合成、各种类型萜类化合物、生物碱类化合物以及黄酮类成分生物合成途径中的unigene分别有214，104，152，55，36个；不同产地样本间差异表达基因的富集性比较显示不同产地样本间存在明显差异；此外，在72 997条unigene中共检测到9 939个SSR序列，其中二核苷酸重复的SSR标记占2075%。研究的结果不仅为挖掘“凤丹”次生代谢物生物合成关键基因提供了基础数据信息，也为药用牡丹的遗传多样性研究和分子标记开发奠定了分子基础。

[关键词]牡丹皮； 转录组； 次生代谢； 差异基因； 简单重复序列

Next generation sequencing and transcriptome analysis of

root bark from Paeonia suffruticosa cv Feng Dan

XIE Dongmei1，2， YU Nianjun1*， HUANG Luqi2*， PENG Daiyin1， LIU Congbin1， ZHU Yuejian3， HUANG Hao4

（1 Institute of Traditional Chinese Medicine Resources Protection and Development， Anhui Academy of Chinese

Medicine， Anhui University of Chinese Medicine， Hefei 230012， China；

2 State Key Laboratory of Daodi Herbs， National Resource Center for Chinese Materia Medica， China

Academy of Chinese Medical Sciences， Beijing 100700， China；

3 Anhui Jiren Pharmaceutical Co， Ltd， Bozhou 236800， China；

4 Beijing Tongrentang Anhui Traditional Chinese Medicinal Materials Co， Ltd， Tongling 244000， China）

[Abstract]Moutan Cortex is an important traditional Chinese medicine， “Fengdan Pi” was known as Daodi herbs from the root bark of Paeonia suffruticosa cv Feng Dan for its extracted various active components However， the genetic basis for their activity is virtually unknown The transcriptome of the root bark from “Fengdan” was sequenced using the Illumina HiSeq 4000 sequencing platform The clean reads were then de novo assembled into 72 997 unigenes Among them， the number of unigenes which could been annotated by dataset Nr and GO was 41 139 and 34 592. The 20 016 unigenes could been annotated by KEGG dataset， which were involved in 5 major categories， 34 middle categories， and 352 metabolism pathways. The number of unigenes which were mapped to the phenylpropanoid biosynthesis pathway， terpenoid backbone biosynthesis pathway， terpenoid biosynthesis pathway， alkaloid biosynthesis pathway， and flavonoid biosynthesis pathway was 214， 104， 152， 55 and 36 respectively， suggesting that they are involves in these pathways of pharmaceutically important Furthermore， there also showed remarkable differences in groups which enrichment ratio of the different expressed gene compared. In addition， a total of 9 939 SSRs were identified from the sequence of 72 997 unigenes This study not only provides many valuable basal data which was important gene in the synthesis pathway of secondary metabolites with gene searching， but also has important significance to find molecular marker in germplasm for breeding and improvement

[Key words]Moutan Cortex； transcriptome； secondary metabolism； different expressed gene； simple sequence repeat

中药牡丹皮Moutan Cortex来源于毛茛科植物牡丹Paeonia suffruticosa Andr的干燥根皮，具有清热凉血，活血化瘀的功效[1]。现代中药化学和药理研究表明，牡丹皮主要化学成分为酚类、酚苷类、单萜及单萜苷类以及三萜、甾醇及其苷类、黄酮、有机酸、香豆素等，具有抗炎抗菌、降血糖以及心血管系统保护、中枢神经保护、增强免疫、止血、凝血等药理作用[23]。……