CN113897415B - 一种鉴别大黄药材三个基原物种的方法与应用 - Google Patents
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Abstract
本发明涉及大黄药材三个基原物种的鉴别及其应用,具体包括基于叶绿体基因组筛选高变区,以筛选的高变区rps16‑trnQ、psaA‑ycf3、psbE‑petL、ndhF‑rpl32与trnT‑trnL序列为模板设计引物1F/1R、2F/2R、3F/3R、4F/4R和5F/5R,并对42份样本进行PCR扩增,Sanger测序后利用MEGA7.0评价所筛选序列的物种鉴定能力,确定能够用于鉴定唐古特大黄、药用大黄与掌叶大黄的特异DNA条形码。本发明公开可有效扩增高变区rps16‑trnQ、psaA‑ycf3、psbE‑petL、ndhF‑rpl32与trnT‑trnL的5对通用引物,可用于大黄药材三个基原物种原植物、种子及药材的鉴定,为大黄药材三个基原物种分类鉴定、保证大黄药材临床用药安全及规范大黄药材市场提供依据。
Description
技术领域
本发明属于中药材来源品种鉴定技术领域,具体涉及基于叶绿体基因组开发鉴别大黄药材三个基原物种的特异DNA条形码。
背景技术
《中国药典》收载药材大黄Rhei Radix et Rhizoma为蓼科植物掌叶大黄Rheumpalmatum L.、唐古特大黄Rh.tanguticum Maxim.ex Balf.和药用大黄Rh.officinaleBaill.的干燥根及根茎。大黄具有广泛药效作用主要与其含有大量活性成分有关,大黄主要化学成分为蒽醌类、蒽酮类、二苯乙烯类、鞣质类等化合物。不同基原的大黄药材在化学物质上存在差别,在不同功效上表现出专长,研究表明掌叶大黄专长于“清热泻火、利湿退黄”,而唐古特大黄专长于“泻下攻积、逐瘀通经”。因此大黄基原物种鉴别对于大黄临床精准用药具有重要意义。
大黄基原物种鉴定方法主要有性状鉴定、显微鉴定、理化鉴定(HPLC、UPLC等)及DNA条形码(ITS2、psbA-trnH及matK等)。大黄药材三个基原物种区别主要在于叶片分裂程度,药用大黄掌状浅裂,呈大齿形或宽三角形;掌叶大黄叶片浅裂至半裂,呈较窄三角形;唐古特大黄叶深裂,裂片狭长呈三角状披针形或狭长形。性状和显微鉴定对大黄基原植物或药材特征的完整性要求较高,鉴别过程需依靠长期经验积累,且经过深加工的大黄饮片及中成药,形态特征破坏会增加基原物种鉴定难度。理化鉴定依据中药材主要成分进行鉴定,但操作较为繁琐。与传统鉴定方法相比,DNA分子能为物种鉴定提供丰富信息,通过ITS2、psbA-trnH及matK等DNA条形码能够有效鉴别正品大黄与华北大黄Rh.franzenbachiiMunt.、河套大黄Rh.hotaoense C.Y.Cheng et Kao等混伪品,但难以精确到基原物种。与传统DNA条形码相比,叶绿体基因组具有更丰富变异位点和更强物种分辨率,因此可基于叶绿体基因组高变区开发有效鉴别唐古特大黄、掌叶大黄和药用大黄的特异DNA条形码,以期用于大黄基原植物、药材、饮片及相关中成药的基原鉴定。
发明内容
本发明第一个目的是筛选大黄药材三个基原物种叶绿体基因组高变区,开发有效鉴别唐古特大黄、掌叶大黄和药用大黄的特异DNA条形码。
本发明第二个目的是以筛选的高变区序列为模板设计引物,提供用于鉴别大黄药材三个基原物种的5对引物,记为1F/1R、2F/2R、3F/3R、4F/4R和5F/5R,本发明所述引物的核苷酸序列如下表1所示:
表1用于鉴别大黄药材三个基原物种的5对引物
本发明提供的大黄药材三个基原物种鉴别方法,包括如下步骤:
步骤1.大黄药材三个基原物种特异DNA条形码的筛选;
应用mVISTA对唐古特大黄、药用大黄、掌叶大黄叶绿体基因组进行全局比对分析,利用DnaSP软件检测3条叶绿体基因组的核苷酸变异程度(Pi),最后以筛选的高变区序列为模板设计引物。
步骤2.大黄药材三个基原物种特异DNA条形码的获得;
取大黄药材三个基原物种的新鲜叶片、种子及根约50~100mg,液氮研磨,提取基因组DNA。以基因组DNA为模板,利用表1所述的5对引物分别进行PCR扩增。
步骤3.大黄药材三个基原物种特异DNA条形码的鉴定分析。
将扩增产物在ABI3730XL测序仪上进行Sanger测序。利用CodonCode Aligner软件对测序所得峰图文件进行序列拼接。利用MEGA 7.0软件进行序列比对、计算K2P距离和构建NJ系统进化树。
以下将结合附图对本发明作进一步说明,以充分说明本发明的目的、技术特征和技术效果。
附图说明
图1所示为大黄药材三个基原物种叶绿体基因组mVISTA全局比对。
图2所示为大黄药材三个基原物种叶绿体基因组DnaSP变异分析图(Pi>0)。
图3所示为用于鉴别大黄药材三个基原物种的5对引物扩增电泳图。
图4所示为基于特异DNA条形码序列构建大黄药材三个基原物种与其混伪品的NJ系统进化树。
具体实施方式
下面结合具体的实施例对本发明作进一步说明,而并非对发明的限定,依照本领域公知的现有技术,本发明的实施方式并不限于此,因此凡依照本公开内容所做出的本领域的等同替换,均属于本发明的保护范围。
实施例1大黄药材三个基原物种特异DNA条形码的筛选
以注释过的掌叶大黄叶绿体基因组为参考,利用mVISTA将唐古特大黄Rh.tanguticum、药用大黄Rh.officinale及掌叶大黄Rh.palmatum的叶绿体基因组进行全局比对(图1)。三个物种叶绿体基因组序列基因间区变异大于基因区,如rps16-trnQ、trnT-psbD、psaA-ycf3、trnT-trnL、psbE-petL、ndhF-rpl32、trnN-ycf1、ccsA-ndhD等,这些高变区可能是唐古特大黄、药用大黄和掌叶大黄物种鉴定的潜在分子标记。
使用DnaSP计算三个物种叶绿体基因组所有基因和基因间区的核苷酸变异值(Pi),进行高变区检测和序列差异水平分析(图2)。三个物种叶绿体基因组的基因间区Pi平均值(0.004)变异大于基因Pi平均值(0.0008)。结合Pi值与mVISTA结果分析,本发明筛选rps16-trnQ、psaA-ycf3、psbE-petL、ndhF-rpl32与trnT-trnL为鉴别大黄药材三个基原物种的特异DNA条形码。
实施例2大黄药材三个基原物种特异DNA条形码的获得
2.1实验材料
本实施例共收集42份样品,包括大黄药材三个基原物种唐古特大黄Rheumtanguticum、药用大黄Rh.officinale及掌叶大黄Rh.palmatum,以及易混伪品食用大黄Rh.rhaponticum、波叶大黄Rh.rhabarbarum Linnaeus、苞叶大黄Rh.alexandrae Batal.、塔黄Rh.nobile Hook.f.et Thoms.及皱叶酸模Rumex crispus各一份,具体信息见表2。
表2大黄药材三个基原物种及其混伪品样品信息表
2.2基因组提取
分别取表2样品约50~100mg,液氮研磨,其余步骤参照植物基因组DNA提取试剂盒标准操作方法进行。
2.3 PCR扩增
以筛选的高变区rps16-trnQ、psaA-ycf3、psbE-petL、ndhF-rpl32与trnT-trnL序列为模板设计引物1F/1R、2F/2R、3F/3R、4F/4R和5F/5R,详见表1,由苏州金唯智生物科技有限公司合成,引物用灭菌双蒸水溶解并稀释至2.5μM。
25μL反应PCR扩增体系:2×Taq MasterMix 12.5μL,上下游引物(2.5μM)各1μL,DNA模板约50ng,加灭菌双蒸水补足体积至25μL。
PCR反应条件参数:94℃,5min;(94℃,30s,55℃,30s,72℃,45s),35个循环;72℃,10min。
2.4 PCR产物测序
PCR产物在ABI3730XL测序仪上进行Sanger双向测序,测序引物同PCR扩增引物。
2.5序列拼接
本实施例采用软件CodonCode Aligner进行序列拼接及校对。首先利用该软件去除序列两端的引物区,然后去除测序结果两端的低质量部分,并对剩余部分进行质量评估。进行序列拼接后,获得大黄药材三个基原物种及相关混伪品的特异DNA条形码序列。
实施例3大黄药材三个基原物种特异DNA条形码的鉴定分析
3.1 K2P距离
根据MEGA7.0计算测序所获高质量序列的种内及种间K2P距离。在本发明中,种间最小遗传距离大于种内最大遗传距离,不存在重叠,表明所获序列符合DNA条形码的要求(表3)。
3.2 NJ系统进化树
根据MEGA7.0构建NJ系统进化树,设置bootstrap的重复值为1000。在本发明中,唐古特大黄单独聚为一支,药用大黄单独聚为一支,掌叶大黄单独聚为一支,表明rpsl6-trnQ、psaA-ycf3、psbE-petL、ndhF-rpl32与trnT-trnL可以作为特异DNA条形码用于大黄药材三个基原物种的鉴定(图4)。psaA-ycf3、ndhF-rpl32与trnT-trnL的NJ系统发育树显示唐古特大黄PS2904MT01~PS2904MT06单独聚为一支,采集地点为四川红原;PS2904MT07~PS2904MT12单独聚为一支,采集地点为青海林川,表明psaA-ycf3、ndhF-rpl32与trnT-trnL可作为潜在特异DNA条形码用于同一物种不同产区的鉴定。
以上所述仅是本发明的优选实施方式,应当指出对于本技术领域的技术人员来说,在不脱离本发明技术原理的前提下,还可以做出若干改进,这些改进也应视为本发明的保护范围。
Claims (3)
1.一种扩增特异DNA条形码的引物用于大黄三个基原物种鉴别的用途,其特征在于,所述大黄药材三个基原物种为唐古特大黄、药用大黄和掌叶大黄,所述特异DNA条形码为rps16-trnQ、psaA-ycf3、psbE-petL、ndhF-rpl32与trnT-trnL,所述引物的核苷酸序列如下:
;
所述用途包括,以大黄药材基因组DNA为模板,利用所述引物进行PCR扩增并测序,根据MEGA7.0计算测序所获高质量序列的种内及种间K2P距离,构建NJ系统进化树;
其中,唐古特大黄单独聚为一支,药用大黄单独聚为一支,掌叶大黄单独聚为一支。
2.根据权利要求1所述的用途,其特征在于,PCR扩增体系:2×TaqMasterMix 12.5μL,2.5 μM上下游引物各1 μL,DNA模板50 ng,加灭菌双蒸水补足体积至25 μL。
3.根据权利要求1所述的用途,其特征在于,PCR反应条件参数:94℃,5min;94℃,30s,55℃,30s,72℃,45s,35个循环;72℃,10min。
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CN116426671B (zh) * | 2023-03-03 | 2024-05-10 | 中国医学科学院药用植物研究所 | 基于时珍法的中药鉴定方法及应用 |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN112379034A (zh) * | 2020-12-14 | 2021-02-19 | 华润三九(雅安)药业有限公司 | 大黄药材基原的鉴别模型及其构建方法,鉴别方法 |
CN113308567A (zh) * | 2021-07-21 | 2021-08-27 | 北京中医药大学 | 一种基于大黄叶绿体基因组多态性区域制备的条形码及其应用 |
-
2021
- 2021-10-21 CN CN202111223899.XA patent/CN113897415B/zh active Active
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN112379034A (zh) * | 2020-12-14 | 2021-02-19 | 华润三九(雅安)药业有限公司 | 大黄药材基原的鉴别模型及其构建方法,鉴别方法 |
CN113308567A (zh) * | 2021-07-21 | 2021-08-27 | 北京中医药大学 | 一种基于大黄叶绿体基因组多态性区域制备的条形码及其应用 |
Non-Patent Citations (5)
Title |
---|
AFLP分析唐古特大黄、掌叶大黄和药用大黄的亲缘关系研究;索风梅等;中草药(第02期);第132-136页 * |
Comparative Chloroplast Genome Analysis of Rhubarb Botanical Origins and the Development of Specific Identification Markers;Yuxin Zhou等;Molecules;第23卷(第11期);第2811-2823页 * |
Complete chloroplast genome sequence determination of Rheum species and comparative chloroplast genomics for the members of Rumiceae;Tao Zhou等;Plant Cell Reports;第39卷;摘要、材料与方法、讨论 * |
Yuxin Zhou等.Comparative Chloroplast Genome Analysis of Rhubarb Botanical Origins and the Development of Specific Identification Markers.Molecules.2018,第23卷(第11期),第2811-2823页. * |
多基原药材大黄叶绿体matK基因序列分析及鉴定研究;张晓芹等;药学学报(第11期);第98-104页 * |
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