龙血竭（Resina Draconis），我国珍稀名贵中药，素有“活血圣药”之称，具有活血散瘀，定痛止血，敛疮生肌的功效。国家药品标准规定其来源为百合科植物剑叶龙血树Dracaena cochinchinensis ( Lour.) S.C.Chen的含脂木材经提取得到的树脂，而早在1991年剑叶龙血树就被定为国家二级濒危保护植物，因此，目前我国龙血竭药材的提取原料完全依赖老挝、越南等东南亚国家进口，且随着人们对龙血竭需求量的不断攀升，龙血竭原料资源在国外亦日益匮乏。因多种龙血树属植物在自然条件或人工诱导后均可形成红色树脂，药材收购商常将几种龙血树的含脂木材混在一起销售，更有甚者甚至将龙血树属以外其它科属植物的红色含脂木材也混在龙血竭提取原料中，严重影响了龙血竭的临床用药安全与正常的市场流通。因此，龙血竭药材基原的准确鉴定意义重大，然而，目前关于龙血竭药材基原的分子鉴定研究尚属空白。此外，我国及老挝、越南等东南亚地区分布着一种被国内习称为岩棕D. sp的龙血树属植物，该植物分布较广、数量较多，且可形成红色树脂，常作为龙血竭的提取原料被广泛应用，然而，对其药材品质评价方面的研究未见报道。 基于以上情况，本研究首先采用常规DNA条形码片段对龙血树属植物进行鉴定效率评价研究，之后利用叶绿体基因组对龙血竭基原及其同属植物进行super-barcode鉴定研究，在此基础上筛选合适的DNA条形码片段，对龙血竭基原及其同属植物进行专一性barcode研究。其次，本研究比较了岩棕与剑叶龙血树所产龙血竭的主要活性成分含量差异，并采用代谢组学方法比较两种龙血竭及原植物茎部的整体化学组成差异性；最后，通过高通量转录组测序初步分析龙血竭的形成机制。主要研究结果如下： 1. 常规DNA条形码片段（ITS2、psbA - trnH、rbcL、matK）对龙血树属植物鉴定效率极低，matk是其中鉴定效率最高的条形码片段，但其鉴定效率仅为42.86 %。采用叶绿体基因组作为super - barcode对龙血树属植物进行鉴定研究，结果显示，super - barcode可很好的区分龙血树属植物。在叶绿体基因组测序基础上，对所筛选的4条高变区间片段（trnP - psaJ、psbK - psbI、trnT - trnL和clpP）进行鉴定效率比较研究，结果显示仅clpP片段可对7种龙血树属植物进行准确鉴定，但其片段较长，扩增及测序成功率较低；采用联合序列片段“psbK - psbI + trnP - psaJ”可对龙血树属植物进行准确的分子鉴定，且扩增及测序成功率高，可作为龙血竭基原及龙血树属植物分子鉴定的最佳DNA条形码片段。此外，依据叶绿体基因组及DNA条形码片段所构建的系统进化树，我们认为剑叶龙血树D. cochinchinensis与海南龙血树D. cambodiana不宜合并处理，应单独列种。 2. 5种主要化学成分（龙血素A、龙血素B、白藜芦醇、紫檀茋、7,4′ - 二羟基黄酮）中，除龙血素A的其余4种成分在两种龙血竭中含量均有显著性差异。以剑叶龙血树为基原的龙血竭（以下简称正品龙血竭）中白藜芦醇、7 - 4′ - 二羟基黄酮含量较高，而岩棕所产龙血竭（以下简称岩棕龙血竭）中龙血素B、紫檀茋含量较高，其中岩棕龙血竭的龙血素B含量明显高于正品龙血竭（9.4倍），因此，岩棕龙血竭有作为正品龙血竭代用品的开发潜力。此外，正品龙血竭的龙血素A含量远远高于龙血素B（26倍），且二者均为龙血竭活血化瘀的主要活性成分，鉴于目前多数正品龙血竭药材的龙血素B含量无法达到国家药品标准规定的最低限量（0.4 %），因此，作者建议相关部门在修订龙血竭药品标准时，将龙血素A含量或龙血素A、B含量之和作为龙血竭质量控制的含量指标。 3. 建立了一种基于超高效液相色谱 - 四级杆 - 静电场轨道阱联用质谱技术（UPLC - Q - Orbitrap - MS）的代谢组学方法，用于比较岩棕龙血竭与正品龙血竭的化合物组成差异性。OPLS-DA分析结果显示，岩棕龙血竭与正品龙血竭的化合物组成存在明显差异（R2Y = 0.999，Q2 = 0.977）。最终,共鉴定（或临时鉴定）差异化合物154个，其中岩棕龙血竭中含量较高的有93个，正品龙血竭中含量较高的化合物有61个，差异化合物并无明显的类别差异性。归一化峰面积比较分析结果显示，胺类、酚酸类、生物碱类、萜类、有机酸类、甾体类化合物在岩棕龙血竭中含量较高，而苯丙素类、黄酮类化合物在正品龙血竭中含量较高，主要活性成分黄酮类化合物在正品龙血竭中含量较高。最后，我们筛选出8个化合物作为区分岩棕龙血竭与正品龙血竭的差异标志物。 4. 运用代谢组学方法比较岩棕与剑叶龙血树健康茎部的化学组成差异性。OPLS-DA分析结果显示，两种龙血树健康茎部的化学组成有明显差异（R2Y = 0.998，Q2 = 0.954）。在所分析的所有特征值（feature）中，同时满足VIP > 1.0且P < 0.01的共有619个，比伤害诱导后的差异化合物（feature）少了470个，说明岩棕与剑叶龙血树茎部的化学组成在伤害诱导前就存在一定的差异性，而这种差异性在伤害诱导后变的愈发明显。最终，共鉴定（或临时鉴定）化合物79个。归一化峰面积比较分析结果显示，剑叶龙血树健康茎部含有较高的氨基酸、有机酸类化合物，而岩棕的茎部则含有较高的黄酮类、生物碱类、甾体类、胺类化合物。 5. 对剑叶龙血树伤害诱导后的茎部样品进行转录组测序，组装后的Unigene number为63,244，得到注释的Unigene共有27,912个。差异表达基因统计结果表明，剑叶龙血树伤害诱导后前3d是其胁迫反应的最活跃时期。组间差异表达基因GO term富集及KEGG pathway统计结果显示，在伤害诱导后3d内，植株中多个与生长发育、器官发育、生殖系统发育等相关的生理进程处于下调阶段，而与多种次生代谢产物的生物合成途径、植株光合作用、抗胁迫反应等相关途径进入上调阶段。而在伤害诱导3d后，多种次生代谢产物生物合成途径及抗胁迫反应途径逐渐进入下调阶段。与黄酮类化合物（龙血竭主要活性成分）相关的代谢途径在伤害诱导后的前3d，并没有被差异表达基因显著富集到，而在伤害诱导3d后，多个差异表达基因富集到相关途径。分析结果显示，剑叶龙血树与海南龙血树在龙血竭形成过程的各个阶段，其差异基因数量及所富集的代谢途径存在明显差异，证明其形成机制有所差异。

Resina Draconis, a rare and precious traditional medicine in China, is known as the "holy medicine for promoting blood circulation". It has the functions of promoting blood circulation and removing blood stasis, calming pain and hemostasis, astringent sore and generating muscle. According to the national drug standard, it's derived from the resin extracted from the wood of D. cochinchinensis (Lour.) s.c.chen, a Liliaceae plant, which has been regarded as the second level endangered plant in China since 1991. At present, the extraction raw materials of dragon's blood in China are completely dependent on Laos, Vietnam and other Southeast Asian countries, and with the increasing of demand, the resource of Resina Draconis in foreign countries is also increasingly scarce. A variety of Dracaena plants can form red resin when suffered from physical injury. The pharmacopolist often mixed the resin wood of several Dracaena plant and supplied the medicinal materials market, and some even mixed the red resin wood of other families and genera species. The safety of clinical medication and normal market circulation of Resina Draconis were seriously affected. Therefore, the accurate identification of Resina Draconis is of great significance. However, the research of molecular identification of Resina Draconis is still blank. In addition, according to our survey, there is a kind of Dracaena plant (named Yan-zong) in China, Laos, Vietnam and other Southeast Asian areas, which has a large number of distribution and can form red resin. It is often used as the extraction material of Resina Draconis. However, the quality of its medicinal materials is unknown. In addition, the standard of medicinal materials stipulates that the resin wood of D. cochinchinensis is the genuine extraction material of Resina Draconis. Based on the above, we first evaluated the identification efficiency of traditional DNA barcode fragments for Dracaena. spp, then the chloroplast genome was used to identify the original plant of Resina Draconis. On this basis, the suitable candidate DNA barcodes were selected to study the specific barcode of Dracaena. spp plant. Secondly, the study compared the main chemical components of Resina Draconis which derived from different Dracaena species (D. cochin- chinensis and D. sp), then compared the overall chemical components used metabonomics method in order to evaluate the quality differences. After that, the differences in chemical composition of the healthy stem wood of the original plant were compared using the metabonomics. Finally, the formation mechanism of Resina Draconis was analyzed by high-throughput transcriptome sequencing. The main results are as follows: 1. The efficiency of routine DNA barcodes (ITS2, psbA-trnH, rbcL, matk) for identification of Dracaena plants is very low, and among them, the most efficient bar- code sequence is matk, which its efficiency is only 42.86 %. The results showed that super-barcode can distinguish Dracaena plants very well using the chloroplast genome as super-barcode. On the basis of chloroplast genome sequencing, the identification effi- ciency of four highly variable fragments （trnP-psaJ、psbK-psbI、trnT-trnL and clpP） was compared. The results showed that only clpP fragments could identify 7 Dracaena species accurately, but the sequence were longer, and the expansion and sequencing success rate were lower. “psbK-psbI + trnP-psaJ” can be used for accurate molecular identification of Dracaena plant, and has high success rate of amplification and sequenc- ing. It can be used as the best DNA barcode for molecular identification of dragon's blood and Dracaena. spp. In addition, according to the phylogenetic tree constructed by chloroplast genome and DNA barcode, we thought that D. cochinchinensis and D. cam- bodiana should not be merged into one species, but should be listed separately. 2. There were significant differences in the contents of the other four components except for loureirin A. The Resina Draconis derived from D. cochinchinensis contains higher content of resveratrol and 7-4 '- dihydroxyflavone, and the dragon's blood derived from D. sp contains higher content of loureirin B and pterostilbene. The content of loureirin B is higher in the Resina Draconis derived from D. cochinchinensis（9.4 times）. the author thinks that Resina Draconis derived from D. sp has the potential to be considered as a substitute of the genuine Resina Draconis. In addition, the content of Loureirin A is much higher than that of Loureirin B (26 times), and both of them are the main active components of Resina Draconis to promote blood circulation and remove blood stasis. Considering that the content of Loureirin B in most of the Resina Draconis material can not meet the National Drug Standard (0.4 %), therefore, it is suggested that the relevant departments should further improve the drug standard of dragon's blood, and take the content of Loureirin A or the sum of Loureirin A and B as the content index of Resina Draconis quality control. 3. A metabonomics method based on UHPLC-Q-Orbitrap-MS was established to study the difference of the chemical composition of Resina Draconis derived from D. cochinchinensis and D. sp. The OPLS-DA analysis results showed that there were significant differences in the chemical composition of Resina Draconis of different sources. Finally, a total of 154 different compounds were identified (or temporarily identified), including 93 compounds with high content in the Resina Draconis derived from D. sp, and 61 compounds with high content in Resina Draconis derived from D. cochinchinensis. The results showed that the contents of amines, phenolic acids, alkaloids, terpenes, organic acids and steroids were higher in Resina Draconis derived from D. sp, while the contents of phenylpropanoids and flavonoids were higher in Resina Draconis derived from D. cochinchinensis. In addition, eight compounds were selected as the differential markers to distinguish the two sources of Resina Draconis. 4. Comparing the chemical composition difference of healthy stem wood of D. cochinchinensis and D. sp using the metabonomics. The results of OPLS - DA analysis showed that there were significant differences in chemical composition between the two species (R2Y=0.998，Q2=0.954). Among the 21,955 features analyzed, there are 619 that meet the requirements of VIP > 1.0 and P < 0.01 (t-test), which is 470 less than the difference compounds after injury induction, which indicated that there was a certain difference in the chemical composition of the stem wood of D. cochinchinensis and D. sp before injury induction, and this difference becomes more and more obvious after injury induction. Finally, we identified (or temporarily identified) 79 compounds. The results of normalized peak area analysis showed that there were higher amino acids and organic acids in the healthy stem wood of D. cochinchinensis, and higher flavonoids, alkaloids, steroids and amines in the healthy stem wood of D. sp. 5. We sequenced the transcriptome of the stem samples of D. cochinchinensis after injury induction. The results showed that the number of Unigene was 63,244, and 27,912 Unigenes were annotated. The digital expression patterns of different induction time points were constructed and verified by qPCR. The statistical results of differentially expressed genes showed that the first three days after injury induction was the most active period of stress response. The results of GO term enrichment and KEGG pathway showed that the physiological processes related to growth and development, organ development and reproductive system development were in the down-regulation stage within three days after injury induction, while the processing of various secondary metabolites biosynthesis, photosynthesis and stress resistance were in the up-regulation stage. However, the up-regulated processing of biosynthesis pathway and stress resis- tance pathway was gradually down-regulated in three days after injury induction. The metabolic pathways related to flavonoids (the main active components of dragon's blood) were not significantly enriched by differentially expressed genes at first three days of injury induction, but multiple differentially expressed genes were enriched to related pathways after three days of injury induction.