[Correlation between flavonoids contents in Hippophae rhamnoides subsp. inensis leaf and ecological factors, and ecological suitability analysis of H. rhamnoides subsp. sinensis].

The study aims at providing a new suitable way to promote artificial cultivation, solving the problem of resources increasingly endangered wild medicine, and protecting the wild resources of Tibetan medicine. The content of quercetin,kaempferol and isorhamnetin was determined by HPLC. The correlation between flavonoids components and ecological factors was analyzed using partial least-squares regression (PLSR). Based on Maxent model combining using ArcGIS software, suitable regionalization for H.rhamnoides subsp. sinensis was studied.The results showed that the difference of quercetin,kaempferol and isorhamnetin content in samples from different regions were obvious. The main factors effecting quercetin content accumulation were the altitude andthe average monthly precipitation in January and August. The main factors effecting kaempferol accumulation were the altitude andthe average monthly precipitation in the coldest quarter and December. The main factors effecting isorhamnetin accumulation were the average monthly precipitation in August, January and the coldest quarter.The regional distribution suitability index for H.rhamnoides subsp. sinensis was 0-0.708. The suitable area 590 500 km², accounting for 6.13% of the total area. The preferably suitable area was 552 500 km², accounting for 5.73% of the total area.The methods used in the study is simple and feasible, the result is reliable which provide a new approach for Tibetan medicine resources sustainable exploitation and utilization.

[Potential distribution of the traditional Tibetan herb Pterocephalus hookeri by Maxent model].

In order to study the ecology suitability of Pterocephalus hookeri, and provide a reference for GAP planting location and regional development, the Maxent model and GIS technology were used to investigate ecology suitability regions for P. hookeri based on the distribution points collected from Chinese virtual herbarium, the references and field trips. The potential distribution areas mainly concentrated in the eastern Tibet, western Sichuan, southern Qinghai, northwest Yunnan, and southern Gansu. There were 7 major environmental factors to have obvious influence on ecology suitability distributions of P. hookeri, including altitude (contribution rate of 62%), precipitation of warmest quarter (contribution rate of 14.4%), coefficient of variation of precipitation seasonality (contribution rate of 7.2%), mean temperature of driest quarter (contribution rate of 3.5%), the electrical conductivity of top and sub-soil (contribution rate of 3%), the total exchangeable bases in the top- and subsoil (contribution rate of 2.4%) and SD of temperature seasonality (contribution rate of 2.2%). The study of the ecological suitability regionalization of P. hookeri based on Maxent model can provide scientific basis for the selection of artificial planting base and GAP planting location.

The Chemical and Genetic Characteristics of Szechuan Pepper (Zanthoxylum bungeanum and Z. armatum) Cultivars and Their Suitable Habitat.

Szechuan peppers, famous for their unique sensation and flavor, are widely used as a food additive and traditional herbal medicine. Zanthoxylum bungeanum and Z. armatum are both commonly recognized as Szechuan peppers, but they have different tastes and effects. The chemical components, genetic characteristics, and suitable habitat of six cultivars were analyzed in this study. The results indicated that Z. armatum contained a larger proportion of volatile oil, whereas Z. bungeanum produced a more non-volatile ether extraction. The average content of volatile oil and non-volatile ether extract of Z. armatum were 11.84 and 11.63%, respectively, and the average content of volatile oil and non-volatile ether extract of Z. bungeanum were 6.46 and 14.23%, respectively. Combined with an internal transcribed spacer 2 (ITS2) sequence characters and chemical PCA results, six cultivars were classified into their own groups, for the two species in particular. The temperature in January and July were the most significant ecological factors influencing the contents of the Z. armatum volatile oil. However, annual precipitation, temperature in January and relevant humidity had a significant positive correlation with the content of non-volatile ether extract in Z. bungeanum. Thus, the most suitable areas for producing Z. bungeanum cultivars ranged from the Hengduan Mountains to the Ta-pa Mountains, and the regions suitable for Z. armatum cultivars were found to be in the Sichuan Basin and Dalou-Wu mountains. The predicted suitable habitat could be used as a preliminary test area for Szechuan pepper cultivar production.

[Potential distribution of Rhodiola crenulata in Tibetan Plateau based on Maxent model].

Wildlife tending and artificial cultivation is an important way to protect the wild resources of Rhodiola crenulata. It is a study hotspot at present. The distribution information of R. crenulata was collected by query data and field survey, the ecological suitability regionalization was conducted based on maximum entropy model combine with ecological factors, including climate, soil and altitude. To provide the reference for production layout, suitable planting area and the selection of artificial planting base by studying the ecological suitability regionalization of R. crenulata. The potential distribution areas mainly concentrated in the easen Tibet, western Sichuan, southern Qinghai, and Gansu Gannan Tibetan Autonomous Prefecture, Yunnan Diqing Tibetan Autonomous Prefecture. There were 5 major environmental factors to have obvious influence on ecology suitability distributions of R. crenulata, including altitude (contribution rate of 61.8%), precipitation of warmest quarter (contribution rate of 19%), the coefficient of variation of precipitation seasonality (contribution rate of 4.7%), the SD of temperature seasonality (contribution rate of 4%), mean temperature of driest quarter (contribution rate of 2.5%). The AUCs of ROC curve were both above 0.9, indicating that the predictive results with the Maxent model were highly precise. The study of the ecological suitability regionalization of R. crenulata based on Maxent can provide a scientific basis for the selection of artificial planting base.

[Research progress on resources and quality evaluation of Tibetan medicine in Qinghai-Tibet Plateau].

With the development of Tibetan medicine industry, the demands for Tibetan medicine were rising sharply. In addition, with the eco-environment vulnerability of Qinghai-Tibet Plateau region and the phenomenon of synonymies and homonymies in Tibetan medicine, there were a lack of resources and varieties in the clinical application of Tibetan medicine. At present, the shortage of Tibetan medicine and the inadequacy of its quality standard have become the two major problems that seriously restricted the sustainable development of Tibetan medicine industry. Therefore, it is important to develop the resources investigation and quality evaluation for Tibetan medicine, which were contribute to its resources protection and sustainable utilization. In this paper, current status of resources investigation, quality standardization, artificial breeding and germplasm resources of Tibetan medicine were presented by the integrated application of the new technologies, such as DNA barcoding and 1H-NMR, which provided a reference information for resources protection, sustainable utilization, variety identification and quality standardization of Tibetan medicine resources in Qinghai-Tibet Plateau.

[Ecology suitability regions and ecological characteristics of Panax notoginseng (Burk.) F.H. Chen based on maximum entropy model].

The ecology suitability and ecological characteristics of Panax notoginseng (Burk.) F. H. Chen were studied to provide a reference for its artificial introduction and cultivation. The maximum entropy model (MaxEnt) and geographic information system (GIS) were used to investigate the global ecology suitability regions for Panax notoginseng (Burk.) F. H. Chen based on its 67 distribution points collected from global biodiversity information facility (GBIF), Chinese virtual herbarium(CVH) and the related references. The results showed that the possible ecological suitable regions of Panax notoginseng (Burk.) F. H. Chen were located in Yunnan, Guangxi, Guangdong, Guizhou, Hainan, Sichuan, Fujian and Chongqing provinces. The areas with ecological similarity higher than 60% were about 89 571.3 square kilometers in total, mainly distributing in Yunnan and Guangxi provinces and small portion was located in Guangdong and Guizhou provinces. The areas with ecological similarity between 40% and 60% were about 155 172 square kilometers, mainly in Yunnan,Guangxi, Guangdong, Guizhou, Hainan, Sichuan provinces. The distribution areas were about 329 952.8 square kilometers with ecological similarity between 20% and 40%, mainly in Yunnan, Guangxi, Guangdong, Guizhou, Hainan, Sichuan, Fujian and Chongqing. The climate factors mainly affecting the distribution of Panax notoginseng (Burk.) F. H. Chen were precipitation of warmest quarter, SD of temperature seasonality, altitude, isothermality, coefficient of variation of precipitation seasonality, mean temperature of monthly, precipitation of driest month, reference bulk density of soil and soil texture.

[Spatial Distribution and Global Potential Suitability Regions of Artemisia annua].

OBJECTIVE: To study the spatial distribution and potential climatic suitability regions of Artemisia annua around the world. METHODS: The spatial distribution and climatic characteristics were researched by factor analysis based on Global Biodiversity Information Facility Database and World Climate Database. The global potential suitability regions of Artemisia annua were analyzed by ArcGIS. RESULTS: Artemisia annua distributed in three longitude zones, including 90. 55 °W - 77. 14 °W, 2. 03 °E - 11. 75 °E and 98. 27 °E - 111. 05 °E,which were respectively in North America, Europe and Asia. The latitude range was mainly 29. 15 °N - 51. 36 ° N. 80% of Artemisia annua were in the regions which elevation range was 22. 00 - 491. 00 m, annual precipitation was 492. 30 ~ 1 366. 70 mm, annual average temperature was from 8. 10 to 17. 27 °C. The potential suitability regions of Artemisia annua with 95% ~ 100% climate similarity were mainly in 30 °S and 30 °N regions, centered around the equator axis. Conclusion: Latitude is closely related to the distribution of Artemisia annua, the key affecting climatic factors are annual precipitation, the wettest season precipitation, the warmest season precipitation and the highest temperature in the warmest month, the average temperature of the warmest season, as well as the average temperature of the wettest season. The potential suitability regions of Artemnisia annua are in eastern North America, western Europe and eastern Asia.

Chemical Constituents, Quality Control, and Bioactivity of Epimedii Folium (Yinyanghuo).

Epimedii Folium (Yinyanghuo in Chinese) is one of the most commonly used traditional Chinese medicines. Its main active components are flavonoids, which exhibit multiple biological activities, such as promotion of bone formation and sexual function, protection of the nervous system, and prevention of cardiovascular diseases. Flavonoids also show anti-inflammatory and anticancer effects. Various effective methods, including genetic and chemical approaches, have been developed for the quality control of Yinyanghuo. In this review, the studies conducted in the last decade about the chemical constituents, quality control, and bioactivity of Yinyanghuo are summarized and discussed.

Determination of production regions for pollution-free Chinese medicinal materials by geographic information system: Achyranthes bidentata (Niu Qi) in Tianjin as an example.

BACKGROUND: The land and ecology suitability for producing pollution-free Chinese medicinal materials should be evaluated based on Geographic Information System (GIS). This study aims to determine production regions for pollution-free Achyranthes bidentata in Tianjin as a case to illustrate the workflow based on GIS. METHODS: The slopes, land usage, residential areas and roads were selected to evaluate the land suitability, to avoid the potential pollution sources. The ecology suitability evaluation was performed based on the soil type and nine climate factors, such as active accumulated temperature, mean January temperature, mean July temperature, the lowest temperature in January, the highest temperature in July, mean annual temperature, sunshine duration, relative humidity, annual precipitation, affecting the natural growth of A. bidentata. RESULTS: The best production regions for pollution-free A. bidentata in Tianjin, with a total area of approximately 575 km(2), were found in Jinghai County, Ninghe County, Wuqing District, and Dagang District. CONCLUSION: This study illustrated a workflow based on GIS for determining the production regions in Tianjin for pollution-free A. bidentata.

Natural Resource Monitoring of Rheum tanguticum by Multilevel Remote Sensing.

Remote sensing has been extensively applied in agriculture for its objectiveness and promptness. However, few applications are available for monitoring natural medicinal plants. In the paper, a multilevel monitoring system, which includes satellite and aerial remote sensing, as well as ground investigation, was initially proposed to monitor natural Rheum tanguticum resource in Baihe Pasture, Zoige County, Sichuan Province. The amount of R. tanguticum from images is M = S*ρ and S is vegetation coverage obtained by satellite imaging, whereas ρ is R. tanguticum density obtained by low-altitude imaging. Only the R. tanguticum which coverages exceeded 1 m(2) could be recognized from the remote sensing image because of the 0.1 m resolution of the remote sensing image (called effective resource at that moment), and the results of ground investigation represented the amounts of R. tanguticum resource in all sizes (called the future resource). The data in paper showed that the present available amount of R. tanguticum accounted for 4% to 5% of the total quantity. The quantity information and the population structure of R. tanguticum in the Baihe Pasture were initially confirmed by this system. It is feasible to monitor the quantitative distribution for natural medicinal plants with scattered distribution.

[Research on spectral reflectance characteristics for Glycyrrhizae Radix].

In order to study the spectral reflectance differences of Glycyrrhizae Radix under different growth conditions and lay the foundation for quantitative monitoring of Glycyrrhizae Radix remote sensing images, spectra of Glycyrrhiza species under different growth period and different varieties and different regions were measured by a portable spectrometer. The results showed that the reflectivity of annual G. uralensis was obviously higher than that of the two years plant in the visible light band own to the contents of crown layer chlorophyll. The reflectivity of two years G. pallidiflora was higher than that of G. uralensis in the near infrared band own to the leaf area index and the content of leaf water. The red edge spectrum of annual plant fluctuated largely than that of two years plant due to vegetation coverage and leaf area index. G. pallidiflora grew well than G. uralensis. Under different regions of the Glycyrrhiza species, spectral data analysis showed that within a certain range, the average annual precipitation and average annual evaporation were the major factors to affect the differences of Glycyrrhiza species spectral data under different regions owe to the leaf water content, the higher leaf water content, the lower spectral reflectance. The principal component analysis and continuum-removed method of the spectral data under different regions found that, within a certain range, the average annual precipitation and average annual evaporation were the major factors caused by the differences of Glycyrrhiza species spectral data under the different regions, Glycyrrhiza species spectral similarity related to the spatial distance.

[Study on traceability system of genuine medicinal materials].

Genuine medicinal materials with special characteristics of Traditional Chinese Medicine (TCM), is recognized as high quality medicine. Both ancient records and modern research considered that the origin is an important reason for the formation of genuine medicinal materials. However, blindly transplanting of genuine medicinal materials has led to the quality decline and counterfeit medicines appeared in production or sale progress, which may increase the risk of accidents in TCM. Frequent accidents emerged in Chinese herbal affects its export. What's more, it is a great threat to the medication safety in TCM clinical. There is an urgent need to implement traceability systems of TCM, which could provide convenient information record and traceability of TCM circulation. This paper reviews a variety of technical methods for genuine medicinal materials traceability, and proposed the establishment of genuine medicinal materials traceability system based on two-dimensional code and network database.

[RNA-Seq and genuine traditional Chinese medicine].

RNA-Sequencing (RNA-Seq) is a newly-developed method in transcriptome research, it can afford more accurate transcription information and be more quickly by using Next-generation Sequencing (NGS) technology. RNA-Seq has been widely used in various biological fields. Genuine traditional Chinese medicines (TCM), with good quality and therapeutic effect, were always praised highly and used by famous physicians. The geo-herbalism formation of TCM is based on the product of the gene expression at specific space and time. So it has been a research hotspot to analyze the mechanism of biosynthesis through RNA-Seq in the study on the secondary metabolism of medicinal plant. This article mainly illustrates the RNA-Seq and its advantages, it also discusses the potential application in genuine TCM, and it can provide useful information for other researchers.

[Quality variation and ecotype division of Panax quinquefolium in China].

Quality variation and ecotype classification of Chinese herbal medicine are important scientific problems in Daodi herbal medicine research. The diversity of natural environmental conditions has led to form unique multi-Daodi, multi-product areas that produce particular Chinese herbal medicine. China is one of three big American ginseng (Panax quinquefolium L.) producing areas worldwide, with over 300 years of application and 40 years of cultivation history. Long-term production practice has led to the formation of three big advocate produce areas in China: Northeast province, Beijing and Shandong. P. quinquefolium L. grown under certain environmental conditions will develop long-term adaptations that will lead to more stable strains (different ecotypes). P. quinquefolium L., can vary greatly in quality; however, the ecological mechanisms causing this variation are still unclear. Root samples were collected from four-year-old cultivated P. quinquefolium L. plants in the three major genuine (Daodi) American ginseng-producing areas of Northeast province, Beijing and Shandong province, China. Ultra-performance liquid chromatography was used to analyze the contents of eight ginsenosides (Rg1, Re, Rb1, Rb2, Rb3, Rc, Rd, Rg2). Data for nine ecological factors, including temperature, moisture and sunlight, were obtained from the ecological database of Geographic Information System for Traditional Chinese Medicine. Soil samples from the sampling sites were collected. Effective boron and iron, available nitrogen and potassium, as well as other trace elements and soil nutrients, were determined by conventional soil physicochemical property assay methods. Analytical methods of biostatistics and numerical taxonomy were used to divide ecotypes of the three main Panax quinquefolium L. producing areas in China based on ginsenoside content, climate, soil and other ecological factors. To our knowledge, this is the first time that ecological division of P. quinquefolium L. producing areas in China has ever been conducted. The results show that there are two chemoecotypes of P. quinquefolium L. in China: ginsenoside Rb1-Re from outside Shanhaiguan, and ginsenoside Rg2-Rd from inside Shanhaiguan. Similarly, there are two types of climatic characteristics: inside Shanhaiguan (Beijing, Shandong) and outside Shanhaiguan (Northeast). This suggests that the formation and differentiation of chemoecotypes of P. quinquefolium L. is closely related to variability of the climatic and geographical environment. Additionally, ecological variation of the three main producing areas, characteristics of two climatic ecotypes, and soil characteristics are also discussed and summarized. These results provide experimental scientific evidence of the quality variation and ecological adaptation of P. quinquefolium L. from different producing areas. They also deepen our understanding of the biological nature of Daodi P. quinquefolium L. formation, and offer novel research models for other multi-origin, multi-Daodi Chinese herbal medicines ecotypes. In addition, the results demonstrate the critical need for improving quality, appropriate ecological regionalization and promoting industrialized development of P. quinquefolium L.

Transcriptome analysis of Bupleurum chinense focusing on genes involved in the biosynthesis of saikosaponins.

BACKGROUND: Bupleurum chinense DC. is a widely used traditional Chinese medicinal plant. Saikosaponins are the major bioactive constituents of B. chinense, but relatively little is known about saikosaponin biosynthesis. The 454 pyrosequencing technology provides a promising opportunity for finding novel genes that participate in plant metabolism. Consequently, this technology may help to identify the candidate genes involved in the saikosaponin biosynthetic pathway. RESULTS: One-quarter of the 454 pyrosequencing runs produced a total of 195, 088 high-quality reads, with an average read length of 356 bases (NCBI SRA accession SRA039388). A de novo assembly generated 24, 037 unique sequences (22, 748 contigs and 1, 289 singletons), 12, 649 (52.6%) of which were annotated against three public protein databases using a basic local alignment search tool (E-value ≤1e-10). All unique sequences were compared with NCBI expressed sequence tags (ESTs) (237) and encoding sequences (44) from the Bupleurum genus, and with a Sanger-sequenced EST dataset (3, 111). The 23, 173 (96.4%) unique sequences obtained in the present study represent novel Bupleurum genes. The ESTs of genes related to saikosaponin biosynthesis were found to encode known enzymes that catalyze the formation of the saikosaponin backbone; 246 cytochrome P450 (P450s) and 102 glycosyltransferases (GTs) unique sequences were also found in the 454 dataset. Full length cDNAs of 7 P450s and 7 uridine diphosphate GTs (UGTs) were verified by reverse transcriptase polymerase chain reaction or by cloning using 5' and/or 3' rapid amplification of cDNA ends. Two P450s and three UGTs were identified as the most likely candidates involved in saikosaponin biosynthesis. This finding was based on the coordinate up-regulation of their expression with ß-AS in methyl jasmonate-treated adventitious roots and on their similar expression patterns with ß-AS in various B. chinense tissues. CONCLUSIONS: A collection of high-quality ESTs for B. chinense obtained by 454 pyrosequencing is provided here for the first time. These data should aid further research on the functional genomics of B. chinense and other Bupleurum species. The candidate genes for enzymes involved in saikosaponin biosynthesis, especially the P450s and UGTs, that were revealed provide a substantial foundation for follow-up research on the metabolism and regulation of the saikosaponins.

[Quantitative study on ecological suitability of Chinese herbal medicine based on GIS].

The quality of Chinese herbal medicine is closely related to its producing region. In order to apply mathematical models to do a quantitative study on the suitability of Chinese herbal medicine, it is necessary to study on the ecological factors and the interpolation of climatic data, which influence the Chinese herbal medicine growth. The paper firstly studied the judgment standard of ecological index from the points of ecology and statistics, and how to calculate the optimum range values and the weight of each ecological factor. Secondly, meteorological element data is essential data in analyzing the suitable region of Chinese herbal medicine, and the spatial distribution of meteorological elements is closely related to terrain environment, so, in order to make the results close to true value by the greatest degree. The paper adopted multiple linear regression interpolation method which based on DEM. The paper distinguished the factor system of suitable region and interpolation on the point of datumization, and made a study on it about some key issues.

Species identification of medicinal pteridophytes by a DNA barcode marker, the chloroplast psbA-trnH intergenic region.

Medicinal pteridophytes are an important group used in traditional Chinese medicine; however, there is no simple and universal way to differentiate various species of this group by morphological traits. A novel technology termed "DNA barcoding" could discriminate species by a standard DNA sequence with universal primers and sufficient variation. To determine whether DNA barcoding would be effective for differentiating pteridophyte species, we first analyzed five DNA sequence markers (psbA-trnH intergenic region, rbcL, rpoB, rpoC1, and matK) using six chloroplast genomic sequences from GeneBank and found psbA-trnH intergenic region the best candidate for availability of universal primers. Next, we amplified the psbA-trnH region from 79 samples of medicinal pteridophyte plants. These samples represented 51 species from 24 families, including all the authentic pteridophyte species listed in the Chinese pharmacopoeia (2005 version) and some commonly used adulterants. We found that the sequence of the psbA-trnH intergenic region can be determined with both high polymerase chain reaction (PCR) amplification efficiency (94.1%) and high direct sequencing success rate (81.3%). Combined with GeneBank data (54 species cross 12 pteridophyte families), species discriminative power analysis showed that 90.2% of species could be separated/identified successfully by the TaxonGap method in conjunction with the Basic Local Alignment Search Tool 1 (BLAST1) method. The TaxonGap method results further showed that, for 37 out of 39 separable species with at least two samples each, between-species variation was higher than the relevant within-species variation. Thus, the psbA-trnH intergenic region is a suitable DNA marker for species identification in medicinal pteridophytes.

TCMGIS-II based prediction of medicinal plant distribution for conservation planning: a case study of Rheum tanguticum.

BACKGROUND: Many medicinal plants are increasingly endangered due to overexploitation and habitat destruction. To provide reliable references for conservation planning and regional management, this study focuses on large-scale distribution prediction of Rheum tanguticum Maxim. ex Balf (Dahuang). METHODS: Native habitats were determined by specimen examination. An improved version of GIS-based program for the distribution prediction of traditional Chinese medicine (TCMGIS-II) was employed to integrate national geographic, climate and soil type databases of China. Grid-based distance analysis of climate factors was based on the Mikowski distance and the analysis of soil types was based on grade division. The database of resource survey was employed to assess the reliability of prediction result. RESULTS: A total of 660 counties of 17 provinces in China, covering a land area of 3.63 × 106 km2, shared similar ecological factors with those of native habitats appropriate for R. tanguticum growth. CONCLUSION: TCMGIS-II modeling found the potential habitats of target medicinal plants for their conservation planning. This technology is useful in conservation planning and regional management of medicinal plant resources.

Mapping the potential distribution of high artemisinin-yielding Artemisia annua L. (Qinghao) in China with a geographic information system.

BACKGROUND: Artemisia annua L. is an important source for artemisinin, a potent drug for treating malaria. This study aims to map and predict the potential geographic distribution of A. annua L. in China. METHODS: The Geographic Information System for traditional Chinese medicine (TCM-GIS) was developed and used to map the potential geographic distribution of A. annua L. RESULTS: Climatic, edaphic and topographic characteristics of A. annua L. microhabitats in Youyang County were mapped to find distribution patterns. The maps identified that certain habitats in the Chongqing region and some potential regions, especially in Guizhou Province, possess similarity indices of >/=98%. In particular, high quality microhabitats A. annua L. were found in the Wuling mountains region. CONCLUSION: The present study demonstrates a GIS approach to predict potential habitats for A. annua L. TCM-GIS is a powerful tool for assessing bioclimatic suitability for medicinal plants.

Using DNA barcoding to identify species within Euphorbiaceae.

In this study, we tested the applicability of four DNA regions (rbcL, matK, ITS, and ITS2) as barcodes for identifying species within Euphorbiaceae. Based on assessments of the specific genetic divergence, the DNA barcoding gap, and the ability for species discrimination, the present results affirmed that ITS/ITS2 is a potential barcode for the Euphorbiaceae species. This study also provided a large-scale test to evaluate the effectiveness of ITS/ITS2 for differentiating species within Euphorbiaceae. Of the 1183 plant samples collected from 871 species in 66 diverse genera, ITS/ITS2 successfully identified > 90% and 100% of them at the species and genus levels, respectively. Therefore, our research indicates that use of the ITS/ITS2 region is a powerful technique for Euphorbiaceae identification.