| Ethnic pharmacy is a discipline about resources, identification, quality standards, the material basis of efficacy, pharmacodynamics mechanism, processing, preparation, basic theory and medication experience in differfent countries home and abroad. The unique national drug effects attract more and more attention.Folk medicine and some foreigh herbals are at an early stage of development. Variety systematization is the foundation of resources, basic identification, quality standards, the material basis of efficacy studies and so on.Brazil is the largest country in Latin America, with a vast farmland, vast forests, and abundant plant resources. Single herbal medicine products have been traded in Brazil and generate considerable economic benefits.Many herbs, as Brazil hematoxylin, stevia and other ones, have been widely used in domestic and foreign health care products and clinical medicine. The study ofBrazil herbal medicine’s development will be in favor of the introduction and rich resources of Traditional Chinese Medicine. Brazilian government also has a strong interest in Chinese medicine. Trade cooperation and scientific Communicationwill benefit both China and Brazil, in favor of China and Pakistan’s economic mutual benefit, the sharing of resources and medical standards and common progress. Brazil already use traditional herbal medicine, but the study of herbal medicine is still in infancy.Botany and herbal research is not deeply enough.Research on the origin of the Brazilian herb is of great significance to ensure the accuracy of the use and exploitation ofBrazil herbs.Traditional methods of variety systematization were literature study combination with folk investigations and identification of original plant, which was the basic method of variety systematization.But Traditional methods cost a lot of time and consumed manpower and resources.With the development of molecular biology, DNA barcoding method for species identification provided a new evidence. More and more attention was paid to DNA barcoding for accuraty, efficience and time-saving advantages. Internal transcribed spacer (ITS) sequences were considered to be the high efficiency to identify, which had been widely used in species identification. But the ambiguous identification standards, the imperfect database and poor quality of DNA confined the application. This study fully combined the advantages of traditional methods with molecular biology techniques, and proposed a newway tovariety systematization--"DNA barcoding-origin-morphological analysis "united method. In that way we didvariety systematization of 10 kinds ofBrazilianherbs.The molecular identification of threshold indicators was established in this study based on GenBank database in NCBI.DNA barcode technology in combination with origin analysis and morphological analysis established medicinal materialsvariety systematization methodto achieve the purpose of the Brazilian herbal varietysystematization collected in Brazilmarket. This study provided the basis for the development and application of Brazilian herbal medicine.The main results are as follows:(1) Analysis methods research was conclueded by comparing BLAST analysis and phylogenetic tree analysis of the 30 kinds of plant specimens in the accuracy, operational efficiency and conditions. The results showed that BLAST analysis was superior to the phylogenetic tree analysis. BLAST analysis accuracy reached 73%, the phylogenetic tree analysis 53%. But the advantage of phylogenetic tree analysis was to rule out some error sequences and we could get more accurate information; BLAST analysisaccuracy reached 73%, compared with the combination of the two methods was only 7% lower, and BLAST analysis was faster and more convenient. Therefore, BLAST analysis was used in the follow-up study. BLAST analysis relied on a powerful database. When the database was incomplete data, in order to avoid errors, the threshold should be set to define the minimum degree of similarity of the same species, below the threshold means that the species were not registered in the database.(2) 220 original species (more than 4 sequences) in the 2010 edition of "Chinese Pharmacopoeia" were registered in the GenBank database. Download all ITS sequences and calculate the similarity of each species, and statistics minimum similarity of the most species (70%). The result was use as the threshold. ITS sequences of 30 various plant specimens was used to test the accuracy of threshold. The results showed that most of the species of the minimum degree of similarity was 97%. BLAST analysis of 30 kinds of plant specimens, all the species could be correctly identified based on 97% except for 2 ones. (3) ITS sequences could be obtained by optimizing the DNA extraction and PCR amplification method. Kit methods, nuclear separate liquid pretreatment, classic CTAB extraction method and segment amplification were used to improve the efficiency of ITS sequences. The results showed that 65% of the medicinal material could be extracted by optimizing DNA kit Act; nuclear separation liquid pretreatment to remove impurities failed to improve the efficiency of DNA amplification; classic CTAB extraction method and segment amplification method could improve the ITS sequences efficiency.(4)Medicinal parts of thespecimens were takenas the Chinese herbal medicinematerials.According to BLAST analysis of the origin of molecular, the value 97%to define the minimum degree of similarity of the same species, we had established "DNA barcoding-Origin-morphological analysis"Variety systematizationmethod.(5) The study studied the 10 kinds of Brazilian herbals’s sourceand medicicinal partsbased on DNA barcoding-Origin-morphological analysis variety systematization method. First, ITS sequences were achieved by DNA barcoding to finish molecular identification of the 10 kinds of unknown Brazil herbals. To identify the species,97%could be the threshold with the highest similarity of species identification results. When no sequence was available in the NCBI nucleic Acid database, it meantthe database was unregistered the corresponding sequences. Then 90% could be the threshold with the highest similarity of species identification results to identify the genus. Second, the results of the DNA barcoding identification results were analyzed according to the book "flora of Brazil" and other medical literature analysis whether it was grown in Brazil. If distributed, it might be the variety. If not, the next higher similarity species would be analyzied until the name of the sequence was also shown in Brazil and the similarity was above 97%. If no anwers were got, the genus information was identified. At last,the DNA barcoding and orgin analysis resultswould be checked by character identification and microscopic identification, and the medical parts could be confermed. It provides the basis for Brazil’s herbal variety systematization, and gives a reference to found the new treatment of Chinese traditional medicine effect. The study has great significance to expand the resources of Chinese materia medica industry.
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