Variability in the genetic information DNA tracking individuals carry is easy to detect using molecular markers. In the thesis we examined related Dracaena species. For the study of genetic diversity in the genus Dracaena are used mainly noncoding regions of cpDNA, spacer trnH -- psbA, regions trnL -- trnF and trnS -- trnG -- trnG, which are more variable than coding regions. Also used in this work were coding regions of matK and rbcL. In the genus Dracaena belongs xerophytic species that are characterized by typical shaped treetop. 14 representatives of species were used in thesis growing in tropical regions of the African continent and adjacent islands and the southeastern part of the Arabian peninsula. They are monocotyledonous trees with atypical abilities of secondary thickness of trunk, which I find interesting. The massive trunk is very strong and there is potencial to used it for wood. Trees are very significant for their red plant sap which flowing from demaged trunk. The sap is very precious resource which is used in many areas of industry, for example pharmacy, traditional medicine, dye making etc. Determination of genetic affinity was based on an amplification of cpDNA template of individual Dracaena samples with primers for the studied regions. Data was obtained and evaluated by Multiple alignment program ClustalX and BioEdit after their sequencing. Evalueted data was used to create dendograms affinity. According the resulting phylogenetic tree we find out similarities and identified relationship of the monitored species of the genus Dracaena. The main purpose of research was to get answers to understand phylogenetic relationship between group of Dracaena forestry used trees. The thesis was made in cooperation of Department of Forest Botany, Dendrology and Geobiocoenology, Faculty of Forestry and Wood Technology of Mendel University in Brno.

As técnicas de DNA barcoding (código de barras de DNA) têm como objetivo principal a identificação taxonômica de organismos através da amplificação e análise de sequências de DNA curtas, padronizadas e previamente definidas. Apesar do sucesso relativo desta abordagem em animais usando um único locus, a aplicação deste método em plantas apresenta menor capacidade de identificar espécies usando uma única região gênica, levando a necessidade de utilização de múltiplos loci. Além disso, ainda há certo debate sobre qual região gênica seria mais apropriada para o DNA barcoding em plantas, embora as regiões plastidiais rbcL, matK e o espaçador intergênico trnH-psbA juntamente com o espaçador intergênico nuclear do RNA ribossomal (ITS) sejam as mais comumente utilizadas até então. As tribos Sisyrinchieae e Tigridieae da família Iridaceae foram testadas de acordo com diferentes métodos e marcadores indicados para o DNA barcoding em plantas. Os resultados indicaram uma alta universalidade para membros da tribo Sisyrinchieae, mas também revelaram uma capacidade de identificação de espécies considerada baixa. Apesar disto, os espaçadores ITS foram indicados como a melhor sequência para DNA barcoding em Sisyrinchieae. Em Tigridieae, problemas inerentes ao sequenciamento impediram a utilização dos ITS em nossas análises. Assim, apenas marcadores plastidiais foram utilizados na tentativa de identificar espécies, apresentando novamente resultados modestos. A região gênica que atingiu maior capacidade de identificação em Tigridieae foi o gene matK. A incapacidade de se alcançar maiores taxas de identificação provavelmente está relacionada à complexa história evolutiva apresentada pelos grupos em análise. Este trabalho forneceu o primeiro conjunto significativo de dados de DNA barcoding aplicados a dois importantes grupos de Iridaceae de considerável biodiversidade no Brasil. As tribos em análise apresentam espécies consideradas filogeneticamente próximas e são de difícil identificação devido a sua morfologia homogênea, principalmente em estado vegetativo, justificando plenamente o uso de métodos molecularespara a identificação taxonômica. / The main objective of DNA barcoding methods is the taxonomic identification of organisms by amplifying and analyzing short, standardized and previously defined DNA sequences. In spite of the relative success of this approach in animals using a single locus, the application of this method in plants has less ability to identify species using a single gene region, leading to the need of using multiple loci. Furthermore, there is still some debate concerning which gene region would be more suitable for DNA barcoding in plants, although the plastid regions rbcL, matK and the trnH-psbA intergenic spacer along with the nuclear intergenic spacer of ribossomal DNA (ITS) are the most commonly regions used thus far. The tribes Sisyrinchieae and Tigridieae of the family Iridaceae were tested according to different methods and markers used for DNA barcoding in plants. The results indicated a great universality for members of tribe Sisyrinchieae, but also showed a low ability to identify species. Nevertheless, ITS was imputed as the best sequence for DNA barcoding in Sisyrinchieae. In Tigridieae, problems inherent of ITS sequencing prevented its use in our analysis. Thus, only plastid markers were used in an attempt to identify species, showing modest results once again. The gene region that reached higher identification ability in Tigridieae was matK. The inability to achieve higher identification levels is probably related to the complex evolutionary history presented by the groups in question. This work provided the first large data set of DNA barcoding applied to two important groups of Iridaceae with significant biodiversity in Brazil. The tribes in question present species considered phylogenetically related and are difficult to identify due to their homogeneous morphology, especially in vegetative stage, fully justifying the use of molecular methods for taxonomic identification.

As técnicas de DNA barcoding (código de barras de DNA) têm como objetivo principal a identificação taxonômica de organismos através da amplificação e análise de sequências de DNA curtas, padronizadas e previamente definidas. Apesar do sucesso relativo desta abordagem em animais usando um único locus, a aplicação deste método em plantas apresenta menor capacidade de identificar espécies usando uma única região gênica, levando a necessidade de utilização de múltiplos loci. Além disso, ainda há certo debate sobre qual região gênica seria mais apropriada para o DNA barcoding em plantas, embora as regiões plastidiais rbcL, matK e o espaçador intergênico trnH-psbA juntamente com o espaçador intergênico nuclear do RNA ribossomal (ITS) sejam as mais comumente utilizadas até então. As tribos Sisyrinchieae e Tigridieae da família Iridaceae foram testadas de acordo com diferentes métodos e marcadores indicados para o DNA barcoding em plantas. Os resultados indicaram uma alta universalidade para membros da tribo Sisyrinchieae, mas também revelaram uma capacidade de identificação de espécies considerada baixa. Apesar disto, os espaçadores ITS foram indicados como a melhor sequência para DNA barcoding em Sisyrinchieae. Em Tigridieae, problemas inerentes ao sequenciamento impediram a utilização dos ITS em nossas análises. Assim, apenas marcadores plastidiais foram utilizados na tentativa de identificar espécies, apresentando novamente resultados modestos. A região gênica que atingiu maior capacidade de identificação em Tigridieae foi o gene matK. A incapacidade de se alcançar maiores taxas de identificação provavelmente está relacionada à complexa história evolutiva apresentada pelos grupos em análise. Este trabalho forneceu o primeiro conjunto significativo de dados de DNA barcoding aplicados a dois importantes grupos de Iridaceae de considerável biodiversidade no Brasil. As tribos em análise apresentam espécies consideradas filogeneticamente próximas e são de difícil identificação devido a sua morfologia homogênea, principalmente em estado vegetativo, justificando plenamente o uso de métodos molecularespara a identificação taxonômica. / The main objective of DNA barcoding methods is the taxonomic identification of organisms by amplifying and analyzing short, standardized and previously defined DNA sequences. In spite of the relative success of this approach in animals using a single locus, the application of this method in plants has less ability to identify species using a single gene region, leading to the need of using multiple loci. Furthermore, there is still some debate concerning which gene region would be more suitable for DNA barcoding in plants, although the plastid regions rbcL, matK and the trnH-psbA intergenic spacer along with the nuclear intergenic spacer of ribossomal DNA (ITS) are the most commonly regions used thus far. The tribes Sisyrinchieae and Tigridieae of the family Iridaceae were tested according to different methods and markers used for DNA barcoding in plants. The results indicated a great universality for members of tribe Sisyrinchieae, but also showed a low ability to identify species. Nevertheless, ITS was imputed as the best sequence for DNA barcoding in Sisyrinchieae. In Tigridieae, problems inherent of ITS sequencing prevented its use in our analysis. Thus, only plastid markers were used in an attempt to identify species, showing modest results once again. The gene region that reached higher identification ability in Tigridieae was matK. The inability to achieve higher identification levels is probably related to the complex evolutionary history presented by the groups in question. This work provided the first large data set of DNA barcoding applied to two important groups of Iridaceae with significant biodiversity in Brazil. The tribes in question present species considered phylogenetically related and are difficult to identify due to their homogeneous morphology, especially in vegetative stage, fully justifying the use of molecular methods for taxonomic identification.

As técnicas de DNA barcoding (código de barras de DNA) têm como objetivo principal a identificação taxonômica de organismos através da amplificação e análise de sequências de DNA curtas, padronizadas e previamente definidas. Apesar do sucesso relativo desta abordagem em animais usando um único locus, a aplicação deste método em plantas apresenta menor capacidade de identificar espécies usando uma única região gênica, levando a necessidade de utilização de múltiplos loci. Além disso, ainda há certo debate sobre qual região gênica seria mais apropriada para o DNA barcoding em plantas, embora as regiões plastidiais rbcL, matK e o espaçador intergênico trnH-psbA juntamente com o espaçador intergênico nuclear do RNA ribossomal (ITS) sejam as mais comumente utilizadas até então. As tribos Sisyrinchieae e Tigridieae da família Iridaceae foram testadas de acordo com diferentes métodos e marcadores indicados para o DNA barcoding em plantas. Os resultados indicaram uma alta universalidade para membros da tribo Sisyrinchieae, mas também revelaram uma capacidade de identificação de espécies considerada baixa. Apesar disto, os espaçadores ITS foram indicados como a melhor sequência para DNA barcoding em Sisyrinchieae. Em Tigridieae, problemas inerentes ao sequenciamento impediram a utilização dos ITS em nossas análises. Assim, apenas marcadores plastidiais foram utilizados na tentativa de identificar espécies, apresentando novamente resultados modestos. A região gênica que atingiu maior capacidade de identificação em Tigridieae foi o gene matK. A incapacidade de se alcançar maiores taxas de identificação provavelmente está relacionada à complexa história evolutiva apresentada pelos grupos em análise. Este trabalho forneceu o primeiro conjunto significativo de dados de DNA barcoding aplicados a dois importantes grupos de Iridaceae de considerável biodiversidade no Brasil. As tribos em análise apresentam espécies consideradas filogeneticamente próximas e são de difícil identificação devido a sua morfologia homogênea, principalmente em estado vegetativo, justificando plenamente o uso de métodos molecularespara a identificação taxonômica. / The main objective of DNA barcoding methods is the taxonomic identification of organisms by amplifying and analyzing short, standardized and previously defined DNA sequences. In spite of the relative success of this approach in animals using a single locus, the application of this method in plants has less ability to identify species using a single gene region, leading to the need of using multiple loci. Furthermore, there is still some debate concerning which gene region would be more suitable for DNA barcoding in plants, although the plastid regions rbcL, matK and the trnH-psbA intergenic spacer along with the nuclear intergenic spacer of ribossomal DNA (ITS) are the most commonly regions used thus far. The tribes Sisyrinchieae and Tigridieae of the family Iridaceae were tested according to different methods and markers used for DNA barcoding in plants. The results indicated a great universality for members of tribe Sisyrinchieae, but also showed a low ability to identify species. Nevertheless, ITS was imputed as the best sequence for DNA barcoding in Sisyrinchieae. In Tigridieae, problems inherent of ITS sequencing prevented its use in our analysis. Thus, only plastid markers were used in an attempt to identify species, showing modest results once again. The gene region that reached higher identification ability in Tigridieae was matK. The inability to achieve higher identification levels is probably related to the complex evolutionary history presented by the groups in question. This work provided the first large data set of DNA barcoding applied to two important groups of Iridaceae with significant biodiversity in Brazil. The tribes in question present species considered phylogenetically related and are difficult to identify due to their homogeneous morphology, especially in vegetative stage, fully justifying the use of molecular methods for taxonomic identification.

Herbal medicines are growing in popularity in the Western world and are becoming more stringently regulated under new EU legislation. Within the arena of herbal medicines, St. John’s Wort (SJW), Hypericum perforatum, is a top ten best seller with clinical evidence to support its use as an anti-depressant. A fundamental requirement of the new legislation is to prove the identity of the plant material in question. This is currently achieved via morphological and chemical methods, neither of which are ideal. A wide range of DNA based methods have been applied to this arena, standardisation is required to realise the potential of DNA based techniques. The DNA barcoding initiative aims to produce sequence data for all plant species, capable of species identification. The proposal is to use these data to design fast and effective DNA based methods of identification. For assay design, the putative barcode region nrITS was selected as a platform. Three assays were designed; • A PCR assay designed to hyper variable sequences within a barcode region. This assay is capable of distinguishing SJW from other closely related species. • A quantitative qPCR assay designed to measure total DNA and specific SJW DNA within a mixed sample. • A multiplex PCR incorporating fluorescently labelled primers, allowing amplicon detection by capillary electrophoresis. This assay identifies four separate Hypericum species, including SJW, with a mixed sample in one reaction. The suitability of the nrITS and three other barcode regions is assessed based on sequence data generated for 32 vouchered samples of different Hypericum species, and a Lithuanian sample set of 22 and 16 H. perforatum and H. maculatum samples respectively. The matK is currently unusable, the rbcL highly conserved, trnH-psbA problematically variable and the nrITS proved to be ideal for assay design.

Herbal medicines (HMs) have been used to supplement, maintain, and treat health conditions, and have inspired the development of many Western pharmaceuticals. Migrant South Asian (SA) communities in the UK have brought with them their own traditional forms of medicine, yet little is known about their current use of HMs in the UK. Consuming HMs alongside conventional Western medicines could affect pharmacological treatment and lead to herb-drug interactions; hence, healthcare professionals (HCPs) should be aware of their patients’ use of HMs. The import of HMs to the UK raises concerns over the quality, safety and regulation of HMs. Deoxyribonucleic acid (DNA) barcoding can be used to discriminate between different species, and identify contaminants and adulterants, thus can be used for the authentication of HMs. The South Asian Traditional Medicines (SATMED) questionnaire explored the knowledge and use of HMs by diasporic SA communities in the UK. It uncovered a vast range of HMs which were used by participants, where ingredients were sourced from, the concurrent use of herbal and Western medicines, and how minor ailments were treated. An online survey designed to investigate UK based practitioners’ views of HMs revealed that HCPs claimed to lack sufficient knowledge of HMs. HCPs said they needed more training on HMs to help them make better informed decisions. Tulsi (Ocimum tenuiflorum L.) was identified as a culturally and commercially valuable plant, which was used for molecular analysis. A variety of tulsi samples were collected for authentication: community samples from SA families in the UK, commercial samples, and referenced specimens. Both ITS and trnH-psbA regions were successfully used to distinguish between several Ocimum species, and identify a potential species substitution. This research represents the first time that DNA based methods have been used to authenticate medicinal plants species used by migrant SA communities living in the UK. The results of this multi-disciplinary study provide a unique contribution to the evolving discipline of ethnopharmacology.