Optimization of parameters for obtaining callus, suspension, and root cultures of meadowsweet (filipendula ulmaria) to isolate the largest number of biologically active substances with geroprotective properties / Otimização de parâmetros para obtenção de culturas de calos, suspensão e raiz de Ulmária (Filipendula ulmaria) para isolar o maior número de substâncias biologicamente ativas com propriedades geroprotetoras

The study of biologically active substances-secondary metabolites of plants that exhibit geroprotective properties is an actual and popular direction in medicine to prevent early aging. This work aims to select the cultivation parameters for obtaining in vitro cell cultures of meadowsweet containing the largest amount of biologically active substances (BAS) for their further extraction as candidate substances for geroprotectors. To specify the effectiveness of the selected cell culture cultivation parameters, biomass growth for callus and root cultures, growth index, specific growth rate, and viability for suspension cultures was carried out. The study results made it possible to select the nutrient media for the cultivation of cell cultures of meadowsweet. It has been found that the greater the antioxidant activity of the extracts, the greater the antimicrobial properties it exhibits. In this study, cell cultures in vitro and alcohol extracts from the plant Filipendula ulmaria were considered as raw materials rich in candidate substances for geroprotectors. According to the data obtained, the plant is rich in hydroxybenzoic and salicylic acids, spireoside, avicularin, and hyperoside.

O estudo de substâncias biologicamente ativas - metabólitos secundários de plantas que apresentam propriedades geroprotetoras - é uma tendência atual e popular no campo da medicina para a prevenção do envelhecimento precoce. O objetivo deste trabalho foi selecionar os parâmetros de cultivo para obtenção de culturas celulares in vitro de Ulmária contendo a maior quantidade de substâncias biologicamente ativas (SBA), para sua posterior extração como substâncias candidatas a serem geroprotetoras. Para especificar a eficácia dos parâmetros selecionados de cultivo em cultura de células, foi realizada a análise de crescimento de biomassa para culturas de calos e raízes, índice de crescimento, taxa de crescimento específica e viabilidade para culturas em suspensão. Os resultados do estudo possibilitaram a seleção do meio nutriente para o cultivo de células de Ulmária. Verificou-se que, quanto maior a atividade antioxidante dos extratos, maiores eram as propriedades antimicrobianas exibidas. Neste estudo, culturas celulares in vitro e extratos alcoólicos da planta Filipendula ulmaria foram considerados matérias-primas ricas em substâncias candidatas a serem geroprotetoras. De acordo com os dados obtidos, a planta é rica em ácidos hidroxibenzoico e salicílico, espirosídeo, avicularina e hiperosídeo.

Physiological and transcriptional responses to heat stress in a typical phenotype of Pinellia ternata / 中国天然药物

Pinellia ternata is an important medicinal plant, and its growth and development are easily threatened by high temperature. In this study, comprehensive research on physiological, cytological and transcriptional responses to different levels of heat stress were conducted on a typical phenotype of P. ternata. First, P. ternata exhibited tolerance to the increased temperature, which was supported by normal growing leaves, as well as decreased and sustained photosynthetic parameters. Severe stress aggravated the damages, and P. ternata displayed an obvious leaf senescence phenotype, with significantly increased SOD and POD activities (46% and 213%). In addition, mesophyll cells were seriously damaged, chloroplast thylakoid was fuzzy, grana lamellae and stroma lamellae were obviously broken, and grana thylakoids were stacked, resulting in a dramatically declined photosynthetic rate (74.6%). Moreover, a total of 16 808 genes were significantly differential expressed during this process, most of which were involved in photosynthesis, transmembrane transporter activity and plastid metabolism. The number of differentially expressed transcription factors in MYB and bHLH families was the largest, indicating that these genes might participate in heat stress response in P. ternata. These findings provide insight into the response to high temperature and facilitate the standardized cultivation of P. ternata.

DUS testing guidelines for new varieties of Chinese medicinal plants / 中国中药杂志

A rich diversity of wild medicinal plant resources is distributed in China, but the breeding of new plant varieties of Chinese medicinal plants started late and the breeding level is relatively weak. Chinese medicinal plant resources are the foundation for new varieties breeding, and the plant variety rights(PVP) are of great significance for the protection and development of germplasm resources. However, most Chinese medicinal plants do not have a distinctness, uniformity, and stability(DUS) testing guideline. The Ministry of Agriculture and Rural Affairs has put 191 plant species(genera) on protection lists, of which only 30 are medicinal species(genera). At the same time, only 29 of 293 species(genera) plants in the Protection List of New Plant Varieties of the People's Republic of China(Forest and Grass) belong to Chinese medicinal plants. The number of PVP applications and authorization of Chinese medicinal plants is rare, and the composition of variety is unreasonable. Up to now, 29 species(genera) of DUS test guidelines for Chinese medicinal plants have been developed. Some basic problems in the breeding of new varieties of Chinese medicinal plants have appeared, such as the small number of new varieties and insufficient utilization of Chinese medicinal plant resources. This paper reviewed the current situation of breeding of new varieties of Chinese medicinal plants and the research progress of DUS test guidelines in China and discussed the application of biotechnology in the field of Chinese medicinal plant breeding and the existing problems in DUS testing. This paper guides the further application of DUS to protect and utilize the germplasm resources of Chinese medicinal plants.

Application of tissue culture technology of medicinal plants in sustainable development of Chinese medicinal resources / 中国中药杂志

Chinese medicinal resources are the cornerstone of the sustainable development of traditional Chinese medicine industry. However, due to the fecundity of species, over-exploitation, and limitations of artificial cultivation, some medicinal plants are depleted and even endangered. Tissue culture, a breakthrough technology in the breeding of traditional Chinese medicinal materials, is not limited by time and space, and can allow the production on an annual basis, which plays an important role in the protection of Chinese medicinal resources. The present study reviewed the applications of tissue culture of medicinal plants in the field of Chinese medicinal resources, including rapid propagation of medicinal plant seedlings, breeding of novel high-yield and high-quality cultivars, construction of a genetic transformation system, and production of secondary metabolites. Meanwhile, the current challenges and suggestions for the future development of this field were also proposed.

Comportamiento fenológico ex situ de Lippia integrifolia "incayuyo" (Verbenaceae) de una población de base genética amplia / Ex situ phenological behavior of Lippia integrifolia "incayuyo" (Verbenaceae) from a population with a broad genetic base

Abstract: Lippia integrifolia "incayuyo" is an aromatic, sub - woody shrub used in popular medicine, aperitit drinks and compound herb s. Its choleretic, antispasmodic, biocidal, antibacterial and larvicidal activity has been proven. The objective of the work was to register the phenology of a sample of 70 genotypes from a population with a broad genetic base. The phenophases studied were : vegetative growth, flower bud, flowering and fruiting fortnightly for two years. The initiation, intensity and prolongation of the phenophases were evaluated. The moment of full bloom occurs during the second half of December. Taking this date as a refer ence, a differentiated beginning of flowering was evidenced. The results of two campaigns were compared, observing that 70% of the specimens had a similar behavor, standing out some genotypes for presenting an early flowering and longer duration. The recor ded variability suggests that much of it could be due to intrinsic factors of the plant, and therefore, feasible to be selected .

Resumen: Lippia integrifolia "incayuyo" es un arbusto aromático, subleñoso empleado en la medicina popular, bebidas aperitivas y yerbas compuestas. Se ha comprobado su actividad colerética, antiespasmódica, biocida, antibacteriana y larvicida. El objetivo del trabajo fue registrar la fenología d e una muestra de 70 genotipos de una población de base genética amplia. Las fenofases estudiadas fueron: crecimiento vegetativo, botón floral, floración y fructificación quincenalmente durante dos años. Se evaluó inicio, intensidad y prolongación de las fe nofases. El momento de plena floración ocurre durante la segunda quincena de diciembre. Tomando esta fecha como referencia, se evidenció un inicio de floración diferenciada. Se compararon los resultados de dos campañas, observando que el 70% de los ejempla res tuvo un compartimiento semejante, destacándose algunos genotipos por presentar una floración temprana y de prolongación superior. La variabilidad registrada sugiere que gran parte de ésta podría deberse a factores intrínsecos de la planta, y por ello, factible de ser seleccionados.

Molecular identification and efficacy analysis of herbs at Orussey Herbal Market, Phnom Penh, Cambodia / 中国中药杂志

Cambodia is rich in medicinal plant resources. One hundred and thirty-three medicinal material samples, including the hole herb, root, stem/branch, leaf, flower, fruit, seed, and resin, were collected from the Orussey Herbal Market in Phnom Penh, Cambodia, and then authenticated by ITS and psbA-trnH. A total of 46 samples were identified based on ITS sequences, belonging to 24 families, 40 genera, and 42 species. A total of 100 samples were identified by psbA-trnH sequences to belong to 42 families, 77 genera, and 84 species. A total of 103 samples were identified by two DNA barcodes. According to the morphological characteristics of the medicinal materials, 120 samples classified into 50 species, 86 genera, and 86 families were identified, and the majority of them were from Zingiberaceae, Fabaceae, and Acanthaceae. Such samples have been commonly used in traditional Cambodian medicine, Ayurvedic medicine, Unani medicine, traditional Chinese medicine, and ethnomedicine, but different medical systems focus on different functional aspects of the same medicinal material. The results of this study have demonstrated that DNA barcoding has a significant advantage in identifying herbal products, and this study has provided basic data for understanding the traditional medicinal materials used in Cambodia.

Genetic analysis of Penthorum chinense Pursh by improved RAPD and ISSR in China

Background: Penthorum chinense Pursh (P. chinense) is a well-known traditional Chinese medicine (TCM) plant, which has long been used for the prevention and treatment of hepatic diseases. This study aimed to genetically characterize the varieties of P. chinense from different geographic localities of China by random amplification of polymorphic DNA (RAPD)-PCR technique and verified with inter-simple sequence repeat (ISSR) markers. Results: The P. chinense samples were collected from nine different geographic localities. Previously improved RAPD and ISSR markers were utilized for genetic analysis using DNA amplification. The genetic relationship dendrogram was obtained by conducting cluster analysis to the similarity coefficient of improved RAPD and ISSR markers. Improved RAPD yielded 185 scorable amplified products, of which 68.6% of the bands were polymorphic, with an average amplification of 9.25 bands per primer. The ISSR markers revealed 156 alleles with 7.8 bands per primers, where 59.7% bands were polymorphic. Furthermore, the similarity coefficient ranges of RAPD and ISSR markers were 0.71­0.91 and 0.66­0.89, respectively. Conclusions: This study indicated that improved RAPD and ISSR methods are useful tools for evaluating the genetic diversity and characterizing P. chinense. Our findings can provide the theoretical basis for cultivar identification, standardization, and molecular-assisted breeding of P. chinense for medicinal use.

Genetic characterization and authentication of Gardenia jasminoides in different regions of China by using improved RAPD analysis.

Gardenia jasminoides is a common garden medicinal plant known for its anticancer, anti-inflammatory, anti-thrombic, anti-fibrotic, antiviral, hepatoprotective, lung-protective, renal-protective, retina-protective and neuroprotective activities. It is found in several regions of the world, including China, but information about its genetic characteristics is limited. Here, we employed an improved method of random amplified polymorphic DNA (RAPD) analysis (with increased RAMP time) to investigate the genetic link between G. jasminoides samples collected from six different regions of Southern China. Total 26 RAPD primers were selected randomly, among which 23 primers generated reproducible polymorphic amplification bands. A total of 174 bands were obtained, where each primer had amplified 5-13 bands with an average of 7.56 bands per primer. The band size ranged approximately 150-2200 bp. Cluster dendrogram was obtained based on the improved RAPD amplification profiles, which showed that the similarity coefficients among six varieties of G. jasminoides ranged 0.67-0.88. To our knowledge, this is the first report of genetic characterization of G. jasminoides using improved RAPD analysis, which may be useful for the preservation of genetic diversity and identification of Gardenia population.

In vitro regeneration and ploidy level analysis of Eulophia ochreata Lindl.

Various parameters including explant-type, medium compositions, use of phytohormones and additives were optimized for direct and indirect regeneration of E. ochreata, a medicinal orchid under threat. Protocorm-like-bodies (PLBs) proved to be the best explants for shoot initiation, proliferation and callus induction. Murashige and Skoog’s (MS) medium containing 2.5 mg L-1 6-benzylaminopurine (BAP), 1.0 mg L-1 kinetin (Kin) and additives (adenine sulfate, arginine, citric acid, 30 mg L-1 each and 50 mg L-1 ascorbic acid) was optimal for shoot multiplication (12.1 shoots and 7.1 PLBs per explant with synchronized growth), which also produced callus. Shoot number was further increased with three successive subcultures on same media and ~40 shoots per explant were achieved after 3 cycles of 30 days each. Additives and casein hydrolysate (CH) showed advantageous effects on indirect shoot regeneration via protocorm-derived callus. Optimum indirect regeneration was achieved on MS containing additives, 500 mg L-1 CH, 2.5 mg L-1 BAP and 1.0 mg L-1 Kin with 30 PLBs and 6 shoots per callus mass (~5 mm size). The shoots were rooted (70% frequency) on one by fourth-MS medium containing 2.0 mg L-1 indole-3-butyric acid, 200 mg L-1 activated charcoal and additives. The rooted plantlets were hardened and transferred to greenhouse with 63% survival rate. Flow-cytometry based DNA content analysis revealed that the ploidy levels were maintained in in vitro regenerated plants. This is the first report for in vitro plant regeneration in E. ochreata.

Medicinal plants used in Rondônia, Western Amazon, Brazil / Plantas medicinais usadas em Rondônia, Amazônia Ocidental, Brasil

This study refers to the use of medicinal plants by populations in the Western Amazon and provides information that can be used in phytochemical studies. It draws upon the traditional knowledge regarding the use of medicinal plants in five regions of the state of Rondônia, in the Brazilian Amazon, focusing on native species. The field research was carried out in five municipalities of the state of Rondônia: Ariquemes, Buritis, Candeias do Jamari, Cujubim and Itapoa do Oeste, characterized by primary economic sectors: agriculture, cattle farming, plant extraction and mineral exploration. Structured interviews were applied to 227 persons chosen because of their prestige in the communities in relation to the knowledge and use of medicinal plants, identifying the therapeutic purpose, parts of the plant used and methods of preparation. The species were taxonomically identified. The ethnobotanic knowledge (inferred by the number of uses of medicinal plants per person) was correlated with the Brazilian region of origin, age, and gender of the interviewees. According to the collected data, 34 botanical families and 53 native species were identified. Of the 53 species, only 7 occur exclusively in the Amazon Forest: Theobroma grandiflorum (Willd. ex Spreng.) K. Schum., Psidium densicomum Mart. ex DC, Piper cavalcantei Yunck., Pilocarpus microphyllus Stapf ex Wardlew., Euterpe oleracea Mart., Croton cajucara Benth., Baccharis altimontana G. Heiden. The most common disorders treated with the plants were kidney problems, influenza, generalized infections and inflammations, malaria and high blood pressure. Leaves were the most used parts in preparations. Barks, fruits, roots, flowers, stems, seeds, oils, buds, tubercles, and rhizomes were also mentioned. Thirteen forms of preparations were recorded, and infusion and decoction were the most used. Syrups, juices, flour, sap, oil and parts of the plant blended with milk, honey and coffee or flamed, macerated and in the form of poultice, were also found. Persons from the Southeast region had more information compared with persons from the other regions; persons in the age groups between 50-59, 60-69 and 70-79 had more information in comparison with the other age groups; and the female interviewees mentioned significantly more uses of medicinal plants than the male ones. The relatively small number of native Amazon species identified can be the result of the loss of knowledge about medicinal plants in the Amazon because of internal migration, extinction of local indigenous groups, increasing urbanization and consequent globalization of the lifestyles.

Este estudo se refere ao uso de plantas medicinais por populações da Amazônia Ocidental, fornece informações passíveis de utilização em estudos fitoquímicos e resgata o conhecimento tradicional a respeito do uso de plantas medicinais em cinco regiões do estado de Rondônia na Amazônia Brasileira com foco nas espécies nativas. A pesquisa de campo foi realizada nos cinco municípios de Rondônia: Ariquemes, Buritis, Candeias do Jamari, Cujubim e Itapoã do Oeste, caracterizados por atividades econômicas primárias: agricultura, pecuária e extrativismo vegetal e mineral. Entrevistas estruturadas foram aplicadas a 227 pessoas escolhidas por seu prestígio nas comunidades em relação ao conhecimento e uso de plantas medicinais, identificando a finalidade terapêutica, as partes das plantas utilizadas, e os métodos de preparação. As espécies foram taxonomicamente identificadas. O conhecimento etnobotânico (inferido a partir do número de citações por entrevistado) foi correlacionado com a região de origem, idade e gênero dos entrevistados. De acordo com os dados coletados, 34 famílias botânicas e 53 espécies nativas foram identificadas. Dentre as 53 espécies, apenas sete ocorrem exclusivamente na Amazônia: Theobroma grandiflorum (Willd. ex Spreng.) K.Schum., Psidium densicomum Mart. ex DC, Piper cavalcantei Yunck., Pilocarpus microphyllus Stapf ex Wardlew., Euterpe oleracea Mart., Croton cajucara Benth., Baccharis altimontana G. Heiden. As afecções mais comuns tratadas com plantas foram: problemas de fígado, gripe, infecções e inflamações generalizadas, malária, e hipertensão arterial. As folhas são as partes mais utilizadas nas preparações. Também foram mencionadas cascas, frutos, raízes, flores, caules, sementes, óleos, brotos, tubérculos e rizomas. Treze formas de preparação foram registradas sendo mais comuns as infusões e decoctos. Outras formas de preparação observadas foram xarope, suco, farinha, seiva, óleo, partes vegetais misturadas com leite, mel ou café, flambada, macerada e em forma de cataplasma. Pessoas da região Sudeste apresentaram maior número de informações do que as outras regiões; faixas etárias entre 50-59, 60-69 e 70-79 apresentaram maior número de citações que os outros grupos etários; e as mulheres mencionaram significativamente mais usos medicinais em comparação com os homens. O número relativamente pequeno de espécies nativas amazônicas identificadas pode ser um resultado da perda de conhecimento sobre plantas medicinais na Amazônia, devido à migração interna, extinção dos grupos indígenas locais, crescente urbanização e consequente globalização dos estilos de vida.

Estudo do sistema de reprodução da fava-d'anta (Dimorphandra mollis Benth. ) / Determination of the reproductive system of fava-d'anta (Dimorphandra mollis Benth. )

Entre as plantas nativas de uso medicinal do Cerrado brasileiro encontra-se a fava-d'anta (Dimorphandra mollis Benth) com alto potencial econômico por possuir inúmeras potencialidades medicinais e fitoquímicas. A indústria extrai dos frutos os princípios ativos rutina, quercetina, e ramnose, dentre outros, usados na fabricação de medicamentos e cosméticos, principalmente no exterior. O conhecimento do sistema reprodutivo da espécie é fundamental para sua conservação e manejo. O presente trabalho objetivou determinar as características morfométricas das flores e caracterizar o sistema reprodutivo da D. mollis em área de Cerrado inalterada antropomorficamente, no município de Olhos D'água - MG. Para as características morfométricas, diâmetro da flor, comprimento da flor, do ovário, e da antera, foram utilizadas cinco flores em pré-antese. Para a determinação do sistema reprodutivo utilizou-se a razão pólen:óvulo (P:O), em 40 flores. As flores da D. mollis apresentaram comprimento da flor de 3,00 mm, diâmetro da flor de 2,00 mm, comprimento do óvulo de 2,60 mm, comprimento da antera de 1,57 mm, e o número de óvulos e de anteras foram 20 e 5,8, respectivamente. A razão P:O foi 765,030, sendo que esse índice não é afetado pelas características morfométricas. O sistema reprodutivo da D. mollis foi classificado como alógama facultativa.

Among the native medicinal plants of the Brazilian Cerrado, we can find the fava-d'anta (Dimorphandra mollis Benth) with a high economic potencial due to its numerous possivbilities for medicinal and phytochemical use. The industry extracts form the fruits the active ingredients rutin, quercetin and rhamnose, among others, used to manufacture medications and cosmetic products, especially abroad. Knowing the reproductive system of the species is important for its conservation and management. This study aimed to determine the morphometric characteristics of flowers and characterize the reproductive system of D. mollis in a Cerrado area that was anthropomorphically unchanged, in the municipality of Olhos D'água - MG. For the morphometric characteristics, flower diameter, length of the flower, of the ovary and the anther, five pre-anthesis flowers were used. To determine the reproductive system, the pollen:ovule (P:O) ratio was used in 40 flowers. The D. mollis flowers presented a flower length of 3.00 mm, flower diameter of 2.00 mm, ovule length of 2.60 mm, anthers length of 1.57 mm, and the number of anthers and ovules were 20 and 5.8, respectively. The P:O ratio was 765,030, and this index is not affected by the morphometric characteristics. The reproductive system of D. mollis was classified as facultative allogamous.

Determinação de óleos essenciais de alfavaca (Ocimum gratissimum L. ), orégano (Origanum vulgare L. ) e tomilho (Thymus vulgaris L.) / Determination of essential oils of basil (Ocimum gratissimum L. ), oregano (Ocimum gratissimum L. ) and thyme (Thymus vulgaris L.)

O presente trabalho teve como objetivo a caracterização de plantas frescas e secas (comerciais) de alfavaca, orégano e tomilho, a obtenção dos óleos essenciais através do método de arraste a vapor e a quantificação dos compostos químicos por CG/EM. As plantas frescas e as secas comerciais foram submetidas às análises de umidade, extrato etéreo, proteína, fibra bruta, cinzas, extrato não nitrogenado, valor calórico, teor de óleo essencial e identificação dos compostos majoritários através da cromatografia gasosa-espectrometria de massas. Dentre a caracterização obtida os resultados na base seca mostraram-se promissores, sendo o teor de proteína e de cinzas na alfavaca seca comercial com 17,34 g 100 g-1 e 8,12 g 100 g-1, respectivamente; a fibra bruta no orégano seco comercial com 15,65 g 100 g-1; o extrato etéreo, o extrato não nitrogenado e o valor calórico no tomilho seco comercial com 9,30 g 100 g-1, 52,72 g 100 g-1 e 356,74 Kcal 100 g-1, respectivamente. Obteve-se o maior rendimento de óleo essencial na alfavaca seca comercial com 1,02%, enquanto a alfavaca fresca apresentou o menor rendimento, com apenas 0,13%. Na alfavaca fresca encontrou-se 87,38% de eugenol e 6,27% de timol, enquanto na alfavaca seca comercial observou-se redução no eugenol (71,12%) e aumento do timol (13,28%). No orégano fresco foram quantificados quatro picos o γ-terpineno (33,45%), 4-terpineol (25,59%), timol (14,21%) e carvacrol (2,30%). Já no óleo essencial de orégano seco comercial houve redução no γ-terpineno (28,73%) e aumento no 4-terpineol (27,58%), timol (19,71%) e carvacrol (3,67%). No óleo essencial do tomilho fresco foram quantificados três picos o borneol (66,66%), timol (13,41%) e linalol (3,24%). Por outro lado, no óleo essencial do tomilho seco comercial houve redução no borneol (37,90%) e aumento no timol (20,61%) e linalol (10,34%). Pode-se concluir que as folhas secas comerciais analisadas de alfavaca, orégano, e tomilho apresentam potencial para o enriquecimento dos alimentos ou para a obtenção dos óleos essenciais.

This study aimed to characterize commercial fresh and dry medicinal plants (basil, oregano and thyme), to obtain essential oil by the steam distillation method and to quantify chemical compounds by means of GC/MS. The fresh and dry plants were subjected to the following analyses moisture, ether extract, protein, crude fiber, ash, non-nitrogenous extract, caloric value, essential oil content and identification of major compounds by gas chromatography-mass spectrometry. Considering the obtained characterization, the following results on dry basis proved promising: protein and ash content in commercial dry basil with 17.34 g 100 g-1 and 8.12 g 100 g-1, respectively; crude fiber in commercial dry oregano with 15.65 g 100 g-1; ether extract, non-nitrogenous extract and caloric value in commercial dry thyme with 9.30 g 100 g-1, 52.72 g 100 g-1 and 356.74 Kcal 100 g-1, respectively. The highest essential oil yield was obtained for commercial dry basil with 1.02% and the lowest yield was obtained for fresh basil with only 0.13%. Chromatography indicated 87.38% eugenol and 6.27% thymol in fresh basil. For commercial dry basil, the chromatogram showed a reduction in eugenol (71.12%) and an increase in thymol (13.28%). Four peaks were quantified for fresh oregano the γ-terpinene (33.45%), 4-terpineol (25.59%), thymol (14.21%) and carvacrol (2.30%). For the essential oil of commercial dry oregano, there was a decrease in γ-terpinene (28.73%) and an increase in 4-terpineol (27.58%), thymol (19.71%) and carvacrol (3.67%). In the chromatogram of the essential oil of fresh thyme, three peaks were quantified: borneol (66.66%), thymol (13.41%) and linalool (3.24%). On the other hand, in the chromatogram of the essential oil of commercial dry thyme, there was a decrease in borneol (37.90%) and an increase in thymol (20.61%) and linalool (10.34%). It can be concluded that commercial dry leaves of basil, oregano and thyme are feasible to enrich foods or to obtain essential oils.

Propagação vegetativa de Brosimum gaudichaudii Tréc. (mama-cadela) por estacas de raízes / Vegetative propagation of Brosimum gaudichaudii Tréc. (mama-cadela) by root cuttings

Brosimum gaudichaudii Tréc. (mama-cadela) é uma planta medicinal nativa do Cerrado, utilizada na medicina tradicional. O objetivo deste trabalho foi verificar a possibilidade de propagação de mama-cadela por meio de estacas de raízes, sob o efeito de reguladores de crescimento e de diferentes substratos. No experimento 1 foram avaliados os efeitos da aplicação de ácido indol-butírico (AIB) e ácido naftaleno-acético (ANA) na concentração de 1000 mg L-1 e três substratos (S1- areia; S2 - 75 por cento de areia + 25 por cento de substrato comercial; S3 - 50 por cento de areia + 50 por cento de substrato comercial). No experimento 2 foram avaliadas 4 doses de AIB: 0 - testemunha; 250 mg L-1 (1,3426 mM ); 500 mg L-1 (2,6853 mM) e 1000 mg L-1 (5,3706 mM); e 4 doses de ácido naftaleno-acético (ANA): 0 - testemunha; 250 mg L-1 (1,2295 mM); 500 mg L-1 (2,458 mM); e 1000 mg L-1 (4,918 mM). O delineamento experimental utilizado foi de blocos ao acaso com três repetições de seis estacas por parcela. No experimento 1, os substratos compostos por areia e areia (75 por cento) + substrato comercial (25 por cento) proporcionaram os maiores Índices de pegamento. A aplicação de AIB (1000 mg L-1) proporcionou aumentos relativos de 30,8 por cento e 51,3 por cento, no IMP quando comparada com a testemunha e a aplicação de ANA, respectivamente. No experimento 2, observou-se resposta quadrática significativa da aplicação de AIB sobre o IMP. A dose de 500 mg L-1 (2,6853 mM) promoveu maior IMP. Não houve efeito significativo das doses de ANA para os parâmetros avaliados. Não houve efeito significativo relevante de substratos ou hormônios sobre os demais parâmetros avaliados. Estes resultados evidenciam o potencial de utilização de estacas de raiz de mama-cadela para a produção de mudas clonais desta espécie.

Brosimum gaudichaudii Tréc. (mama-cadela) is a medicinal plant native to Cerrado and largely used in traditional medicine. The aim of this study was to verify the propagation of mama-cadela by means of root cutting under the effect of plant growth regulators and different substrates. In experiment 1, the effects of indole butyric acid (IBA) and naphthylacetic acid (NAA) at the concentration of 1000 mg L-1 were evaluated together with three substrates (S1 - sand; S2 - 75 percent sand + 25 percent commercial substrate; S3 - 50 percent sand + 50 percent commercial substrate). In experiment 2, 4 IBA levels were evaluated: 0 - control; 250 mg L-1 (1.3426 mM ); 500 mg L-1 (2.6853 mM) and 1000 mg L-1 (5.3706 mM), as well as 4 NAA levels: 0 - control; 250 mg L-1 (1.2295 mM); 500 mg L-1 (2.458 mM) and 1000 mg L-1 (4.918 mM). The adopted experimental design was in randomized blocks with three replicates of six cuttings per plot. In experiment 1, substrates containing sand and sand (75 percent) + commercial substrate (25 percent) promoted the highest average rooting indexes. Application of IBA (1000 mg L-1) led to increases of 30.8 percent and 51.3 percent in the average rooting indexes compared to control and NAA application, respectively. In experiment 2, there was a significant quadratic response of IBA application on the average rooting index. The level of 500 mg L-1 (2.6853 mM) promoted the highest average rooting index. There was no significant effect of NAA levels for the evaluated parameters. Similarly, there was no significant effect of substrates or hormones on the remaining parameters evaluated. These results show the potential use of mama-cadela root cuttings for the production of clonal seedlings of this species.

Adaptações fisiológicas e anatômicas de melissa officinalis L. (Lamiaceae) cultivadas sob malhas termorrefletoras em diferentes intensidades luminosas / Physiological and morphological adaptations of melissa officinalis L. (Lamiaceae) cultivated under thermo-reflector shading nets at different luminous intensities

Objetivou-se, com a realização da pesquisa, avaliar modificações fisiológicas e anatômicas em plantas de melissa, cultivadas sob malhas termorrefletoras (Aluminet®), em diferentes níveis de sombreamento, visando conhecer a plasticidade fenotípica em resposta de adaptação a diferentes quantidades de luz. Os tratamentos foram caracterizados por plantas submetidas a pleno sol e a 20 e 60 por cento de intensidade luminosa, e arranjados conforme o delineamento inteiramente casualizado (DIC). As quantificações de clorofila foram feitas em quatro repetições, as medições das epidermes e parênquimas foram repetidas 15 vezes e utilizou-se 10 repetições para as avaliações das características de cloroplastos e grãos de amido destes. Plantas submetidas a 20 por cento de intensidade luminosa apresentaram maior quantidade de clorofila a e, portanto, maior razão clorofila a/b. Comparativamente, as folhas de melissa a pleno sol e a 60 por cento de luz apresentaram células da epiderme adaxial mais espessas, mas as células da epiderme abaxial mostraram características encontradas em folhas de sombra, ou seja, mais finas. Quanto maior a intensidade luminosa, maior o número de cloroplastos, porém, a pleno sol mostraram-se mais finos e com menor área. Os grãos de amido de plantas cultivadas sob ambientes sombreados tiveram maior área e ocuparam maior parte nos cloroplastos de plantas a 60 por cento de intensidade luminosa. Assim, plantas de melissa, quando submetidas ao sombreamento, tiveram plasticidade fenotípica.

The aim of this study was to evaluate physiological and anatomical modifications in lemon balm plants, cultivated under thermo-reflector nets (Aluminet®) at different levels of shading, in order to understand the phenotypic plasticity in adaptation response to different light quantities. The treatments were characterized by plants subjected to full sun and 20 and 60 percent of luminous intensity, and arranged in completely randomized design (CRD). The quantifications of chlorophylls were done in four replicates, the measurements of epidermis and parenchymas were repeated 15 times and 10 replicates were used to evaluate characteristics of chloroplasts and their starch grains. Plants subjected to 20 percent of luminous intensity showed higher quantity of chlorophyll a and, therefore, higher chlorophyll a/b ratio. Lemon balm leaves under full sun and 60 percent of light showed thicker adaxial epidermis cells, but the abaxial epidermis cells showed characteristics found in shaded leaves, i.e., they were slender. The higher the light intensity, the larger the number of chloroplasts; however, under full sun, they were slender and had smaller area. The starch grains of leaves grown under shaded environments showed larger area and, at 60 percent of luminous intensity, occupied the largest part of chloroplasts. Thus, lemon balm plants, subjected to shading conditions, showed phenotypic plasticity.

Siparuna guianensis Aublet (negramina): uma revisão / Siparuna guianensis Aublet (negramina): a review

As pesquisas realizadas a partir de recursos genéticos de espécies medicinais nativas somente serão aplicadas se o material genético estiver seguro quanto à sobrevivência e disponibilidade. O objetivo deste artigo foi realizar uma revisão bibliográfica adotando como critério a escolha de artigos publicados em conceituadas revistas nacionais e internacionais, que permitissem melhor entendimento dos aspectos etnobotânicos, ecológicos e químicos de importante planta medicinal e aromática: Siparuna guianensis Aublet. Como tem sido apontada como uma das espécies prioritárias de conservação para a região do cerrado brasileiro, os estudos da propagação devem ser incentivados para que este recurso genético esteja disponível às gerações futuras.

Studies with genetic resources of native medicinal species will only be applied if their genetic material is safe regarding survival and availability. The aim of this study was to do a literature review adopting as criterion the choice of papers published in renowned national and international journals to allow better understanding on ethnobotanical, ecological and chemical aspects of an important medicinal and aromatic plant, Siparuna guianensis Aublet. Since the latter has been considered one of the major species for the conservation of Brazilian cerrado, studies on its propagation should be encouraged in order to make this genetic resource available to future generations.

Antagonistic properties of some microorganisms isolated from Brazilian tropical savannah plants against Staphylococcus coagulase-positive strain

Endophytic microorganisms are relatively unstudied as potential sources of novel natural products for medical and commercial exploitation. The aim of this work was to investigate some Brazilian tropical savannah trees Cassia leptophylla and Prunus spp. in order to isolate the endophytic microorganisms associated with these plants. The samples were disinfected to eliminate the epiphytic population. Colonies were diluted and displayed as drops in media and growing colonies were inactivated. Staphylococcus coagulase-positive strain was used as indicator microorganism and subjected to the antibioses test. Data showed that the microorganisms isolated from Cassia leptophylla had no inhibition against Staphylococcus. On the other hand, microorganisms isolated from Prunus spp. leaves showed antibacterial activity and inhibited Staphylococcus when cultivated in peptone agar as well as in yeast extract agar. Investigation proceeds in order to classify the isolated microorganisms presenting bioactive substance and exploit the potential of the compounds produced to inhibit the indicator bacteria. Other bioactive properties will be investigated.

Molecular characterization of Ephedra species found in Pakistan

Ephedra, also known as “ma huang”, is a dioecious, drought- and frost-resistant, perennial, evergreen shrub with compelling medicinal value. The genus is represented by 42 species around the world, 9 of which were provisionally reported from Pakistan. Species of the genus have a controversial taxonomy due to their overlapping morphological features. Conventional tools alone are not sufficient for characterizing the species. The objective of present study was to assess the genetic variability present in different biotypes of Ephedra growing in Pakistan using molecular markers. A total of six genotypes collected from diverse geographic zones of Pakistan were used. The DNA of all genotypes was amplified using nine randomly amplified polymorphic DNA (RAPD) primers to study genetic variability at the molecular level. The dissimilarity coefficient matrix based on the data of 9 RAPD primers was used to construct a dendrogram which was then used to group the genotypes in clusters. Based on the dendrogram and dissimilarity coefficient matrix, the RAPD markers used here revealed a moderate to high level of genetic polymorphism (6 to 49%) among the genotypes. It was found that the collection of genotype accessions from Swat Valley in northwestern Pakistan was most distantly related to the other five collections. More molecular markers including functional genes and ribosomal spacer regions are suggested to find a better estimate of the genetic diversity present in Ephedra growing in Pakistan. The information provided here is useful for identifying valuable Ephedra variants which will be used for medicinal purposes and earning foreign currency.

A standardized protocol for genomic DNA isolation from Terminalia arjuna for genetic diversity analysis

For studying genetic diversity in natural populations of Terminalia, a medicinal plant, our attempts to isolate high quality DNA using several previously reported protocols and even modifications were unsuccessful. We therefore combined CTAB based isolation, and column based purification step, to isolate DNA from Terminalia arjuna. The DNA isolated using this standardized protocol was high in quality and suitable for restriction digestion and generation of random amplification of polymorphic DNA (RAPD) and amplified fragment length polymorphism (AFLP).

Atividade mutagênica de plantas medicinais / Mutagenic activity of medicinal plants

Os estudos de genotoxicidade com plantas têm crescido juntamente com o aumento do uso terapêutico e com o interesse de comprovação da eficácia das mesmas nas mais diversas finalidades farmacológicas.Isso deve-se ao fato de muitas das plantas utilizadas por um grande número de pessoas,apesar de possuírem propriedades farmacológicas, também podem causar alterações no DNA.Os riscos são ainda maiores quando o uso de tratamentos clínicos alternativos dá-se de forma não controlada, sem a devida atenção quanto a identificação correta da planta, a parte do vegetal a ser utilizada e a forma de preparo e administração.Nessa revisão são abordados aspectos da atividade mutagênica de algumas plantas medicinais.

Effects of Pterocaulon polystachyum DC. (Asteraceae) on onion (Allium cepa) root-tip cells

In Brazil leaf infusions of the plant Pterocaulon polystachyum DC (Asteraceae) are used to treat digestive problems. We used plants from six P. polystachyum populations to prepare fresh aqueous leaf infusions at the concentration normally used in Brazil (2.5 g L-1) and at twice (5 g L-1) and four times (10 g L-1) this concentration. We rooted onion bulbs in a water control or one of the infusions for 24 h and used the in vivo onion (Allium cepa) root-tip cell test to investigate the potential mutagenicity of the infusions by calculating the mitotic index for the control and the experimental treatments and comparing them using the Chi-squared test (chi2, p = 0.05). There was a highly significant decrease in the mitotic index of root-tip cells treated with infusion as compared to those exposed to water only. These results indicate that P. polystachyum infusions present cytotoxic and anti-proliferative activity and therefore have therapeutic potential.