Re-evaluation of the phylogenetic relationships and species delimitation of two closely related families (Lamiaceae and Verbenaceae) using two DNA barcode markers.

The families Lamiaceae and Verbenaceae comprise several closely related species that possess high morphological synapomorphic traits. Hence, there is a tendency of species misidentification using only the morphological characters. Herein, we evaluated the discriminatory power of the universal DNA barcodes (matK and rbcL) for 53 species spanning the two families. Using these markers, we inferred phylogenetic relationships and conducted species delimitation analysis using four delimitation methods: Automated Barcode Gap Discovery (ABGD), TaxonDNA, Bayesian Poisson Tree Processes (bPTP) and General Mixed Yule Coalescent (GMYC). The phylogenetic reconstruction based on the matK gene resolved the relationships between the families and further suggested the expansion of the Lamiaceae to include some core Verbanaceae genus, e.g., Gmelina. The rbcL marker using the TaxonDNA method displayed high species delimitation resolutions, while the ABGD, GMYC, and bPTP generated different number of Operational Taxonomic Units/genetic clusters. Our results underscored the efficiency of the matK and rbcL genes as reliable markers for resolving phylogenetic relationships and species delimitation of both families, respectively. The current study provides insights into the DNA barcode applications in these families, at the same time contributing to the current understanding of genetic divergence patterns in angiosperms.

Pharmaceutical development of tablets containing a spray-dried optimized extract from Lippia origanoides H. B. K.: influence of excipients and toxicological assessment

ABSTRACT Lippia origanoides is a honey shrub which has showed hypotensive potential assessed by in vivo studies. The aim of this work is the development of a pharmaceutical formulation composed by an optimized extract obtained from aerial parts of L. origanoides. The quantification of the naringenin marker in the dry extract and tablets developed was performed, as well as the assessment of the oral acute toxicity in rats. The hydroalcoholic extract of L. origanoides was spray-dried with the addition of colloidal silicon dioxide (Lo-HAE/CSD), and then applied in the preparation of eight different lots of tablets. The influence of the diluent (cellulose or babassu mesocarp), the presence of binder, and the percentage of lubricant, as well as organoleptic and physicochemical characteristics were screened. For the quantification of the marker content both in Lo-HAE/CSD and in the tablets, an analytical curve of the naringenin standard was fitted, and the samples were then analyzed in UFLC. The toxicological assessment was performed in female Wistar rats according to the Acute Toxic Class Method from OECD. The developed tablets produced meet acceptable macroscopic characteristics, and the presence of babassu as diluent provided improved physicochemical properties. The best content of Lo-HAE/CSD in the tablet (100.27%) was identified for the lot containing babassu, composed by 1.0% magnesium stearate, without PVP binder in its formulation. Moreover, Lo-HAE/CSD showed no signs of toxicity. Therefore, the babassu mesocarp powder is a promising pharmaceutical excipient for the development of herbal tablets containing the Lippia origanoides extract.

Potencial do óleo essencial de erva-luísa (Aloysia citriodora Palau) no controle de Fusarium sp. in vitro / Essential oil potential herb-luisa (Aloysia citriodora Palau) in the control of Fusarium sp. in vitro

RESUMO O presente trabalho teve como objetivo principal avaliar o potencial do uso do óleo essencial de Aloysia citriodora no controle in vitro de Fusarium sp., isolado de plântulas de beterraba infectadas com o mesmo. O trabalho foi realizado por meio de dois experimentos: um sobre o efeito do óleo essencial no crescimento micelial, e outro sobre o efeito na germinação de conídios do fungo. No primeiro trabalho, avaliou-se em delineamento inteiramente ao acaso, o efeito das concentrações 0,0155%; 0,0315%; 0,0625%; 0,125%; 0,250% e 0,500% do óleo essencial de A. citriodora em placas de Petri® com meio de cultivo BDA, mais a testemunha, com meio BDA puro. Cada placa foi considerada uma repetição, as quais foram incubadas a 24ºC±1ºC e submetidas a fotoperíodo de doze horas. Avaliou-se o crescimento radial do patógeno em função do crescimento micelial do patógeno quando este atingia as bordas da primeira placa. No segundo experimento, as mesmas concentrações foram testadas, nas mesmas condições de incubação, no entanto, em lâminas de microscopia com meio BD. Utilizou-se delineamento inteiramente ao acaso, sendo considerada como unidade experimental cada lâmina utilizada. Avaliou-se 24 horas após a incubação, em microscópio óptico, a germinação dos primeiros vinte conídios visualizados a partir do canto esquerdo superior para o direito. Os resultados do segundo experimento foram expressos em porcentagem de germinação de conídios. Os resultados obtidos permitiram concluir que este óleo essencial possui efeito fungistático e fungicida sobre o crescimento micelial e na germinação de conídios de Fusarium sp.. Além disso este efeito é maior em função do aumento da concentração do óleo essencial.

ABSTRACT This study aimed to evaluate the potential use of Aloysia citriodora essential oilisolated from beet seedlings infected with it. The study was performed by conducting two experiments evaluating the effect of the essential oil on mycelial growth and fungus conidia germination. In the first study, in a completely randomized design, the effect concentrations (0.0155%; 0.0315%; 0.0625%; 0.125%; 0.250% and 0.500%) of essential oil of A. citriodorawas evaluated on Petri® dish with a PDA cultivation medium, plus the control, with half pure PDA. Each plate was taken as a repeat, and incubated at 24°C±1°C and a photoperiod of twelve hours. The radial growth of the pathogen, when the first plate was hit by the mycelial growth of the pathogen on its edges was evaluated. In the second experiment, the same concentrations were tested under the same incubationconditions, however, on microscope slides with half PD medium. The design was used completely randomized, each microscope slide used was considered as one experimental unit. Germination of the first 2 conidia strains, viewed from the upper left to the right was evaluated24 hours after incubation, using an optical microscope. The results of the second experiment were expressed as a percentage of conidia germination. The results obtained showed that this essential oil has fungistatic and fungicidal effect on the mycelial growth and at the conidia germination of Fusarium sp., which has a greater effect with increasing concentrations.

Population genetic structure in Phyla scaberrima from Mexico and Colombia assessed by AFLP markers and implications for conservation.

Phyla scaberrima (Verbenaceae) is a herbaceous perennial species that is distributed from Mexico (center of origin) to Colombia, growing in forest and swamp edges or grasslands from sea level up to an altitude of 1800 m. The chemical properties and uses in popular medicine have drastically affected the population size of this species. In this study, we investigated genetic variability in populations of P. scaberrima using AFLP markers. Three AFLP primer combinations rendered a total of 997 markers in a sample of 131 individuals from five populations, including two populations from Mexico and three from Colombia. The average percentage of polymorphic loci, gene diversity and Shannon-Wiener index were 46.62, 0.0695, and 0.119, respectively. Analysis of molecular variance showed that the distribution of the genetic variability within populations (85.41%) was higher than between groups (8.11%) and between populations (6.48%). Principal coordinate analysis and Bayesian analysis for the K number of clusters showed that the individuals were dispersed in five (K= 5) clusters. The low levels of genetic diversity observed in these populations demonstrated that the populations from Mexico and Colombia need urgent management to recover their genetic variability.

Anesthetic activity and bio-guided fractionation of the essential oil of Aloysia gratissima (Gillies & Hook.) Tronc. in silver catfish Rhamdia quelen.

This work aimed to determine the efficacy of the essential oil of A. gratissima as anesthetic for silver catfish, and to perform the bio-guided fractionation of essential oil aiming to isolate compounds responsible for the noted effects. Fish were submitted to anesthesia bath with essential oil, its fractions and isolated compounds to determine time of anesthetic induction and recovery. Eugenol (50 mg L(-1)) was used as positive control. Essential oil of A. gratissima was effective as an anesthetic at concentrations of 300 to 900 mg L(-1). Fish presented involuntary muscle contractions during induction and recovery. The bio-guided fractionation of essential oil furnished E-(-)-pinocamphone, (-)-caryophyllene oxide, (-)-guaiol and (+)-spathulenol. E-(-)-pinocamphone caused the same side effects observed for essential oil. (-)-Caryophyllene oxide, (-)-guaiol and (+)-spathulenol showed only sedative effects at proportional concentrations to those of the constituents in essential oil. (+)-Spathulenol (51.2 mg L(-1)) promoted deep anesthesia without side effects. A higher concentration of (+)-spathulenol, and lower or absent amounts ofE-(-)-pinocamphone could contribute to increase the activity and safety of the essential oil of A. gratissima. (+)-Spathulenol showed potent sedative and anesthetic activities in silver catfish, and could be considered as a viable compound for the development of a new anesthetic.

Evidences that human disturbance simplify the ant fauna associated a Stachytarpheta glabra Cham. (Verbenaceae) compromising the benefits of ant-plant mutualism.

Interaction among species, like ants and plants through extrafloral nectaries (EFNs), are important components of ecological communities' evolution. However, the effect of human disturbance on such specific interactions and its ecological consequences is poorly understood. This study evaluated the outcomes of mutualism between ants and the EFN-bearing plant Stachytarpheta glabra under anthropogenic disturbance. We compared the arthropod fauna composition between two groups of twenty plant individuals, one in an area disturbed by human activities and one in a preserved area. We also check the plant investment in herbivory defense and the consequential leaf damage by herbivore. Our results indicate that such disturbances cause simplification of the associated fauna and lack of proper ant mutualist. This led to four times more herbivory on plants of disturbed areas, despite the equal amount of EFN and ant visitors and low abundance of herbivores. The high pressure of herbivory may difficult the re-establishment of S. glabra, an important pioneer species in ferruginous fields, therefore it may affect resilience of this fragile ecological community.

Evidences that human disturbance simplify the ant fauna associated a Stachytarpheta glabra Cham. (Verbenaceae) compromising the benefits of ant-plant mutualism / Evidências de que os impactos antrópicos simplificam a fauna de formiga associada a Stachytarpheta glabra (Verbenaceae) comprometendo os benefícios do mutualismo formiga-planta

Interaction among species, like ants and plants through extrafloral nectaries (EFNs), are important components of ecological communities’ evolution. However, the effect of human disturbance on such specific interactions and its ecological consequences is poorly understood. This study evaluated the outcomes of mutualism between ants and the EFN-bearing plant Stachytarpheta glabra under anthropogenic disturbance. We compared the arthropod fauna composition between two groups of twenty plant individuals, one in an area disturbed by human activities and one in a preserved area. We also check the plant investment in herbivory defense and the consequential leaf damage by herbivore. Our results indicate that such disturbances cause simplification of the associated fauna and lack of proper ant mutualist. This led to four times more herbivory on plants of disturbed areas, despite the equal amount of EFN and ant visitors and low abundance of herbivores. The high pressure of herbivory may difficult the re-establishment of S. glabra, an important pioneer species in ferruginous fields, therefore it may affect resilience of this fragile ecological community.

As interações entre espécies, como por exemplo formigas e plantas através de nectários extraflorais (NEFs), são importantes componentes na evolução das comunidades. Entretanto, pouco é conhecido sobre os efeitos dos impactos antrópicos em interações específicas e suas consequências ecológicas. Este estudo avaliou os resultados do mutualismo entre formigas e NEF em Stachytarpheta glabra em área impactada pela atividade humana. Nós comparamos a composição e estrutura da fauna de artrópodes, em quarenta plantas de dois grupos, um impactado por atividades humanas e o outro preservado. Nós também avaliamos o investimento da planta em defesas contra herbívoros e os danos foliares causados por herbívoros. Nossos resultados indicam que os distúrbios causam a simplificação da fauna associada e a ausência de uma formiga mutualista anti-herbívoros. Isto leva a quatro vezes mais herbivoria nas plantas da área impactada, a despeito da mesma quantidade encontrada para NEF e formigas visitantes. A grande pressão de herbivoria pode dificultar o reestabelecimento de S. glabra, uma espécie pioneira importante nos campos ferruginosos, dessa forma, afetando a resiliência dessa comunidade ecológica ameaçada.

Biomassa e teor de óleo essencial em Aloysia triphylla (l'hérit) Britton submetida a diferentes níveis de reposição hídrica e à variação sazonal das condições ambientais / Biomass and essential oil content in Aloysia triphylla (L'Herit) Britton subjected to different levels of water reposition and season alchanges of environmental conditions

RESUMOA variação nos elementos meteorológicos ao longo do ano, configurando as quatro estações, proporciona diferentes condições de crescimento e desenvolvimento das plantas, alterando o metabolismo e a constituição dos tecidos vegetais. Assim, o presente trabalho teve o objetivo de determinar a influência da sazonalidade em parâmetros fenométricos e na produção de óleo essencial em Aloysia triphylla (L'Hérit) Britton, submetida a diferentes níveis de reposição hídrica cultivada em ambiente protegido. As avaliações foram realizadas no dia que marcou a metade de cada estação do ano (verão, outono, inverno e primavera), sendo coletado todo o material vegetativo 20 cm acima do solo. A reposição hídrica foi baseada na evapotranspiração de referência (ETo) para o ambiente externo em função dos dados de entrada da equação de Penman-Monteith a qual apresenta parametrizações para dados obtidos em estações meteorológicas em padrões oficiais, sendo assim aplicados 125, 100, 75 e 50% da ETo em turno de rega de dois dias. A produção de massa fresca, seca e de parte aérea, bem como a área foliar, apresentaram maiores valores na estação primavera, sendo o inverno e o verão responsáveis pelos menores valores. O maior teor de óleo essencial foi observado nas estações de menor produção de biomassa, ou seja, verão e inverno. A reposição hídrica que proporcionou maior produção de material vegetativo foi a de 125% da ETo; no entanto, o maior teor de óleo foi observado nas menores reposições hídricas (75 e 50 % da ETo, respectivamente). A produção de óleo foi maior na primavera, a qual não sofreu influência da reposição hídrica ao longo do ano.

ABSTRACTThe variation in weather elements throughout the year, setting the four seasons, provides different conditions for growth and development of plants, altering the metabolism and the structure of plants` tissues. This study aimed to determine the influence of seasonality on fenometrics parameters and on the oil production in Aloysia triphylla (L'Herit) Britton, subjected to different levels of water availability cultivated in greenhouse conditions. The evaluations were made on the day that marked the middle of each season (summer, autumn, winter and spring), and all plant material were collected 20 cm above ground. The water reposition was based on the evapotranspiration reference (ETo) to the external environment being applied 125, 100, 75 and 50% of ETo in an irrigation interval of two days. The production of fresh, dry and aerial part mass, as well as leaf areamass, showed higher values in the spring season, while winter and summer were responsible for lower values. The highest content of essential oil was observed at seasons with lower biomass production. The water availability that provided higher production of plant material was the one with 125% of ETo, however, the highest oil content was observed at lower water availability (75% and 50% of ETo, respectively). The oil production was higher in the spring and was not affected throughout the year by the water availability.

Uso popular de espécies medicinais da família Verbenaceae no Brasil / Popular use of medicinal species of the Verbenaceae family in Brazil

RESUMO Com o presente trabalho objetivou-se realizar um levantamento bibliográfico acerca das espécies da família Verbenaceae utilizadas na medicina popular nas diversas regiões do Brasil. Os trabalhos acadêmicos utilizados para elaboração desta pesquisa foram extraídos de revistas indexadas junto ao Scielo publicadas no período de 2000 a 2012, sendo também realizada busca no Google acadêmico. Os dados foram compilados e organizados em ordem alfabética por espécies, seguidas do nome popular, forma de uso, parte usada, indicações e área de estudo. Foram encontrados um total de 85 publicações indicando 55 espécies distribuídas em oito gêneros. A espécie com maior número de citações foi Lippia alba (Mill.) N.E.Br., com 56 registros. Das plantas descritas na literatura, a folha foi o órgão mais citado como parte usada e o chá, o modo de preparo mais utilizado. Dentre as categorias de indicações medicinais, as plantas que mereceram destaque foram às relacionadas ao tratamento das doenças do sistema respiratório e digestório. Constatou-se que o Rio de Janeiro foi o estado que contou com o maior número de publicações na área. O uso de plantas medicinais está cada vez mais presente entre as diversas comunidades. Com isso, as informações contidas neste trabalho servirão para auxiliar na elaboração de banco de dados sobre o modo de uso das plantas pertencentes à família Verbenaceae com potencial medicinal encontradas nos Estados brasileiros.

ABSTRACT The current work aimed to conduct a literature review about the Verbenaceae family species used in popular medicine in different regions of Brazil. The academic papers employed for the preparation of this research were taken from journals indexed by the Scielo published from 2000 to 2012, and a research in Google scholar tool was also performed. The data were compiled and organized alphabetically by species, followed by the popular name, manner of use, employed part, indications and study area. A total of 85 publications indicating 55 species were found and distributed into eight genera. The species with the highest number of citations was the Lippia alba (Mill.) N.E.Br., with 56 records. From the plants described in the literature, the leaf was the most cited part of the vegetables, and the tea was the most widely used preparation method. Among the categories of medicinal recommendations, the highlighted plants were the ones related to the treatment of diseases of the respiratory and the digestive systems. It was observed that Rio de Janeiro was the state in which the largest number of publications in the area was concentrated. The use of medicinal plants is increasingly present among several communities. Thus, the information in this work will be applied to assist in database development on how to use the plants belonging to the family Verbenaceae with medicinal potential found in Brazilian states.

[Natural regeneration response in mangroves of the gulf of Urabá (Colombia) to the environmental and intra-annual climate variability]. / Respuesta de la regeneración natural en manglares del Golfo de Urabá (Colombia) a la variabilidad ambiental y climática intra-anual.

The natural regeneration process allows the mangrove forests remain over time. Both, biological and physical factors can affect the establishment and early stages along the development of trees. This study examined the response of natural regeneration of mangroves in the Turbo River delta and El Uno bay (Urabá Gulf, Colombia) to intra-annual environmental variability. We quantified mortality, survival and recruitment of seedlings of three mangrove species, seasonally during one year, in 72 semi-permanent sub-plots of 1 m2. In the sub-plots, the total height and the diameter at the base of the stem of all mangrove seedlings with basal diameter less than 2.5cm were measured. Damage by herbivores was also recorded to each seedling. While Laguncularia racemosa recorded the highest rates of mortality, Rhizophora mangle showed the highest survival rate during the study period, although Avicennia germinans dominated the natural regeneration. Through a Redundancy Analysis these processes were associated to environmental variables such as: Direct Site Factor-DSF (sunlight), sediments input rate, herbivory, distances from mangroves to the river, to inner lakes, and to the coastline. These variables explained 43% of the natural regeneration variation; sedimentation rate was the most important variable, while light was very representative for the R. mangle survival. Based on historical records of precipitation, Turbo River flow rate and associated sediment loads, it was established that during the highest precipitation peak, the survival of all species decreases and during the dry season, when the conditions of flooding and sediments input were lower, it was improved. The results indicated that the sediments input rates and sunlight play an important role in the survival of natural regeneration of evaluated mangrove species.

Respuesta de la regeneración natural en manglares del Golfo de Urabá (Colombia) a la variabilidad ambiental y climática intra-anual / Natural regeneration response in mangroves of the Gulf of Urabá (Colombia) to the environmental and intra-annual climate variability

The natural regeneration process allows the mangrove forests remain over time. Both, biological and physical factors can affect the establishment and early stages along the development of trees. This study examined the response of natural regeneration of mangroves in the Turbo River delta and El Uno bay (Urabá Gulf, Colombia) to intra-annual environmental variability. We quantified mortality, survival and recruitment of seedlings of three mangrove species, seasonally during one year, in 72 semi-perma-nent sub-plots of 1m². In the sub-plots, the total height and the diameter at the base of the stem of all mangrove seedlings with basal diameter less than 2.5cm were measured. Damage by herbivores was also recorded to each seedling. While Laguncularia racemosa recorded the highest rates of mortality, Rhizophora mangle showed the highest survival rate during the study period, although Avicennia germinans dominated the natural regeneration. Through a Redundancy Analysis these processes were associated to environmental variables such as: Direct Site Factor-DSF (sunlight), sediments input rate, herbivory, distances from mangroves to the river, to inner lakes, and to the coastline. These variables explained 43% of the natural regeneration variation; sedimentation rate was the most important variable, while light was very representative for the R. mangle survival. Based on historical records of precipitation, Turbo River flow rate and associated sediment loads, it was established that during the highest precipitation peak, the survival of all species decreases and during the dry season, when the conditions of flooding and sediments input were lower, it was improved. The results indicated that the sediments input rates and sunlight play an important role in the survival of natural regeneration of evaluated mangrove species.

En los bosques de mangle, tanto factores biológicos como físicos, afectan el establecimiento y los estados tempranos de desarrollo de los árboles y determinan la distribución espacial de las especies. Se evaluó la respuesta de la regeneración natural de los manglares del delta del río Turbo y la bahía El Uno a la variabilidad ambiental y climática intra-anual. Se cuantificaron la mortalidad, la sobrevivencia y el reclutamiento de las plántulas de tres especies, estacionalmente durante un año. Avicennia germinans dominó la regeneración natural, Rhizophora mangle presentó la mayor la supervivencia y Laguncularia racemosa la mayor mortalidad. Estos procesos se asociaron principalmente con el aporte de sedimentos aluviales, la disponibilidad de luz, la herbivoría, la distancia al río, a las lagunas internas y a la línea de costa, las cuales explicaron el 43% de la variación en la regeneración. La sedimentación fue la variable más importante para el reclutamiento de A. germinans, mientras la luz fue muy representativa para la supervivencia de R. mangle. Con base en registros históricos de precipitación, caudales y carga de sedimentos, se estableció que en la época con mayor precipitación se presenta un descenso en la supervivencia, y en la época seca se favorece el balance entre el reclutamiento y la mortalidad. Los resultados indicaron que la precipitación, el aporte de sedimentos y la luz desempeñan un papel importante en el proceso de la regeneración de las especies evaluadas.

Chemical composition and antimicrobial activity of the essential oil from the leaves and flowers of Aloysia gratissima / Composição química e atividade antimicrobiana do óleo essencial de folhas e flores de Aloysia gratissima

Volatile oils from leaves and flowers of Aloysia gratissima were investigated for their chemical composition and antimicrobial activity against the bacteria Bacilus subtilis, Staphylococcus aureus, Salmonella choleraesuis, Pseudomonas aeruginosa, Streptococcus pneumoniae and the Candida albicans yeast. The minimum inhibitory concentrations (MIC) of the oils were determined by the micro-dilution method, while the chemical composition was determined by GC-MS (gas chromatography mass spectrometry). The fresh leaves and inflorescence were subjected to hydrodistillation for 120 min using a Clevenger-type apparatus, and the essential oil was tested against microorganisms. High concentrations of sesquiterpenes were observed for the inflorescence, and monoterpenes were observed for the leaves. The main compounds of the inflorescence essential oil were E-caryophyllene, germacrene B, guaiol and bulnesol, while in the leaves the main compounds were trans-pinocamphone, trans-pinocarveyl acetate, and guaiol. The essential oil from the leaves showed an effect against P. aeruginosa and S. pneumonia, and the essential oil of the inflorescence showed an effect against P. aeruginosa, S. pneumonia, and Candida albicans.

O óleo essencial de folhas e de flores de Aloysia gratissima foi avaliado quanto à composição química e ação antimicrobiana contra as bactérias Bacilus subtilis, Staphylococcus aureus, Salmonella choleraesuis, Pseudomonas aeruginosa, Streptococcus pneumoniae, e a levedura Candida albicans. A concentração mínima inibitória (MIC) dos óleos essenciais foi determinada pelo método da microdiluição e a composição química determinada por CG-EM (Cromatografia Gasosa acoplada a Espectrômetro de Massas). Folhas e inflorescências frescas foram hidrodestiladas por 120 minutos em aparelho Clevenger sendo o óleo essencial testado contra microorganismos. Para as flores foi observada maior concentração de sesquiterpenos, enquanto que as folhas apresentaram maior concentração de monoterpenos. Os principais constituintes do óleo essencial da flor foram: E-cariofileno, germacreno B, guaiol e bulnesol; e das folhas foram: trans-pinocamfona, acetato de trans-pinocarveol e guaiol. O óleo essencial da folha mostrou atividade contra P. aeruginosa e S. pneumoniae, e o óleo essencial da flor mostrou atividade contra P. aeruginosa, S. pneumoniae e Candida albicans.

Evolution of morphological traits in Verbenaceae.

PREMISE OF THE STUDY: A new infrafamilial circumscription of the Verbenaceae with eight tribes: Casselieae, Citharexyleae, Duranteae, Lantaneae, Neospartoneae, Petreeae, Priveae, and Verbeneae, has been recently proposed, on the basis of molecular phylogenetic studies. Two genera, Dipyrena and Rhaphithamnus, remain unplaced. The aim of this work is to reconstruct the evolutionary history of morphological characters traditionally employed in the classification of the Verbenaceae, with special attention to tribes Verbeneae and Lantaneae. METHODS: Twenty-one characters, related to habit and vegetative morphology, inflorescence and floral morphology, ovary and fruit morphology, as well as chromosome number, were optimized over a molecular phylogeny of Verbenaceae. KEY RESULTS: All tribes are supported by at least one morphological trait except tribes Duranteae and Citharexyleae. Suffrutescent habit, sessile flowers, and four cluses are synapomorphies for tribe Verbeneae. Gynoecium with short style and entire stigma are synapomorphic traits for tribe Lantaneae. Sessile flowers and unicarpellate ovaries are morphological synapomorphies for the new tribe Neospartoneae. Suffrutescent habit is a synapomorphic trait for tribe Priveae. Homothetic pleiobotrya and absence of the adaxial staminode are synapomorphic traits for tribe Casselieae. Undivided fleshy fruits are probably a synapomorphic trait for tribe Petreeae. Putative plesiomorphies for the ancestor of the Verbenaceae are discussed as well as synapomorphic traits within other Verbenaceae clades. CONCLUSIONS: Many of the characters traditionally employed in classification have proven to be very homoplastic, or have been shown not to support relationships within the family. Moreover, traditional assumptions concerning character polarity have in some cases been shown to be incorrect.

[DNA bacording of Verbenaceae medicinal plant by using ITS2 and psbA-trnH region].

OBJECTIVE: To evaluate the ability of several hotspot candidate sequence of DNA barcodes for identifying medicinal plant species in Verbenaceae. METHOD: Using universal primers, three chloroplast sequences, psbA-trnH, rbcL, matK, two nuclear ribosomal DNA ITS2 and ITS were amplified and sequenced. PCR amplification and sequencing efficiency, intra- and inter-specific variation, barcoding gap and identification efficiency (with BLAST 1 and Nearest Distance methods) were used to evaluate these loci. RESULT: The rate of successful amplification using matK was too low to further analyze, and the rate of successful amplification and sequencing using psbA-trnH, ITS, and ITS2 and rbcL sequence was 83.6%, 83.6%, 96.4%, 98.2%, respectively. The rate of successful identification using psbA-trnH, ITS, and ITS2 was 100% at the species level except that rbcL was 77.8%, 75.9% for 55 samples belonging to 32 species, but ITS2 did better in intra- and inter-specific variation, barcoding gap than the other loci. The rate of successful identification of ITS2 was 89.5%, 87.6% even when joining the date of 165 samples from GenBank. CONCLUSION: It proposes that the combination of ITS2 and psbA-trnH senquence is promising for the identification of the species in Verbenaceae.

Growth rate and ring width variability of teak, Tectona grandis (Verbenaceae) in an unmanaged forest in East Timor.

Teak (Tectona grandis) is one of the most valuable timbers in international trade and an important species for tropical forestry. Teak is found on the island of East Timor but no information is available on teak growth from this region. A pure stand planted in 1940-50 in the North of East Timor and left unmanaged was studied. Fifteen trees were sampled in October-November 2003 and stem discs taken at three height levels of its height (1.7m, 9.5m and 18.7m), and cores were collected at DBH. Transverse surfaces of the discs and cores were polished for ring identification. Core cross sections were first digitized and disc cross sections were observed under the microscope. Three randomly selected radii were analyzed in each disc. Ring width measurement and ring counting were done using image analysis software. The distinction between heartwood and sapwood was performed macroscopically by colour difference, and heartwood radius and sapwood width were measured. The relationship between stem and heartwood radius was studied for each disc and heartwood percentage by radius was determined. Radial ring width curves are presented for the different axial positions within the stem, and ring width variability was analyzed. Growth rates were calculated and age-radius relationships were estimated using cumulative growth curves. Growth rings were large and well defined in the juvenile phase, reflecting the specie's fast-growing character. The year-to-year variation of ring width showed a similar pattern among trees. Mean ring width ranged between 4.3-7.3mm for the first 20 years and 3.3-5.1mm for 30 to 45 years. Pith eccentricity was evident in the lower part of the stem and ring wedging occurred. On average, heartwood represented 84% of the radius and sapwood contained 6 to 11 rings. The age-related variation of ring width and the occurrence in the lower part of the tree stems of eccentricity and wedging rings, highlights the importance of appropriate stand management, particularly regarding basal density distribution over time, whenever optimized timber production is envisaged.

Growth rate and ring width variability of teak, Tectona grandis (Verbenaceae) in an unmanaged forest in East Timor

Teak (Tectona grandis) is one of the most valuable timbers in international trade and an important species for tropical forestry. Teak is found on the island of East Timor but no information is available on teak growth from this region. A pure stand planted in 1940-50 in the North of East Timor and left unmanaged was studied. Fifteen trees were sampled in October-November 2003 and stem discs taken at three height levels of its height (1.7m, 9.5m and 18.7m), and cores were collected at DBH. Transverse surfaces of the discs and cores were polished for ring identification. Core cross sections were first digitized and disc cross sections were observed under the microscope. Three randomly selected radii were analyzed in each disc. Ring width measurement and ring counting were done using image analysis software. The distinction between heartwood and sapwood was performed macroscopically by colour difference, and heartwood radius and sapwood width were measured. The relationship between stem and heartwood radius was studied for each disc and heartwood percentage by radius was determined. Radial ring width curves are presented for the different axial positions within the stem, and ring width variability was analyzed. Growth rates were calculated and age-radius relationships were estimated using cumulative growth curves. Growth rings were large and well defined in the juvenile phase, reflecting the specie’s fast-growing character. The year-to-year variation of ring width showed a similar pattern among trees. Mean ring width ranged between 4.3-7.3mm for the first 20 years and 3.3-5.1mm for 30 to 45 years. Pith eccentricity was evident in the lower part of the stem and ring wedging occurred. On average, heartwood represented 84% of the radius and sapwood contained 6 to 11 rings. The age-related variation of ring width and the occurrence in the lower part of the tree stems of eccentricity and wedging rings, highlights the importance of appropriate stand management, particularly regarding basal density distribution over time, whenever optimized timber production is envisaged.

La madera de teca (Tectona grandis L.f.) es una de las más valiosas en el comercio internacional y una especie importante para la silvicultura tropical. La teca se encuentra en la isla de Timor Leste, pero no existe información disponible sobre su crecimiento en esta región. Se estudió una plantación pura no manejada que fue establecida entre 1940 y 1950 en el Norte de Timor Leste. Entre Octubre- Noviembre 2003 se cosecharon 15 árboles y al tronco se les extrajeron discos a tres niveles de altura (1.7m, 9.5m y 18.7 m), y se recolectaron núcleos a la altura del pecho (DAP). El conteo y la medición de los anillos se realizaron mediante el software de análisis de imagen. La distinción entre el duramen y la albura se realizó macroscópicamente por la diferencia de color, y se midieron el radio del duramen y el ancho de la albura. Para cada disco se estudió la relación entre el tallo y radio del duramen y el porcentaje del duramen. Las curvas de variación del ancho de los anillos se presentan para las diferentes posiciones axiales dentro del tronco y se analizó la variabilidad del ancho de los anillos. Las tasas de crecimiento fueron calculadas y las relaciones de radio con la edad se calcularon usando las curvas de crecimiento acumulado. Los anillos de crecimiento eran grandes y bien definidos en la fase juvenil, lo que refleja el carácter de crecimiento rápido de esta especie. La variación en la anchura de los anillos año a año mostró un patrón similar entre los árboles. La anchura media del anillo osciló entre 4.3-7.3mm para los primeros 20 años y 3.3-5.1mm para los de 30 a 45 años. La excentricidad de la médula fue evidente en la parte inferior del tallo y se observó la formación de anillos sobrepuestos. En promedio, el duramen representó el 84% del radio y la albura mostró entre 6 y 11 anillos. La variación del ancho de los anillos con la edad, la presencia de excentricidad en la parte inferior del árbol y los anillos sobrepuestos pone de relieve la importancia del manejo forestal con particular atención a la distribución de densidad basal en el tiempo, cuando se pretende la producción optimizada de la madera.

Folk medicinal uses of Verbenaceae family plants in Bangladesh.

Folk medicinal practitioners form the first tier of primary health-care providers to most of the rural population of Bangladesh. They are known locally as Kavirajes and rely almost solely on oral or topical administration of whole plants or plant parts for treatment of various ailments. Also about 2% of the total population of Bangladesh are scattered among more than twenty tribes residing within the country's borders. The various tribes have their own tribal practitioners, who use medicinal plants for treatment of diseases. The objective of the present survey was to conduct an ethnomedicinal survey among the Kavirajes and tribal practitioners to determine which species of plants belonging to the Verbenaceae family are used by the practitioners. The Verbenaceae family plants are well known for constituents having important bio-active properties. The present survey indicated that 13 species belonging to 8 genera are used by the folk and tribal medicinal practitioners of Bangladesh. A comparison of their folk medicinal uses along with published reports in the scientific literature suggests that the Verbenaceae family plants used in Bangladesh can potentially be important sources of lead compounds or novel drugs for treatment of difficult to cure debilitating diseases like malaria and rheumatoid arthritis.

Phenotypic divergence during the invasion of Phyla canescens in Australia and France: evidence for selection-driven evolution.

Rapid adaptive evolution has been advocated as a mechanism that promotes invasion. Demonstrating adaptive evolution in invasive species requires rigorous analysis of phenotypic shifts driven by selection. Here, we document selection-driven evolution of Phyla canescens, an Argentine weed, in two invaded regions (Australia and France). Invasive populations possessed similar or higher diversity than native populations, and displayed mixed lineages from different sources, suggesting that genetic bottlenecks in both countries might have been alleviated by multiple introductions. Compared to native populations, Australian populations displayed more investment in sexual reproduction, whereas French populations possessed enhanced vegetative reproduction and growth. We partitioned evolutionary forces (selection vs. stochastic events) using two independent methods. Results of both analyses suggest that the pattern of molecular and phenotypic variability among regions was consistent with selection-driven evolution, rather than stochastic events. Our findings indicate that selection has shaped the evolution of P. canescens in two different invaded regions.

An empirical demonstration of using pentatricopeptide repeat (PPR) genes as plant phylogenetic tools: phylogeny of Verbenaceae and the Verbena complex.

The pentatricopeptide repeat (PPR) gene family, with hundreds of members in land plant genomes, has been recognized as a tremendous resource for plant phylogenetic studies based on publicly available genomic data from model organisms. However, whether this appealing nuclear gene marker system can be readily applied to non-model organisms remains questionable, particularly given the potential uncertainties in designing specific primers to only amplify the locus of interest from the sea of PPR genes. Here we demonstrate empirically the use of PPR genes in the family Verbenaceae and the Verbena complex. We also lay out a general scheme to design locus-specific primers to amplify and sequence PPR genes in non-model organisms. Intergeneric relationships within the family Verbenaceae were fully resolved with strong support. Relationships among the closely related genera within the Verbena complex and among some species groups within each genus were also well resolved, but resolution among very closely related species was limited. Our results suggest that PPR genes can be readily employed in non-model organisms. They may be best used to resolve relationships in a spectrum from among distantly related genera to among not-so-closely related congeneric species, but may have limited use among very closely related species.