Analysis of genetic diversity of Hyptis pectinata (L.) Poit. plants using ISSR markers.

Hyptis pectinata, popularly known as 'sambacaitá' or 'canudinho', is a medicinal and aromatic species widely used in the Brazilian Northeast. In Sergipe, the excessive extraction of natural resources may reduce the genetic variability of native plants. Thus, molecular markers have frequently been applied to the characterization of genetic diversity as the basis for germplasm conservation and breeding programs. The objective of the present study was to evaluate the genetic diversity of H. pectinata plants collected in different municipalities of the State of Sergipe using ISSR molecular markers. Thirty-four primers were tested, nine of which were selected for providing reproducible and analyzable amplification products, resulting in 67 polymorphic bands. The expected heterozygosity ranged from 0.32 to 0.45, with a mean of 0.39. Polymorphism information content was of 0.49, which classifies the markers as moderately informative. A dendrogram was constructed using unweighted pair group method with arithmetic mean, forming three clusters: Cluster I (79 plants); Cluster II (4 plants); and Cluster III (2 plants). Jaccard's similarity coefficients ranged from 0.06 to 0.98. The plants SAM-117 and SAM-119 presented greater similarity. Conversely, SAM-107 and SAM-171 were the most genetically distant. In general, H. pectinata plants collected in the State of Sergipe presented low to moderate genetic diversity.

Assessment of genetic diversity of a native population of Eplingiella fruticosa: a plant with therapeutic potential.

Eplingiella fruticosa (Salzm. ex Benth.) Harley & J.F.B. Pastore, ex Hyptis fruticosa Salzm. ex Benth. is an aromatic and medicinal plant of the family Lamiaceae, found mainly in regions with intense anthropic activity. Information on the genetic diversity of this species is scarce. However, it can be assessed using molecular markers that identify the level of diversity among phenotypically identical individuals. The present study aimed to characterize the genetic diversity of a native population of E. fruticosa from the State of Sergipe using ISSR molecular markers. Samples of 100 plants were collected in 11 municipalities of the State of Sergipe and analyzed using eight ISSR primers, resulting in 72 informative bands. The cluster analysis obtained using the neighbor joining method resulted in three groups: Group I consisted of 50 plants, mainly from the municipalities of Areia Branca, Estância, Japaratuba, Moita Bonita, Pirambu, and Salgado; Group II was formed by 21 plants, with nine representatives from the municipality of Itaporanga d'Ajuda and 13 representatives from other municipalities; Group III was composed by 29 plants, being represented mainly by the municipalities of Malhada dos Bois and São Cristóvão. The smallest genetic distance occurred between plants EPF94 and EPF96 (0.250), and the greatest distance occurred between plants EPF50 and EPF96 (0.9778). The Shannon index had a mean value of 0.42, and diversity was considered moderate. Heterozygosity had a mean value of 0.267 and was considered low. Polymorphic information content (0.253) was considered moderately informative. Genetic diversity of E. fruticosa plants was intermediate, and the results of the present study can assist in the conservation and use of the genetic resources of this species.

Genetic diversity of native populations of Croton tetradenius Baill. using ISSR markers.

Brazil has about 300 Croton species in different types of vegetation. Croton tetradenius Baill., which is endemic to the Northeast region and predominant in the Caatinga vegetation, stands out among the several species of this genus. Considering the importance of knowing the genetic variability of a species, the objective of this study was to analyze the genetic diversity of the genotypes of natural populations of C. tetradenius in the State of Sergipe, using ISSR molecular markers. Forty individuals were sampled in four natural populations of the State of Sergipe, Brazil. Thirteen primers were used for DNA amplification using ISSR-PCR, totaling 77 amplified fragments, of which 94.8% were polymorphic. Results of the cluster analysis obtained by the Jaccard's similarity index, using the UPGMA method, resulted in the formation of six distinct clusters. Analysis of molecular variance (AMOVA), used to estimate the genetic variability among populations, revealed significant genetic variance (P < 0.01) between and within the studied populations, and most of the genetic diversity was found (87%) within populations. According to the Jaccard's similarity index, none of the studied plants was genetically identical. CTE210 and CTE305 presented high similarity index (0.76), while CTE105 presented low similarity index (<0.16) with all related individuals. ISSR markers were efficient and allowed the formation of a molecular profile, and had sufficient polymorphism to estimate the genetic variability between the accessions of the studied populations.

Analysis of genetic diversity of a native population of Myrcia lundiana Kiaersk. plants using ISSR markers.

Myrcia lundiana Kiaersk. is a tree of the family Myrtaceae found in tropical and subtropical areas of the southern hemisphere that produces essential oil. The aim of this study was to characterize the genetic diversity of M. lundiana plants from a native population of Parque Nacional de Itabaiana, using inter-simple sequence repeat molecular markers. Thirty-five primers were tested, 20 of which were polymorphic, resulting in 135 polymorphic and informative bands. Results of the cluster analysis, obtained using the unweighted pair group method with arithmetic mean, grouped plants into three clusters: Cluster I - MLU001, MLU002, MLU003, MLU004, MLU005, MLU006, MLU018, MLU019, MLU020, MLU021, MLU022; MLU008, MLU011, MLU012, MLU014, MLU015, MLU017, MLU026, and MLU028; Cluster II - MLU007, MLU009, MLU010, MLU013, and MLU016; and Cluster III - MLU023, MLU024, MLU025, and MLU027. Jaccard similarity coefficients for pair-wise comparisons of plants ranged between 0.15 and 0.87. MLU014 and MLU015 presented low genetic diversity, with a similarity index of 0.87. Conversely, MLU007 and MLU019 presented high diversity, with a similarity index of 0.15. According to the structure analysis, three distinct clusters were formed. Genetic diversity of M. lundiana plants was intermediate, and expansion of its genetic diversity is necessary. MLU026 and MLU028 are the most suitable for selection in breeding programs, since they clearly represent all of the diversity present in these plants. Moreover, these results provide important information on the existing genetic variability, highlighting the importance of Parque Nacional de Itabaiana for the conservation of this species.

Genetic diversity of Lippia sidoides Cham. and L. gracilis Schauer germplasm.

The conservation of plants in germplasm banks ensures the characterization and availability of these resources for future generations. The present study used DNA markers to obtain genetic information about germplasm collections of Lippia sidoides and L. gracilis, which are maintained in an Active Germplasm Bank (AGB). Genetic variability of samples in the AGB was assessed using 12 combinations of amplified fragment length polymorphism (AFLP) primers (EcoRI/MseI). Twenty simple sequence repeat primers designed for L. alba were tested to determine their transferability in L. sidoides and L. gracilis. The AFLP markers generated 789 markers. The assessed loci exhibited a moderate Shannon diversity index (I = 0.42) in both species, suggesting that the conserved accessions possess an intermediate level of genetic diversity. Twelve microsatellite loci amplified satisfactorily, and nine loci were polymorphic in each species. A total of 23, 22, and 36 alleles, with an average of 2.5, 2.4, and 3.27 alleles per locus were identified for L. sidoides and L. gracilis accessions in the AGB, and Lippia sp sampled plants, respectively. Analyses of genetic structure permitted the identification of three different groups using both sets of markers, of which two were representative of L. sidoides. The information generated in this study may help to create, expand, and maintain collections of these species and may assist in genetic-breeding programs.

Genetic diversity analysis of Varronia curassavica Jacq. accessions using ISSR markers.

Varronia curassavica Jacq. is a medicinal and aromatic plant from Brazil with significant economic importance. Studies on genetic diversity in active germplasm banks (AGB) are essential for conservation and breeding programs. The aim of this study was to analyze the genetic diversity of V. curassavica accessions of the AGB of Medicinal and Aromatic Plants of the Federal University of Sergipe (UFS), using inter-simple sequence repeat molecular markers. Twenty-four primers were tested, and 14 were polymorphic and informative, resulting in 149 bands with 97.98% polymorphism. The UPGMA dendrogram divided the accessions into Clusters I and II. Jaccard similarity coefficients for pair-wise comparisons of accessions ranged between 0.24 and 0.78. The pairs of accessions VCUR-001/VCUR-503, VCUR-001/VCUR-504, and VCUR-104/VCUR-501 showed relatively low similarity (0.24), and the pair of accessions VCUR-402/VCUR- 403 showed medium similarity (0.78). Twenty-eight accessions were divided into three distinct clusters, according to the STRUCTURE analysis. The genetic diversity of V. curassavica in the AGB of UFS is low to medium, and it requires expansion. Accession VCUR-802 is the most suitable for selection in breeding program of this species, since it clearly represents all of the diversity present in the AGB.

Analysis of genetic diversity of Laeliinae (Orchidaceae) in the State of Sergipe using ISSR markers.

The Orchidaceae represent one of the largest and most diverse families on the planet. However, this family is constantly threatened by predators and by the advancement of urban centers over its natural habitats. The objective of this study was to use inter-simple sequence repeat markers to evaluate the genetic diversity between orchid accessions of the Laeliinae subtribe, which comprise part of the Orchidaceae study collection at the Department of Agronomic Engineering of the Federal University of Sergipe. DNA was extracted from each specimen by using an adapted 2% cetyltrimethyl ammonium bromide protocol. Similarity between individuals was calculated using the Jaccard method. Clustering was carried out by the unweighted pair group method with arithmetic mean method, with resampling and 10,000 bootstraps. Eighty-seven fragments were obtained, all of which were polymorphic, revealing high variability between accessions. The mean similarity was 35.77% between Encyclia sp individuals, and 35.90% between specimens of Cattleya tigrina. For Epidendrum secundum, a relationship between geographic and genetic distances was observed, and the accession collected in the southern part of the State of Sergipe (Serra de Itabaiana National Park) was more divergent than that of the other parts of the state. The data generated in this study will guide further research aimed at the ex situ conservation of these materials.

Molecular characterization of patchouli (Pogostemon spp) germplasm.

Patchouli [Pogostemon cablin (Blanco) Benth.] is an aromatic, herbaceous plant belonging to the Lamiaceae family native to Southeast Asia. Its leaves produce an essential oil regularly used by the perfume and cosmetics industries. However, since patchouli from the Philippines and India were described and named Pogostemon patchouli, there has been a divergence in the identity of these species. The objective of the current study was to study the genetic diversity of patchouli accessions in the Active Germplasm Bank of Universidade Federal de Sergipe using microsatellite and inter simple sequence repeat markers. The results of both types of molecular markers showed that there are two well-defined clusters of accessions that harbor exclusive alleles. It was observed that these two clusters are genetically distant, suggesting that they belong to two different species. Based on the results, two accessions were classified as Pogostemon heyneanus and the remaining accessions were classified as P. cablin.

Influence of storage time and temperature on the chemical composition of the essential oil of Hyptis pectinata L. Poit / Influência do tempo de armazenamento e da temperatura na composição química do óleo essencial de Hyptis pectinata L. Poit

ABSTRACT Hyptis pectinata, popularly known as “sambacaitá”, is a native medicinal herb used by local people for treating disease. Its pharmacological, antimicrobial, and medicinal properties have been reported in several studies. The essential oil of the leaves is rich in terpene compounds responsible for its properties. Because there are factors that can influence the chemical composition of essential oils, the objective of this study was to evaluate the influence of storage on Hyptis pectinata essential oil for 360 days at two different temperatures. The essential oil was distilled from dried leaves of seven-month-old plants using the hydrodistillation method, and the chemical composition was determined using GC-MS and GC/FID. The effects of storage on the essential oil Hyptis pectinata were tested over the course of a year (0, 15, 30, 60, 90, 120, 150, 180, 240, 300, and 360 days), using two temperatures: room (±32ºC) and freezer (-20ºC). The predominant compounds in the essential oil are β–caryophyllene, caryophyllene oxide, and germancrene D. The concentration of the chemical constituents of the essential oil varied depending on the storage temperature, and over the 360 days. The storage of the essential oil at room temperature over one year resulted in higher concentrations of β-elemene, α-copaene, germacrene D, caryophyllene oxide, and (E,E)-α-farnesene and lower content of α-humulene and β-caryophyllene, compared to the results obtained from storing the essential oil in a freezer. These results indicate that the essential oil should be stored in a freezer, which provides greater stability to the concentration of the chemical constituents.

RESUMO Hyptis pectinata, conhecida popularmente como Sambacaitá é uma erva medicinal nativa utilizada pela população local para o tratamento de enfermidades, cujas propriedades farmacológicas, antimicrobianas e medicinais foram relatadas em diversos estudos. O óleo essencial das folhas é rico em compostos terpênicos responsáveis pelas suas propriedades. Devido à existência de fatores que podem influenciar a composição química dos óleos essenciais, o objetivo deste trabalho foi avaliar a influência do armazenamento do óleo essencial Hyptis pectinata durante 360 dias em duas temperaturas diferentes. O óleo essencial foi destilado a partir de folhas secas de plantas de sete meses de idade, utilizando o método de hidrodestilação e a composição química foi determinada usando CG-EM e CG-DIC. Os efeitos do armazenamento no óleo essencial Hyptis pectinata foram testados ao longo de um ano (0, 15, 30, 60, 90, 120, 150, 180, 240, 300 e 360 dias), utilizando duas temperaturas: ambiente (±32ºC) e freezer (-20ºC). Os compostos predominantes no óleo essencial foram o β-cariofileno, óxido de cariofileno e germancreno D e houve variação na concentração dos constituintes químicos do óleo essencial, dependendo da temperatura de armazenamento, bem como ao longo dos 360 dias. O armazenamento do óleo essencial, à temperatura ambiente ao longo de um ano resultou em maiores concentrações de β-elemeno, α-copaeno, germacreno D, óxido de cariofileno e (E, E) -α-farneseno e menores teores de α-humuleno e β-cariofileno, quando comparado com os resultados obtidos a partir de armazenamento do óleo essencial no freezer. Estes resultados indicam que o óleo essencial deve ser estocado no freezer, o que propicia maior estabilidade na concentração dos constituintes químicos.

Molecular and chemical characterization of vetiver, Chrysopogon zizanioides (L.) Roberty, germplasm.

Due to the economic interests in vetiver, Chrysopogon zizanioides (L.) Roberty, molecular and chemical studies are essential to generate information for its sustainable exploitation. The aim of this study was to undertake a molecular and chemical characterization of vetiver accessions of the active germplasm bank of the Universidade Federal de Sergipe. The molecular characteristics of the accessions were studied using amplified fragment length polymorphism markers, with a total of 14 primer combinations that generated 442 loci, allowing us to observe that these accessions have similar genomes. The vetiver accessions were divided into three distinct groups, where accession UFS-VET005 was the most differentiated and accession UFS-VET004 had the lowest essential oil content (0.70%). The content of the chemical constituents of the essential oils was observed to vary, with a predominance of khusimol, which ranged from 18.97 to 25.02%. It was possible to divide the vetiver accessions into two groups based on chemical composition, and these groups do not correlate with the molecular grouping. Therefore, it is necessary to perform molecular and chemical analyses to characterize vetiver accessions.

Cross-species transferability of microsatellite markers in the genus Lippia.

The cross-species transferability of 20 microsatellite markers was tested in the genus Lippia. Eleven markers were polymorphic after screening 19 accessions of Lippia sidoides and Lippia gracilis maintained in the Active Germplasm Bank (AGB) from Universidade Federal de Sergipe. Additionally, 40 accessions of Lippia spp were collected in Sergipe to increase the germplasm bank. A total of 23, 22, and 36 alleles were identified, with an average of 2.3, 2.2, and 3.27 alleles per locus, respectively, for each group. The markers that were used were efficient tools to access genetic diversity in the germplasm bank and will be useful for further research aiming at the conservation and management of these important aromatic species.

Molecular characterization of bromeliads from northeast Brazil.

Bromeliaceae is an important botany family that includes many species with economic value; demand for members of this family is increasing. However, illegal collection frequently occurs, drastically reducing the species populations; thus, it is necessary to collect and store Bromeliaceae genetic material. In this study, we identified and quantified genetic variability of the Bromeliad family using dominant markers to create the first Germplasm Bank in the northeast region of Brazil. Molecular tools were used to characterize the collected accessions. The combination of 11 inter-simple sequence repeats and 13 random amplified polymorphic DNA markers were used to detect the genetic variability of wild bromeliad accessions.

Development and characterization of novel microsatellite markers in Hyptis pectinata (Lamiaceae).

A microsatellite-enriched library was constructed and a set of 19 SSR markers were developed to characterize a germplasm collection of Hyptis pectinata (L.) Poit., maintained at the Universidade Federal de Sergipe (UFS). Fifteen markers of 19 ranged from moderately to highly polymorphic. A total of 113 alleles were identified, with a mean of 7.52 alleles per locus. The mean HO and HE were 0.582 and 0.657, respectively. The primers developed were efficient tools for accessing the genetic diversity of the germplasm collection analyzed and may also be useful for other studies involving this species and other species in the genus Hyptis.

Cytotoxic effects of essential oils from three Lippia gracilis Schauer genotypes on HeLa, B16, and MCF-7 cells and normal human fibroblasts.

This study aimed to evaluate the chemical composition of the essential oils from three genotypes of Lippia gracilis Schauer (Verbenaceae) and investigate the cytotoxic activities of these oils. Essential oils were extracted from the leaves using a Clevenger-type apparatus, and chemical analysis was performed using a gas chromatograph coupled to a mass spectrometer and flame ionization detector. 3T3, MRC5, B16, HeLa, and MCF-7 cell lines were used to study the in vitro cytotoxicity of the essential oils, and the level of cell death was determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide test with three replicates. The cytotoxic activity was expressed as the concentration that inhibited 50% of cell growth. The main compound in the essential oil of LGRA-106 was thymol (40.52%), while LGRA-109 and LGRA-201 contained 45.84 and 32.60% carvacrol, respectively, as their major compound. The essential oils of L. gracilis showed cytotoxic activity against both normal and tumor cells at concentrations below 100 µg/mL; this demonstrated the antitumor potential of these essential oils, which should be further investigated.

Estabelecimento in vitro e aclimatização de Lippia alba (Mill. ) N. E. Brown / In vitro establishment and acclimatization of Lippia alba (Mill. ) N. E. Brown

O presente trabalho teve por objetivo estabelecer in vitro a espécie Lippia alba (Mill.) N. E. Brown e promover a aclimatização de mudas dessa espécie. Para isso, foi testada a influência de diferentes concentrações e tempos de imersão em hipoclorito de sódio na assepsia dos explantes. Segmentos nodais foram imersos em hipoclorito de sódio nas concentrações 0,4; 0,6; 0,8 e 1,0 % e nos tempos 8, 12 e 16 minutos. Após 30 dias avaliou-se a contaminação bacteriana (%), número de brotos, número de folhas, e a taxa de sobrevivência (%). A concentração de 1% de hipoclorito de sódio foi a mais eficiente no controle da contaminação. Ápices caulinares de L. alba foram estabelecidos in vitro em meio MS suplementado com diferentes doses de BAP (0; 0,5; 1,0; 1,5 mg L-1). Após 140 dias avaliou-se a contaminação (%), a taxa de sobrevivência (%), a oxidação (%) e o número de brotos. Os melhores resultados foram obtidos para a dose 1,5 mg L-1 deste regulador. Para a aclimatização foram testados quatro tipos de substratos: pó de coco + calcário (1 g L-1), Plantmax® + calcário (1 g L-1), vermiculita + calcário (1 g L-1) e pó de coco + Plantmax® + vermiculita (1:1:1) + calcário (1 g L-1). Avaliou-se a taxa de sobrevivência (%), comprimento da parte aérea (cm), comprimento da raiz (cm), massa fresca da parte aérea e da raiz (g), e a massa seca da parte aérea e da raiz (g). Os melhores resultados foram obtidos quando com o substrato comercial Plantmax®.

The present study aimed to establish the species Lippia alba (Mill.) N. E. Brown in vitro and promote the acclimatization of the seedlings of this species. Therefore, we tested the effect of different concentrations and immersiog times in sodium hypochloritetfor the disinfection of explants. Nodal segments were immersed in sodium hypochlorite at the different concentrations of ,.4; ,.6; ,.8 and ,.0% during the different times of 8, 12 and 16 minutes. After 30 days, bacterial contamination (%), number of shoots, number of leaves and survival rate (%) were evaluated. The 1% concentration of sodium hypochlorite was more effective in controlling contamination. Shoot apices of L. alba were established in vitro on MS medium supplemented with different concentrations of BAP (;, ,.5; ,.0; ,.5 mg L-1). After 140 days, we evaluated the contamination (%), survival rate (%), oxidation (%) and number of shoots. The best results were obtained when we used the dose of ,.5 mg L-1 of this regulator. For acclimatization, we tested four types of substrates: coconut powder + lime (1 g L-1), Plantmax® + lime (1 g L-1), vermiculite + lime (1 g L-1) and coconut powder + Plantmax® + vermiculite (1:1:1) + lime (1 g L-1). We evaluated the survival rate (%), shoot length (cm), root length (cm), fresh weight of shoot and root (g) and dry weight of shoot and root (g). The best results were obtained when using the substrate Plantmax®.

Development and characterization of microsatellite primers in Pogostemon cablin (Lamiaceae).

Microsatellite primers were developed and optimized for patchouli (Pogostemon cablin) to characterize the patchouli Active Germplasm Bank of Universidade Federal de Sergipe. Creation of a genomic library for patchouli enabled the design of 12 microsatellite primers. Six of these microsatellites were polymorphic, revealing two well-defined groups of individuals that possess exclusive alleles. The data allowed us to characterize the patchouli active Germplasm Bank, identify its genetic diversity, and provide new information for researching this species.

Óleos essenciais de plantas medicinais e aromáticas no controle do carrapato rhipicephalus microplus / Essential oils of medicine and aromatic plants not controlled by carrapato rhipicephalus microplus

O carrapato bovino Riphicephalus microplus (Acari: Ixodidae) constitui um dos grandes problemas enfrentado pelos pecuaristas de várias partes do mundo. No Brasil este ectoparasita causa grandes prejuízos à bovinocultura devido ao processo de hematofagia e pela transmissão de agentes patogênicos nos animais hospedeiros. Diante dessa situação os produtores normalmente aplicam carrapaticidas organossintéticos de forma indiscriminada. Este método normalmente resulta em: intoxicação dos animais e dos aplicadores, resíduos de carrapaticidas nos produtos de origem animal, resistência dos carrapatos aos carrapaticidas químicos e poluição ambiental. Uma alternativa para diminuir esses problemas é uso de biocarrapaticidas de origem botânica. Os pesticidas botânicos são produtos naturais derivados do metabolismo secundário das plantas. Esse metabolismo produz uma grande diversidade de compostos com ação carrapaticida que podem ser explorados através do uso de óleos essenciais ou como modelo para síntese de pesticidas sintéticos. A necessidade de métodos mais seguros, menos agressivos ao homem e ao meio ambiente, tem estimulado a busca de novos carrapaticidas a partir de produtos vegetais. Assim, acredita-se que o uso de óleos essenciais de plantas medicinais e aromáticas de uma forma isolada ou associada pode reduzir: a velocidade do desenvolvimento da resistência dos carrapatos aos carrapaticidas, os resíduos nos produtos de origem animal e a toxicidade para os animais, aplicadores e meio ambiente.

Micropropagação e aclimatização de gerânio (Pelargonium graveolens L. ) / Micropropagation and acclimatization of geranium (Pelargonium graveolens L. )

O gerânio (Pelargonium graveolens L.) é uma planta medicinal e aromática nativa do sul da África, cujo óleo essencial é amplamente empregado nas indústrias de perfumaria e cosmético, além de ser usado como terapêutico. Avaliou-se o efeito da luminosidade e dos reguladores de crescimento BAP e ANA na multiplicação in vitro e o uso de diferentes substratos na aclimatização de mudas micropropagadas. Foram avaliadas as concentrações 0; 0,5; 1,0 e 2,0 mg L-1 de BAP, 0; 0,1 e 0,5 mg L-1 de ANA e duas condições de luminosidade (ausência e presença de luz), em esquema fatorial 4x3x2; para a aclimatização foram utilizados quatro substratos, pó de coco + Biosafra® (3-12-6) (12 g L-1) + calcário (1 g L-1) (PCBC); pó de coco + Biosafra® (3-12-6) (12 g L-1) + calcário (1 g L-1) + vermiculita (1:1) (PCBCV 1:1); pó de coco + Biosafra® (3-12-6) (12 g L­-1) + calcário (1 g L-1) + vermiculita (PCBCV 2:1) e vermiculita e adição semanal de sais MS (VS). Para a micropropagação de P. graveolens a utilização de 1,3 mg L-1 de BAP e 0,5 mg L-1 de ANA é eficiente na regeneração direta de plantas de gerânio, sendo a condição de escuro a mais indicada por proporcionar o maior número de brotos por explante. Para a aclimatização das mudas de gerânio, os melhores resultados foram obtidos em mudas aclimatizadas no substrato vermiculita com adição semanal de sais de MS seguido do substrato PCBCV 1:1.

Geranium (Pelargonium graveolens L.) is a medicinal and aromatic plant native to the south of Africa and whose essential oil is widely used by perfume and cosmetic industries, as well as in therapeutics. The effect of luminosity and growth regulators BAP and NAA on in vitro multiplication was evaluated, together with the use of different substrates on the acclimatization of micropropagated seedlings. The evaluated concentrations were 0, 0.5, 1.0 and 2.0 mg L-1 BAP; 0, 0.1 and 0.5 mg L-1 NAA; and two luminosity conditions (absence and presence of light), in a 4 x 3 x 2 factorial arrangement. For acclimatization, four substrates were tested: coconut dust + Biosafra® (3-12-6) (12 g L-1) + limestone (1 g L-1) (CDBL); coconut dust + Biosafra® (3-12-6) (12 g L-1) + limestone (1 g L-1) + vermiculite (1:1) (CDBLV 1:1); coconut dust + Biosafra® (3-12-6) (12 g L-1) + limestone (1 g L-1) + vermiculite (CDBLV 2:1); and vermiculite with weekly addition of MS salts (VS). For P. graveolens micropropagation, the use of 1.3 mg L-1 BAP and 0.5 mg L-1 NAA is efficient for the direct regeneration of geranium plants, and the dark condition is the most suitable since it provides a larger number of shoots per explant. For the acclimatization of geranium seedlings, the best results were obtained with seedlings acclimatized in the substrate vermiculite with weekly addition of MS salts, followed by the substrate CDBLV 1:1.

Produção de mudas de dois genótipos de alecrim-de-tabuleiro (Lippia gracilis Schauer) em função de fertilizante mineral, calcário, substratos e recipientes / Seedling production of two "alecrim-de-tabuleiro" (Lippia gracilis Schauer) genotypes under mineral fertilizer, limestone, substrates and containers

Lippia gracilis Schauer popularmente conhecida como alecrim-de-tabuleiro possui moderada atividade antibacteriana, antimicrobiana e antiséptica. Objetivando avaliar o efeito de doses de fertilizante mineral, calcário, substratos e recipientes na produção de mudas de dois genótipos de alecrim-de-tabuleiro realizou-se experimentos com estacas de dois genótipos de L. gracilis. No experimento 1, foram utilizadas estacas apicais distribuídas em bandejas de poliestireno expandido de 128 células com o substrato pó-de-coco + areia (1:1), três repetições e oito estacas por repetição. O delineamento experimental foi em blocos ao acaso, em esquema fatorial 4x2x2, sendo quatro doses do fertilizante (6-24-12+micronutrientes) (1, 2, 3 e 4 g L-1), duas doses de calcário (0 e 1 g L-1) e dois genótipos (LGRA106 e LGRA201). Aos 35 após plantio, foram avaliadas a sobrevivência ( por cento), enraizamento ( por cento), comprimento de raiz (cm) e massa seca de raiz (mg). No experimento 2, foram utilizadas três repetições com oito estacas por repetição. O delineamento experimental foi em blocos ao acaso, em esquema de parcelas subdivididas onde foram testados nas parcelas dois recipientes (bandeja de poliestireno expandido de 128 alvéolos e tubetes de 110 cm³, na sub parcela foram testadas as combinações de quatro substratos (PC - pó de coco; PCA (1:1) - pó de coco + areia (1:1); PCA (2:1) - pó de coco + areia (2:1) e PCA (3:1) - pó de coco + areia (3:1) e dois genótipos (LGRA106 e LGRA201). Aos 35 dias após o plantio foram analisadas as mesmas variáveis do experimento 1, além de altura de parte aérea (cm) e massa seca de parte aérea (mg). A utilização de 1 g L-1 do fertilizante na ausência de calcário foi efetivo para sobrevivência de plantas e enraizamento de estacas de alecrim-de-tabuleiro. Com base nesses experimentos, concluiu-se que o genótipo LGRA106 é superior ao LGRA201 nas variáveis analisadas e recomenda-se a bandeja de poliestireno expandido para produção de mudas de L. gracilis via estaquia.

Commonly known as "alecrim-de-tabuleiro", Lippia gracilis Schauer (Verbenaceae) has moderate antibacterial, antimicrobial and antiseptic activity. This study aimed to evaluate the effect of doses of mineral fertilizer, limestone, substrates and containers on seedling production of two "alecrim-de-tabuleiro" genotypes, using cuttings. In experiment 1, apical cuttings were distributed on expanded polystyrene trays of 128 cells containing coir + sand (1:1), using three replicates of eight cuttings each. Experimental design was in randomized blocks, in a 4x2x2 factorial arrangement, i.e. four doses of fertilizer (6-24-12 + micronutrients) (1, 2, 3 and 4 g L-1), two doses of limestone (0 and 1 g L-1) and two genotypes (LGRA106 and LGRA201). At 35 days after planting, we evaluated survival ( percent), rooting ( percent), root length (cm) and root dry matter (mg). In experiment 2, three replicates of eight cuttings each were used. Experimental design was in randomized blocks, in a split-plot scheme. In the plots, we tested two containers (expanded polystyrene tray of 128 cells and 110 cm³ tubets). In the subplots, we tested four substrate combinations [coir, coir + sand (1:1), coir + sand (2:1) and coir + sand (3:1)] and two genotypes (LGRA106 and LGRA201). At 35 days after planting, the same variables of experiment 1 were evaluated, besides shoot length (cm) and shoot dry matter (mg). The use of 1 g L-1 fertilizer without limestone was effective for plant survival and cutting rooting. Based on these experiments, the genotype LGRA106 is superior to LGRA201 as to the evaluated variables and the expanded polystyrene tray is recommended for the production of L. gracilis seedlings through cuttings.

Hyptis pectinata essential oil: chemical composition and anti-Streptococcus mutans activity.

OBJECTIVES: The aim of the present study was to evaluate the anti-Streptococcus mutans activity of Hyptis pectinata essential oil, and present its promising potential against oral diseases. MATERIALS AND METHODS: The essential oil of H. pectinata was obtained by hydrodistillation from dried leaves and analyzed by GC / MS. The effectiveness of this essential oil regarding the antimicrobial activity against several S. mutans strains was investigated by the agar diffusion and microdilution methods, and chlorohexidine was used as a standard control. RESULTS: The H. pectinata essential oil exhibited considerable inhibitory effect against either all the clinical isolates obtained from patients' saliva or the ATCC strains tested, with minimum inhibitory and bactericidal concentrations of 200 microg ml(-1). The study also compared the efficiency of the emulsifying agents Tween 20, Tween 80, dimethyl sulfoxide and propylene glycol in H. pectinata essential oil when tested against S. mutans. The data obtained confirmed the better inhibitory effect of the oil when using all tested diluents, although Tween 80 seemed to be more suitable for emulsification. CONCLUSION: According to our results, H. pectinata essential oil can be considered a promising alternative to chlorhexidine for the control of oral bacteria-related diseases and hygiene.