Chemical characterization and antimicrobial activity of essential oils from Croton grewioides Baill. accessions on the phytopathogen Xanthomonas campestris pv. campestris.

Croton grewioides Baill. is an aromatic species with proven bioactive properties. Considering the potential of the species, the aim of this study was to chemically characterize and evaluate the in vitro antibacterial activity of the essential oils of C. grewioides on Xanthomonas campestris pv. campestris. The essential oils of the accessions of C. grewioides were extracted by the hydrodistillation method and analyzed by gas chromatography - mass spectrometry. For determination of the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the essential oils and of the compound eugenol, the microdilution method was used at concentrations that ranged from 125 to 4000 µg.mL-1. Streptomycin sulfate was used as a positive control (12.5 to 100 µg.mL-1). Growth kinetics and the membrane permeability trial were evaluated for the concentrations 2×, 1×, 1/2×, 1/4×, and 1/8× MIC of the essential oil CGR-108. The major compounds identified in the essential oils were eugenol, methyl eugenol, and methyl chavicol. The essential oil of the accession CGR-108 had a lower MIC (> 500 and < 1000 µg.mL-1) and MBC equal to <2000 µg.mL-1. For eugenol, MIC was obtained with contractions >250 and < 500 µg.mL-1 and MBC with >500 µg.mL-1 and < 1000 µg.mL-1. A loss of cell viability of the bacteria was observed after 30 min of exposure to the essential oil of the accession CGR-108 at the concentrations of 2× and 1× MIC, which was proven by the fluorescence intensity with propidium iodide. The essential oils of Croton grewioides Baill. and the compound eugenol show antibacterial potential on Xanthomonas campestris pv. campestris.

Antibacterial activity of Lippia alba, Myrcia lundiana and Ocimum basilicum essential oils against six food-spoiling pathogenic microorganisms / Actividad antibacteriana de los aceites esenciales de Lippia alba, Myrcia lundiana y Ocimum basilicum contra seis microorganismos patógenos que estropean los alimentos

The aim of this study was to undertake a screening experiment on essential oils (EO) of Myrcia lundiana, Ocimum basilicum and Lippia alba against six food-spoiling pathogenic bacteria. Seventy-two (72) samples were initially analyzed fo antimicrobial activity based on the agar diffusion test. The minimum inhibitory (MIC) and bactericidal (MBC) concentrations were determined for the 12 samples which showed greatest antimicrobial potential in this stage. Two samples of L. alba, three samples of M. lundiana and seven samples of O. basilicum showed a MIC of 0.12-125 µL/mL for the six tested bacteria. Of these, the EO of O. basilicum cultivar Maria Bonita stood out with the lowest MIC and MBC. Thus, a mixture simulating this essential oil was prepared from commercial standards of the compounds (±)-linalool, geraniol and 1,8-cineole. Significantly higher MIC and MBC were detected in the simulation compared to the respective EO, suggesting a synergistic effect between compounds.

El objetivo de este estudio fue realizar un experimento de detección en aceites esenciales (AE) de Myrcia lundiana, Ocimum basilicum y Lippia alba contra seis bacterias patógenas que estropean los alimentos. Setenta y dos (72) muestras fueron analizadas inicialmente para la actividad antimicrobiana basada en la prueba de difusión en agar. Se determinaron las concentraciones mínimas inhibitoria (CMI) y bactericida (CMB) para las 12 muestras que mostraron el mayor potencial antimicrobiano en esta etapa. Dos muestras de L. alba, tres muestras de M. lundiana y siete muestras de O. basilicum mostraron un CMI de 0.12-125 µL/mL para las seis bacterias analizadas. De estos, el AE de O. basilicum cultivar Maria Bonita se destacó con el CMI y CMB más bajos. Por lo tanto, se preparó una mezcla que simula este aceite esencial a partir de los estándares comerciales de los compuestos de (±)-linalol, geraniol y 1,8-cineol. Se detectaron CMI y CMB significativamente más altos en la simulación en comparación con el AE respectivo, lo que sugiere un efecto sinérgico entre los compuestos.

Influence of storage temperatures and storage time of dry leaves on patchouli [Pogostemon cablin (Blanco) Benth. ] essential oil / Influência do tempo e temperaturas de armazenamento das folhas secas no óleo essencial de patchouli [Pogostemon cablin (Blanco) Benth. ]

Patchouli [Pogostemon cablin (Blanco) Benth.] is a plant of the family Lamiaceae, widely used as an essential oil in the cosmetics and perfumery industry. This study aimed to evaluate the influence of storage time and temperature of dry leaves on the patchouli essential oil content and chemical composition. The experiment was performed in a completely randomized design in a 6x2x2 factorial scheme, testing storage time (0, 1, 2, 4, 8, and 16 weeks) and temperature (28°C and 33°C) of dry leaves of two patchouli genotypes (POG-015 and POG-021). The variables essential oil content and chemical composition, and the identification of fungus during storage were evaluated. Results showed that the storage significantly influenced the essential oil content. Patchoulol was identified as the major compound in both genotypes, ranging from 55.05% to 68.77% (POG-15) and from 52.83% to 64.06% (POG-021). Based on the results of patchoulol, dry leaves of both genotypes (POG-015 and POG-021) can be stored for up to eight weeks at 28- 33°C without altering the essential oil quality.

Patchouli [Pogostemon cablin (Blanco) Benth.] é uma planta pertencente à família Lamiaceae, e seu óleo essencial é utilizado nas indústrias de perfumes e cosméticos. O objetivo do trabalho foi avaliar a influência do tempo e temperaturas de armazenamento das folhas secas no teor e na composição química do óleo essencial de patchouli. O delineamento experimental foi o inteiramente casualizado em esquema fatorial 6x2x2, testando tempo de armazenamento (0, 1, 2, 4, 8 e 16 semanas), temperatura de armazenamento (28°C e 33°C) de folhas secas de dois genótipos (POG-015 e POG-021) de patchouli. As variáveis avaliadas foram o teor e a composição química do óleo essencial e a identificação dos fungos que se desenvolveram durante o armazenamento. Os resultados mostraram que o armazenamento influenciou significativamente o teor de óleo essencial. Dos compostos identificados, o patchoulol foi o composto majoritário nos dois genótipos, variando de 55,05% a 68,77% (POG-15) e 52,83% a 64,06% (POG-021). Baseado no patchoulol, folhas secas dos genótipos (POG-015 e POG-021) de patchouli podem ser armazenadas por um período de até oito semanas em temperatura de 28 e 33°C, sem alterar a qualidade do óleo essencial.

Morphoagronomic characterization of Lantana camara L. germplasm / Caracterização morfoagronômica de germoplasma de Lantana camara L

The medicinal and aromatic species Lantana camara is a bush that receives a lot of interest regarding ornamental use. However, due to its condition of invasive plant in natural environments, it suffers constant eradication attempts, which causes genetic erosion of the species. To work around this problem, a sample of the genetic variability of L. camara was saved in the Active Germplasm Bank of medicinal and aromatic plants of the Federal University of Sergipe (UFS). This study aimed to evaluate morphoagronomically 22 accessions of L. camara, collected in 15 municipalities in the state of Sergipe. To carry out morphoagronomic characterization, morphological and agronomic traits were analyzed. Phenotypic variability among the accessions of the L. camara collection for all the analyzed morphological quantitative and agronomic characteristics was observed. The means ranged from 4.91 to 6.90 cm for leaf length; from 2.81 to 5.03 cm for leaf width; from 1.32 to 1.87 for the leaf length/width ratio; from 9.59 to 22.98 mm2 for leaf area; from 99.00 to 215.00 cm for crown diameter; from 85.67 to 214.33 for plant height; and from 1.08 to 2.43 cm for stem diameter. Most accessions presented erect form. The mean number of flowers per inflorescence varied from 12 to 38 flowers. For the agronomic traits, we observed that the dry mass ranged from 49.86 to 649.04 g.plant-1. The mean content of essential oil ranged from 0.13 to 0.26% and yield between 0.10 and 1.55 mL.plant-1. The most divergent phenotypic accessions were LAC-001 (São Cristóvão) and LAC-038 (Moita Bonita) and the ones with the lowest divergence were LAC-004 (Itaporanga D'ajuda) and LAC-019 (Siriri). Thus, it was observed that the variables were efficient to differentiate the accessions and suggest the existence of genetic variability among them.

A espécie aromática e medicinal Lantana camara, é um arbusto muito usado para ornamentação. Devido à sua condição de planta invasora em ambientes naturais, sofre com tentativas de erradicação constantes, o que ocasiona erosão genética da espécie. Para contornar essa problemática, uma amostra da variabilidade genética da L. camara está sendo conservada no Banco Ativo de Germoplasma (BAG) de Plantas Medicinais e Aromáticas da Universidade Federal de Sergipe (UFS). Assim, objetivou-se com o presente trabalho avaliar morfoagronomicamente 22 acessos de L. camara, coletados em 15 municípios do estado de Sergipe. Para caracterização morfoagronômica, variáveis morfológicas e agronômicas foram analisadas. Observou-se variabilidade fenotípica entre os acessos da coleção de L. camara para a maioria das características morfológicas quantitivas e agronômicas. A média para comprimento de folha variou de 4,91 e 6,90 cm; largura de folha de 2,81 a 5,03 cm; relação comprimento/largura de 1,32 a 1,87; área folia de 9,59 a 22,98 mm²; diâmetro da copa de 99, 00 a 215,00 cm; altura de planta de 85,67 a 214,33 cm; diâmetro do caule de 1,08 a 2,43 cm; A maioria dos acessos apresentou porte ereto. O número médio de flores por inflorescências variou de 12 a 38. Para as características agronômicas observou-se que a massa seca variou de 49,86 a 649,04 g.planta-¹. A média para teor de óleo essencial variou de 0,13 a 0,26% e o rendimento entre 0,10 a 1,55 mL.planta-¹. Os acessos mais divergentes fenotipicamente foram LAC-001 (São Cristóvão) e LAC-038 (Moita Bonita) e os que apresentaram a menor divergência foram LAC-004 (Itaporanga D'ajuda) e LAC-019 (Siriri). Dessa forma, nota-se que as variáveis foram eficientes para diferenciar os acessos e sugerem a existência de variabilidade genética entre eles.

Toxicity of the essential oil of basil cultivars and hybrids and its repellent effect on stored grain pests / Toxicidade do óleo essencial de cultivares e híbridos de manjericão e seu efeito repelente sobre pragas de grãos armazenados

Essential oils have emerged as an alternative to synthetic insecticides in the control of stored grain pests. The toxicity and repellency of the essential oils of four basil cultivars and three basil hybrids and the monoterpenes linalool, citral, and (E)-methyl cinnamate were evaluated in the stored grain pests Callosobruchus maculatus and Sitophillus zeamais. The essential oils of the cultivar Genovese and the hybrid 'Genovese' x 'Maria Bonita' were more toxic to C. maculatus. Conversely, the essential oils of the cultivar Sweet Dani and the hybrid 'Cinnamom' x 'Maria Bonita' were more toxic to S. zeamais. Among the monoterpenes, (E)-methyl cinnamate was the most toxic to both pests, taking 0.14 and 0.34 µ L.mL-1 to kill 50% of the C. maculatus and S. zeamais populations, respectively. All essential oils from cultivars, hybrids, and monoterpenes were repellent to S. zeamais, except for (E)-methyl cinnamate. For C. maculatus, this effect was lower, being citral the most repellent compound. Results demonstrate the insecticidal potential of the essential oil of O. basilicum and its monoterpenes in the control of stored grain pests.

Os óleos essenciais surgem como alternativa aos inseticidas sintéticos no controle das pragas de grãos armazenados. A toxicidade e a repelência dos óleos essenciais de quatro cultivares e três híbridos de manjericão e dos monoterpenos linalol, citral e (E)-cinamato de metila foram avaliadas nas pragas de grãos armazenados Callosobruchus maculatus e Sitophillus zeamais. Os óleos essenciais da cultivar Genovese e do híbrido 'Genovese' x 'Maria Bonita' foram mais tóxicos para C. maculatus. Já para S. zeamais, os óleos essenciais das cultivar Sweet Dani e do híbrido 'Cinnamom' x 'Maria Bonita' apresentaram maior toxicidade. Dentre os monoterpenos, o (E)-cinamato de metila foi o mais tóxico para ambas as pragas. Foram necessários 0,14 e 0,34 µ L.mL-1 para matar 50% da população de C. maculatus e S. zeamais. Todos os óleos essenciais das cultivares, dos híbridos e dos monoterpenos foram repelentes a S. zeamais, com exceção do (E)-cinamato de metila. Já para C. maculatus, este efeito foi reduzido, sendo o citral o composto mais repelente. Nos resultados demonstram o potencial inseticida dos óleos essenciais de O. basilicum e seus monoterpenos para o controle de pragas de grãos armazenados.

Morpho-agronomic characterization of Varronia curassavica germplasm conservated "Ex situ" / Caracterização morfoagronômica de germoplasma de Varronia curassavica conservado "Ex situ"

Varronia curassavica Jacq. is a medicinal and aromatic plant native to Brazil. The essential oil of this species is valued by the pharmaceutical industry due to its bioactive substances containing anti-inflammatory properties. This study aimed to morpho-agronomically characterize 27 accessions of the collection of V. curassavica of the Active Germplasm Bank of Medicinal and Aromatic Plants of the Federal University of Sergipe. The experiment consisted of a randomized block design with three replications. The morphological characterization occurred in the field, by evaluating the variables plant height; canopy width; canopy shape; stem diameter; stem color; leaf blades width, length, and length/width ratio; leaves, petals, and sepals color. The agronomic characterization was performed using the essential oils extracted from dried leaves by hydrodistillation in a Clevenger apparatus. Leaves dry matter yield per plant and essential oil yield and content were also evaluated. The accessions showed great variation for leaves color, with different shades of green, which allowed separating them into dark green leaf and light green leaf plants. Plant height values ranged from 101.33 cm (VCUR-801) to 345.33 cm (VCUR-701). The clustering analysis of the traits revealed seven distinct groups. The accessions VCUR-103 and VCUR-001 were the most divergent, whereas the accessions VCUR-401 and VCUR-404 were the most similar. The morpho-agronomic characterization of V. curassavica accessions provides a correct description of the species by pointing out perspectives for genetic improvement, besides optimizing the conservation process. KEYWORDS: Cordiaceae. Medicinal plant. Aromatic plant. Essential oil. Domestication. Genetic breeding INTRODUCTION Brazil is rich in plant biodiversity (SOUZA et al., 2010). However, in recent years, its biodiversity has undergone accelerated destruction,mainly affecting the natural vegetation. This fact is due to agricultural expansion, fires, logging, extractivism, and uncontrolled urban growth. All these changes have posed a great threat to several plant species in the country (OLIVEIRA, 2010). An alternative for the maintenance and conservation of endangered species is the formationof collections in Active Germplasm Banks, which holds base collections for the conservation of a broad plant genetic variability, enabling biodiversity maintenance (SILVA et al., 2012). Varronia curassavica Jacq. (ex Cordia verbenacea DC.)] is a medicinal and aromatic plant native to Brazil, belonging to the Cordiaceae family (GASPARINO; BARROS, 2009). It occurs from Central America to southern Brazil (LORENZI; MATOS, 2008) in a wide range of habitats, such as beaches, restinga, forests, and cerrado (WANDERLEY et al., 2002). V. curassavica has stood out both in folk medicine and in the pharmaceutical industry, mainly due to the anti-inflammatory substances present in its essential oil (PASSOS et al., 2007; PARISSOTO et al., 2012; PIMENTEL et al., 2012). The effectiveness of V. curassavica is recognized by the National Sanitary Surveillance Agency (ANVISA). The plant is included in the List of the Unified Health System (SUS) and the National List of Medicinal Plants of Interest to the Unified Health System (RENISUS) (BRASIL, 2011). Due to its therapeutic properties, several studies have been developed aimed at the selection Received: 14/05/18 Accepted: 20/11/19

Varronia curassavica Jacq. é uma planta medicinal e aromática nativa do Brasil. O óleo essencial desta espécie é utilizado pela indústria farmacêutica por conter substâncias bioativas com propriedades anti-inflamatórias. Objetivou-se caracterizar morfoagronomicamente 27 acessos da coleção de V. curassavica do Banco Ativo de Germoplasma de Plantas Medicinais e Aromáticas da Universidade Federal de Sergipe. O delineamento experimental foi em blocos casualizados com três repetições. A caracterização morfológica foi realizada em campo, por meio da avaliação da altura da planta; largura da copa; formato da copa; diâmetro do caule a 10 cm do solo; cor do caule; largura, comprimento e relação comprimento/largura das lâminas foliares; e cor das folhas, pétalas e sépalas. A caracterização agronômica foi realizada a partir dos óleos essenciais extraídos de folhas secas por hidrodestilação em aparelho Clevenger. Foi avaliado o rendimento de massa seca das folhas por planta e o teor e rendimento do óleo essencial. Grande variação foi observada entre os acessos para a coloração das folhas, as quais apresentaram diferentes tonalidades de verde, sendo possível separá-las entre plantas com folhas verde-escuro e plantas com folhas verde-claro. Para a variável altura de planta, os valores variaram de 101,33 cm (VCUR-801) a 345,33 cm (VCUR-701). Sete grupos distintos foram formados na análise de agrupamento realizada a partir dos caracteres avaliados. Os acessos VCUR-103 e VCUR-001 foram os que se mostraram mais divergentes, enquanto que os mais semelhantes foram os acessos VCUR-401 e VCUR-404. A caracterização morfoagronômica dos acessos de erva-baleeira permitirá a realização de uma correta descrição da espécie, apontando perspectivas para o melhoramento genético, além de possibilitar a otimização do processo de conservação.

Antifungal activity of essential oils of Myrcia ovata chemotypes and their major compounds on phytopathogenic fungi / Atividade antifúngica de óleos essenciais de quimiotipos de Myrcia ovata e seus principais compostos em fungos fitopatogênicos

This work evaluated the antifungal activity of essential oils of Myrcia ovata chemotypes (MYRO-175, MYRO-156, MYRO-154, MYRO-165, and MYRO-015) and their major compounds (linalool, geraniol, citral, and (E)-nerolidol) on the phytopathogenic fungi Fusarium pallidoroseum (which causes melon postharvest rot) and Colletotrichum musae (which causes anthracnose in banana). The essential oils were obtained by hydrodistillation and analyzed by GCMS/FID. To evaluate the antifungal activity, the essential oils and their major compounds were tested at different concentrations (0.1; 0.3; 0.4; 0.5; 0.7; 1.0; 3.0, and 5.0 mL/L). The major compounds found in the essential oils were nerolic acid, linalool, geraniol, citral, and (E)-nerolidol. The essential oils of the plants MYRO-154, MYRO-165, and MYRO-015 had the minimum inhibitory concentration (MIC) (0.3 mL/L) for F. pallidoroseum and the lowest minimum fungicidal concentration (MFC) (0.7 mL/L), for C. musae. Geraniol and citral had the lowest MFC (0.5 mL / L) for the two fungi tested. For F. pallidoroseum, the essential oils of the chemotypes were more effective than their major compounds. Conversely, the major compounds geraniol of the chemotype MYRO-156 (74.37%) and citral were more effective than their respective essential oils for C. musae. (E)-nerolidol and geraniol of the chemotype MYRO-015 (33.15%) were responsible for the antifungal activity of the essential oils of their respective chemotypes.

No presente trabalho avaliou-se a atividade antifúngica de óleos essenciais de quimiotipos de Myrcia lundiana (MYRO-175, MYRO-156, MYRO-154, MYRO-165, and MYRO-015) e seus compostos majoritários (linalol, geraniol, citral e (E)-nerolidol) sobre os fungos fitopatogênicos Fusarium pallidoroseum(causa podridão em frutos de melão) e Colletotrichum musae (causa antracnose em frutos de banana). Os óleos essenciais foram obtidos hidrodestilação e analisados por CGEM/DIC. Para avaliação da atividade antifúngica foram testados os óleos essenciais e os compostos majoritários nas concentrações: 0,1; 0,3; 0,4; 0,5; 0,7; 1,0; 3,0 e 5,0 mL/L. Os principais compostos presentes nos óleos essenciais foram o ácido nerólico, o linalol, o geraniol, o citral e o (E)-nerolidol. Os óleos essenciais das plantas MYRO-154, MYRO-165 e MYRO-015 apresentaram CIM de 0,3 mL/L e a planta MYRO-015 apresentou a menor concentração fungicida mínima (CFM) (1,0 mL/L). O geraniol e o citral foram os compostos que apresentaram o menor valor de CFM, 0,5 mL/L, frente aos dois fungos testados. O óleo essencial dos quimiotipos testados foram mais promissores que seus componentes majoritários puros, frente o F. pallidoroseum. Já para o C. musae, os componentes majoritários geraniol do quimiotipo MYRO-156 (74,37%) e o citral foram mais promissores que seus respectivos óleos essenciais. Já o (E)-nerolidol e o geraniol do quimiotipo MYRO-015 (33,15%) foram os responsáveis pela atividade antifúngica apresentada pelos óleos essenciais dos respectivos quimiotipos.

Essential oils of basil chemotypes: Major compounds, binary mixtures, and antioxidant activity.

The antioxidant potential of the essential oils (EO) of 24 basil genotypes was assessed by 4 distinct in vitro evaluation methods. Different combinations of the major compounds found in the EO were also tested to identify those combinations responsible for the antioxidant activity of the volatile oils and verify the occurrence of synergism or antagonism between them. Results indicate that 9 EO exhibited promising antioxidant potential, with at least 52.68% of inhibition of the linoleic acid peroxidation at 10 µL/mL and 76.34% of inhibition of the DPPH radical at 1 µL/mL. The major compound eugenol had the highest antioxidant activity. The antioxidant activity of these EO cannot be explained solely by the presence of the major compounds. Despite the influence of eugenol, the antioxidant activity is also related to the synergism between other minor compounds found in the EO. This fact confers a potent antioxidant activity to some basil EO.

In vitro conservation and acclimatization of epidendroideae (Orchidaceae) from Sergipe, Brazil / Conservação in vitro e aclimatização de Epidendroideae (Orchidaceae) de Sergipe, Brasil

Orchids are known for the beauty, exuberance, and color of their flowers and therefore represent one of the most coveted ornamental plants. Due to their high commercial value, several illegal practices of removal and commercialization have led to the verge of extinction. This study aimed to develop protocols for in vitro conservation under slow growth and acclimatization of species of the subfamily Epidendroideae (Orchidaceae), from the state of Sergipe, Brazil. Two experiments were conducted: the first one tested four concentrations of MS (MURASHIGE and SKOOG, 1962) (25%, 50%, 75%, 100%), three orchid species (Catasetum macrocarpum, Oeceoclades maculata, and Polystachya estrellensis), and two temperatures (18 and 25ºC); the second one tested three combinations of carbon sources and osmotic regulators (20 g L-1 sucrose; 10 g L-1 sucrose + 5 g L-1 de mannitol; 10 g L-1 sucrose + 5 g L-1 sorbitol) and the same three species and two temperatures. For the in vitro conservation of C. macrocarpum, O. maculata, and P. estrellensis over 450 days, the use of 25% of MS salts or 20 g.L-1 sucrose at 25ºC is recommended. C. macrocarpum seedlings were acclimatized using sand + shredded pine bark + worm castings at a ratio of 2:2:1 (w:w:w). The use of sand alone is recommended for the acclimatization of O. maculata

As orquídeas são conhecidas pela beleza, exuberância e cores de suas flores e por isso representam uma das plantas ornamentais mais cobiçadas. Devido ao alto valor comercial, muitas são as práticas ilegais de retirada e comercialização de orquídeas, o que leva as espécies à beira de extinção. O objetivo deste estudo foi desenvolver protocolos para a conservação in vitro sob crescimento lento e aclimatação de espécies da subfamília Epidendroideae (Orchidaceae) do Estado de Sergipe, Brasil. Dois experimentos foram realizados: o primeiro ensaio testou quatro concentrações dos sais MS (MURASHIGE e SKOOG, 1962) (25, 50, 75 e 100%), três espécies (Catasetum macrocarpum, Oeceocladesmaculata e Polystachya estrellensis) e duas temperaturas (18 e 25ºC); o segundo ensaio testou três combinações de fontes de carbono e reguladores osmóticos (20 g L-1 de sacarose; 10 g L-1 de sacarose + 5 g L-1 manitol; 10 g L-1 de sacarose + 5 g L-1 sorbitol), três espécies (C. macrocarpum, O. maculata e P. estrellensis) e duas temperaturas (18 e 25ºC). Para executar a aclimatação, oito substratos foram testados para as espécies C. macrocarpum e O. maculata. Para realizar a conservação in vitro de C. macrocarpum, O. maculata e P.estrellensis ao longo de 450 dias, recomenda-se a utilização 25% dos sais de MS ede 20 g L-1 de sacarose a 25°C. Mudas de C. macrocarpum foram aclimatizadas utilizando areia + casca de pinus + húmus de minhoca (2:2:1). Recomenda-se a utilização de areia para aclimatização de O. maculata.

Chemical characterization of the essential oil from leaves of basil genotypes cultivated in different seasons / Caracterización química del aceite esencial de hojas genotipos de albahaca cultivados en diferentes estaciones del año

The aim of this study was to evaluate the concentration and chemical composition of the essential oil the leaves of basil cultivars and hybrids cultivated in different cropping seasons: dry season and rainy season. The variables evaluated were the content and composition of essential oils in the two seasons. The essential oil content ranged from 0.66% to 3.21% in the dry season and from 0.80% to 4.20% in the rainy season. The major compounds found among the genotypes were linalool, methyl chavicol, neral, geranial, eugenol, and methyl (E)- cinnamate, defining the formation of five groups in each season, classified in the following chemotypes: methyl chavicol (Group 1), citral (neral+geranial) (Group 2), methyl cinnamate (Group 3), linalool (Group 4), and intermediate linalool (Group 5). All the traits evaluated had heritability (h ) greater than 95% and high CVg/CVe ratio values. The cropping season affected the content and chemical compositions of basil essential oil.

El objetivo de este estudio fue evaluar la concentración y composición química del aceite esencial las hojas de cultivares de albahaca e híbridos cultivados en diferentes temporadas de cultivo: estación seca y estación lluviosa. Las variables evaluadas fueron la contenido y la composición de los aceites esenciales en las dos estaciones. La contenido de aceite esencial varió de 0.66% a 3.21% en la estación seca y de 0.80% a 4.20% en la estación lluviosa. Los principales compuestos encontrados entre los genotipos fueron linalool, metilchavicol, neral, geranial, eugenol y metil (E)-cinamato, definiendo la formación de cinco grupos en cada estación, clasificados en los siguientes quimiotipos: metil chavicol (Grupo 1), citral (neral + geranial) (Grupo 2), cinamato de metilo (Grupo 3), linalool (Grupo 4) y linalol intermedio (Grupo 5). Todos los rasgos evaluados mostraron una heredabilidad (h ) mayor que el 95% y altos valores de relación CVg/CVe. La temporada de cultivo afectó la contenido y las composiciones químicas del aceite esencial de albahaca.

Chemical diversity of essential oils from Hyptis pectinata (L. ) Poit / Diversidade química de óleos essenciais de Hyptis pectinata (L. ) Poit

The essential oils are secondary metabolites formed by several chemical compounds that confer to these substances great social and economic importance. This diversity of compounds is generally determined by the genetic constitution of the plant, although environmental factors may also influence the type, amount, and concentrations of the compounds present in the essential oil. The aim of this work was to analyze the chemical diversity of the essential oils of native Hyptis pectinata plants collected in the state of Sergipe. The essential oils of 24 plants were obtained by hydrodistillation and analyzed by GC-MS/FID, revealing 30 compounds. Two clusters were formed by the cluster analysis. Cluster I consisted of 18 plants, and presented -elemene (2.46-25.77%), -caryophyllene (16.20-60.95%), germacrene D (0.00-21.59%), and caryophyllene oxide (5.38-42.21%) as major compounds. Cluster II consisted of six plants, and presented -caryophyllene (5.68-15.57%), (Z)--guaiene (2.18-7.31%), caryophyllene oxide (1.58-22.89%), and calamusenone (23.12-64.36%) as major compounds. Strong correlation was observed between pcymene and -terpinene (r=0.94), and between (E)--guaiene and lepidozene (r=0.95). Results of the present study indicate variation in the essential oil content, and show that the compounds -elemene, -caryophyllene, germacrene D, (Z)--guaiene, caryophyllene oxide and calamusenone were detected in greater proportions in native plants of H. pectinata of the state of Sergipe. The knowledge of the chemical diversity found in H. pectinata plants can assist in the selection of plants of specific interest.

Os óleos essenciais são metabólitos secundários formados por diversos compostos químicos que atrelam a estas substâncias grande importância social e econômica. Essa diversidade de compostos geralmente é determinada pela constituição genética da planta, embora fatores ambientais também possam influenciar quanto ao tipo, quantidade e concentrações dos compostos presentes no óleo essencial. O objetivo deste trabalho foi analisar a diversidade química dos óleos essenciais de plantas nativas de H. pectinata coletadas no Estado de Sergipe. O óleo essencial de 24 plantas foi obtido por hidrodestilação e analisados por GC/MS-FID. Foram detectados 30 compostos no óleo essencial. Pela análise de agrupamento, foi observada a formação de dois grupos. Grupo 1 foi constituído por 18 plantas, e apresentou o -elemeno (2,46-25,77%), -cariofileno (16,20-60,95%), germacreno-D (0,00-21,59%) e óxido de cariofileno (5,38-42,21%) como compostos majoritários. Grupo 2 foi constituído por 6 plantas com -cariofileno (5,68-15,57%), Z-- guaieno (2,18-7,31%), óxido de cariofileno (1,58-22,89%) e calamusenona (23,12-64,36%) como compostos majoritários. Uma forte correlação foi observada entre os compostos p-cimeno e -terpineno (r=0,94) e entre (E)--guaieno e lepidozeno (r=0,95). Os resultados do presente estudo indicam que existe variação no teor do óleo essencial, e que os compostos - elemeno, -cariofileno, germacreno-D, (Z)--guaieno, óxido de cariofileno e calamusenona, foram detectados em maiores proporções nas plantas nativas de H. pectinata do Estado de Sergipe. O conhecimento da diversidade química encontrada nas plantas de H. pectinata pode auxiliar na seleção de plantas de interesse específico.

Distillation methods affect the chemical composition of Varronia curassavica Jacq. essential oil? / Métodos de destilação afetam a composição química do óleo essencial de Varronia curassavica Jacq?

The objective of this work was to evaluate the chemical composition of essential oil from Varronia curassavica Jacq. obtained by microwave (MI) and hydrodistillation (HD) extraction methods. The MI method tested three powers (500, 600, and 700W), three distillation times (20, 30, and 40 min.), and three water volumes (0, 25, and 50 mL per sample). The HD method tested three distillation times (100, 120, and 140 min.) and three water volumes (1.0, 1.5, and 2.0 L per 3-liter flask). The essential oils were analyzed by GC/MS-FID. The optimal condition for the essential oil extraction by the MI method was 700W for 40 min. (3.28%), regardless of the volume of water. In its turn, the best condition for essential oil extraction by the HD method was 120 min. with 1.0 L of water per flask (3.34%). The most abundant compounds for MI (700 W for 40 min. without water) were shyobunol (26.53%) and bicyclogermacrene (4.96%); and the most abundant compounds for HD (120 min. with 1.0 L of water/flask) were shyobunol (24.00%) and germacrene D-4-ol (10.23%). Methyl farnesoate (2E, 6E) and farnesyl acetate (2Z, 6E) were not detected in the essential oil extracted by HD; however, they were identified by the MI method. By increasing the distillation time and/or volume of water in HD, a reduction was observed for the content of the chemical compounds -elemene (from 1.23 to 0.97%), Ecaryophyllene (from 5.49 to 4.35%), -humulene (from 1.80 to 1.43%), alloaromadendrene (from 1.78 to 1.44%), bicyclogermacrene (from 5.63 to 4.55%), and germacrene D-4-ol (from 11.40 to 9.86%). Power, extraction time, and their interactions influenced the content of essential oil obtained by microwave extraction (MI). Within each power, the highest essential oil content was extracted at the longest distillation time (40 min.), except for 600W, where no significant difference was detected between 30 and 40 min. The optimal essential oil contents for both extraction methods were statically similar by the t-test for dependent samples. However, the MI method presents advantages, such as shorter distillation time and less energy and water consumption.

O objetivo deste trabalho foi avaliar a composição química do óleo essencial de Varronia curassavica Jacq. obtido pelos métodos de extração micro-ondas (MI) e hidrodestilação (HD). Para MI, foram testadas três potências (500, 600 e 700W), três tempos de destilação (20, 30 e 40 min.) e três volumes de água (0, 25 e 50 mL por amostra). Para HD, foram testados três tempos de destilação (100, 120 e 140 min.) e três volumes de água (1,0; 1,5 e 2,0 L por balão de 3 litros). Os óleos essenciais foram analisados por CG/EM-FID. Maiores teores de óleo essencial foram obtidos nas condições de 700 W por 40 min. (3.28%), independente do volume de água para MI, e 120 min. com 1,0 L de água por balão para HD (3,34%). Os compostos mais abundantes para MI (700W, por 40 min., sem água) foram o shyobunol (26,53%) e biciclogermacreno (4,96%) e para HD (120 min. com 1,0 L de água /balão) foram shyobunol (24,00%) e germacreno D -4 -ol (10,23%). Metil farnesoato (2E, 6E) e farnesil acetato (2Z, 6E) não foram detectados no óleo essencial extraído por HD, porém, foram detectados nas amostras extraídas por MI. Com o aumento do tempo de destilação e/ou do volume de água em HD, houve redução no conteúdo dos constituintes químicos -elemeno (de 1,23 para 0,97%), E-cariofileno (de 5,49 para 4,35%), -humuleno (1,80 para 1,43%), aloaromadendreno (de 1,78 para 1,44%), biciclogermacreno (de 5,63 para 4,55%) e germacreno D-4-ol (de 11,40 para 9,86%). A potência, o tempo de extração e suas interações influenciaram no teor de óleo essencial obtido na extração por micro-ondas (MI). Dentro de cada potência, o maior teor de óleo essencial foi obtido no tempo mais longo de extração (40 min.), exceto para 600 W, que não apresentou diferença significativa entre 30 e 40 min. Nas condições ótimas de extração, os teores de óleo essencial obtidos foram estatisticamente semelhantes pelo teste t para amostras dependentes. No entanto, a extração por micro-ondas apresenta algumas vantagens em relação a HD, como menor tempo de destilação e menor consumo de energia e água.

Influence of N-P-K fertilization at the acclimatization stage on micropropagated seedlings of Tillandsia bulbosa Hook / Influência da adubação com N-P-K na fase de aclimatização em mudas micropropagadas de Tillandsia bulbosa Hook

Bromeliads are known worldwide for their ornamental potential. In Brazil, species of the genus Tillandsia occur in the Atlantic rainforest, Amazon rainforest, and rocky fields. This work aimed to evaluate the influence of nitrogen, phosphorus, and potassium fertilization on micropropagated seedlings of Tillandsia bulbosa, at the acclimatization stage, and their leaf anatomy. The experiment was carried out in a completely randomized design, in a 4x4+1 factorial scheme, using coconut coir: earthworm humus: sand mixture as substrate (2:1:1). Urea, single superphosphate, and potassium chloride were used as nitrogen, phosphorus and potassium sources, respectively, at proportions of 50, 100, 200, and 400% of the dose recommended. The doses were distributed in four applications, testing total application at planting (1); ½ application at planting and ½ at 80 DAP (2); application at planting, at 50 DAP, and at 100 DAP (3); and » application at planting, » at 30 DAP, » at 60 DAP, and » at 120 DAP (4); and a control (without fertilization). Leaves anatomy was analyzed at 180 days after planting. Fertilization did not significantly influence the development of seedlings during acclimatization. The doses of 50, 100, and 200% provided thicker parenchyma of chlorophyll and aquifer and leaf blade. Tillandsia bulbosa can be acclimatized without fertilizer application.

As bromélias são conhecidas mundialmente por seu potencial ornamental. No Brasil, as espécies de Tillandsia podem ser encontradas na Mata Atlântica, Floresta Amazônica e campos rupestres. O objetivo deste trabalho foi verificar a influência de adubação com nitrogênio, fósforo e potássio em plântulas micropropagadas de Tillandsia bulbosa, na fase de aclimatização e na sua anatomia foliar. O experimento foi implantado em delineamento inteiramente casualizado, em esquema fatorial 4x4+1, utilizando como substrato a mistura de pó de coco: húmus de minhoca: areia (2:1:1). Ureia, super fosfato simples e cloreto de potássio foram utilizados como fontes de nitrogênio, fósforo e potássio, nas proporções de 50, 100, 200 e 400% da dose recomendada. As doses foram distribuídas em quatro aplicações, testando aplicação total no plantio (1); ½ no plantio e ½ aos 80 DAP (2); no plantio, aos 50 DAP e aos 100 DAP (3) e » no plantio, » aos 30 DAP, » aos 60 DAP e » aos 120 DAP (4), e uma testemunha (sem adubação). Foi realizado o estudo anatômico das folhas aos 180 dias. A adubação não influenciou significativamente o desenvolvimento das plântulas durante a aclimatização. As doses de 50, 100 e 200% proporcionaram maior espessura de parênquimas clorofiliano e aquífero e do limbo foliar. A aclimatização de Tillandsia bulbosa pode ser realizada sem a necessidade de adubação.

Cropping season affect the performance of basil cultivars and hybrids / Época de plantio afeta o desempenho de cultivares e híbridos de manjericão

The objective of this work was to evaluate the performance of basil cultivars and hybrids in two cropping seasons. The experiments were conducted in a randomized block design, with three replications. Twenty-four basil genotypes were tested (20 commercial cultivars and four hybrids), in the dry season (oct.-dec./2015) and in the rainy season (apr.-jun./2016). The evaluated variables were: plant height (cm plant-1), canopy width (cm plant-1), leaf dry weight (g plant-1), and essential oil yield (mL plant-1). The data of each period were subject to analysis of variance and the means were grouped by the Scott-Knott test (p0.05). The individual and joint analyses of variance were performed for the experiments in both seasons. The estimated parameters were: coefficient of genetic variation (CVg), coefficient of environmental variation (CVe), CVg/CVe ratio, and heritability (h2). Leaf dry weight values per plant ranged from 6.23 to 75.00 g plant-1 (dry season) and from 9.17 to 31.34 g plant-1 (rainy season). The hybrid Cinnamon x Maria Bonita (1.50 mL plant-1) and the cultivar Mrs. Burns (1.44 mL plant-1) presented higher essential oil yield in the dry season. All the evaluated variables showed high heritability (h2) (> 50%) and CVg/CVe ratio (> 1.0), in both cropping seasons, indicating a favorable condition for selection. The cropping season influences the biomass and essential oil yield of basil.

O objetivo do presente trabalho foi avaliar o desempenho de cultivares e híbridos de manjericão em duas épocas de plantio. Para o ensaio, utilizou-se o delineamento experimental em blocos casualizados, com três repetições. Foram avaliados 24 genótipos de manjericão, sendo 20 cultivares comerciais e quatro híbridos cultivados na época seca (out.-dez./2015) e chuvosa (abr.-jun./2016). As variáveis avaliadas foram: altura de planta (cm planta-1), largura da copa (cm planta-1), massa seca de folhas (g planta-1) e rendimento (mL planta-1) de óleo essencial. Os dados de cada época foram submetidos à análise de variância, e as médias foram agrupadas pelo teste Scott-Knott (p0,05). Realizou-se a análise de variância individual e conjunta para os experimentos das duas épocas. Foram estimados os seguintes parâmetros: coeficiente de variação genética (CVg), coeficiente de variação ambiental (CVe), razão CVg/CVe e a herdabilidade (h2). As médias deferiram significativamente em relação à época de plantio para a maioria das cultivares. Os valores de massa seca de folha variaram de 6,23 a 75,00 g planta-1 (época seca) e 9,17 a 31,34 g planta-1 (época chuvosa). O híbrido Cinnamon x Maria Bonita (1,50 mL planta-1) e a cultivar Mrs. Burns (1,44 mL planta-1) apresentaram maior rendimento de óleo essencial na época seca. Na época chuvosa foi de 1,29 mL planta-1 (Mrs. Burns). Todos os caracteres avaliados apresentaram alta herdabilidade (h2) (>50%) e razão CVg/CVe (>1,0), nas duas épocas, indicando condição favorável para seleção em um programa de melhoramento. A época de plantio influencia a produção de biomassa e óleo essencial de manjericão.

Essential Oils of Hyptis pectinata Chemotypes: Isolation, Binary Mixtures and Acute Toxicity on Leaf-Cutting Ants.

Leaf-cutting ants are pests of great economic importance due to the damage they cause to agricultural and forest crops. The use of organosynthetic insecticides is the main form of control of these insects. In order to develop safer technology, the objective of this work was to evaluate the formicidal activity of the essential oils of two Hyptis pectinata genotypes (chemotypes) and their major compounds on the leaf-cutting ants Acromyrmex balzani Emery and Atta sexdens rubropilosa Forel. Bioassays of exposure pathways (contact and fumigation) and binary mixtures of the major compounds were performed. The major compounds identified in the essential oils of H. pectinata were ß-caryophyllene, caryophyllene oxide and calamusenone. The essential oils of H. pectinata were toxic to the ants in both exposure pathways. Essential oils were more toxic than their major compounds alone. The chemotype calamusenone was more toxic to A. balzani in both exposure pathways. A. sexdens rubropilosa was more susceptible to the essential oil of the chemotype ß-caryophyllene in both exposure pathways. In general, the binary mixtures of the major compounds resulted in additive effect of toxicity. The essential oils of H. pectinata is a raw material of great potential for the development of new insecticides.

Anti-cryptococcal activity of ethanol crude extract and hexane fraction from Ocimum basilicum var. Maria bonita: mechanisms of action and synergism with amphotericin B and Ocimum basilicum essential oil.

CONTEXT: Ocimum basilicum L. (Lamiaceae) has been used in folk medicine to treat headaches, kidney disorders, and intestinal worms. OBJECTIVE: This study evaluates the anti-cryptococcal activity of ethanol crude extract and hexane fraction obtained from O. basilicum var. Maria Bonita leaves. MATERIALS AND METHODS: The MIC values for Cryptococcus sp. were obtained according to Clinical and Laboratory Standards Institute in a range of 0.3-2500 µg/mL. The checkerboard assay evaluated the association of the substances tested (in a range of 0.099-2500 µg/mL) with amphotericin B and O. basilicum essential oil for 48 h. The ethanol extract, hexane fraction and associations in a range of 0.3-2500 µg/mL were tested for pigmentation inhibition after 7 days of treatment. The inhibition of ergosterol synthesis and reduction of capsule size were evaluated after the treatment with ethanol extract (312 µg/mL), hexane fraction (78 µg/mL) and the combinations of essential oil + ethanol extract (78 µg/mL + 19.5 µg/mL, respectively) and essential oil + hexane fraction (39.36 µg/mL + 10 µg/mL, respectively) for 24 and 48 h, respectively. RESULTS: The hexane fraction presented better results than the ethanol extract, with a low MIC (156 µg/mL against C. neoformans T444 and 312 µg/mL against C. neoformans H99 serotype A and C. gattii WM779 serotype C). The combination of the ethanol extract and hexane fraction with amphotericin B and essential oil enhanced their antifungal activity, reducing the concentration of each substance needed to kill 100% of the inoculum. The substances tested were able to reduce the pigmentation, capsule size and ergosterol synthesis, which suggest they have important mechanisms of action. CONCLUSIONS: These results provide further support for the use of ethanol extracts of O. basilicum as a potential source of antifungal agents.

In vitro conservation and leaf anatomy of different chemotypes of Lippia alba (Mill.) N. E. BR / Conservação in vitro e anatomia foliar de diferentes quimiotipios de Lippia alba (Mill.) N. E. BR

The aim of this work was to develop an in vitro conservation protocol for Lippia alba that involved mineral oil and to study the leaf anatomy of L. alba genotypes in in vitro and ex vitro environments. In vitro conservation involved five genotypes; LA-13 and LA-57 (carvone chemotype); LA-22 (linalool chemotype); and LA-29 and LA-44 (citral chemotype). Each genotype was treated with mineral oil, with water, and a control at 18°C and 23°C. Survival (%), shoot height (cm) and leaf color were assessed at 180 and 270 days. The genotypes with the best in vitro conservation outcomes (LA-13 and LA-57) were used to study the anatomy of the leaves. Midrib thickness, leaf blade thickness, adaxial and abaxial cuticles and the number of glandular and tector trichomes were assessed. A temperature of 18°C was optimal for the conservation of genotypes of the carvone chemotypes. The explants were short and had green leaves. This was especially true for LA-57, whose height did not exceed 2.0 cm. At 270 days, the LA-13 and LA-57 genotypes continued to have green and viable leaves, especially LA-57, which had the lowest mean height. Field plants of the carvone chemotype had thicker midribs, leaf blades, and cuticles and had more trichomes. The LA-57 genotype had the most glandular and tector trichomes.

O objetivo deste estudo foi desenvolver um protocolo de conservação in vitro com óleo mineral e estudar a anatomia foliar de genótipos de L. alba nos ambientes in vitro e ex vitro. A conservação in vitro constou de cinco genótipos LA-13 e LA-57 (quimiotipo carvona); LA-22 (quimiotipo linalol); LA-29 e LA-44 (quimiotipo citral), três tratamentos (com óleo mineral, com água e controle) e duas temperaturas 18 e 23°C. Aos 180 e 270 dias, avaliou-se a sobrevivência (%), altura da parte aérea (cm) e coloração das folhas. Os genótipos com os melhores resultados da conservação in vitro (LA-13 e LA-57) foram utilizados para estudos de anatomia. Avaliou-se espessura da nervura central, espessura do limbo foliar, cutículas adaxial e abaxial e quantidade de tricomas glandulares e tectores. A temperatura a 18°C foi a ideal para a conservação dos genótipos do quimiotipo carvona, com folhas verdes e baixos valores para altura, destacando-se LA-57 cuja altura não ultrapassou 2,0 cm. Aos 270 dias LA-13 e LA-57 permaneceram com folhas verdes e viáveis, com destaque para LA-57 com menores médias de altura. A anatomia do quimiotipo carvona mostrou que as plantas de campo apresentaram maiores espessuras da nervura, limbo e cutículas, e maiores quantidades de tricomas. O genótipo LA-57 obteve a maior quantidade de tricomas glandulares e tectores.

Chemical Diversity in Lippia alba (Mill.) N. E. Brown Germplasm.

The aim of this study was to perform chemical characterization of Lippia alba accessions from the Active Germplasm Bank of the Federal University of Sergipe. A randomized block experimental design with two replications was applied. The analysis of the chemical composition of the essential oils was conducted using a gas chromatograph coupled to a mass spectrometer. The chemical composition of the essential oils allowed the accessions to be allocated to the following six groups: group 1: linalool, 1,8-cineole, and caryophyllene oxide; group 2: linalool, geranial, neral, 1,8-cineol, and caryophyllene oxide; group 3: limonene, carvone, and sabinene; group 4: carvone, limonene, g-muurolene, and myrcene; group 5: neral, geranial, and caryophyllene oxide; and group 6: geranial, neral, o-cymene, limonene, and caryophyllene oxide.

Acaricidal activity of essential oils from Lippia alba genotypes and its major components carvone, limonene, and citral against Rhipicephalus microplus.

The goal of the present study was to evaluate the acaricidal potential of Lippia alba essential oil, citral chemotypes (LA-10 and LA-44 genotypes) and carvone chemotypes (LA-13 and LA-57 genotypes), as well as purified citral and enantiomers of carvone and limonene. Efficacy against Rhipicephalus microplus was assessed by the larval packet and the engorged female immersion tests. Citral chemotypes had greater larvicidal activity than carvone chemotypes, and this was further supported by larvicidal and adulticidal activity of purified citral with LC50 values of 7.0 and 29.8 mg/mL, respectively. While purified enantiomers of carvone exhibited greater larvicidal activity than those of limonene, enantioselectivity of limonene was observed with R-(+) displaying significantly higher efficacy (LC50 of 31.2mg/mL) than S-(-) (LC50 of 54.5mg/mL). The essential oils and purified compounds were much less toxic toward engorged adult females, with the exception of citral, and this may be due to limited cuticular penetration.

Chemical diversity in basil (Ocimum sp.) germplasm.

The present study aimed to chemically characterize 31 accessions and seven cultivars of basil. The percentage composition of the essential oils of the accessions and cultivars was based on the 14 most abundant constituents: 1,8-cineole, linalool, methyl chavicol, neral, nerol, geraniol, geranial, methyl cinnamate, ß-bourbonene, methyl eugenol, α-trans-bergamotene, germacrene-D, epi-α-cadinol, and δ-cadinene. The genetic materials were classified into eight clusters according to the chemical composition of the essential oils: Cluster 1--mostly linalool and 1,8-cineole; Cluster 2--mostly linalool, geraniol, and α-trans-bergamotene; Cluster 3--mostly linalool, methyl chavicol, methyl cinnamate, and ß-bourbonene; Cluster 4--mostly linalool, methyl chavicol, epi-α-cadinol, and α-trans-bergamotene; Cluster 5--mainly linalool, methyl eugenol, α-trans-bergamotene, and epi-α-cadinol; Cluster 6--mainly linalool, geraniol, and epi-α-cadinol; Cluster 7--mostly linalool and methyl chavicol; Cluster 8--mainly geranial and neral.