Production of rosmarinic acid and correlated gene expression in hairy root cultures of green and purple basil (Ocimum basilicum L.).

Rosmarinic acid (RA) is an active constituent of Ocimum basilicum. It has been shown that hairy root production (measured as dry weight) improves when green basil (O. basilicum "Cinnamon") is cultured under the light. In contrast, purple basil (O. basilicum "Purpurascens") shows greater hairy root production when cultured under dark conditions. The level of gene expression was highest in hairy roots of green basil under dark conditions for up to 1 week. Transcript levels were highest in hairy roots of purple basil under both dark and light conditions after 2 weeks of culturing. After 3 weeks of culture under light conditions, green basil had accumulated 1.9-fold higher RA content than that of purple basil, which in turn was fivefold higher than that of the natural roots (42.86 µg/mg). Tyrosine aminotransferase showed a higher transcript level when compared to the other phenylpropanoid pathway genes (phenylalanine ammonia-lyase, cinnamate 4-hydroxylase, and coenzyme-A ligase) in both dark and light conditions and in all-time regimens. RA accumulation was higher in the cultured hairy roots of green basil than those of purple basil under both light and dark conditions.

Influence of water stress and storage time on preservation of the fresh volatile profile of three basil genotypes.

The main goal of the present study was to describe the volatile profile of three different basil genotypes (Genovese and Green and Purple Iranian), and the impact that water stress (75% and 50% field capacity) and storage time (up to 7days) have under mild refrigerated conditions. The chromatographic profile pointed to three different chemotypes: linalool/eugenol, neral/geranial, and estragol, for Genovese, Green, and Purple genotypes, respectively. Water stress depleted the volatile profile of these three landraces, due to a reduction in the absolute concentrations of some of the components related to fresh aroma (linalool, nerol, geraniol and eugenol). The stability of the basil volatile profile during storage varied depending on the water stress that had been applied. Concentration reductions of close to 50% were quantified for most of the components identified in the Purple genotype.

Evaluation of basil extract ( Ocimum basilicum L. ) on oxidative, anti-genotoxic and anti-inflammatory effects in human leukocytes cell cultures exposed to challenging agents

ABSTRACT Ocimum is one of the most important genera of the Lamiaceae family. Several studies about basil and its popular use reveal many characteristics of the herb, including its use as antioxidant, anti-aging, anti-inflammatory, anti-carcinogenic, anti-microbial, and cardiovascular agents, among others. In this paper, we evaluated genotoxic, oxidative, and anti-inflammatory parameters from the extract of Ocimum basilicum in different concentrations, using human leukocytes cultures exposed to challenging agents. Our results confirm that the O. basilicum extract acts as an antioxidant and effectively reverts or subjugates the effects of high oxidizing agents such as hydrogen peroxide. These actions are attributed to its composition, which is rich in polyphenols and flavonoids as well as compounds such as rosmarinic acid, all of which have well-known antioxidant activity. We also show that our basil extract presents anti-inflammatory properties, the mechanism of which is a composed interaction between the inhibition of pro-inflammatory mediator and the stimulation of anti-inflammatory cytokines. Although pharmacodynamics studies are necessary to evaluate the activities in vivo, our results demonstrated that basil could act as an antioxidant and anti-inflammatory and a possible alternative for medicinal treatment.

Efeito da adubação nitrogenada e da sazonalidade na produtividade de Ocimum basilicum L / Effect of nitrogen fertilization on the production and productivity of Basil, red basil (Ocimum basilicum L. ) in spring and autumn seasons.

RESUMO O manjericão (Ocimum basilicum L.) atualmente encontra-se distribuído por todos os continentes e foi introduzido no Brasil com a chegada da colonização italiana. As principais partes utilizadas para comercialização no Brasil são as folhas frescas ou secas. O presente trabalho teve como objetivo avaliar o efeito de doses da adubação nitrogenada, durante o cultivo na primavera e outono, nas características produtivas e na produtividade de manjericão, Alfavaca basilicão vermelho, em casa de vegetação. O delineamento utilizado foi em blocos casualizados, em esquema fatorial 6 x 2, com cinco repetições, onde o primeiro fator foi constituído de cinco doses de nitrogênio e uma testemunha (0,0; 45,0; 90,0; 135,0; 180,0 e 225,0 kg ha-1 de nitrogênio) e o segundo, pelas épocas de cultivo (primavera e outono). Para as características produtivas da cultivar de manjericão Alfavaca Basilicão vermelho, verificou-se que as doses de nitrogênio entre 90,0 a 135,0 kg ha-1 foram mais adequadas, elevando todas as características produtivas avaliadas, do cultivo de primavera. Quando observado o cultivo no outono, teve-se ajuste apenas para a projeção de copa, massa fresca de parte aérea e área foliar. Quando se obteve ajuste significativo, a dose de aproximadamente 110,0 kg ha-1 de nitrogênio foi a que promoveu maior desenvolvimento da cultura. Em relação às épocas de cultivo na primavera recomenda-se o uso de adubação mineral nitrogenada, porém quando esse cultivo é realizado outono, a adubação nitrogenada não é responsiva.

ABSTRACT The basil (Ocimum basilicum L.) currently is distributed to all continents and was introduced in Brazil with the arrival of Italian colonization. The main parts used for commercialization in Brazil are the fresh or dried leaves. This study aimed to evaluate the effect of doses of nitrogen fertilization for cultivation in spring and fall, the yield characteristics and the productivity of basil, Basil red in the greenhouse. The experimental design was a randomized block, in a factorial 2 x 6 with five replications, where the first factor constituent of five nitrogen doses and one control (0.0; 45.0; 90.0; 135.0; 180.0 and 225.0 kg ha-1 of nitrogen) and by the second, the growing seasons (Spring and Autumn). For the productive characteristics of the cultivar red Basil, it was found that the nitrogen rates between 90.0 to 135.0 kg ha-1 were better, bringing all tested productive characteristics, the spring crop. When observed growing in the autumn, was only fit for the crown projection, fresh weight of shoot and leaf area. When there was significant adjustment, the dose of approximately 110.0 kg ha-1 of nitrogen was that promoted further development of culture. Regarding the growing seasons in the spring we recommended the use of mineral nitrogen fertilization, but when this crop is performed autumn, nitrogen fertilization is not responsive.

Adubação foliar no desenvolvimento e produção de óleo essencial de manjericão / Foliar fertilization in the development and production of essential basil oil

RESUMO O objetivo do trabalho foi avaliar o desenvolvimento e a produção de óleo essencial de manjericão com a aplicação de adubo foliar em Londrina-PR. O delineamento experimental utilizado foi o de blocos casualizados com três doses de adubo foliar (0, 2 e 4 L.ha-1) e sete repetições. Foram realizadas três colheitas durante o desenvolvimento do experimento. Os cortes ocorreram no início do florescimento, a altura do oitavo nó aos 40, 80 e 120 dias após o transplantio. Avaliou-se a altura, área foliar, produção de massa seca de folhas e rendimento de óleo essencial. A aplicação de 4 L ha-1 do adubo foliar proporcionou os maiores acréscimos na produção de massa seca de folhas e no rendimento de óleo essencial até o segundo corte das plantas de manjericão. A renovação da cultura se faz necessária a partir do terceiro corte.

ABSTRACT The aim was to evaluate the development and production of basil essential oil with the application of foliar fertilizer in Londrina-PR. The experimental design was a randomized block with three doses of foliar fertilizer (0, 2 and 4 L.ha-1) and seven replications. Three harvests were made during the development of the trial. The cuts occurred at flowering, at the height of the eighth node and at 40, 80 and 120 days after the transplanting. We evaluated the height, leaf area, dry matter production of leaves and essential oil yield. The application of 4 L ha-1 foliar fertilizer provided greater rise in production of dry matter of leaves and essential oil yield until the second harvest of basil plants. The renewal of the cultivation is needed from the third cut.

A novel in vitro whole plant system for analysis of polyphenolics and their antioxidant potential in cultivars of Ocimum basilicum.

Plants are an important source for medicinal compounds. Chemical screening and selection is critical for identification of compounds of interest. Ocimum basilicum (Basil) is a rich source of polyphenolics and exhibits high diversity, therefore bioprospecting of a suitable cultivar is a necessity. This study reports on the development of a true to type novel "in vitro system" and its comparison with a conventional system for screening and selection of cultivars for high total phenolics, individual polyphenolics, and antioxidant content. We have shown for the first time using online acidic potassium permanganate chemiluminescence that extracts from Ocimum basilicum showed antioxidant potential. The current study identified the cultivar specific composition of polyphenolics and their antioxidant properties. Further, a distinct relationship between plant morphotype and polyphenolic content was also found. Of the 15 cultivars examined, "Holy Green", "Red Rubin", and "Basil Genovese" were identified as high polyphenolic producing cultivars while "Subja" was determined to be a low producer. The "in vitro system" enabled differentiation of the cultivars in their morphology, polyphenolic content, and antioxidant activity and is a cheap and efficient method for bioprospecting studies.

Chloroplast DNA phylogeography of holy basil (Ocimum tenuiflorum) in Indian subcontinent.

Ocimum tenuiflorum L., holy basil "Tulsi", is an important medicinal plant that is being grown and traditionally revered throughout Indian Subcontinent for thousands of years; however, DNA sequence-based genetic diversity of this aromatic herb is not yet known. In this report, we present our studies on the phylogeography of this species using trnL-trnF intergenic spacer of plastid genome as the DNA barcode for isolates from Indian subcontinent. Our pairwise distance analyses indicated that genetic heterogeneity of isolates remained quite low, with overall mean nucleotide p-distance of 5 × 10(-4). However, our sensitive phylogenetic analysis using maximum likelihood framework was able to reveal subtle intraspecific molecular evolution of this species within the subcontinent. All isolates except that from North-Central India formed a distinct phylogenetic clade, notwithstanding low bootstrap support and collapse of the clade in Bayesian Inference. North-Central isolates occupied more basal position compared to other isolates, which is suggestive of its evolutionarily primitive status. Indian isolates formed a monophyletic and well-supported clade within O. tenuiflorum clade, which indicates a distinct haplotype. Given the vast geographical area of more than 3 million km(2) encompassing many exclusive biogeographical and ecological zones, relatively low rate of evolution of this herb at this locus in India is particularly interesting.

Teor e composição do óleo essencial de quatro acessos e duas cultivares de manjericão (Ocimum basilicum L. ) / Essential oil composition and content of different accessions and cultivars of basil (Ocimum basilicum L. )

O objetivo desse estudo foi avaliar o teor e a composição química dos óleos essenciais de quatro acessos silvestres de manjericão (Ocimum basilicum L.), provenientes de diferentes regiões geográficas, e duas cultivares comerciais cultivados sob as mesmas condições edafoclimáticas. As sementes dos acessos foram coletadas nos município de Gurupi-TO (GUR) e de Monte Alegre-GO (MAL, MVE e FPS) e as sementes da cultivar comercial manjericão roxo (MR) foram adquiridas no comércio de Gurupi e da cultivar Maria Bonita (MB) foram fornecidas pelo Horto de Plantas Medicinais da Universidade Federal de Lavras. O plantio foi realizado em vasos com capacidade de dez litros utilizando-se como substrato: solo de barranco e esterco bovino na proporção 2:1. O óleo essencial foi obtido por hidrodestilação em aparelho tipo Clevenger e a identificação e a porcentagem relativa dos compostos do óleo essencial foi realizada por Cromatografia à Gás acoplada a Detector de Espectrometria de Massas. Foram identificados trinta e nove substâncias nos óleos essenciais obtidos, abrangendo monoterpenos, sesquiterpenos e fenilpropanoides. Nas cultivares MB e MR o composto majoritário foi o linalol, enquanto nos acessos silvestres (MAL, GUR, FPS e MVE) o composto em maior quantidade foi o (E)-cinamato de metila. O maior teor de óleo essencial foi obtido na cultivar Maria Bonita (3,77%).

The objective of this study was to evaluate the essential oil chemical composition and content of four wild accessions of basil (Ocimum basilicum L.), from different geographical regions, and two commercial cultivars grown under the same edaphoclimatic conditions. The seeds of the accessions were collected in the municipality of Gurupi, state of Tocantins (GUR), and Monte Alegre, state ofGoiás (MAL, MVE and FPS), and the seeds of the commercial cultivar of purple basil (MR) were acquired in the Gurupi market and the seeds of the commercial cultivar Maria Bonita (MB) were provided by the Garden of Medicinal Plants of the Federal University of Lavras. The seeds were planted in pots with a capacity of ten liters using as substrate soil from earth works and cattle manure in the proportion of 2:1. The essential oil was obtained by hydrodistillation using a Clevenger-type apparatus, and the identification and the relative percentage of the compounds of the essential oil were performed by Gas Chromatography coupled to Mass Spectrometry Detector. Thirty-nine compounds were identified in the essential oils obtained, which were divided into monoterpenes, sesquiterpenes and phenylpropanoids. In the cultivars MB and MR, the major compound was linalool, and in the wild accessions (MAL, GUR, FPS and MVE) it was the (E)-methyl cinnamate. The highest content of essential oil was obtained from the cultivar Maria Bonita (3.77%).

Quantification of character-impacting compounds in Ocimum basilicum and 'Pesto alla Genovese' with selected ion flow tube mass spectrometry.

Basil (Ocimum basilicum) is an important flavourant plant which constitutes the major ingredient of the pasta sauce 'Pesto alla Genovese'. The characteristic smell of basil stems mainly from a handful of terpenoids (methyl cinnamate, eucalyptol, linalool and estragole), the concentration of which varies according to basil cultivars. The simple and rapid analysis of the terpenoid constituents of basil would be useful as a means to optimise harvesting times and to act as a quality control process for basil-containing foodstuffs. Classical analytical techniques such as gas chromatography/mass spectrometry (GC/MS) are, however, slow, technically demanding and therefore less suitable for routine analysis. A new chemical ionisation technique which allows real-time quantification of traces gases, Selected Ion Flow Tube Mass Spectrometry (SIFT-MS), was therefore utilised to determine its usefulness for the assay of terpenoid concentrations in basil and pesto sauce headspace. Trace gas analysis was performed using the NO(+) precursor ion which minimised interference from other compounds. Character-impacting compound concentration was measured in basil headspace with good reproducibility and statistically significant differences were observed between cultivars. Quantification of linalool in pesto sauce headspace proved more difficult due to the presence of interfering compounds. This was resolved by careful selection of reaction product ions which allowed us to detect differences between various commercial brands of pesto. We conclude that SIFT-MS may be a valid tool for the fast and reproducible analysis of flavourant terpenoids in basil and basil-derived foodstuffs.

Evaluation of bioactivity of linalool-rich essential oils from Ocimum basilucum and Coriandrum sativum varieties.

Essential oils from Ocimum basilicum L. and Coriandrum sativum L. varieties originating from Turkey were investigated for their antimicrobial properties. The antimicrobial effects of the oil varieties were evaluated by the disc diffusion and minimum inhibitory concentration (MIC) methods against eight bacteria and three fungi. The compositions of the essential oils were analyzed and identified by GC and GC-MS. O. basilicum, C. sativum var. macrocarpum and var. microcarpum oils revealed the presence of linalool (54.4%), eugenol (9.6%), methyl eugenol (7.6%); linalool (78.8%), gamma-terpinene (6.0%), nerol acetate (3.5%); and linalool (90.6%), and nerol acetate (3.3%) as the major components, respectively. The oils exhibited antibacterial activity ranging from 1.25 to 10 microL disc(-1) against the test organisms with inhibition zones of 9.5-39.0 mm and minimal inhibitory concentrations values in the range 0.5- > or =1 microL/L. Linalool, eugenol, and methyl eugenol at 1.25 microL disc(-1) had antimicrobial effects on all microorganisms, giving inhibition zones ranging from 7 to 19 mm.

Influência do cultivo em consórcio na produção de fitomassa e óleo essencial de manjericão (Ocimum basilicum L. ) e hortelã (Mentha x villosa Huds. ) / Influence of intercropping on phytomass and essential oil production in basil (Ocimum basilicum L. ) and mint (Mentha x villosa Huds. )

As plantas medicinais produzem seus princípios ativos de acordo com os estímulos do ambiente, por isso a presença de outras plantas pode comprometer a produção dos metabólicos secundários. Assim, o presente trabalho teve como objetivo avaliar o efeito do cultivo consorciado sobre a produção de fitomassa e o teor de óleo essencial de manjericão (Ocimum basilicum L.) e hortelã (Mentha x villosa Huds.). O experimento foi conduzido na área experimental do Núcleo de Ciências Agrárias da Universidade Federal de Minas Gerais, Campus Regional de Montes Claros- MG. O delineamento utilizado foi o de blocos casualizados, com doze repetições, compreendendo seis tratamentos: manjericão + cenoura, manjericão + alface, manjericão solteiro, hortelã + cenoura, hortelã + alface e hortelã solteira. A produção de fitomassa e óleo essencial da hortelã não foram influenciados pelos tratamentos. Para o manjericão, apenas a massa fresca variou significativamente com os tratamentos.

Medicinal plants produce their active principles according to environmental stimuli; thus, the presence of other plants may affect the synthesis of secondary compounds. The present work aimed to evaluate the effect of intercropping on phytomass and essential oil production in basil (Ocimum basilicum L.) and mint (Mentha x villosa Huds.). The study was carried out in the experimental field of the Center for Agriculture, Federal University of Minas Gerais, Montes Claros Campus, Minas Gerais State, Brazil. The experimental design was in randomized blocks, with twelve replicates and six treatments: basil + carrot, basil + lettuce, sole crop of basil, mint + carrot, mint + lettuce, and sole crop of mint. Phytomass and essential oil production in mint were not affected by the treatments. As regards basil, only fresh matter significantly varied with treatments.

Evolution of Cinnamate/p-coumarate carboxyl methyltransferases and their role in the biosynthesis of methylcinnamate.

Methylcinnamate, which is widely distributed throughout the plant kingdom, is a significant component of many floral scents and an important signaling molecule between plants and insects. Comparison of an EST database obtained from the glandular trichomes of a basil (Ocimum basilicum) variety that produces high levels of methylcinnamate (line MC) with other varieties producing little or no methylcinnamate identified several very closely related genes belonging to the SABATH family of carboxyl methyltransferases that are highly and almost exclusively expressed in line MC. Biochemical characterization of the corresponding recombinant proteins showed that cinnamate and p-coumarate are their best substrates for methylation, thus designating these enzymes as cinnamate/p-coumarate carboxyl methyltransferases (CCMTs). Gene expression, enzyme activity, protein profiling, and metabolite content analyses demonstrated that CCMTs are responsible for the formation of methylcinnamate in sweet basil. A phylogenetic analysis of the entire SABATH family placed these CCMTs into a clade that includes indole-3-acetic acid carboxyl methyltransferases and a large number of uncharacterized carboxyl methyltransferase-like proteins from monocots and lower plants. Structural modeling and ligand docking suggested active site residues that appear to contribute to the substrate preference of CCMTs relative to other members of the SABATH family. Site-directed mutagenesis of specific residues confirmed these findings.

Rapid classification of basil chemotypes by various vibrational spectroscopy methods.

The potential of vibrational spectroscopy methods (attenuated total reflectance/Fourier-transform-infrared (ATR/FT-IR), FT-Raman and near infrared (NIR) spectroscopy) for the identification and quantification of valuable as well as carcinogenic substances in different basil chemotypes is described. It is shown that all main volatile components occurring in different basil accessions can be reliably determined in the isolated essential oils or solvent extracts but also in the air-dried herbs. While NIR data can be interpreted only by chemometric methods, IR and Raman spectra present characteristic key bands of the individual volatiles; therefore, in the latter case, a discrimination of basil chemotypes is frequently possible without applying chemometric algorithms. NIR calibrations are successfully established for various terpenoids and phenylpropanoids; on the basis of these data, the content of the two carcinogenic compounds methyleugenol (range: 2-235 microg/100 g) and estragole (range: 34-138 microg/100 g) can be reliably predicted in air-dried basil leaves (R (2) (coefficient of determination) = 0.951; SECV (standard error of cross validation) = 19.1 microg/100 g and R (2) = 0.890; SECV = 12.8 microg/100 g, respectively). The described methods were found to be very useful tools for the efficient selection of special basil single plants, adapted to the new demands set by the legislator and the consumer. Furthermore, they can be applied in industry to very easily control the purifying, blending, and redistilling processes of basil oil.