Hernandulcin in hairy root cultures of Lippia dulcis

The hairy root culture of Lippia dulcis Trev., Verbenaceae, was established by transformation with Agrobacterium rhizogenes A4. The transformed roots grew well in Murashige and Skoog medium containing 2% sucrose. The roots turned light green when they were cultured under 16 h/day light. The green hairy roots produced the sweet sesquiterpene hernandulcin (ca. 0.25 mg/g dry wt) together with 20 other mono- and sesquiterpenes, while no terpenes were detected in the nontransformed root cultures. The growth and hernandulcin production in the hairy root cultures were influenced by the addition of auxins to the medium. The addition of a low concentration of chitosan (0.2 – 10.0 mg / l) enhanced the production of hernandulcin 5-fold.Abbreviations Cht chitosan - IAA indole-3-acetic acid - NAA 1-naphthaleneacetic acid - 2,4-D 2,4-dichlorophenoxyacetic acid - MS Murashige and Skoog(1962)     

Trypanocidal Activity of Dysphania ambrosioides,Lippia alba,and Tetradenia riparia Essential Oils against Trypanosoma cruzi

Despite the current treatments against Chagas Disease (CD), this vector-borne parasitic disease remains a serious public health concern. In this study, we have explored the in vitro and/or in vivo trypanocidal and cytotoxic activities of the essential oils (EOs) obtained from Dysphania ambrosioides (L.) Mosyakin & Clemants (Amaranthaceae) (DA-EO), Lippia alba (Mill.) N.E. Brown (Verbenaceae) (LA-EO), and Tetradenia riparia (Hochst.) Codd (Lamiaceae) (TR-EO) grown in Brazil Southeast. DA-EO was the most active against the trypomastigote and amastigote forms in vitro; the IC₅₀ values were 8.7 and 12.2 μg mL⁻¹, respectively. The EOs displayed moderate toxicity against LLCMK2 cells, but the DA-EO showed high selectivity index (SI) for trypomastigote (SI=33.2) and amastigote (SI=11.7) forms. Treatment with 20 mg/kg DA-EO, LA-EO, or TR-EO for 20 days by intraperitoneal administration reduced parasitemia by 6.36 %, 4.74 %, and 32.68 % on day 7 and by 12.04 %, 27.96 %, and 65.5 % on day 9. These results indicated that DA-EO, LA-EO, and TR-EO have promising trypanocidal potential in vitro, whereas TR-EO has also potential trypanocidal effects in vivo.     

Toxicity,behavior impairment,and repellence of essential oils from pepper‐rosmarin and patchouli to termites

Plant essential oils are potential sources of insecticidal compounds, but have rarely been explored for their effect on termites. In the present study, we assessed the chemical composition of essential oils of Lippia sidoides Cham. (pepper‐rosmarin; Verbenaceae) and Pogostemon cablin (Blanco) Benth. (patchouli; Lamiacaeae) and evaluated their toxicity, behavioral impairment, and repellence to termite species of the genera Amitermes and Microcerotermes (Isoptera: Termitidae: Termitinae). The main components of essential oils of L. sidoides and P. cablin were thymol (44.6%) and patchouli alcohol (36.6%), respectively. The essential oil of P. cablin was most potent against Amitermes cf. amifer Silvestri and had the lowest LD₅₀ (0.63 μg mg^?1). There was no difference in toxicity for Microcerotermes indistinctus Mathews between the essential oils of L. sidoides (LD₅₀ = 1.49 μg mg^?1) and P. cablin (LD₅₀ = 1.67 μg mg^?1). Pogostemon cablin essential oil was the most toxic to M. indistinctus (LC₅₀ = 0.32 μl ml^?1) and A. cf. amifer (LC₅₀ = 0.29 μl ml^?1). The essential oils analyzed exhibited high toxicity and repellence to the termites, in addition to reducing behavioral interactions among individuals, thus constituting potential termiticides.     

Chemical composition and antifungal activity of the essential oils of Lippia rehmannii from South Africa

Lippia rehmannii H.Pearson (Verbenaceae) is an aromatic bush, indigenous to the northern parts of South Africa. As far as could be ascertained, the essential oil composition has not been previously reported and forms the subject of this investigation. Aerial parts of the shrub were collected from two localities in Gauteng, South Africa, and the isolated essential oils were analysed by gas chromatography. Citral, a mixture of the E- and Z-isomers, was found to be the main constituent of the oils, while borneol, camphor, neryl acetate, isocaryophyllene, p-cymene, β-caryophyllene and β-caryophyllene oxide were other major compounds present. Oil compositions, within and between the two localities, did not differ significantly. The in vitro antifungal activity of L. rehmannii essential oil was compared to that of Cympopogon citratus (lemongrass) and pure citral, against a number of pre- and postharvest fungal food pathogens. At a concentration of 3000 µL/L, lemongrass oil and pure citral caused complete growth inhibition of all the pathogens tested. Lippia rehmannii, containing less citral than lemongrass oil, was effective at this concentration against the majority of pathogens, but only partially restricted the growth of Lasiodiplodia theobromae and Botrytis cinerea. This finding suggests that citral may be largely responsible for the observed antifungal activities. Essential oil from L. rehmannii appears to be a good candidate for the in vitro control of Fusarium oxysporum and Rhizoctonia solani and application of these oils in the field should be investigated.     

The essential oil of Lippia alba: analysis of samples from French overseas departments and review of previous works

This paper contains new data on the chemical composition of the essential oil of Lippia alba (Mill.) N. E. Brown, as well as an overview of the available literature. Although the composition of the essential oil of this species is well-documented, discrepancies between the reported results suggest that many chemotypes and morphotypes exist. The analysis of essential oils obtained from the leaves of samples from three different locations in the French Overseas Departments (French Guiana, Martinique, and two different places in Guadeloupe) have shown that the composition of each one is quite different. This new data, along with a review and discussion of published studies, enabled us to establish seven distinct chemotypes. Possible connections between these chemotypes and morphotypes already described were also investigated.     

Morphology and histochemistry of the glandular trichomes of Lippia scaberrima (Verbenaceae)

BACKGROUND AND AIMS: Lippia scaberrima, an aromatic indigenous South African plant, with medicinal application, potentially has economic value. The production of essential oil from this plant has not been optimized, and this study of the chemico-morphological characteristics was aimed at determining the location of oil production within the plant. Furthermore, the locality of other secondary metabolites important in medicinal applications needed to be ascertained. This information would be useful in deciding the protocol required for isolation of such compounds. METHODS: The morphology of the glandular trichomes was investigated using a combination of scanning electron and light microscopy. Concurrently, the chemical content was studied by applying various chemical reagents and fluorescence microscopy. KEY RESULTS: Three types of trichomes were distinguished on the material investigated. Large, bulbous peltate glands containing compounds of terpenoid nature are probably the main site of essential oil accumulation. Small glands were found to be both peltate and capitate and fluorescent stain indicated the possible presence of phenolic compounds. The third type was a slender tapered seta with an ornamented surface and uniseriate base, and evidently secretory in nature. CONCLUSIONS: This study linking the chemical content and morphology of the glandular trichomes of L. scaberrima has contributed to the knowledge and understanding of secretory structures of Lippia spp. in general.     

Lippia dulcis shoot cultures as a source of the sweet sesquiterpene hernandulcin

The axenic shoot culture of Lippia dulcis Trev., Verbenaceae, was established on hormone-free Murashige-Skoog solid medium containing 3% sucrose. Shoots were cultured in various liquid or solid media. Woody Plant liquid medium was best for shoot multiplication, but the production of hernandulcin was relatively low. The highest hernandulcin content (2.9% dry wt) was obtained after 28 days of culture on Murashige-Skoog solid medium containing 2% sucrose. The addition of chitosan to the culture media enhanced the growth of shoots as well as the production of hernandulcin, especially with the liquid medium.Abbreviations MS(2%) Murashige-Skoog medium containing 2 % sucrose - MS(3%) Murashige-Skoog medium containing 3 % sucrose - 1/2MS half strength Murashige-Skoog medium containing 2% sucrose - B5 Gamborg B5 medium containing 2% sucrose - WP Woody Plant medium containing 2% sucrose     

Chromosome banding and essential oils composition of Brazilian accessions of <Emphasis Type="Italic">Lippia alba</Emphasis> (Verbenaceae)

The essential oil components and a karyotypic analysis of five Lippia alba (Verbenaceae) accessions from Brazil were performed with the objective of investigating the variation among different populations. The chemistry analysis allowed the grouping of the accessions in two main chemotypes: neral chemotype (LaCat, LaJF and LaRJ) and linalool chemotype (LaGua and LaVC). However, large karyotypic differences, verified by different chromosome banding techniques, were not detected. The results presented the same chromosome number for all accessions (2n = 30) with 10 metacentric chromosomes and 5 submetacentric. The chromosome banding showed great blocks of constitutive heterochromatin (C-bands) around the centromeric region, which was rich in AT bases (DAPI₊), while the CMA bands were observed only in terminal regions of six chromosomes. Through Ag-NOR techniques, only two active pairs of NORs were detected on the three pairs of secondary constrictions (the NOR activity is discussed). This work relates the pattern of heterochromatin for Lippia alba for the first time.     

Bioactivity of selected essential oils and some components on Listeria monocytogenes biofilms

Listeria monocytogenes is a Gram-positive bacterium, able to survive and grow in water, soil, agricultural products, various foods and the food-processing environment. The ubiquitous nature of the organism, coupled with its ability to colonise food-processing surfaces by forming biofilms, causes it to be of a major concern to the industry. Increased foodborne pathogen resistance and negative consumer perceptions regarding the use of synthetic bacteriocides, has resulted in natural antimicrobials being sourced from the plant kingdom. The listerial antibiofilm activities of Syzygium aromaticum (clove), Mentha spicata (spearmint), Lippia rehmannii and Cymbopogon citratus (lemongrass) essential oils and their major components were evaluated using the crystal violet assay and confocal scanning laser microscopy. Listerial biofilms treated with S. aromaticum or M. spicata essential oils, or the pure compounds nerol and citral, exhibited a similar biofilm biomass to the positive control. However, the essential oils of lemongrass, L. rehmannii, eugenol and R-carvone caused biofilm enhancement, rather than inhibition. L. rehmannii and lemongrass essential oils did not display any antibiofilm properties. Results obtained were confirmed by microscopic observations indicating either a reduction (inhibition) or an increase (enhancement) in biofilm biomass when exposed to the essential oils or pure compounds. The present study revealed that M. spicata and S. aromaticum essential oils as well as pure citral and nerol are good candidates for further development of ecofriendly disinfectants.