Claudia Giuliani

Insight into the structure and chemistry of glandular trichomes of Labiatae, with emphasis on subfamily Lamioideae

Glandular trichomes of Labiatae are among the most investigated secretory structures. Most species studied belong to subfamily Nepetoidae, including plants with aromatic properties, while so far a few species of subfamily Lamioideae were examined. In this work, we studied the micromorphology, ultrastructure, type and release of secretion of the glandular trichomes present on leaves and flowers of several species belonging to subfamily Lamioideae, (Stachys alopecuros (L.) Bentham subsp. alopecuros, S. officinalis (L.) Trevisan subsp. officinalis, S. germanica L. subsp. germanica, S. germanica L. subsp. salviifolia (Ten.) Gams, S. sylvatica L., S. heraclea All., S. plumosa Griseb., S. annua L., Prasium majus L., Sideritis romana L.) and one to the sister group Scutellarioideae (Scutellaria galericulata L.). Besides the well-known peltate and small capitate trichomes, widely described in the literature, other types of glandular trichomes were encountered; stalked peltate hairs and large capitate hairs. In particular, a new type of capitate trichome, exclusive of calices and corollas, which presents a mode and release of secretion never described before, is reported.

Forest plant diversity is threatened by Robinia pseudoacacia (black-locust) invasion

The effects of black-locust invasion on plant forest diversity are still poorly investigated. Vascular plants are likely to be influenced by increasing nutrient availability associated with the nitrogen-fixing activity of black-locust, whereas it is not clear if, along with stand aging, black-locust formations regain forest species. The main aim of the present study was to test whether the increase of black-locust stand age promoted a plant variation in mature stands leading to assemblages similar to those of native forests. Therefore, plant richness and composition of stands dominated by native trees were compared with pure black-locust stands of different successional stages. Our study confirmed that the replacement of native forests by pure black-locust stands causes both plant richness loss and shifts in species composition. In black-locust stands plant communities are dominated by nitrophilous species and lack many of the oligothrophic and acidophilus species typical of native forests. Plant communities of native forests are more diverse with respect to pure black-locust stands, suggesting that black-locust invasion also causes a homogenization of the plant forest biota. We did not detect differences across the successional gradient of black-locust stands, and mature stands do not recover the diversity of plant species which are lost by the replacement of the native forests by black-locust. Accordingly some efforts in reducing the negative impacts of black-locust invasion on plant forest biota should be focused at least in those areas where conservation is among management priorities, such in the case of habitats included in the Habitat Directive (92/43 ECE).

Soil and plant changing after invasion: the case of Acacia dealbata in a Mediterranean ecosystem.

Acacia dealbata Link (Fabaceae) is one of the most invasive species in the Mediterranean ecosystems of Europe, Africa and America, where it has been proved to exert strong effects on soil and plant communities. In Italy A. dealbata has been largely used for ornamental and forestry purpose and is nowadays spreading in several areas. The present study was addressed to evaluate the impacts on soil chemical properties, soil microbial communities and understory plant communities and to assess the relationships among these compartments after the invasion of A. dealbata in a typical Mediterranean shrubland. Towards these aims, a soil and vegetation sampling was performed in Elba Island where A. dealbata is invading the sclerophyllous native vegetation. Three levels of invasion status were differentiated according to the gradient from invaded, to transitional and non-invaded vegetation. Quantitative and qualitative alterations of soil chemical properties and microbial communities (i.e. bacterial and fungal communities) and above-ground understory plant communities were found. In particular, the invaded soils had lower pH values than both the non-invaded and transitional ones. High differences were detected for both the total N and the inorganic fraction (NH4(+) and NO3(-)) contents, which showed the ranking: invaded>transitional>non-invaded soils. TOC and C/N ratio showed respectively higher and lower values in invaded than in non-invaded soils. Total plant covers, species richness and diversity in both the non-invaded and transitional subplots were higher than those in the invaded ones. The contribution of the nitrophilous species was significantly different among the three invasion statuses, with a strong increase going from native to transitional and invaded subplots. All these data confirm that A. dealbata modifies several compartments of the invaded ecosystems, from soil chemical properties to soil and plant microbial communities determining strong changes in the local ecosystem processes.

Characterization of Secondary Metabolites, Biological Activity and Glandular Trichomes of Stachys tymphaea Hausskn. from the Monti Sibillini National Park (Central Apennines, Italy)

Stachys tymphaea (Lamiaceae) is a perennial herb growing in forest openings and dry meadows of central and southern Italy. It was investigated for the first time here, determining the content of secondary metabolites, the micromorphology of glandular trichomes, the histochemical localization of secretion, and the biological activity of the volatile oil, namely, the cytotoxic, antioxidant, and antimicrobial properties. The plant showed a peculiar molecular pattern, being rich of biophenolic compounds as flavonoids, phenylethanoid glycosides, and caffeoylquinic acid derivatives, but poor of iridoids, which are known as marker compounds of the genus Stachys. The essential oil was characterized by GC‐FID and GC/MS analyses, revealing a high percentage of sesquiterpene hydrocarbons (54.6%), with germacrene D (30.0%) and (E)‐β‐farnesene (12.4%) as the most abundant compounds, while other main components were representatives of the diterpenes (19.2%), represented mainly by (E)‐phytol (11.9%). This composition supported the taxonomic relationships in the genus Stachys, which comprises oil‐poor species producing essential oils rich in hydrocarbons, with germacrene D as one of the predominant components. The micromorphological study revealed three types of glandular hairs, i.e., Type A peltate trichomes, being the primary sites of essential oil biosynthesis, Type B short‐stalked trichomes, typical mucopolysaccharide producers, and Type C long capitate trichomes, secreting a complex mixture of both lipophilic and hydrophilic substances, with a major phenolic fraction. Moreover, the MTT assay revealed the potential of the volatile oil to inhibit A375, HCT116, and MDA‐MB 231 tumor cells lines (IC50 values of 23.9–34.4 μg/ml).

Glandular trichomes as further differential characters between Stachys subgenus Betonica (L.) Bhattacharjee and Stachys subgenus Stachys

Abstract The genus Stachys L. has always presented great taxonomic and nomenclatural problems; the identification of two different genera, Betonica and Stachys, or of a single genus, Stachys, with the subgenera Betonica and Stachys, is still debated. In this work, different types of glandular trichomes (peltate and capitate) occurring in species belonging to both subgenera Betonica and Stachys are described. Observations were carried out on living plants growing in Italy, using scanning electron microscopy (SEM), to characterise trichome structure and distribution on vegetative and reproductive organs, and light microscopy (LM) to characterise their type of secretion, by means of histochemical methods. Stachys subgenus Betonica presents only peltate hairs, while Stachys subgenus Stachys has different types of large capitate hairs lacking in the subgenus Betonica. Furthermore, the peltate trichomes of the subgenus Betonica show an unusual secretion composed of flavonoids and essential oils. A thorough examination of numerous exsiccata belonging to subgenera Betonica and Stachys, stored in the Herbarium Centrale Italicum of Florence (FI), confirmed observations made on living material. The recorded characters allowed a clear distinction between the species of the subgenus Betonica and those of the subgenus Stachys. Consequently, a higher taxonomic rank than subgenus should be recommended for these taxa.

Phytochemistry, micromorphology and bioactivities of Ajuga chamaepitys (L.) Schreb. (Lamiaceae, Ajugoideae): Two new harpagide derivatives and an unusual iridoid glycosides pattern.

Ajuga chamaepitys (L.) Schreb, well-known as Camaepitium or Ground Pine, is an annual herb typical of the Mediterranean area accounting several uses in the traditional medicine. In this work we have, analyzed the plant iridoid fraction together with the essential oil composition and study of the plant indumentum. Finally, we assayed the polar extracts and essential oil obtained from the aerial parts for antioxidant activity and cytotoxicity on tumor cells. The analysis of the monoterpene glycosides allowed us to isolate from roots and aerial parts and to structurally elucidate by NMR and MS the following compounds: ajugoside (1), reptoside (2), 8-O-acetylharpagide (3), harpagide (4), 5-O-β-d-glucopyranosyl-harpagide (5), asperulosidic acid (6), deacetyl asperulosidic acid (7) and 5-O-β-d-glucopyranosyl-8-O-acetylharpagide (8), among which 5 and 8 were two new natural products. Chemotaxomic relevance of these constituents was discussed. The chemical analysis of A. chamaepitys essential oil by GC-FID and GC-MS showed ethyl linoleate (13.7%), germacrene D (13.4%), kaurene (8.4%), β-pinene (6.8%), and (E)-phytol (5.3%) as the major volatile components. The micromorphological and histochemical study showed that iridoids and essential oil are mainly produced in the type III capitates and peltate trichomes of leaves and flowers. Biological evaluations of A. chamaepitys polar extracts and essential oil showed that the former were more potent as radical scavengers than the latter. MTT assay revealed that essential oil and ethanolic extracts were moderately cytotoxic on tumor cells with IC50 of 36.88 and 59.24μg/mL on MDA-MB 231 cell line, respectively, and IC50 of 60.48 and 64.12μg/mL on HCT116, respectively.

A checklist of the alien flora of the Tuscan Archipelago (Italy)

Biological invasions represent one of the major threats to biodiversity. Their impact is considered to be even higher in the island ecosystems, especially in Mediterranean ones, which host a very high diversity. Moreover nowadays the global changes and the local changes in the socio-economic drivers of these island enhanced human impact on the insular biota, also leading to an increase in alien species introductions and in a shift in the introduction pathways. In this framework, checklists of alien species represent a valuable tool for monitor the invasion process at a local scale. Aim of this paper is to provide an updated Checklist of the alien flora of the Tuscan Archipelago, also reporting some important information such year of first and more recent record and invasion status for each island. Intense field surveys were made to verify the presence of the reported species and detect the presence of new arrivals. The Checklist includes a total of 141 taxa belonging to 101 genera in 49 families. 31 taxa are considered invasive, 62 naturalized and 48 casual. Most of the species were introduced for ornamental purpose, according to socio-economical changes which nowadays see the tourism as the main economic driver in the Archipelago.

Congruence of Phytochemical and Morphological Profiles along an Altitudinal Gradient in Origanum vulgare ssp. vulgare from Venetian Region (NE Italy)

Plants of Origanum vulgare L. ssp. vulgare from the Veneto region (NE Italy) were selected to study the variability of the essential‐oil composition from leaves and inflorescences throughout an elevation gradient. We investigated also the morphology of non‐glandular and glandular trichomes, their distribution on the vegetative and reproductive organs, as well as the histochemistry of the secreted products, with special focus on the terpenoidic fraction.

Chemical differences in volatiles between Melittis melissophyllum L. subsp. melissophyllum and subsp. albida (Guss) P. W. Ball (Lamiaceae) determined by solid-phase microextraction (SPME) coupled with GC/FID and GC/MS.

Melittis melissophyllum (Lamiaceae) is a perennial herb, typical of woody places, occurring in Italy with two subspecies, i.e., melissophyllum and albida. So far, the classification of these two taxa was only based on morphology, i.e., the presence of glandular trichomes, the dimension of the leaves, and the number of teeth on each side as the main discriminant characters. To find marker compounds to chemically discriminate the subsp. melissophyllum with respect to the subsp. albida, a solid‐phase microextraction SPME analysis coupled with GC/FID (=flame ionization detector) and GC/MS was carried out. SPME proved to be a chemotaxonomically useful technique that permitted a clearly differentiation of the two subspecies at headspace level. The subsp. melissophyllum was characterized by high amount of the mushroom alcohol oct‐1‐en‐3‐ol and the phenolic coumarin, whilst the subsp. albida exhibited a high content in monoterpenes and sesquiterpenes, α‐pinene, sabinene, and (E)‐caryophyllene being the major compounds. Multivariate chemometric techniques, such as cluster analysis (CA) and principal‐component analysis (PCA), were used to support chemical data and characterize the population according to the taxonomy. In addition, the micromorphology and distribution of glandular trichomes of both subspecies were studied by scanning electron microscopy (SEM).

Two alien invasive acacias in Italy: Differences and similarities in their flowering and insect visitors

In Italy, alien acacias have been introduced for ornamental and reforestation purposes, and some species became invasive occupying patches of the Mediterranean landscape. On the Island of Elba (Central Italy), Acacia dealbata and A. pycnantha form dense stands at short distance, showing an impressive massive flowering at the end of the winter/early spring. Our aim was to investigate the behaviour of the two species in relation to the flowering features, from phenology to floral characteristics, and their replay to the observed flower visitors. Differences between the two species emerged on all the parameters considered. A. pycnantha peak of flowering occurred later than A. dealbata and showed larger flower heads (FHs), more flowers/head, stamens/flower and polyads. On A. dealbata, we recorded longer racemes and more FHs/raceme, determining a more flower-dense crown. Even if contacts with flower visitors were generally low on both species, A. dealbata showed a more heterogeneous visitor assemblage. Both acacias species interacted with local generalist pollinators, as bumblebees and honey bees. Flower handling and resource collection strategy by the honey bee indicate a long-term relationship between the bee and the acacias, with bees investing longer time on the larger A. pycnantha flower heads.