GNPL#1709193, VOL 0, ISS 0 Essential oil compositions of Teucrium fruticans, T. scordium subsp. scordioides and T. siculum growing in Sicily and Malta Rossella Gagliano Candela, Vincenzo Ilardi, Natale Badalamenti, Maurizio Bruno, Sergio Rosselli, and Filippo Maggi QUERY SHEET This page lists questions we have about your paper. The numbers displayed at left are hyperlinked to the location of the query in your paper. The title and author names are listed on this sheet as they will be published, both on your paper and on the Table of Contents. Please review and ensure the information is correct and advise us if any changes need to be made. In addition, please review your paper as a whole for typographical and essential corrections. Your PDF proof has been enabled so that you can comment on the proof directly using Adobe Acrobat. 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Please correct if this is inaccurate. 1 NATURAL PRODUCT RESEARCH https://doi.org/10.1080/14786419.2019.1709193 2 3 4 5 Essential oil compositions of Teucrium fruticans, 6 T. scordium subsp. scordioides and T. siculum growing 7 8 in Sicily and Malta 9 Rossella Gagliano Candelaa, Vincenzo Ilardib, Natale Badalamentia, Maurizio 10 Q5 Brunoa, Sergio Rossellic and Filippo Maggid 11 12 aDepartment of Biological, Chemical and Pharmaceutical Sciences and Technologies (STEBICEF), b 13 University of Palermo, Palermo, Italy; Department of Earth and Marine Sciences (DISTeM), University of Palermo, Palermo, Italy; cDepartment of Agricultural and Forest Sciences (SAF), University of 14 Palermo, Palermo, Italy; dSchool of Pharmacy, University of Camerino, Camerino, Italy 15 16 ABSTRACT ARTICLE HISTORY 17 In the present study, the chemical composition of the essential Received 7 December 2019 18 oils from the aerial parts of Teucrium fruticans L. collected in Sicily Accepted 15 December 2019 19 and Malta, Teucrium scordium subsp. scordioides (Schreb.) Arcang. and Teucrium siculum (Raf.) Guss., collected in Sicily, were eval- KEYWORDS 20 Teucrium uated by GC-MS. The main volatile components of both T. fruti- ssp.; Volatile 21 components; Germacrene D; cans collections were germacrene D (29.4% and 50.0%), (E)- b b (E)- -Caryophyllene; 1- 22 -caryophyllene (19.6% and 21.9%), and 1-octen-3-ol (19.7% and Octen-3-ol; 23 7.4%); T. scordium subsp. scordioides essential oil was rich in car- Chemotaxonomy yophyllene oxide (25.8%), a-pinene (19.4%) and b-pinene (8.5%); 24 T. siculum essential oil was rich in (E)-b-caryophyllene (30.9%), 1- 25 octen-3-ol (9.0%), a-humulene (8.6%) and germacrene D (8.0%). 26 The chemotaxonomic relationship with other members of the 27 respective botanical sections was discussed. 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 CONTACT Sergio Rosselli [email protected] ß 2019 Informa UK Limited, trading as Taylor & Francis Group 2 R. GAGLIANO CANDELA ET AL. 46 1. Introduction 47 Teucrium is a plant genus of the Lamiaceae family, represented mostly by perennial, 48 bushy or herbaceous plants, living commonly in sunny habitats. The genus consists of 49 415 accepted taxa, including species, subspecies, varieties, forms and hybrids (World 50 Checklist), that are divided in ten botanical sections (McClintock and Epling 1946; 51 Tutin and Wood 1972). 52 The species of the genus Teucrium grow in moderate climate zones, particularly in 53 the Mediterranean basin and Central Asia. As the species can be found in southern, 54 south-western and south-eastern part of Europe, the continent is regarded as one of 55 the main centre of differentiation of the genus. A significant number of species have 56 been also observed in south-western Asia, north-western Africa, southern North 57 America and south-western South America. As for Australia, the species of genus 58 Teucrium are distributed in both southern parts of the continent and certain nearby 59 islands (Mesuel et al. 1978, Hollis and Brummitt 1992) 60 In Sicily the genus Teucrium is represented by ten taxa: Teucrium fruticans L., 61 Teucrium campanulatum L., Teucrium siculum (Raf.) Guss., Teucrium scordium subsp. 62 scordioides (Schreb.) Arcang., Teucrium spinosum L., Teucrium chamaedrys L., Teucrium 63 flavum L., Teucrium montanum L., Teucrium luteum (Mill.) Degen, and Teucrium capita- 64 65 tum L. (Pignatti 2017) 66 Teucrium fruticans is a Mediterranean species occurring in Southern Europe and 67 North Africa growing mainly on calcareous rocks near the seaside. Due to its blue 68 flowers with its evergreen foliage, it is largely used as an ornamental plant. In Italy, it 69 grows along the Tyrrhenian coasts up to Tuscany, Sicily and Sardinia, and in almost all 70 the minor islands. It also grows in Maltase archipelago (Euro Check List). In Tuscany, “ ” 71 where it is known as Erba di Santa Lucia the infusion of the leaves is used as depura- 72 tive and diuretic (Maccioni et al. 1997), whereas on Mt Etna (Sicily), where it is known “ ” 73 as Erba ricottara , the leaves are used to treat hemorrhoids (Tuttolomondo et al. 74 2014). Phytochemical investigations on the extracts of this species showed that it is 75 quite rich of diterpenoids. Researches, performed on the aerial parts of a Sicilian acces- 76 sion, allowed to identify three new neo-clerodanes and a new seco-neo-clerodane 77 derivative (Savona et al. 1978a; 1978b; Bruno et al. 1992), whereas the analysis of its 78 roots showed the presence of three new rearranged abietane diterpenoids (Bruno 79 et al. 1990; Cuadrado et al. 1992). Other metabolites isolated from the aerials parts of 80 the same population were triterpenoids (Passannanti et al. 1983), an unusual fatty acid 81 ester (Fontana et al. 1999) and several flavonoids (Kisiel et al. 2014). The investigation 82 of plants collected in Barcellona (Spain), showed the occurrence, apart from the previ- 83 ously detected metabolites, of nine neo-clerodanes. All these compounds were tested 84 for their antifeedant activity against Spodoptera littoralis, showing for some of them a 85 moderate activity (Coll and Tandron 2004; 2005). Similar antifeedant studies on natural 86 and synthetic derivatives indicated a good potency of isofruticolone (Bruno et al. 87 1999). Recently, the investigation of the aerial parts of T. fruticans, cultivated in China, 88 showed the occurrence of other seven new neo-clerodane diterpenoids. All the diter- 89 penes were evaluated for their cytotoxic activities on three human cancer cell lines, 90 and for their ability to inhibit LPS-induced nitric oxide production in RAW 264.7 mac- rophages. None of the compounds displayed cytotoxic activities on the cancer cell NATURAL PRODUCT RESEARCH 3 91 lines, and only 11-hydroxyfruticolone showed weak NO inhibitory activity (Lv et al. 92 2015). The methanol extract of the leaves of T. fruticans collected in Northern Morocco 93 was tested for its antioxidant and antibacterial activities. The results indicated moder- 94 ate antioxidant properties and poor activity against Staphylococcus spp. and B. subtilis 95 (Boudkhili et al. 2012). Also, the ethanol extract of the inflorescences of plants col- 96 lected on Mt. Etna (Sicily) inhibited the growth of several bacterial strains, with better 97 activity against Gram þ bacteria (Acquaviva et al. 2018). To the best of our knowledge, 98 only one paper on the analysis of the essential oil of T. fruticans has been published 99 and regards plants collected in Tuscany (Italy) (Flamini et al. 2001). 100 Teucrium scordium subsp. scordiodes [Bas.: Teucrium scordioides Schreb.; sinonymus: T. 101 scordium L. subsp. scordioides (Schreb.) Maire & Petitm.; Syn.: T. scordioides var. lanugino- 102 sum (Hoffmanns. & Link) Lojac.] is a perennial species distributed in southern and south- 103 eastern Europe, Asia Minor and Morocco. It grows mainly on wet soil and swamps 104 (Horvat et al. 1974). Chemical investigation on a Serbian accession of this taxa showed 105 the presence of several flavonoids in the methanol extract (Kundakovicetal.2011). In 106 the ethanol extract of T. scordium ssp. scordioides collected in Spain many free flavone 107 aglycones and glycosides were identified (Harborne et al. 1986). The cyclohexane and 108 dichloromethane extracts of T. scordium subsp. scordiodes have shown to possess high 109 cytotoxicity against MDA-MB-361 and MB-453 cell lines (Kundakovicetal.2011). Further 110 studies indicated for this taxa antioxidant, proapoptotic and cytotoxic activity against 111 HeLa and K562 cancer cell lines (Stankovicetal.2011; 2012; 2015). With regard to the 112 analysis of the essential oil only one report has been published and concerns a Serbian 113 population of T. scordium subsp. scordiodes (Radulovicetal.2012). 114 Teucrium siculum [Bas.: Scorodonia sicula Raf., T. scorodonia subsp. siculum (Raf.) 115 Nyman, T. scorodonia subsp. crenatifolium (Guss.) Arcang., T. scorodonia L. var. crenati- 116 folium Guss., T. pseudo-scorodonia Ces., Passer. & Gibelli non Desf.] is an herbaceous 117 perennial shrub. It is widely spread in Middle and Southern peninsular Italy, as well as 118 in Sicily. Moreover, small disjointed areas of the latter species are known on the 119 Euganean hills and in Vicenza and Trento outskirts (Servettaz et al.