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Lamiaceae) Species from Iran Received: May 15, 2013; Accepted: July 15, 2013

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Lamiaceae) Species from Iran Received: May 15, 2013; Accepted: July 15, 2013 Progress in Biological Sciences Vol. 3, Number. 2, Summer/Fall 2013/81-98 Identification of flavonoids in leaves of seven wild growing Salvia L. (Lamiaceae) species from Iran Received: May 15, 2013; Accepted: July 15, 2013 Navaz Kharazian1* 1- Department of Botany, Faculty of Sciences, University of Shahrekord, Shahrekord, Iran ABSTRACT This study documents the flavonoid constituents of seven Salvia species in Iran namely S. atropatana Bunge, S. limbata C. A. Mey, S. sclarea L., S. ceratophylla L., S. multicaulis Vahl., S. hydrangea Dc. ex Benth., and S. eremophila Boiss. The studied species were collected from their natural habitats in Iran and were analyzed for their flavonoid constituents using two- dimensional thin layer chromatography with silica gel 60F 254 as solid phase. The purification of flavonoid compounds of each species was carried out using column chromatography with sephadex LH20. Based on the results, 53 flavonoid compounds were identified. The most frequent flavonoid subclasses among seven Salvia species were flavones (35.7%) and the least of these were dihydroflavonoles (5.3%). The most important structural variation observed in flavonoid was related to hydroxylation patterns. Among the identification of flavonoid, eight of them were reported for the first time inSalvia species of Iran. The highest numbers of flavonoid compounds were identified in S. multicaulis and S. hydrangea. It can be concluded that the flavonoid constituents seem to be a suitable indicator in chemotaxonomic studies in Salvia genus. Key Words: identification, Iran, Lamiaceae, leaves, flavonoid, Salvia . * Corresponding author: [email protected] Review Article Perspectives in taxonomy and phylogeny of the genus Astragalus (Fabaceae): a review Shahin Zarre*, Nasim Azani Center of Excellence in Phylogeny of Living Organisms and Department of Plant Science, School of Biology, College of Science, University of Tehran, Tehran, Iran Received: November 10, 2012; Accepted: November 24, 2012 Abstract The genus Astragalus L. (Fabaceae) is reviewed from both phyloenetic and taxonomic points of view. As the largest genus of flowering plants it has attracted many researchers, but much work remains to be done. A short taxonomic history with special focus on infrageneric classification of the genus, a list of phylogenetic studies including the applied markers and sampling strategies as well as a short discussion on evolution of morphological characters are presented. Keywords: Astragalus, Fabaceae, Taxonomy, Phylogeny Introduction R.Br. and few small genera (5). The taxonomy and phylogeny of Astragalus are very challenging. In With some 2900 species distributed in both Old- the present paper we will review the background of (1; ca. 2400 spp.) and New World (2; ca. 500 spp.), taxonomic and phylogenetic studies on Astragalus, Astragalus L. is by far the largest genus of the sources of complexity in the taxonomy and flowering plants, following by Bulbophyllum phylogeny of the genus, and future perspectives in Thouars of Orchidaceae with approximately 2032 studies on this genus. species and Psychotria L. of Rubiaceae comprising about 1951 species (1). The plants vary from short living annual herbs (ca. 80 spp.) to perennial rhizomatous or hemicryptophytic herbs (ca. 2500 Taxonomic background spp.) and to cushion forming spiny shrubs (ca. 300 Since its description in volume 2 of ‘Species species) in habit (Fig. 1). Most species grow in Plantarum’ (6), Astraglus has been subjected to semi-arid and arid areas throughout the world, but many taxonomic studies aiming mostly to achieve a few species prefer humid habitats (e.g. a natural subgeneric classification. Among these A. glycyphyllos L.), or are known as weeds. The systems, Bunge’s (7) classification of the genus in plants show the typical papilionaceous flowers and 1868 into eight subgenera and 105 sections, has are characterized by any unique morphological been widely used until recently (8, 9). Based on synapomorphy. As a result of this fact, the detailed morphological studies with emphasis on delimitation of the genus is sometimes very indumentum type (focusing on hair attachment: difficult and the assignment of some species (such basifixed vs. medifixed), Podlech (10) explained as A. annularis Forssk.) to this genus is doubtful the convergent nature of many of the and not supported by phylogenetic studies (2-4). morphological characters used in the delimitation According to recent investigations, the genus is of subgenera as proposed by Bunge (7) and currently placed in the well supported Astrgalean reduced the number of recognized subgenera to clade of tribe Galegeae s. l. close to genera three: subgen. Astragalus, subgen. Cercidothrix Oxytropis DC., Phyllolobium Fisch., Colutea L., Bunge and subgen. Trimeniaeus Bunge. He also Lessertia DC., Swainsona Salisb., Carmichaelia considered a unique position for tragacanthic * Corresponding author: [email protected] ©2013 Progress in Biological Sciences Tel.: +98-21-61112482 Introduction around the world such as antiviral, antitumor, antioxidant, etc. (24). Salvia L. genus with about 1000 species worldwide and 55 species in Iran, is the As Iran is one of the centers of diversity largest genera of Lamiaceae. It is distributed for Salvia species and the flavonoid all around the world, in temperate, compounds of this genus have not been subtropical, arctic and sub- arctic areas identified, there is a need for elucidating as well as the tropical regions of Iran (1, this genetic resource in this country. 2). Some of these species are perennial, Accordingly, this study aims to identify the herbaceous, suffruticose, fruticose and flavonoid constituents from seven Salvia sub shrubby (1, 3). Salvia genus displays species such as S. atropatana Bunge, S. a remarkable range of variation and sclarea L., S. ceratophylla L., S. limbata C. represents an enormous and cosmopolitan A. Mey., S. multicaulis Vahl., S. hydrangea distribution (2). The main speciation Dc. ex Benth. and S. eremophila Boiss. and centers of this taxon are considered to the reveal the chemotaxonomic value of these east of Mediterranean regions, south- west, compounds. western, eastern and central Asia, South Africa and central and South America (2, Materials and methods 4, 5). In recent years, studies on chemical Plant materials compounds of plant species were generally Seven Salvia species were collected from their constrained to the phenols and essential natural habitats in Iran (Table 1). The voucher oils (6). Salvia genus is a rich source of specimens were deposited in the Herbarium of phenolic compounds, essential oils and Shahrekord University (HSU). polysaccharides (6, 7, 8, 9, 10, 11). The flavonoid constituents have been generally Extraction and identification methods identified in some of Salvia species (12, 13, 14, 15, 16, 17, 18, 19, 20, 21). Lu and Foo Extraction of flavonoids was based on the (6) revealed that the flavonoid constituents in protocol suggested by Markham (25). The Salvia genus were mostly present as flavones, flavonoids were extracted from air-dried flavonols and their glycosides. Furthermore, leaf sample (10.5 g) of seven Salvia species B-ring and A-ring substitutions, oxygenation using 85% MeOH at 60°C. The extracts on the A-ring in c6 and/or c6 plus c8 positions were concentrated using a rotary evaporator were detected in Salvia genus, and mono- at 70°C for total solvent removal. The substituted (4') and di-substituted (3',4') separation of chlorophyll was initiated with H O at 60°C, filtered by Whatman paper B-ring are frequent (22). Investigation of 2 the chemical compounds of Salvia extracts and carotenoid pigments were removed can probably help to better understand the from the flavonoid extracts using n-BuOH. biological potential and the taxonomic The flavonoid extracts were separated from relationships among the investigated species n-BuOH using a rotary evaporator at 85°C (23). It is known that Salvia species are used in and solved in 100% MeOH. Subsequently, traditional medicines and natural activity all the crude extract was analyzed by two- dimensional thin layer chromatography Progress in Biological Sciences Vol. 3, Number 2, Summer/ Fall 2013 82 (TLC; 3 µm, 20× 20 cm) on silica gel MeOH 20% mixture) in 100 ml MeOH 60F 254 (15 mg silica gel, 67.5 ml H2O). solution (with increasing MeOH content Silica gel plates with the following solvent 20%, 40%, 60%, 80%, 100% and Acetone) systems were used: 1) BuOH-C2H4O2- and extracted in fractions (the amount of H2O (BAW 3:1:1V/V/V) representing an packing material is 50 ml for each MeOH organic system and 2) H2O- C2H4O2 (WA content 20%, 40%, 60%, 80%, 100% and 85:15V/V) representing an aqueous system. Acetone). The fractions were subjected to Spots' detection with natural product one dimensional thin layer chromatography identifiers (H2SO4/ MeOH solution) was on silica gel plates (3 µm). Identification of performed under UV-366 nm (26). The purified compounds was performed on the purification of flavonoid compounds of basis of their UV spectra (366 nm), MeOH each species was carried out using column solution and shift reagents such as AlCl3, chromatography (65 × 3 cm) with sephadex AlCl3/HCl, NaOAc, NaOAc/H3Bo3 and LH20 Sigma- Aldrich (Sephadex and MeOH. Table 1. The locality of Salvia species in their natural habitats from Iran Table 1. The locality of Salvia species in their natural habitats from Iran Species Locality Height (m) S. hydrangea (114) Fars- Abadeh; 31°08'N (latitude), 52°40'E (longitude); July 2010 1800 S. multicaulis (148) Isfahan- Semirom, Vanak; 31°32'N (latitude), 51°25'E (longitude); July 2010 1950 S. ceratophylla (144) Chaharmahal va Bakhtiari- Bostanshir; 32°05'N (latitude), 50°55'E (longitude); July 2010 2120 S. sclarea (139) Isfahan-Daran, Damane; 33°01'N (latitude), 50°29'E (longitude); August 2010 1856 S. atropatana (142) Kordestan- Marivan; 35°30'N (latitude), 46°25'E (longitude); August 2010 1820 S. limbata (125) Chaharmahal va Bakhtiari- Saman, Horeh; 32°32'N (latitude), 50°45'E (longitude); July 2010 2070 S.
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