Revista Brasileira de Farmacognosia 27 (2017) 143–152

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Original Article

Pharmacobotanical study of thymopsis

Mehmet Tekin

Department of Pharmaceutical Botany, Faculty of Pharmacy, Cumhuriyet University, Sivas, Turkey

a b s t r a c t

a r t i c l e i n f o

Article history: Hypericum thymopsis Boiss., , is an endemic herb which generally grows at the calcareous

Received 16 June 2016

steppe regions of Central Turkey. In flowering stage, the aerial parts of this species are used for wound-

Accepted 19 September 2016

healing and sedation, and its infusions are used against stomach diseases and throat infections by local

Available online 21 October 2016

people. The aim of this study, to examine and to reveal of the morphology, anatomy and histology of

the aerial vegetative and reproductive organs of the H. thymopsis, which are used in popular medicine

Keywords:

and thus contributing to the pharmacognostic evaluation of the species. In comparison with previous

Anatomy

published morphological description of the species, some different findings about height and leaf

Clusiaceae

length were found. In addition, some morphological characteristics such as dimensions of sepals and

Hypericum thymopsis

ovaries, length of filaments, anthers, pistils and pedicels were examined here for the first time. The

Medicinal plant

Morphology anatomical characteristics of stem, leaf, sepal, filament and pistil were studied using light microscopy

Turkey and additionally for stem and leaf using scanning electron microscopy. The stem has the secondary

growth, and circular shape. The leaves are amphistomatic and the mesophyll is dorsiventral. Stomata are

anisocytic and sunken. The glandiferous emergences are present on stem and translucent glands exist in

leaf mesophyll. Type A secretory canals are present in stem, leaf, sepal and petal cross section. While type

B secretory canals are observed in sepal and petal, type C secretory canals are observed solely in ovary

cross section. All the structural features herein found can assist the diagnosis of H. thymopsis.

© 2016 Sociedade Brasileira de Farmacognosia. Published by Elsevier Editora Ltda. This is an open

access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

Introduction and diabetes in Turkish folk medicine (Bingol et al., 2011; Özkan

and Mat, 2013). The taxa of Hypericum characterized by having

Hypericaceae is a small cosmopolite dicot family represented extensively secretory structure such as translucent glands, black

by nine genera and about 500 species of tree, shrub and herb in nodules and secretory canals which synthesizing and accumulating

the world (Simpson, 2006; APG III, 2009; Carine and Christenhusz, biologically active substances, so they have great phytochem-

2010). Hypericum is the largest genus of Hypericaceae by com- ical potential and composition (Siersch, 1927; Metcalfe et al.,

prising about 400 species throughout the world except the poles, 1950; Curtis and Lersten, 1990; Ciccarelli et al., 2001a, 2001b;

deserts and low altitude areas of tropical regions (Lewis, 2003; Onelli et al., 2002). Some of these phytochemical compositions of

Meseguer and Sanmartín, 2012). In Turkey, Hypericum is repre- the Hypericaceae are naphthodianthrones (notedly hypericin and

sented by 96 species, 104 taxa in which 45 of them are endemic pseudohypericin), acylphloroglucinol derivatives (notedly hyper-

with the endemism ratio 43% (Güner et al., 2012). The genus forin and adhyperforin), flavonoids (notedly quercetin, quercitrin,

Hypericum characterized by possess generally pale to dark yellow hyperoside and biapigenin), tannins, n-alkanes, xanthones and

flowers and transparently dotted leaves with red or black col- essential oils (Bombardelli and Morazzoni, 1995; Bruneton, 1995;

ored glands (Davis, 1966, 1988; Güner et al., 2000). The genus Kitanov, 2001; Maggi et al., 2004). Due to their phytochemical

Hypericum is known worldwide with its wide folk medicinal composition and waste usage in traditional folk medicine, many

usage (Avato, 2005; Assadi et al., 2011). Hypericum species are studies were made on biological activities of different species

known under the names “binbirdelik otu”, “kanotu”, “peygamber extracts of Hypericum in Turkey and in the world. For exam-

c¸ ic¸ egi”˘ and by a majority “sarı kantaron”, and have been used as ple, analgesic activity (Öztürk, 2001), wound healing activity

sedatives, antiseptics, anti-inflammatory, anti-astmatic, antispas- (Öztürk et al., 2007), hepatoprotective activity (Öztürk et al., 1992),

modics, wound-healing and to against stomach diseases, ulcers anti-inflammatory activity (Öztürk et al., 2002), anti-Helicobacter

pylori activity (Yes¸ ilada et al., 1999) and antidepressant activity

(Öztürk, 1997) of well-known Hypericum, H. perforatum L. were

studied by different researchers. In addition, antioxidant activ-

E-mail: [email protected] ity of H. venustum Fenzl was studied by Spiteller et al. (2008),

http://dx.doi.org/10.1016/j.bjp.2016.09.004

0102-695X/© 2016 Sociedade Brasileira de Farmacognosia. Published by Elsevier Editora Ltda. This is an open access article under the CC BY-NC-ND license (http:// creativecommons.org/licenses/by-nc-nd/4.0/).

144 M. Tekin / Revista Brasileira de Farmacognosia 27 (2017) 143–152

antimicrobial activity of H. hyssopifolium Chaix var. microcalycinum University, Faculty of Science Herbarium (CUFH), Department

(Boiss. & Heldr.) Boiss. was studied by Toker et al. (2006), and of Biology, Sivas, Turkey. Taxonomical identification was made

myeloperoxidase activity of H. empetrifolium Willd. was studied by according to Flora of Turkey (Davis, 1966) by author.

Kültür (2007). In addition there are some studies focused morphol-

ogy, anatomy and secretory structure of some Hypericum species

(Metcalfe et al., 1950; Toma and Rugina,˘ 1998; Hong-Fei and Zheng- Structural analysis

Hai, 2001; Yaylacı et al., 2013; Perrone et al., 2013a, 2013b).

Hypericum thymopsis Boiss. is an endemic species for Turkey Morphological description of H. thymopsis was made using both

Flora and naturally growing in Sivas, Kayseri, Malatya and fresh and herbarium samples collected from different localities.

Kahramanmaras¸ provinces (Davis, 1966). Essential oil composition Parts of the some fresh material were stored in 70% alcohol–water

of H. thymopsis was studied by Özkan et al. (2009, 2013). Özkan et al. solution for later histo-anatomical procedures. For this reason,

(2013) studied composition of essential oils of five endemic Hyper- hand-made transverse sections of stem, leaf, sepal, petal, filament,

icum species for Turkey (H. uniglandulosum Hausskn. ex Bornm., ovary and style were taken. Also superficial hand-made sections

H. scabroides Robson and Poulter, H. kotschyanum Boiss., H. sal- of adaxial and abaxial surface of the leaf blade were taken with

sugineum Robson and Hub.-Mor. and H. thymopsis). According to razor blade. The sections were stained in 1% Alcian blue (Sigma)

their study, ␣-pinene, baeckeol, limonene and spathulenol were and 1% Safranine O (Sigma), in a ratio 3/2 (Davis and Barnett, 1997).

identified as major components of H. thymopsis. Furthermore, H. The sections were kept about 5 min in the dye. Semi-permanent

thymopsis has greatest potential by having so much more ␣-pinene, slides were mounded using glycerin-gelatine (Jensen, 1962). The

within these five species. ␣-Pinene is one of the active substance structural investigations of vegetative and reproductive parts of H.

which has bronchodilator and anti-inflammatory activity (Russo, thymopsis were made using Olympus BX22 light microscopy. Pho-

2011) and has bacteriostatic and bactericide effect against espe- tomicrographs were taken using Olympus BX51 light microscopy

cially gram (+) and gram (−) bacteria with broad spectrum (Nissen coupled with Olympus DP70 digital camera. The anatomical ter-

et al., 2010). In addition, it has acetylcholinesterase inhibitor activ- minology used is that according to Esau (1965). For secretarial

ity, thus it has effect that strengthens the memory of human (Russo, analyses, the study of Ciccarelli et al. (2001b) was carried out:

2011). ␣-Pinene is also reported as one of the main components (a) Translucent glands. These are pale glands or glandular pockets

of the essential oils of other some Hypericum species (Joulain and (Robson, 1981) and are spherical or oblong glands delimited by two

König, 1998; Santos et al., 1999; Zeng and Zhou, 2001; Gudziˇ c´ et al., layers. (b) Three types of secretory canals. Type A: the lumen of the

2002, 2004). canals is usually surrounded by four (or more in especially flow-

In the field works, by interviewing local peoples under an ers parts) polygonal cells which have very thin wall toward canal

ethnobotanical studies, it was learned that above ground parts lumen. Type B: they have a same structure with translucent glands

of H. thymopsis are used for wound-healing and sedation, and its with their wide lumen, but they seem elongated and pale which

infusions and decoctions as tea are used also against stomach dis- differ from the translucent glands. Type C: type C canals composed

eases and throat infections. By considering great usage of various of wide cavity surrounded by one or more cell layers which have

Hypericum taxa in folk medicine, their anatomical features are densely stained and thin walls.

not well-known except H. perforatum. In Turkey, there are some For scanning electron microscopy (SEM) analyses, the some

anatomical studies on other Turkish Hypericum taxa (Yaylacı et al., plant parts were mounted on aluminum stubs and coated with gold.

2013; Altıntas¸ and Akc¸ in, 2015). The morphological characteristics The micromorphological and anatomical observations were made,

of all Turkish Hypericum taxa are more or less known. But there and micrographs were taken at different magnifications by using

is some deficiency on some endemic species such as H. thymop- LEO 440 SEM.

sis. For example, in Flora of Turkey (Davis, 1966), dimensions of

sepals and ovaries, length of filaments, anthers, pistils and pedicels

Results

of H. thymopsis are not reported. Despite of some essential oil stud-

ies on H. thymopsis (Özkan et al., 2009, 2013), there is no report

Morphological aspects

on anatomical and histological structure of the species. The aim

of this study, revealed entirely morphological characteristics, and

Perennial herbs. Stems 3–14 cm, erect, numerous, glabrous,

anatomical and histological structure in all above ground parts and

scabrid with glandiferous emergences (Fig. 1B). Leaves

their secretory structures which make up biologically active sub-

5–18 × 0.6–1.6 mm, linear, revolute with obtuse or rounded

stances of H. thymopsis.

apex, glabrous, usually papillose at adaxial surface and especially

at margins. Black glands present at the margins of small leaves

which positioned at the base of pedicel on stem. Inflorescence

Materials and methods

corybose, up to 17 cm in fruiting time, soliter or 20 flowered. Sepals

1.9–2.8 × 0.8–1.1 mm, united at base, lanceolate to lanseolate-

Plant material

elliptic with sessile or subsessile black glands at margins and

acute apex (Fig. 1D). Sepals reddish when flowers in bud stage,

The specimens of Hypericum thymopsis Boiss., Hypericaceae,

green in flowering time (Fig. 1A, D). Petals 4.5–7.5 × 2–3.8 mm,

were collected during the flowering and fruiting time from differ-

pentamerous, elliptic-oblanceolate to obovate and without black

ent natural populations in Sivas province of Turkey. Field works

glands (Fig. 1C, E). Filaments 4–5.5 mm, thin and numerous.

were carried out in between the years 2012–2014. Locality 1: B6

◦   ◦  

Anthers 0.35–0.45 mm, dorsifixed and longitudinal dehiscence

Sivas: Ulas¸ district, Ziyarettepe, 1444 m, 39 33 06.7 N; 37 01 11.9

(Fig. 1F). Pistils 4–5 mm, ovaries 1.5–2.2 × 0.9–1.4 mm, 3 loculed

E, 26.06.2013; Locality 2: B6 Sivas: Ulas¸ district, Kurtlukaya vil-

◦   ◦  

and ovoidal shaped. Style 3, free and selender, 2.5–3.2 mm (Fig. 1G).

lage to Bogazdere˘ village, 1458 m, 39 23 02.1 N; 36 55 48.3 E,

Placentation axillar, ovules anatropous. Pedicels 2–8 mm. Fruit

05.06.2014; Locality 3: B6 Sivas: Sivas-Kangal-Gürün road inter-

◦   ◦  

4.5–5.5 × 2.7–3.1 mm, ovoid, reddish-brown, septicidal capsule

section, 1560 m, 39 07 53.1 N; 37 14 32.9 E, 06.08.2012, ibid.

rostrate and with 3 locule, each locule bear usually 2, occasionally

05.06.2014, ibid. 05.07.2014.

1 seed. Seeds 2–2.3 × 0.7–1 mm, light brown, oblong-cylindrical

These were registered under collector numbers M. Tekin 1309,

and covered by densely short thick white hairs (Fig. 1).

1470, 1562, 1566, 1625 and are conserved at the Cumhuriyet

M. Tekin / Revista Brasileira de Farmacognosia 27 (2017) 143–152 145

A

B

100 μm

1 cm

C D E F G

1 mm

2 mm 1 mm 1 mm 1 mm

Fig. 1. Morphology of Hypericum thymopsis. In natural habitat at flowering stage (A); stem under SEM, arrows indicate glandiferous emergences (B); flower (C); calyx, arrow

indicate black glands (D); petal (E); stamen (F); pistil (G).

Anatomical and histological aspects Leaf

Leaf is “3” shaped in transverse section. There are uniseriate epi-

Stem dermises on both surfaces of the leaves. Adaxial epidermis cells are

The stem is in the secondary growth stage and circular shaped very large, oval or rectangular shaped and usually papillose espe-

in transverse section (Fig. 2A, E). The epidermis is uniseriate and cially near lamina margins, conversely, abaxial epidermis cells are

its cells are small, oval or usually irregular shaped, and are sur- small, rectangular or occasionally squarish shaped and are all non-

rounded by thick cutinized wall at all around the cell. Cuticle layer papillose (Fig. 3A, D). Epidermal cells have a smooth cuticle on both

is thick. There are two small wings and glandiferous emergences sides. There is conspicuous epicuticular wax on abaxial epidermis

on stem (Fig. 2A). Cortex consists of 3–7 parenchymatous cell lay- while it is lack of on adaxial surface (Fig. 4C–F). The cell wall of

ers. Cortex cells are oval or circular-shaped and there are some adaxial epidermis is very thick, despite that it is thin in abaxial epi-

intercellular spaces between them. There is a thin collenchyma dermis cells (Fig. 3D). In addition, margins of adaxial epidermis cells

which compose a ring in cortex. Endodermis is uniseriate and cells are distinct and slightly wavy, while margins of abaxial epidermis

are dorsoventrally depressed. The pericycle is conspicuous with cells are obscure. Stomata are anisocytic and sunken, occurring in

thickened laterally cell walls, just beneath the endodermis (Fig. 2B, both on the adaxial and predominantly abaxial sides of the leaf lam-

F). Vascular bundles are located in the major portion of the stem ina (Figs. 3F and 4). The mesophyll is dorsiventral and is formed

transverse section. In phloem, there are lots of type A secretory by 2–3 layers of palisade parenchyma and 2–3 layers of spongy

canals which are composed of four secretory cells. Cambium is parenchyma. While, the palisade parenchyma cells are gener-

indistinguishable. Secondary xylem is ring-porous. Pith rays are ally cylindrical, occasionally rectangular, squarish or ovoid-shaped,

parenchymatous and uniseriate, occasionally biseriate (Fig. 2C, G). the spongy parenchyma cells are usually cylindrical occasionally

Pith cells are parenchymatous and are oval or circular shaped. Size ovoid or irregular-shaped (Fig. 3B, D). There are large translucent

of pith cells enlarge toward center and the cell walls are lignified glands that positioned between just beneath the adaxial epider-

and prominently thickened in this region (Fig. 2D, H). mis and the middle area of spongy parenchyma (Fig. 3A, D). Small

146 M. Tekin / Revista Brasileira de Farmacognosia 27 (2017) 143–152

Fig. 2. Photomicrographs of stem transverse section of Hypericum thymopsis. Under light microscopy (LM) (A–D); under scanning electron microscopy (SEM) (E–H). General

view (A, E), red arrows indicate small wings in A; detail of cortex region (B, F); detail of vascular bundles (C, G); detail of pith region (D, H). Abbreviations: cl, collenchyma;

cx, cortex; ep, epidermis; en, endodermis; ge, glandiferous emergence; pe, pericycle; ph, phloem; pt, pith; pt-lg, pith cells-lignified; pt-nlg, pith cells-nonlignified; pr, pith

ray; sc, secretory canal-type A; sl, secretory cell; ve, vessel element; xy, xylem.

collateral vascular bundles are observed immersed into the meso- collenchyma cap which cells are usually ovoid or circular-shaped

phyll, associated with one or a few type A secretory canals and which make the midrib even more prominent and conspicuous

in the phloem (Fig. 3E). The midrib is oval-shaped, particularly (Fig. 3B). The cambium is clearly distinguishable between xylem

large and protrude from the abaxial surface. The midrib sur- and phloem, so vascular bundle type is open collateral. In vascu-

rounded by a uniseriate parenchymatous bundle sheath. There are a lar bundle, there are numerous type A secretory canals which are

M. Tekin / Revista Brasileira de Farmacognosia 27 (2017) 143–152 147

Fig. 3. Photomicrographs of the leaf trasnverse sections of Hypericum thymopsis. General view (A); detail of midrib region (B); detail of vascular bundle in midrib (C); detail

of lamina with translucent gland (D); detail of small vascular bundle with a type A secretory canal in mesophyll (E); detail of sunken stomata in adaxial epidermis (F).

Abbreviations: ab, abaxial epidermis; ad, adaxial epidermis; as, air space; bs, bundle sheath; ca, cambium; cx, cortex; gc, guard cells; ph, phloem; pp, palisade parenchyma;

sc, secretory canal-type A; sl, secretory cell; sp, spongy parenchyma; st, stoma; tg, translucent gland; vb, vascular bundle; xy, xylem.

composed of four secretory cells and are immersed the phloem Petal

(Fig. 3C). Adaxial and abaxial epidermises are uniseriate in petal trans-

verse section. While abaxial epidermis cells are rectangular or

squarish-shaped, adaxial epidermis cells are usually irregular

Sepal

shaped. The mesophyll consists of 5–8 layered parenchymatous

The transverse section of sepal was taken from calyx tube.

cells which are usually oval-shaped and have intercellular spaces

Epidermis is uniseriate in both sides. Adaxial epidermis cells are

between them (Fig. 6A, C). Vascularization is made by a few small

rectangular or rectangular ovoid-shaped, but they are larger and

vascular bundles in the center of mesophyll. Type A and type B

irregular shaped especially above the main vascular bundles region.

secretory canals may occur in same section, as they can be seen in

Abaxial epidermis cells are rectangular, squarish or oval shaped and

separate sections as in Fig. 6A and C. Type A secretory canals are

are usually larger than adaxial epidermis cells (Fig. 5A). Mesophyll

present in vascular bundles and they consist of four or more secre-

consists of 7–12 parenchymatous cell layers. These cells are usually

tory cells (Fig. 6B). Type B secretory canals occur just beneath the

oval, rarely circular-shaped and they have intercellular spaces. The

abaxial epidermis and they alternate with the veins (Fig. 6C, D).

midrib is almost circular-shaped and lacks bundle sheath (Fig. 5B).

There are type A secretory canals in phloem of vascular bundles and

these canals consist of 4–6 secretory cells (Fig. 5C). Beneath the Filament

abaxial epidermis, there are large type B secretory canals which

Filaments are very thin and triangular-shaped in transverse

alternate with the vascular bundles and immersed in mesophyll

section. Uniseriate epidermal cells are large and usually irregular-

(Fig. 5D).

shaped. Cuticle is thick and rugose. There is small single vascular

148 M. Tekin / Revista Brasileira de Farmacognosia 27 (2017) 143–152

Fig. 4. Photomicrographs of the leaf blade superficial sections on light microscopy (A and B); micrographs of the leaf blade surface on scanning electron microscopy (C–F)

of Hypericum thymopsis; general view of adaxial surface (A, C); detail of adaxial surface (B, D); general view of abaxial surface (E); detail of abaxial surface (F). Abbreviations:

wx, epicuticular wax; adc, adaxial epidermis cell; gc, guard cell; st, stoma; tg, leaf surface view of translucent gland.

bundle in the center of transverse section. Between epidermis and Discussion

vascular bundle, there are irregular-shaped parenchymatous cells.

The members of secretory structure are absent (Fig. 7A). Despite of its usage in folk medicine, the morphology of H. thy-

mopsis is not entirely known. There are some general knowledge

on morphology of the species in Flora of Turkey (Davis, 1966), but

Pistil there are no report on the shape of petals, dimensions of sepals and

The style is rectangular-shaped in transverse section. Epidermis ovaries, length of filaments, anthers, pistils, styles and pedicels of

is uniseriate, with rectangular, squarish or irregular-shaped cells. the species. These features are examined in present study. In Flora

Outer walls of epidermal cells are thick. Cuticle is rugose and thick. of Turkey (Davis, 1966), stems height and petals length of H. thy-

There is single vascular bundle which positioned close to epider- mopsis are given as 3–11 cm and 5–7 mm, respectively. In present

mis. There is pollen tube transmitting tract (PTTT) in the center of study, stems height and petals length of H. thymopsis were found

transverse section. Between the region of epidermis and PTTT are as 3–14 cm and 4.5–7.5 mm, respectively. In addition, black glands

filled by oval, circular or irregular shaped parenchymatous cells. were observed at the margins of small leaves which are positioned

The members of secretory structure are absent in transverse section at the base of pedicel on stem. It is not reported in Flora of Turkey

of style (Fig. 7B). The ovaries have 3 locules (Fig. 7C). Adaxial epider- (Davis, 1966) (Fig. 1).

mis cells of carpels are usually irregular, occasionally rectangular According to Metcalfe et al. (1950), pith rays are uniseriate, the

or squarish shaped. Abaxial epidermis cells of carpels are rectan- xylem and phloem are relatively narrow, whilst there is a greater

gular or squarish shaped. The mesophyll cells are parenchymatous development of pith in stem of herbaceous Hypericum. Conversely,

and usually irregular-shaped. Just beneath the adaxial epidermis, in H. thymopsis stem, phloem and especially xylem fill major part

there are numerous type C secretory canals which immersed in the of the transverse section, pith located in the small region of the

parenchymatic mesophyll of each locules (Fig. 7D). transverse section center (Fig. 2A, E). Pith rays of H. thymopsis are

M. Tekin / Revista Brasileira de Farmacognosia 27 (2017) 143–152 149

Fig. 5. Photomicrographs of calyx tube transverse section of Hypericum thymopsis. General view (A); detail of sepal midrib region with type A and type B secretory canals

(B); detail of vascular bundle and type A secretory canal in sepal (C); detail of type B secretory canal (D). Abbreviations: ab, abaxial epidermis; abw, abaxial epidermis cell; ad,

adaxial epidermis; cu, cuticle; pc, parenchymatous cell; ph, phloem; scA, secretory canal-type A; scB, secretory canal-type B; sl, secretory cell; st, stoma; vb, vascular bundle;

xy, xylem.

Fig. 6. Photomicrographs of petal transverse section of Hypericum thymopsis. General view with type A secretory canals (A) and detail view of vascular bundle and type A

secretory canal (B). General view with type B secretory canals (C) and detail view of type B secretory canal (D). Abbreviations: ab, abaxial epidermis; ad, adaxial epidermis;

cu, cuticle; pc, parenchymatous cell; ph, phloem; scA, secretory canal-type A; scB, secretory canal-type B; sl, secretory cell; vb, vascular bundle; xy, xylem.

150 M. Tekin / Revista Brasileira de Farmacognosia 27 (2017) 143–152

Fig. 7. Photomicrographs of the reproductive parts transverse section of flower of Hypericum thymopsis. General view of filament (A); general view of style (B); general view

of ovary (C); detail of carpel (D). Abbreviations: ab, abaxial epidermis; ad, adaxial epidermis; cp, carpel wall; cu, cuticle; ep, epidermis; pc, parenchymatous cell; ph, phloem;

pt, pollen-tube transmitting tract; scC, secretory canal-type C; sl, secretory cell; vb, vascular bundle; xy, xylem.

commonly uniseriate, occasionally biseriate. In addition, pith ray are absent in H. maculatum and H. hirsutum. In stem of H. thymop-

cells of H. thymopsis are upright and usually oval or rectangular- sis, type A canals and glandiferous emergences are present, but type

shaped, whereas the examined Hypericum species of Metcalfe et al. B canals are absent as well as H. maculatum and H. hirsurum (Fig. 2).

(1950), they are upright and entirely square-shaped (Fig. 2C, G). The However, there are no reports on presence of glandiferous emer-

outlines of stem transverse section are circular-shaped, vascular gences, in stems of their studied four Hypericum species. According

bundles of the H. thymopsis stems always formed a circle which to Gîtea et al. (2011), all examined species have 1–2 layered palisade

is reported for some other Hypericum species by Arda (1989) and parenchyma in leaf mesophyll, whereas it was observed as 2–3 lay-

Tokur and Mısırdalı (1989) (Fig. 2A, E). ered in H. thymopsis (Fig. 3B, D). According to Shields (1950), having

According to Metcalfe et al. (1950), the leaves of Hypericaceae more strongly developed palisade tissue is one of the characteris-

are dorsiventral with hypoderm in certain species of Hypericum and tic of the xerophytic , due to being the xeromophic plant, H.

stomata of Hypericum species are commonly surrounded by three thymopsis has similar feature of mentioned literature.

or more cells. The leaves of H. thymopsis are dorsiventral, with- Recently, Perrone et al. (2013a) were studied the leaf and stem

out hypodermis and stomata are anisocytic (Figs. 3B, D and 4A, B). anatomy of eight Hypericum species (H. aegypticum L., H. androsae-

The level of stomatal guard cells is lower than epidermis cells, so mum L., H. hircinum L., H. perfoliatum L., H. perforatum, H. pubescens

stomata are sunken type which is known as a special feature of Boiss., H. tetrapterum Fr. and H. triquetrifolium Turra) which grow

the xerophytes (Fig. 3F). Prominent epicuticular waxes are present heterogeneous environments of Sicily Island. General anatomical

on abaxial side of the leaf and thus, the capacity of reduce the characteristics of H. thymopsis are similar with those in their study.

water loss is increase (Fig. 4E, F). Furthermore, presence of the At the same time, there are some evident different characteristics

wax reduces mechanical damage and prevents from fungal and for H. thymopsis especially in leaf anatomy. In stem anatomy, there

insect attack (Eglinton and Hamilton, 1967). In addition, accord- are numerous glandiferous emergences outer of the H. thymopsis

ing to Briquet (1919) leaves of certain species of Hypericum growing stem (Figs. 1B and 2A, E). There is no report on the presence of this

arid regions in Brasil have thickened margins consisting of enlarged characteristic in stems of their studied eight species. Transverse

epidermal cells or collenchyma. The margin epidermal cells of H. section shape of the stem and presence of slightly two wings in

thymopsis leaves are enlarged, the cuticle and outer cell walls of H. thymopsis are common characteristics with some species stud-

these cells are more thickened than other epidermal cells (Fig. 3A, ied by Perrone et al. (2013a). Secondary xylem of H. thymopsis is

D) so, the leaves of H. thymopsis show the same feature of men- ring-porous (Fig. 2C, G) such as their studied six species, except H.

tioned knowledge. pubescens and H. aegypticum which have diffuse-porous secondary

Gîtea et al. (2011) were studied four Hypericum (H. perfora- xylem. The leaves of H. thymopsis are dorsiventral (Fig. 3A, B, D) as

tum, H. tetrapterum Fries, H. maculatum Crantz and H. hirsutum with H. perforatum, H. perfoliatum, H. tetrapterum, H. androsaemum

L.) in point of secretory structure of some vegetative organs. In and H. hircinum, but the leaves of H. pubescens, H. triquetrifolium

their study, while type A canals are present in the phloem of all and H. aegypticum are isobilateral (Perrone et al., 2013a). The leaves

four species stems, type B secretory canals are present solely, in of H. thymopsis are amphistomatic (Fig. 4) as with H. pubescens, H.

subepidermic area of H. perforatum and H. tetrapterum, and they tetrapterum, H. triquetrifolium and H. aegypticum while the leaves of

M. Tekin / Revista Brasileira de Farmacognosia 27 (2017) 143–152 151

the H. perforatum, H. perfoliatum, H. androsaemum and H. hircinum glands, type A, type B and type C secretory canals which produce

are hypostomatic. All stomatal complexes are anisocytic in the biologically active substances were shown anatomically and his-

studied eight species by Perrone et al. (2013a), and this feature is tologically in vegetative and reproductive organs of H. thymopsis.

the same for H. thymopsis (Fig. 4A, B). Among their studied eight Therefore, some results given in this research were compared with

species, solely H. triquetrifolium is xerophyte as H. thymopsis and the results of other members of Hypericum in the literature records

they have many common characteristics, such as possess two small and differences between them were stated in detail. All this results

wings on stem, amphistomatic leaves and ring-porous secondary can be used to evaluate of the diagnostic features of H. thymopsis

xylem. One of the prominent differences between two species is as well as the genus Hypericum.

the leaf mesophyll which is dorsiventral in H. thymopsis while it is

isobilateral in H. triquetrifolium.

Conflicts of interest

There are some studies on morphology and anatomy of the Turk-

ish Hypericum (Arda, 1989; Tokur and Mısırdalı, 1989; Erkaya and

The authors declare no conflicts of interest.

Tokur, 2004; Altıntas¸ and Akc¸ in, 2015). Erkaya and Tokur (2004)

were studied morphology and anatomy of H. montbretii Spach.,

References

H. origanifolium Willd. and H. perforatum growing in and around

Eskis¸ ehir province of Turkey. Many anatomical characteristics of

Altıntas¸ , M.Y., Akc¸ in, Ö.E., (M.Sc. thesis) 2015. Ordu ve Giresun’da yayılıs¸ gösteren

stems and leaves of these three species and H. thymopsis are simi-

bazı Hypericum L. (Hypericaceae, Guttiferae) türleri üzerinde morfolojik,

lar such as presence of collenchyma in stem cortex, amphistomatic anatomik ve mikromorfolojik bir c¸ alıs¸ ma. Ordu University, Ordu.

APG III, 2009. An update of the Angiosperm Phylogeny Group classification for the

leaves, anisocytic stomata and translucent glands in leaf mesophyll.

orders and families of flowering plants: APG III. Bot. J. Linn. Soc. 161, 105–121.

However, H. thymopsis has dorsivental leaves while all their studied

Arda, H., (M.Sc. thesis) 1989. Edirne C¸ evresindeki Hypericum L. (Guttiferae) Türleri

three species have equifacial leaves which is the main difference for Üzerinde, Morfolojik, Anatomik ve Karyolojik Aras¸ tırmalar. Trakya University,

Edirne.

H. thymopsis. Yaylacı et al. (2013) were studied anatomical charac-

Assadi, A., Zarrindast, M.R., Jouyban, A., Samini, M., 2011. Comparing of the effects of

teristics of recently described new Hypericum species, H. sechmenii

hypericin and synthetic antidepressants on the expression of morphine-induced

Ocak & Koyuncu for Turkey. According to their study, presence of conditioned place preference. Serv. Iran. J. Pharm. Res. 10, 619–626.

Avato, P., 2005. A Survey of the Hypericum genus: secondary metabolites and bioac-

amphistomatic leaves and xeromorphic stomata in H. sechmenii are

tivity. Stud. Nat. Prod. Chem. 30, 603–634.

the same features as for those in H. thymopsis. But, having dorsiven-

Bingol, U., Cosge, B., Gurbuz, B., 2011. Hypericum species in flora of Turkey. In:

tral leaves, anisocytic stomata (Fig. 4A, B) and 2–3 layered palisade Odabas, M.S., Cirak, C. (Eds.), Hypericum. Medicinal and Aromatic Plant Science

parenchyma (Fig. 3B, D) are distinctive features for H. thymopsis, and Biotechnology, vol. 5 (Special Issue 1), pp. 86–90.

Bombardelli, E., Morazzoni, P., 1995. . Fitoterapia 66, 43–68.

because H. sechmenii has equifacial leaves, anomocytic stomata

Briquet, J., 1919. La structure foliaire des Hypericum à feuilles scléro-marginées. C.

and 1–2 layered palisade parenchyma. In addition while stem pith

R. Soc. Phys. Hist. Nat. Genève 36, 75–90.

region is made of solely xylem elements in H. secmenii, there is Bruneton, J., 1995. Pharmacognosy, Phytochemistry, Medicinal Plants. Lavoisier Pub-

lishing, Paris, pp. 367–370.

clearly parenchymatous pith cells some of which have lignified cell

Carine, M.A., Christenhusz, M.J.M., 2010. About this volume: the monograph of

wall in H. thymopsis (Fig. 2D, H).

Hypericum by Norman Robson. Phytotaxa 4, 1–4.

There are several studies focused on secretory structure of some Ciccarelli, D., Andreucci, A.C., Pagni, A.M., 2001a. The ‘black nodules’ of Hypericum

perforatum L. subsp. perforatum: morphological, anatomical and histochemical

Hypericum species (Ciccarelli et al., 2001a, 2001b; Gîtea et al.,

studies during the course of ontogenesis. Israel J. Plant Sci. 49, 33–40.

2011). One of these, Ciccarelli et al. (2001b) were studied secre-

Ciccarelli, D., Andreucci, A.C., Pagni, A.M., 2001b. Translucent glands and secretory

tory structure of H. perforatum, considering the translucent glands canals in Hypericum perforafum L. (Hypericaceae): morphological, anatomical

and histological studies during the course of ontogenesis. Ann. Bot. 88, 637–644.

and secretory canals of vegetative and floral parts. They observed

Curtis, J.D., Lersten, N.R., 1990. Internal secretory structures in Hypericum (Clusi-

translucent glands, type A and type B secretory canals in sepals

aceae): H. perforatum L. and H. balearicum L. New Phytol. 114, 571–580.

and petals. While, they were observed translucent glands and type Davis, P.H., 1966. Flora of Turkey and the East Aegean Islands, vol. 2. Edinburgh

University Press, Edinburgh, UK, pp. 355.

A secretory canals in leaves, they were observed type B secretory

Davis, P.H., 1988. Flora of Turkey and the East Aegean Islands, vol. 10. Edinburgh

canals with type A secretory canals in stem transverse section. In

University Press, Edinburgh, UK, pp. 96.

addition, there were only type A and type C secretory canals in Davis, A.P., Barnett, J.R., 1997. The leaf anatomy of the genus Galanthus L. (Amaryll-

transverse section of ovaries of H. perforatum. When compare these idaceae J. St.-Hil.). Bot. J. Linn. Soc. 123, 333–352.

Eglinton, G., Hamilton, R.J., 1967. Leaf epicuticular waxes. Science 156, 1322–1335.

results with those in H. thymopsis, there are some differences. There

Erkaya, I.P.,˙ Tokur, S., 2004. Morphological and anatomical investigations on some

are present type A and type B secretory canals in sepals and petals

Hypericum L. species growing naturally in and around Eskis¸ ehir. Trakya Univ. J.

of H. thymopsis, but translucent glands are absent (Figs. 5 and 6). Sci. 5, 97–105.

Esau, K., 1965. Plant Anatomy. John Wiley and Sons, New York.

In ovary, while H. thymopsis has numerous type C secretory canals,

Gîtea, D., S¸ ıpos¸ , M., Tamas¸˘ , M., Pas¸ ca, B., 2011. Secretory structures at species of

type A secretory canals are absent (Fig. 7C, D). In stem transverse

Hypericum genera from Bihor county, Romania. Note I. Vegetative organs. Fama-

section of H. thymopsis, type A canals are present in phloem, but cia 59, 424–431.

Gudziˇ c,´ B., Ðordevic,´ S., Vajs, V., Palic,´ R., Stojanovic,´ G., 2002. Composition and

type B canals are absent. One of the other differences for H. thymop-

antimicrobial activity of the essential oil of Crantz. Flavour

sis is presence of glandiferous emergences (Figs. 1B and 2A, E, B, F)

Fragr. J. 17, 392–394.

which is not reported on the stem of H. perforatum by Ciccarelli et al. Gudziˇ c,´ B., Ðordevic,´ S., Nedeljkovic,´ J., Smelceroviˇ c,´ A., 2004. Essential oil composi-

tion of Boiss. Chem. Ind. 58, 413–415.

(2001b). The secretory structures are absent in filament transverse

Güner, A., Özhatay, N., Ekim, T., Bas¸ er, K.H.C., 2000. Flora of Turkey and the East

section of H. thymopsis (Fig. 7A) and this feature is similar with H.

Aegean Islands, vol. 11. Edinburgh University Press, Edinburgh, UK, pp. 71.

perforatum (Ciccarelli et al., 2001b). Güner, A., Aslan, S., Ekim, T., Vural, M., Babac¸ , M.T. (Eds.), 2012. Türkiye Bitkileri Lis-

tesi (Damarlı Bitkiler). Nezahat Gökyigit˘ Botanik Bahc¸ esi ve Flora Aras¸ tırmaları

As a conclusion, as can be seen from the literature survey, this

Dernegi˘ Yayını, Istanbul.˙

study seems to be the first extensive research on morphology and

Hong-Fei, Lü, Zheng-Hai, Hu, 2001. Comparative anatomy of secretory structures of

first research on anatomy of the stem, leaf, sepal, petal, filament and leaves in Hypericum L. Acta Phytotaxon. Sin. 39, 393–404.

pistil of H. thymopsis. In addition some micromorphological char- Jensen, W.A., 1962. Botanical Histochemistry: Principles and Practice. W.H. Freeman

and Company, London.

acteristics are revealed for the first time. In morphological studies

Joulain, D., König, W.A., 1998. The Atlas of Spectra Data of Sesquiterpene Hydrocar-

some different results and new characters were determined, and

bons. E.B.-Verlag, Hamburg, Germany.

by using these characters, morphological description of the H. thy- Kitanov, G.M., 2001. Hypericin and pseudohypericin in some Hypericum species.

Biochem. Syst. Ecol. 29, 171–178.

mopsis was possibly extended. The anatomy of all aerial parts used

Kültür, S., 2007. Medicinal plants used in Kırklareli Province (Turkey). J. Ethnophar-

in folk medicine and their secretory structures were investigated

macol. 111, 341–364.

in detail. The presence of glandiferous emergences, translucent Lewis, L.W., 2003. Medical Botany. John Wiley and Sons, Inc., Hoboken, NJ, pp. 654.

152 M. Tekin / Revista Brasileira de Farmacognosia 27 (2017) 143–152

Maggi, F., Ferretti, G., Pocceschi, N., Menghini, L., Ricciutelli, M., 2004. Morphological, Perrone, R., Rosa, P.D., Castro, O.D., Colombo, P., 2013b. A further analysis

histochemical and phytochemical investigation of the genus Hypericum of the of secretory structures of some taxa belonging to the genus Hypericum

Central Italy. Fitoterapia 75, 702–711. (Clusiaceae) in relation to the leaf vascular pattern. Turk. J. Bot. 37,

Meseguer, A.S., Sanmartín, I., 2012. Paleobiology of the genus Hypericum (Hyperi- 847–858.

caceae): a survey of the fossil record and its palaeogeographic implications. An. Robson, N.K.B., 1981. Studies in the genus Hypericum L. (Guttiferae). 2. Characters of

Jard. Bot. Madr. 69, 97–106. the genus. Bull. Br. Museum (Nat. Hist. Bot. Ser.) 8, 55–226.

Metcalfe, C.R., Chalk, L., Chattaway, M.M., Hare, C.L., Richardson, F.R., Slatter, E.M., Russo, E.B., 2011. Taming THC: potential Cannabis synergy and

1950. Hypericaceae. In: Metcalfe, C.R., Chalk, L. (Eds.), Anatomy of the Dicotyle- phytocannabinoid–terpenoid entourage effects. Br. J. Pharmacol. 163,

dons, I. Leaves, Stem and Wood in Relation to Taxonomy with Notes on Economic 1344–1364.

Uses. Clarendon Press, Oxford, pp. 165–169. Santos, P.A.G., Figueiredo, A.C., Barroso, J.G., Pedro, L.G., Scheffer, J.J.C., 1999. Compo-

Nissen, L., Zatta, A., Stefanini, I., Grandi, S., Sgorbati, B., Biavati, B., Monti, A., 2010. sition of the essential oil of Aiton from five Azorean Islands.

Characterization and antimicrobial activity of essential oils of industrial hemp Flavour Fragr. J. 14, 283–286.

varieties (Cannabis sativa L.). Fitoterapia 81, 413–419. Shields, L.M., 1950. Leaf xeromorphy as related to physiological and structural influ-

Onelli, E., Rivetta, A., Giorgi, A., Bignami, M., Cocucci, M., Patrignani, G., 2002. Ultra- ences. Bot. Rev. 16, 399–447.

structural studies on the developing secretory nodules of Hypericum perforatum. Siersch, E., 1927. Anatomie und Mikrochemie der Hypericumdruesen (Chemie des

Flora 197, 92–102. Hypericins). Planta 3, 481–489.

Özkan, A.M.G., Demirci, B., Bas¸ er, K.H.C., 2009. Essential oil composition of H. thy- Simpson, M.G., 2006. Plant Systematics, 1st edition. Elsevier-Academic Press,

mopsis Boiss. J. Essent. Oil Res. 21, 149–153. Amsterdam.

Özkan, E.E., Demirci, B., Gürer, C¸ .Ü., Kültür, S¸ ., Mat, A., Bas¸ er, K.H.C., 2013. Composi- Spiteller, M., Özen, T., Smelcerovic, A., Zuehlke, S., Mimica-Dukic, N., 2008. Phenolic

tion of essential oils from five endemic Hypericum species of Turkey. Org. Chem. constituents and the in vitro antioxidant activity of the flowers of H. venustum.

Curr. Res. 2, 1. Fitoterapia 79, 191–193.

Özkan, E.E., Mat, A., 2013. An overview on Hypericum species of Turkey. J. Pharma- Toker, Z., Kızıl, G., Özen, H.C., Kızıl, M., Ertekin, S., 2006. Compositions and antimi-

cogn. Phytother. 5, 38–46. crobial activities of the essential oils of two Hypericum species from Turkey.

Öztürk, Y., Aydın, S., Bas¸ er, K.H.C., Kirimer, N., Kurtar-Özturk, N., 1992. Hepatopro- Fitoterapia 77, 57–60.

tective activity of H. perforatum L. alcoholic extract in rodents. Phytother. Res. 6, Tokur, S., Mısırdalı, H., 1989. Bazı Hypericum Türleri Üzerinde Anatomik C¸ alıs¸ malar

44–46. Anadolu Üniv. Fen Ed. Fak. Derg. 2, 1–8.

Öztürk, Y., 1997. Testing the antidepressant effects of Hypericum species on animal Toma, C., Rugina,˘ R., 1998. Atlas de anatomia plantelor medicinale. Ed. Acad. Rom.

models. Pharmacopsychiatry 30, 125–128. 98, 142–145.

Öztürk, Y., 2001. Possible mechanism of the analgesic effect of St. John’s Wort. Yaylacı, Ö.K., Özgis¸ i, K., Sezer, O., Orhanoglu,˘ G., Öztürk, D., Koyuncu, O., 2013.

Fundam. Clin. Pharmacol. 15, 113–153. Anatomical studies and conservation status of rare endemic Hypericum sech-

Öztürk, B., Apaydın, S., Goldeli, E., Ince,˙ I., Zeybek, U., 2002. H. triquetrifolium Turra. menii Ocak & Koyuncu (Sect: Adenosepalum) from Eskis¸ ehir-Turkey. J. Selc¸ uk

extracts exhibits antiinflammatory activity in the rat. J. Ethnopharmacol. 80, Univ. Nat. Appl. Sci. 2, 1–11.

207–209. Yes¸ ilada, E., Gürbüz, I., Shibata, H., 1999. Screening of Turkish antiulcerogenic

Öztürk, N., Korkmaz, S., Öztürk, Y., 2007. Wound-healing activity of St. John’s Wort folk remedies for anti-Helicobacter pylori activity. J. Ethnopharmacol. 66,

(H. perforatum L.) on chicken embryonic fibroblasts. J. Ethnopharmacol. 111, 289–293.

33–39. Zeng, H., Zhou, P., 2001. Analysis of the chemical constituents of the essential

Perrone, R., Rosa, P.D., Castro, O.D., Colombo, P., 2013a. Leaf and stem anatomy in oils from the leaves of two Hypericum plants. Nat. Sci. J. Xiangtan Univ. 23,

eight Hypericum (Clusiaceae). Acta Bot. Croat. 72, 269–286. 52–54.