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Anatomic Studies on Verbascum pestalozzae Boiss. and Verbascum pycnostachyum Boiss. & Heldr

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Objectives: The genus Verbascum L. (Scrophulariaceae) known as "sığır kuyruğu" in Anatolia is represented by 248 species, 193 of which are endemic. The flowers contain essential oil, mucilage, and glycosides. Some species of Verbascum have some folkloric uses as expectorants, sedatives, and treatment for dysmenorrhea and rheumatalgia. Its use for healing wounds in animal care has also been reported. In this study, the anatomic structures of root, stem, and leaves of Verbascum pestalozzae Boiss. and Verbascum pycnostachyum Boiss. & Heldr. are given for the first time. According to the results, some of the differences between stem and leaf anatomic features of these species were described and the data were supported by detailed photographs. Materials and methods: Specimens were collected from C3 Antalya: Voucher specimens of V. pestalozzae and V. pycnostachyum were deposited in the Herbarium of the Biology Department, Akdeniz University in Antalya, Turkey and Herbarium of the Faculty of Pharmacy, Anadolu University in Eskişehir, Turkey. The materials were identified as V. pestalozzae and V. pycnostachyum using flora of Turkey and the East Aegean islands. For anatomic studies, samples were collected from the natural habitats and kept in 70% alcohol. In the research, root, stem and leaves of mature and flowered plants were used. Investigations were performed on the cross-sections of the root, the flowering stem, and the leaf. The anatomic structures of the species were drawn using a Leitz SM-LUX binocular microscope. All sections were embedded in glycerin gelatin and stained using Sartur reactive, and photographs were taken through a light microscope (Olympus BX51T). Results: The cross sections taken from root, stem, and leaves of V. pestalozzae and V. pycnostachyum were examined and the anatomic features belonging to these plants were determined and compared. Conclusion: The anatomic properties belonging to the two species were generally compatible with findings of Metcalfe and Chalk and others signified in the genus. We believe that our results provide additional evidence for taxonomists to help separate the species. The lack of a taxonomic, morphologic, and anatomic study on the species made it important for the systematic introduction of the research.
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347
ORIGINAL ARTICLE
©Turk J Pharm Sci, Published by Galenos Publishing House.
ÖZ
ABSTRACT
*Correspondence: E-mal: salan@anadolu.edu.tr, Phone: +90 545 299 72 14 ORCID-ID: orcd.org/0000-0002-3594-0364
Receved: 11.08.2016, Accepted: 18.05.2017
Amaç: Anadolu’da “Sığır kuyruğu” olarak adlandırılan Verbascum L. (Scrophulariaceae) cinsi, ülkemizde 193’ü endemik olmak üzere 248 tür ile
temsil edilmektedir. Çiçekleri müsilaj, uçucu yağ ve glikozitler taşır. Bazı Verbascum türleri halk arasında göğüs yumuşatıcı ve balgam söktürücü
olarak, adet sancısını gidermede, yatıştırıcı, romatizma ağrılarını giderici ve ayrıca hayvan yaralarını tedavi etmekte kullanılmaktadır. Verbascum
pestalozzae Boiss. ve Verbascum pycnostachyum Boiss. & Heldr. türlerinin kök, gövde ve yaprak anatomik yapıları ilk kez bu çalışmada verilmiştir.
Araştırmalar sonucunda türlerin gövde ve yaprak anatomik özellikleri belirlenmiş ve veriler detaylı fotoğraflarla desteklenmiştir. Gen merkezi
Anadolu olmasına rağmen Verbascum ile ilgili yeterli çalışmaların bulunmaması ve taşıdığı sekonder metabolitler ile potansiyel bir tıbbi bitki olması
çalışmanın önemini artıracaktır.
Gereç ve Yöntemler: V. pestalozzae ve V. pycnostachyum’a ait örnekler C3 Antalya’dan toplanmış, Akdeniz Üniversitesi Biyoloji Bölümü ve Anadolu
Üniversitesi Eczacılık Fakültesi Herbaryumu’na konulmuştur. Türlerin teşhisi Türkiye florası ve Doğu Ege adaları kullanılarak belirlenmiştir.
Anatomik çalışmalar için, örnekler doğal habitatlardan toplanmış ve %70 alkolde tutulmuştur. Araştırmada, olgun ve çiçekli bitkilerin kök, gövde ve
yaprakları kullanılmıştır. Kök, çiçekli gövde ve yaprak kesitleri üzerinde incelemeler yapılmış. Türlerin anatomik yapılarının çizimleri Leitz SM-LUX
binoküler mikroskop kullanılarak yapılmıştır. Kesitler gliserin jelatin ile daimileştirilip, Sartur reaktifiyle boyanmış ve fotoğraflar ışık mikroskobu
(Olympus BX51T) ile çekilmiştir.
Objectives: The genus Verbascum L. (Scrophulariaceae) known as “sığır kuyruğu” in Anatolia is represented by 248 species, 193 of which are
endemic. The flowers contain essential oil, mucilage, and glycosides. Some species of Verbascum have some folkloric uses as expectorants,
sedatives, and treatment for dysmenorrhea and rheumatalgia. Its use for healing wounds in animal care has also been reported. In this study, the
anatomic structures of root, stem, and leaves of Verbascum pestalozzae Boiss. and Verbascum pycnostachyum Boiss. & Heldr. are given for the first
time. According to the results, some of the differences between stem and leaf anatomic features of these species were described and the data
were supported by detailed photographs.
Materials and Methods: Specimens were collected from C3 Antalya: Voucher specimens of V. pestalozzae and V. pycnostachyum were deposited
in the Herbarium of the Biology Department, Akdeniz University in Antalya, Turkey and Herbarium of the Faculty of Pharmacy, Anadolu University
in Eskişehir, Turkey. The materials were identified as V. pestalozzae and V. pycnostachyum using flora of Turkey and the East Aegean islands. For
anatomic studies, samples were collected from the natural habitats and kept in 70% alcohol. In the research, root, stem and leaves of mature
and flowered plants were used. Investigations were performed on the cross-sections of the root, the flowering stem, and the leaf. The anatomic
structures of the species were drawn using a Leitz SM-LUX binocular microscope. All sections were embedded in glycerin gelatin and stained
using Sartur reactive, and photographs were taken through a light microscope (Olympus BX51T).
Results: The cross sections taken from root, stem, and leaves of V. pestalozzae and V. pycnostachyum were examined and the anatomic features
belonging to these plants were determined and compared.
Conclusion: The anatomic properties belonging to the two species were generally compatible with findings of Metcalfe and Chalk and others
signified in the genus. We believe that our results provide additional evidence for taxonomists to help separate the species. The lack of a taxonomic,
morphologic, and anatomic study on the species made it important for the systematic introduction of the research.
Key words: Scrophulariaceae, Verbascum, anatomy, endemic
1Anadolu University, Faculty of Pharmacy, Pharmaceutical Botany, Eskişehir, Turkey
2Akdeniz University, Faculty of Science, Department of Biology, Antalya, Turkey
Sevim KÜÇÜK
1
*, Melike Belkıs GÖKÇE
1
, Ramazan Süleyman GÖKTÜRK
2
Verbascum pestalozzae Boiss. ve Verbascum pycnostachyum Boiss. & Heldr.
Üzerinde Anatomik Araştırmalar
Anatomic Studies on Verbascum pestalozzae Boiss.
and Verbascum pycnostachyum Boiss. & Heldr.
Turk J Pharm Sci 2018;15(3):347-353
DOI: 10.4274/tjps.16779
348
INTRODUCTION
The genus Verbascum Linnaeus (1753: 177) (excl. Celsia
Linnaeus 1753: 621) (Scrophulariaceae) has about 360 species
from all over the world (Heywood 1993, Mabberley 2008).
Represented with 248 species in Turkey, the genus has been
divided into 13 partly artificial groups with 130 additional
hybrids. Among them, 193 species are endemic to Turkey, with
an endemism percentage of about 80%. The genus Verbascum
L. (Scrophulariaceae) known as “Sığır Kuyruğu” in Anatolia.1-5
Many plant species among the flora of Turkey play important
roles in traditional medicine. There are approximately 9300
plant species in Turkish flora, some of which are widely used in
folkloric medicine due to their antimicrobial and anticarcinogenic
properties.6,7 One of the well-known Verbascum species is V.
thapsus L., which has been used for the treatment of several
diseases including asthma, spasmodic cough, migraine,
and earache. Moreover, V. thapsus, V. fruticulosum Post, V.
undulatum Lam. and V. georgicum Bentham have anti-malarial
and anti-viral effects, which have been investigated in both in
vitro and in vivo studies.6
It is reported that leaves and flowers of Verbascum species have
expectorant, mucolytic, and demulcent properties, and they are
used to treat respiratory disorders such as bronchitis, dry coughs,
tuberculosis, and asthma in Anatolia.8,9 Verbascum species are
also used to treat hemorrhoids, rheumatic pain, superficial
fungal infections, wounds and diarrhea. Furthermore, these
species demonstrate several inhibitory activities against the
murine lymphocytic leukemia and influenza viruses A2 and B.
Macerated oil prepared from the flowers is used for reducing
earache, and applied externally for eczema and other types of
inflammatory skin disorders.10
Verbascum species have some folkloric uses such as a
sedative, and treatment for dysmenorrhea and rheumatalgia. Its
use for healing wounds has also been reported in animal care.
Iridoid and neolignan type glycosides, oleanan type terpenes,
flavonoids, polysaccharides, saponins, steroids and alkaloids
were major compounds isolated from Verbascum species.11 In
several bioactivity studies on Verbascum sp., anti-proliferative,12
anti-inflammatory,13 antioxidant,14,15 anti-histaminic, anti-fungal,
anti-bacterial,16 wound healing,17 anti-microbial18 and anti-
cancer effects19 have been shown in crude extracts of roots,
leaves, flowers, and aerial parts.
Verbascum genus is one of the largest genus with regard to
the number of species in Turkey and is also known for several
problems in diagnosis and taxonomy.
In addition, the anatomic structures of root, stem and leaves
of and Verbascum pestalozzae Boiss. (endemic), “Boz Sığır
Kuyruğu’’ and Verbascum pycnostachyum Boiss. & Heldr. “Eğirdir
Sığır Kuyruğu’’2,4 are given in this study for the first time.
EXPERIMENTAL
Plant material
The flowering aerial parts of Verbascum pestalozzae were
collected from C3 Antalya: Konyaaltı, Doyran, between
Saklıkent and Karçukuru (360 49’ 01’’ N, 300 21’ 54’’ E), on lime
stone rocks, 2100 m, 17.07.2008, ESSE 15069! Göktürk 7338;
Verbascum pycnostachyum were collected from C3 Antalya,
between Korkuteli and Fethiye (37° 02’ 53’’ N, 30° 06’ 26 ‘’ E),
steppe, 1370 m above the sea level, at the end of July 2007,
Eskişehir (ESSE) 14730!, Göktürk 6093; (Figures 1-3). Voucher
specimens of V. pestalozzae and V. pycnostachyum were
deposited in the Herbarium of the Biology Department, Akdeniz
University in Antalya, Turkey and Herbarium of the Faculty of
Pharmacy, Anadolu University in ESSE, Turkey. The materials
were identified as V. pestalozzae and V. pycnostachyum using
flora of Turkey and the East Aegean islands.2
KÜÇÜK et al. Verbascum pestalozzae Boiss. ve Verbascum pycnostachyum Boiss. & Heldr.
Bulgular: V. pestalozzae ve V. pycnostachyum’un kök, gövde ve yapraklarından alınan kesitler incelenmiş ve bu bitkilere ait anatomik özellikler
karşılaştırılmıştır.
Sonuç: İki türe ait anatomik özellikler genellikle Metcalfe ve Chalk ve diğer çalışmalarda belirtilen bulgularla uyumludur. Sonuçlarımızın,
taksonomistlere ek kanıt sağladığına ve türlerin ayrılmasına yardımcı olabileceğine kanısındayız. Türler üzerinde taksonomik, morfolojik ve anatomik
bir çalışma olmaması, araştırmanın sistematik olarak tanıtımı için önemli hale getirmiştir.
Anahtar kelimeler: Scrophulariaceae, Verbascum, anatomi, endemik
Figure 1. Distribution map of V. pestalozzae () and V. pycnostachyum ()
in Turkey
Figure 2. V. pestalozzae
349
Anatomical
For anatomic studies, samples were collected from the natural
habitats and kept in 70% alcohol. In the research, root, stem, and
leaves of mature and flowered plants were used. Investigations
were performed on the cross-sections of the root, the flowering
stem, and the leaf. The anatomic structures of glandular and
covering hairs were drawn using a Leitz SM-LUX binocular
microscope. All sections were embedded in glycerin gelatin
and stained using Sartur reactive, and photographs were taken
through a light microscope (Olympus BX51T).
RESULTS AND DISCUSSION
The cross sections taken from root, stem, and leaves of V.
pestalozzae and V. pycnostachyum were examined and the
anatomic features belonging to these plants were determined
and compared (Figures 4-10, Table 1).
Root
The root in both species is composed of periderm on the
outside and felloderm where 4-5 radial rows are broken
down and felloderm with 2-3 rows of tissue. Outer felloderm
cells are tissue debris of the primary cortex that has been
shattered or crushed in place. Secondary phloem formed of
elliptical-shapeless, round-shaped, irregular-arranged and
4-6 row cells under the periderm is taken part. Cambium is
uncertain. The secondary xylem part covers a large area and
consists of tracheal elements with large and small sizes in a
KÜÇÜK et al. Verbascum pestalozzae Boiss. ve Verbascum pycnostachyum Boiss. & Heldr.
Table 1. Anatomic differences of the species
V. pestalozzae V. pycnostachyum
Root Pith Parenchymatic Parenchymatic
Stem Sclerenchyma 3-6 celled 1-5 celled
Phloem Thick, 3-8 celled Thick, 8-15 celled
Leaf
Abaxial surface is projected under the vascular
bundle Protrusion
Vascular bundles Crescent-shaped Horne shaped
Upper parenchymatic cells 5-6 celled 25-30-celled
Lower epidermis parenchymatic cells 5-10 celled 10-15 celled
Eglandular hairs in the stem and leaf Candelabriform, stellate, multicellular Candelabriform, stellate, multicellular
Glandular hairs in the stem and leaf
Head 1 stalk 2 celled;
Head 2 stalk 1 celled;
Head 3 stalk 2 celled;
Head 1 stalk 3 celled;
Head 2 stalk 3 celled;
pellucid glands
Head 1 stalk 1 celled
Head 2 stalk 1 celled
Head 3 stalk 2 densely
Figure 3. V. pycnostacyum habit (picture: R.S. Göktürk)
Figure 4. V. pestalozzae; cross-section of root
cp: Cortex parenchym, pd: Peridermis, pi: Pith, sph: Secondary phloem, sx: Secondary
xylem
350 KÜÇÜK et al. Verbascum pestalozzae Boiss. ve Verbascum pycnostachyum Boiss. & Heldr.
sclerenchymatic tissue. Medullary rays are 2-3 rows of cells.
The pith region, covering a narrow area, is parenchymatic in V.
pestalozzae and V. pycnostacyum (Figures 4, 7).
Stem
When cross-sections were taken on the stems of two Verbascum
species, secondary growth was observed. The epidermis
is formed by a single-row, thick membrane elliptic or round
cells. The upper and lower walls are thick but the lateral sides
are thin. Its upper surface is covered with cuticle (Figures
5, 8). Covering hair and glandular trichomes were observed.
Covering hairs of V. pestalozzae are candelabriform, stellate, and
multicellular. Glandular trichomes were of 6 types; head 1 stalk
2 celled; head 2 stalk 1 celled; head 3 stalk 2 celled; head 1 stalk
3 celled; head 2 stalk 3 celled; pellucid glands (Figures 11b).
Covering hairs are candelabriform, stellate, and multicellular in
V. pycnostachyum. Its glandular trichomes are of three types;
head 1 stalk 1 celled, head 2 stalk 1 celled, head 3 stalk 2 celled
(dense, Figures 10). Parenchymatic cortex in 5 or 20 rows is
found in both species under the epidermis. Collenchyma cells
under the epidermis in the primary cortex were seen, and
parenchyma cells including oval-shaped chloroplast inside
the epidermis were found. Druse crystals were observed in
parenchymatic cells. The endodermis, consisting of flattened
cells, can hardly be distinguished from the cortex parenchyma.
There were large, small, and discontinuous sclerenchyma
bunches, 3 or 6 rows in secondary phloem of V. pestalozzae,
but 1-5 rows in V. pycnostachyum. Phloem is much narrower
in V. pestalozzae with 3-8 rows, but it is much wider in V.
Figure 5. V. pestalozzae; cross-section of stem
cp: Cortex parenchym, e: Epidermis, ph: Phloem, pi: Pith, pr: Pith ray, s: Scleranchyma,
x: Xylem
Figure 6. V. pestalozzae; cross-section of leaf, respectively
ch: Covering hair, cl: Collenchyma, cu: Cuticle, gh: Glandular hair, p:
Parenchyma, ph: Phloem, ue: Upper epidermis, x: Xylem
Figure 7. V. pycnostachyum; cross-section of root
pi: Pith, pr: Pith ray, sph: Secondary phloem, sx: Secondary xylem
351
KÜÇÜK et al. Verbascum pestalozzae Boiss. ve Verbascum pycnostachyum Boiss. & Heldr.
pycnostachyum with 8-15 rows, but has a circle shape consisting
of flattened, shapeless or oval cells. Cambium is uncertain.
Bunches in both types of secondary xylem narrowed towards
the primary xylem. Sclerenchymatic cells in this part, formed
from trache and tracheids in both types, created regular rows
in a radial direction. Pith bunches were in the form of large
polygon or round- shaped and parenchymatic cells where their
walls were lignified. Druse crystals were clearly found in these
cells (Figures 5, 8).
Leaf
In the cross-section of the main and inter vascular tissues, the
outer and inner layers of V. pestalozzae and V. pycnostachyum
were clearly protrusive. The epidermis includes a single
flattened row, rectangular, round or oval-shaped cells. Outer
epidermal cells are larger than the inner epidermal cells,
covered with a thin cuticle layer, which is curled over. Outer
membranes are thicker than inner and longitudinal membranes,
but the inner membranes of epidermal cells in the middle vein
area became thicker. Covering hairs and glandular hairs were
the same as the stem and seen in both epidermis. Covering hairs
are candelabriform, stellate, and multicellular in V. pestalozzae.
Glandular hairs were of 6 types. The covering hairs of V.
pycnostachyum were candelabriform, stellate, and multicellular.
Glandular hairs were of 3 types. Stoma (amphistomatic) found on
both surfaces of the leaf were much denser on the lower surface.
The transverse cross-section is higher than the epidermal cells
(hygromorphic stoma). In all types of mesophyll was arranged
as two or three-rows under the outer epidermis. It was formed
with plentiful chloroplasted palisade parenchyma and three
or five rows of sponge parenchyma, which was underneath
(bifacial leaf). Vascular bundles were collateral. These were
well developed in a continuous crescent in V. pestalozzae but
interrupted and horn-shaped in V. pycnostachyum. Xylem was
located in the upper epidermis, while the floor was located in
the lower epidermis. In xylem, tracheal elements are arranged
radially and thin-walled parenchymal cells are present. Phloem
is placed under the xylem. Clear parenchymatic cells were
arranged in 2 or 3 rows under the outer epidermis, five or
six rows in V. pestalozzae after collenchyma, and 5 or 6 rows
in V. pycnostachyum. Parenchymatic tissue was found in V.
pycnostachyum with 25-30 rows under the phloem till the
inner epidermis, but much narrower in V. pestalozzae with 5-10
rows. Thick lateral veins in both sides of the middle vein were
arranged till the edge of palm and the middle vein made a deep
outgrowth. The lateral veins had the same anatomical structure
as the middle veins, but vascularity was much more reduced
(Figures 6, 9-12).
Figure 8. V. pycnostachyum; cross-section of stem
cp: Cortex, parenchym, cu: Cuticle, e: Epidermis, ph: Phloem, pi: Pith, pr: Pith ray, s:
Scleranchyma, x: Xylem
Figure 9. V. pycnostachyum; cross-section of leaf, respectively
ch: Covering hair, cl: Collenchyma, cp: Cortex, parenchym, cu: Cuticle, gh: Glandular
hair, le: Lower epidermis, p: Parenchyma, ph: Phloem, pp: Palisade parenchyma, st:
Sp-spongy parenchyma, ue: Upper epidermis, x: Xylem
Figure 10. Hairs of stem and leaf. (A) V. pestalozzae eglanular hairs of; leaf
(B). V. pycnostachyum eglandular hairs of leaf, glandular hairs of stem (C)
g: Glandular hairs
352 KÜÇÜK et al. Verbascum pestalozzae Boiss. ve Verbascum pycnostachyum Boiss. & Heldr.
According to the results, the root, stem, and leaf anatomic
structures of each Verbascum species, V. pestalozzae differs
anatomically from each other. These differences can be
summarized as follows: in the stem, there are narrower
sclerenchymatic cells and phloem than those in the leaf of V.
pestalozzae; having a protrusion below, continuous crescent
vascular bundles, less parenchymatic cells in outer and inner
epidermis, stellate type hairs and differences in the number of
cells in secretion hairs.
CONCLUSIONS
The anatomic properties belonging to the two species are
generally compatible with the findings of Metcalfe and Chalk20
and others signified in the genus.21 -24 We believe that our results
provide additional evidence for taxonomists and can help to
separate the species. The lack of a taxonomic, morphologic,
and anatomic studies on the species made it important for the
systematic introduction of the research.
Conflict of Interest: No conflict of interest was declared by the
authors.
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